Age-related changes in polyamine biosynthesis after fasting and refeeding

The effect of aging on ornithine decarboxylase (ODC) activity and polyamine biosynthesis in the proximal small intestine was studied in two groups of male Fisher 344 rats (young [4-month old] and aged [26- to 27-month old]) using a fasting and refeeding model. In control (nonfasted) rats, levels of polyamines (putrescine, spermidine and spermine) and ODC activity were significantly higher in aged compared with young rats. In aged rats, fasting significantly reduced the levels of putrescine by 41%, spermidine by 23%, and spermine by 11%; however, fasting had no effect on polyamine levels in young rats. ODC activity was decreased 75% in young and 50% in aged rats after fasting compared with the respective age-matched controls. Conversely, 2 h after reinstituting a chow diet increased ODC activity by 17-fold in young rats but only 8-fold in aged rats. Putrescine levels were also increased in both age groups after refeeding; however, similar to ODC activity, these increases were much less in aged rats. In addition, spermidine and spermine levels remained significantly depressed in the aged groups even after 24 h of refeeding. These findings suggest that the normal rigid control of gut polyamine biosynthesis and proliferation noted in young rats is markedly altered with aging.     

Putrescine as a source of instant energy in the small intestine of the rat

Background and aims—It has been suggested thatputrescine acts as a growth factor in the gut, but its exact functionin some aspects of cellular metabolism is still in question. The aim of the present work was to identify some functions of putrescine in smallbowel metabolism.
Animals—Rats (about 80 g), in groups of five, weregiven either phytohaemagglutinin- or lactalbumin-containing diets,fed ad libitum or were fasted for 48 hours and re-fed for six or twelve hours before being killed.
Methods—Uptake of intraperitoneally orintragastrically administered [¹⁴C]putrescine and itsconversion to succinate by the rat small bowel mucosa was measured.Tissue polyamine and succinate contents were measured by highperformance liquid chromatography and amino acid analysis respectively.
Results—Uptake of putrescine by the small bowelmucosa from the systemic circulation and conversion of about 30% ofthis to succinate occurs in the epithelium of the healthy small bowel. Compared with rats given food ad libitum, putrescine uptake was doubledin fasted animals and more than 70% of it was converted to succinate.All these changes returned to control values on refeeding. Using phyto-haemagglutinin induced gut growth as a model, the uptake ofputrescine from the systemic circulation by the serosal side of thesmall intestinal epithelium was increased immediately after growth wasstimulated. During phytohaemagglutinin induced growth of the gut,putrescine was converted to succinate in the same proportion as inthe healthy small bowel.
Conclusions—The experiments identified a novelfunction for putrescine in gut metabolism: it can be used as an instantenergy source when required.

Keywords:putrescine; luminal uptake; phytohaemagglutinin; succinate; basolateral uptake; small bowel

     

Aging and intestinal polyamine metabolism in the rat

Hyperproliferation and delayed expression of enzyme activity occur in small intestinal enterocytes of aging rats, and starvation and refeeding result in impaired control of these processes. Since altered polyamine metabolism may accompany changes in enterocyte proliferation, we studied the effects of nutrient manipulation upon cell numbers, ornithine decarboxylase (ODC) activity and polyamine content in jejunum and ileum of 4- to 5- and 26- to 27-month Fischer rats. In both groups, cell numbers fell during starvation and and increased during refeeding. Crypt cell hyperplasia was found in aging animals. Jejunal putrescine, spermine and spermidine content were greater in older rats, fell during starvation, and rose during refeeding. Ileal ODC activity was 66% greater in the aging rats, but jejunal ODC activity was modestly increased in young animals. Intestinal polyamine content correlates with proliferative changes and polyamine metabolism responds appropriately to nutrient manipulation during aging. Dissociation of ODC activity and polyamine content in aging jejunum probably occurred because enterocyte differentiation was delayed. Investigation of intestinal polyamine metabolism may be useful in elucidating deranged proliferative activities found in the intestine of aging rodents.     

Rapid absorption of luminal polyamines in a rat small intestine ex vivo model

BACKGROUND AND AIM: Not only biosynthesis, but also uptake from the intestinal lumen, are important polyamine sources. However, there has been no information regarding dynamic polyamine transport in the small intestine. We evaluated polyamine uptake from the small intestine using a rat ex vivo model. METHODS: The organ block consisting of the small intestine and blood vessels was used. The isolated small intestine was placed in a warmed saline bath and perfused in a non-circulating manner via the superior mesenteric artery. Radio-labeled putrescine, spermidine or spermine (7.4 x 104 Bq), with 1.0 mL of phosphate buffer saline (pH 7.4) was instilled into the jejunal lumen for 1 min. Blood samples from the portal vein were collected and sample radioactivity was determined. In another experiment, an immunohistochemical study of polyamine was performed. RESULTS: After 14C-polyamine instillation, radioactivity in the portal vein samples immediately increased and then decreased gradually. The absorptive pattern did not differ among the three polyamines. The recovery rates from radioactivity at the portal vein among the three polyamines were approximately 61-76% during the initial 10 min after the administration of 14C-polyamine, and were not different from each other. Aminoguanidine, which inhibits putrescine degradation, significantly suppressed initial putrescine uptake and recovery percentage. The intraluminal administration of spermine caused an increase in the immunoreactivity of the spermine antibody in the intestinal villi. CONCLUSION: Luminal polyamines were rapidly absorbed by the intestinal mucosa and then subsequently transferred into the portal vein using a rat ex vivo model. The prior administration of aminoguanidine significantly inhibited initial putrescine transport into the portal vein.     

Uptake of Extracellular, Dietary Putrescine Is an Important Regulatory Mechanism of Intracellular Polyamine Metabolism DuringCamostate-Induced Pancreatic Growth in Rats

Aim of the present study was to evaluate therole of cellular uptake of dietary [ H]putrescine forthe regulation of pancreatic, hepatic, and smallintestinal polyamine metabolism during normal andcamostate-induced pancreatic growth in rats in vivo. Initiallydose-response and time-course studies of[³H]putrescine uptake were performed. MaleWistar rats were either treated with the synthetictrypsin inhibitor camostate (200 mg/kg body wt orally twice daily),camostate plus the ornithine decarboxylase inhibitoralpha-difluoromethylornithine (DFMO) (2% in drinkingwater plus 3 × 300 mg/kg body wt intraperitoneallyduring daytime) or saline as controls. After 4, 8, 12,24, 36, 48, or 120 hr, five to seven animals per groupwere killed, respectively. Orally fed [ H]putrescine (10nmol/kg body wt. 2 hr prior to death) is rapidly taken up and further metabolized tospermidine in normal growing pancreas, liver, and smallintestine. Feeding of camostate significantly enhanceddietary [³H]putrescine uptake, whilesimultaneous inhibition of de novo synthesis ofintracellular polyamines by DFMO resulted in a highlysignificant further increase in cellular uptake oforally fed [³H]putrescine, which isimmediately metabolized to spermidine. The present in vivo data confirmthe important role of dietary putrescine uptake for themaintenance of intracellular polyamine pool in normaland stimulated pancreatic growth. Furthermore, dietary putrescine uptake is an importantregulatory mechanism to maintain the normal andgrowth-stimulated cellular polyamine pool in thepancreas after potent simultaneous inhibition ofintracellular de novo polyamine synthesis.     

Ethanol-Associated Alterations in the Kinetics of Putrescine Uptake and Metabolism by the Regenerating Liver

Biosynthesis of the polyamines, putrescine, spermidine, and spermine is required for DNA synthesis and liver regeneration after partial hepatectomy. We have previously reported that chronic ethanol consumption impairs polyamine synthesis and significantly retards liver regeneration after partial hepatectomy. In those studies, supplementation with putrescine restored hepatic DNA synthesis in ethanol-fed rats but exerted no effect in pair-fed controls. These differences in the response to putrescine treatment may have resulted from ethanol-associated differences in hepatic uptake, release, or metabolism of putrescine. To resolve these issues and define more completely how putrescine treatment affects DNA synthesis, we now assess the kinetics of putrescine uptake and metabolism after intraperitoneal or intravenous injection of radiolabeled putrescine (1.2 mmol/kg, specific activity 1 microCi/mmol) into rats fed 36% ethanol diets or isocaloric, nonethanol diets for 6 weeks prior to partial hepatectomy. After putrescine treatment, hepatic putrescine concentrations were greater in ethanol-fed rats than controls. Differences in post-treatment hepatic putrescine levels between ethanol and pair-fed groups could not be explained by differences in the rates of hepatic putrescine uptake or excretion into bile; residual de novo synthesis of putrescine from ornithine or metabolism of hepatic putrescine to its polyamine products, spermidine and spermine. Indeed, supplemental putrescine was not appreciably converted to spermidine or spermine in either ethanol or control rats. Hence, these latter polyamines are unlikely to be responsible for the treatment-associated improvement in DNA synthesis that has been noted in ethanol-fed rats. This suggests that putrescine itself acts to restore hepatic DNA synthesis in ethanol-fed rats.     

EGF-stimulated polyamine accumulation in the colon carcinoma cell line, Caco-2

BACKGROUND: Polyamines (putrescine, spermidine and spermine) are ubiquitous molecules indispensable for cell proliferation. In the intestinal lumen they are present in high amounts. Polyamine accumulation in proliferating cells of the intestinal mucosa is high, and it occurs both by enhanced synthesis and by increased uptake from the lumen. AIMS: To study mitogen-induced polyamine accumulation in the gut, we treated proliferating Caco-2 cells with epidermal growth factor (EGF) and measured the activity of ornithine decarboxylase (ODC) and putrescine uptake. Furthermore, we investigated whether EGF-induced changes in the apical membrane could be responsible for the effect of EGF on polyamine uptake in Caco-2 cells. METHODS: Putrescine uptake, ODC activity and intracellular polyamine content were evaluated in the presence of 100 ng/ml EGF. To study the mechanisms of EGF-stimulated polyamine uptake, apical membrane vesicles were isolated, and putrescine uptake into the vesicles measured. Possible enrichment in brush border membrane cytoskeleton proteins (ezrin and villin) was assessed by Western blot. RESULTS: Treatment with EGF induced an increase in ODC activity, which occurred within the first minutes of treatment and reached peak values after 3 h. In contrast, an increase in putrescine uptake was more sustained, with peak levels at 12 h. Both synthesis and uptake contributed to an over 60% increase in intracellular putrescine and spermidine after EGF treatment. There were no detectable changes in apical membrane cytoskeleton (as concluded by the absence of ezrin and villin enrichment in EGF-treated Caco-2 cells). However, in apical membrane vesicles isolated from EGF-pretreated cells, putrescine uptake was enhanced twofold. CONCLUSIONS: EGF stimulates both synthesis and uptake of polyamines in Caco-2 cells. Enhanced synthesis seems to ensure rapid supply with polyamines in the earliest stages of growth, while the uptake is responsible for the maintenance of high polyamine intracellular levels during late growth phases. EGF-stimulated polyamine uptake is apparently not a consequence of structural changes in the apical membrane, but is likely to occur by a distinct EGF-induced alteration of the polyamine transporter itself.     

Increased polyamine levels of normal-appearing mucosa and cancers in DMH induced cancer-bearing colon in rats]

Polyamine levels (putrescine, spermidine, spermine) in normal-appearing colonic mucosa of DMH administrated rats were measured in order to assess their importance as markers of precancerous changes. Mean putrescine, spermidine and spermine levels of normal-appearing mucosa were more than three times mean putrescine, more than twice mean spermidine and more than 1.5 times mean spermine levels of normal colonic mucosa. Mean polyamine levels of colon cancers were higher than those of normal-appearing mucosa but only spermidine level was significantly different between them. The mucosal polyamine levels may be a good biochemical marker to detect precancerous changes. There was no correlation between the polyamine levels and the growth rate of the colon cancers.     

Hyperoxic lung injury and polyamine biosynthesis. Age-related differences

To determine if lung cell replication and repair might be different between younger (30-day-old) and older (60-day-old) rats, we studied polyamine and DNA biosynthesis in rats exposed to 1.0 atm oxygen for 24, 48, 56, or 72 h. By 24 h, no statistically significant changes were observed, but by 48 h, ornithine decarboxylase and putrescine increased; S-adenosylmethionine decarboxylase activity increased by 56 h in the younger rats but not in the older rats. By 72 h, spermidine, [3H]thymidine incorporation, and the labeling index of cells in the alveolar zone had increased only in the younger rats. During the first 56 h, hyperoxia inhibited DNA synthesis. We conclude that hyperoxia initially suppresses lung cell replication but subsequently, if the rat survives, there are increases in polyamine biosynthesis and cell replication that may be important for the development of oxygen tolerance.     

Trophic effects of pentagastrin on gastrointestinal tract in fed and fasted rats

In rats injected with pentagastrin (2-500 micrograms/kg subcutaneously) in saline, peak gastric acid responses occurred after 31 (30-min output) or 63 micrograms/kg (60-min output). Rats were fed or fasted for 48 h and injected every 8 h with 63, 250, or 1000 micrograms/kg pentagastrin. Fasting decreased body weight (30% versus fed), serum gastrin (90%), and weight and protein content of oxyntic and pyloric gland areas, pancreas, small intestine, and colon. Deoxyribonucleic acid content or [3H]thymidine incorporation was decreased in all organs except colon. The lowest dose of pentagastrin significantly increased [3H]thymidine incorporation in the oxyntic gland area of fed rats and the small intestine of fasted rats, although organ weight, protein, or deoxyribonucleic acid content did not increase. These data indicate that short-term fasting has dramatic effects on gastrointestinal organ growth in rats and that pentagastrin reverses some of these changes.     

Prolactin and polyamine catabolism: specific effect on polyamine oxidase activity in rat thymus

Much evidence suggests that prolactin has an immunoregulatory function and that its effects on cells of the immune system depend on the level and specific forms of the receptors present on the target cells. The effect of administration of prolactin on polyamine catabolism was investigated in thymus of male intact rats by measuring the activities of spermidine/spermine N(1)-acetyltransferase and polyamine oxidase, because of the relationships between polyamines (especially putrescine) and the immune system. The administration of prolactin to rats resulted in the rapid induction of spermidine/spermine N(1)-acetyltransferase activity in the thymus (1.6-times the level of control rats, within 4 h), and in a marked decrease in polyamine oxidase activity at 24 h. The changes in enzyme activities were accompanied by an increase in putrescine concentration and a decrease in spermidine and spermine concentrations. In the spleen, prolactin increased SAT activity only 24 h after administration and was ineffective on PAO activity.     

Effects of feeding, fasting, and caerulein treatment on ornithine decarboxylase in rat pancreas

Ornithine decarboxylase (ODC) is the rate-limiting enzyme in polyamine biosynthesis. We examined circadian variations in pancreatic ODC activity and time-course effects of caerulein in fed and fasted rats. Significant circadian variations in amount of ODC activity were observed. The highest values were obtained during the dark period (1855 +/- 406 pmoles CO2/h), and the lowest during the light period (359 +/- 84 pmoles CO2/h). Caerulein treatment induced hypertrophy and hyperplasia of the pancreas in fed rats; increases in pancreatic ODC activity preceded the rise in protein and DNA contents (447 +/- 44 pmoles CO2/h and 5573 +/- 893 pmoles CO2/h, 6 and 12 h after the first injection of caerulein, respectively). In fasted rats, pancreatic ODC activity was very low (149 +/- 37 pmoles CO2/h) and caerulein treatment induced a transient increase in this activity 12 h after the first injection; hypertrophy but not hyperplasia of the pancreas was observed. In caerulein-treated fasted rats, refeeding during the night following a 48 h fasting period was not enough to increase either ODC activity or DNA content. These findings demonstrate that nutritional status is an important factor in the regulation of ODC activity and, thereby, in caerulein-induced pancreatic growth.     

Role of polyamines in isoproterenol-induced cardiac hypertrophy: effects of alpha-difluoromethylornithine, an irreversible inhibitor of ornithine decarboxylase

DFMO is a selective irreversible inhibitor of ornithine decarboxylase (ODC), the initial enzyme in the polyamine biosynthetic pathway. DFMO was utilized to determine the role of polyamines in isoproterenol (ISO)-induced cardiac hypertrophy. Daily subcutaneous administration of 200 mg/kg of DFMO reduced cardiac putrescine levels but did not significantly alter the basal levels of spermidine or spermine, nor was normal cardiac growth affected. ISO-induced cardiac hypertrophy was accompanied by increased putrescine and spermidine levels but spermine was not significantly altered. DFMO reversed the ISO-induced increases in putrescine and inhibited or attenuated both the increases in spermidine content and the cardiac hypertrophy. Although normal ODC activity appears not to be necessary for the maintenance of basal levels of polyamines or for normal cardiac growth, sustained inhibition of ODC interferes with ISO-induced elevations of putrescine, spermidine and heart weight.     

The role of polyamines in growth factor induced DNA synthesis in cultured rat hepatocytes.

BACKGROUND/AIMS: Hepatocyte growth factor and transforming growth factor-alpha are growth factors with important roles in hepatocyte proliferation. The polyamines, putrescine, spermidine, and spermine are widely distributed in many different cells and play an essential role in cell growth and differentiation. The present study examined the role of polyamine in this growth promoting factor-induced hepatocyte proliferation, in primary cultured rat hepatocytes. METHODOLOGY: Hepatocytes were isolated from rats by the collagenase perfusion method. Ornithine decarboxylase and S-adenosylmethionine decarboxylase activities were measured as the release of 14CO2 from L-[-14C]ornithine and S-adenosyl-L-[carboxyl14C]methionine, respectively. The concentration of polyamine was analyzed by high performance liquid chromatography. RESULTS: When transforming growth factor-alpha and hepatocyte growth factor were added to the hepatocyte culture simultaneously, ornithine decarboxylase activity, S-adenosylmethionine decarboxylase activity, polyamine concentration and DNA synthesis increased additively. The increase in DNA synthesis caused by transforming growth factor-alpha, hepatocyte growth factor, or both was completely inhibited by alpha-difluoromethylornithine and methylglyoxal bis(guanylhydrazone). The inhibition was reversed by exogenous spermidine or spermine, but not by putrescine. CONCLUSIONS: Increased spermidine or spermine levels are essential for hepatocyte proliferation in cultured rat hepatocytes.     

Characteristics of putrescine uptake by human brush border membrane vesicles.

Both putrescine and the polyamines spermidine and spermine are essential factors for growth and differentiation in all cells of higher eucaryotes. In principle, increased requirements of polyamines in mucosal cells either can be met by de novo-synthesis or by increasing the uptake from lumen (brush border membrane) or bloodstream (basolateral membrane). We therefore evaluated putrescine uptake in intestinal mucosal cells by using human brush border membrane vesicles (BBMV). Intravesicular uptake of putrescine was shown by osmoplots. This process was not saturable over a substrate range from 1 to 80 microM. Putrescine transport was also found to be independent of temperature (Q10 = 1.23). No differences in putrescine uptake rates were found in the presence or absence of Na+, and there was no evidence for any dependence of putrescine uptake from other cations. Our data indicate that putrescine uptake by human intestinal brush border membrane vesicles occurs by passive diffusion. It is concluded that a formerly described saturable and carrier mediated uptake in isolated intestinal mucosal cells from different species is probably influenced by active transport across the basolateral membranes. Therefore, further studies with isolated basolateral membranes are advocated.     

The influence of plasma concentrations of tri-iodothyronine on the acute increases in insulin and muscle protein synthesis in the refed fasted rat

To examine whether the low plasma levels of triiodothyronine (T3) in fasted rats might limit the recovery of muscle protein synthesis on refeeding, rats were fasted for either 3 or 4 days and refed with or without pretreatment with thyroid hormones. Fasting suppressed T3 levels, plasma insulin and the rate of the translational phase of muscle protein synthesis (KRNA; the rate per unit RNA), especially after the 4-day fast. On refeeding, plasma T3 levels remained low for more than 3 h after the 3-day fast and for more than 8 h after the 4-day fast. Insulin concentrations increased within the first hour of refeeding, eventually achieving supranormal concentrations after the 3-day fast. The KRNA increased within the first hour of refeeding, achieving well-fed control values by 3 h after the 3-day fast or 24 h after the 4-day fast. The increases in KRNA were significantly correlated with the increases in insulin at low insulin concentrations, achieving a plateau value at 150 pmol/l, so that further increases in insulin were not associated with any further increases in protein synthesis. Pretreatment with thyroid hormone induced increased T3 levels which were maintained for up to 8 h of refeeding. This had no effect on the responses of either insulin or protein synthesis to refeeding after the 3-day fast, but did result in an acceleration of the recovery in the KRNA and plasma insulin levels in the rats fasted for 4 days. Analysis of the insulin-KRNA relationship showed no evidence for any increase in the insulin sensitivity of muscle protein synthesis with thyroid pretreatment, the initial stimulation of protein synthesis on refeeding the rats fasted for 4 days reflecting increased insulin secretion. Since in the untreated animals, insulin secretion on refeeding was also correlated with T3 levels, these results are consistent with the previously reported thyroidal dependence of insulin secretion.     

Transepithelial transport of putrescine across monolayers of the human intestinal epithelial cell line, Caco-2

AIM: To study the transepithelial transport characteristics of the polyamine putrescine in human intestinal Caco-2 cell monolayers to elucidate the mechanisms of the putrescine intestinal absorption. METHODS: The transepithelial transport and the cellular accumulation of putrescine was measured using Caco-2 cell monolayers grown on permeable filters. RESULTS: Transepithelial transport of putrescine in physiological concentrations (> 0.5 mM) from the apical to basolateral side was linear. Intracellular accumulation of putrescine was higher in confluent than in fully differentiated Caco-2 cells, but still negligible (less than 0.5%) of the overall transport across the monolayers in apical to basolateral direction.EGF enhanced putrescine accumulation in Caco-2 cells by four fold, as well as putrescine conversion to spermidine and spermine by enhancing the activity of S adenosylmethionine decarboxylase. However, EGF did not have any significant influence on putrescine flux across the Caco-2 cell monolayers. Excretion of putrescine from Caco-2 cells into the basolateral medium did not exceed 50 picomoles, while putrescine passive flux from the apical to the basolateral chamber, contributed hundreds of micromoles polyamines to the basolateral chamber. CONCLUSION :Transepithelial transport of putrescine across Caco2 cell monolayers occurs in passive diffusion, and is not influenced when epithelial cells are stimulated to proliferate by a potent mitogen such as EGF.     

Stimulation of hepatic and renal ornithine decarboxylase activity by selected amino acids

The activity of ornithine decarboxylase (ODC), the rate-limiting enzyme in polyamine synthesis, increases after a protein meal. The effect of amino acid mixtures on hepatic and renal ODC activity and polyamine content was studied in postabsorptive and 72-hour fasted rats. Fasting decreased ODC activity in liver and in kidney by approximately 50%. Hepatic ODC activity increased tenfold 4 hours after intraperitoneal injection of either 1 g/kg of a synthetic mixture of 17 amino acids or of casein hydrolysate to fed rats and about 20-fold in fasted rats. Renal ODC activity increased four- and tenfold respectively. A mixture of glutamate, aspartate, and alanine at concentrations given in the hydrolysate reproduced the full amino acid effect. No amino acid was effective when given alone, nor were mixtures of the other amino acid constituents of the hydrolysate. Glutamate + alanine was ineffective as were glucose or various combinations of arginine, ornithine, aspartate and NH₃. Ornithine + glutamate or aspartate + glutamate were active but stimulated less than aspartate + glutamate + alanine. Hepatic and renal putrescine content increased in parallel with ODC activity. The data suggest that specific amino acids possess the full ODC-stimulating capability of a high quality protein and that polyamine synthesis is linked to urea cycle activity.     

Exercise before or after refeeding prevents refeeding-induced recovery of cell size after fasting with a different pattern of metabolic gene expressions in rat epididymal adipocytes

We investigated the effect of exercise before or after refeeding on cell size and on the expression of several messenger RNAs (mRNAs) involved in lipolysis and lipogenesis in fasted rat epididymal adipocytes. Fasting for 65 hours reduced the diameter of adipocytes to 72.0 microm from 78.4 microm in fed control rats, whereas refeeding for 1 or 2 days restored adipocyte size to 74.0 or 75.8 microm, respectively. Exercise before or after refeeding blocked refeeding-induced restoration of adipocyte size and led to adipocyte size similar to that observed after fasting. Fasting dramatically reduced expression of the fatty acid synthase mRNA, although expression of this gene returned to the control level after refeeding. However, exercise after but not before refeeding inhibited recovery of the expression of fatty acid synthase mRNA resulting from refeeding. In contrast, exercise before but not after refeeding led to enhanced expression of mRNAs encoding the hormone-sensitive lipase and beta(3)-aderenoceptor. Thus, exercise before or after refeeding prevents refeeding-induced restoration of adipocyte size after fasting via different pathways. Exercise before and after refeeding enhanced the expression of lipolytic mRNAs or inhibited the expression of lipogenic mRNAs, respectively.