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Inhibition of TPN-associated intestinal mucosal atrophy with monoacetoacetin.

Total parenteral nutrition (TPN) is associated with intestinal mucosal atrophy. Acetoacetate is oxidized in preference to glucose by both enterocytes and colonocytes and is not present in TPN. The purpose of this study was to determine whether replacement of a portion of glucose calories with monoacetoacetin, the glycerol ester of acetoacetate, could inhibit TPN-associated intestinal atrophy. Male Sprague-Dawley rats (200-250 g) underwent superior vena caval cannulation and were assigned to receive chow ad libitium (CHOW), TPN with 0.86 M monoacetoacetin (ACAC), or TPN with 0.86 M glycerol to control for the glycerol component of monoacetoacetin (GLYC). Nitrogen balance was measured over 7 days after which time the animals were weighed and sacrificed. Jejunal and colonic segments were harvested and the mucosal weight, protein, RNA, and DNA contents measured. All groups showed comparable weight gain. Cumulative nitrogen balance was positive for both TPN groups. Significant decreases in mucosal parameters occurred in both TPN groups compared to the CHOW group, but atrophy was significantly inhibited in both jejunum and colon of the ACAC group compared to the GLYC group. Thus, the substitution of monoacetoacetin for glucose calories in parenteral nutrition solutions inhibited TPN-related atrophy of intestinal mucosa while maintaining normal growth.

Acetoacetates↗

Interaction of ketosis and liver regeneration in the rat.

Monoacetoacetin, the monoglyceride of acetoacetate, was investigated as a nutritional support for the regenerating liver. Following partial hepatectomy, rats were either fed an oral diet ad libitum or administered by total parenteral feeding glucose alone, monoacetoacetin-glucose mixture, or lipid emulsion-glucose for the nonprotein calories. Five rats from each treatment were killed at 6-hr intervals beginning 12 hr after partial hepatectomy and ending at 72 hr. The number of cells synthesizing DNA and the number of cells in mitosis were compared. Rats fed orally or infused with glucose alone or with lipid emulsion had similar parameters throughout. Rats infused with monoacetoacetin had approximately double the number of cells in mitosis and DNA synthesis compared to the other treatments. This stimulation by monoacetoacetin persisted 72 hr. It was concluded from the data that acetoacetate was the agent responsible for increased DNA synthesis and mitosis, but the mechanism for the stimulation was not identified.

3-Hydroxybutyric Acid↗

Substrate alterations in a sarcoma-bearing rat model: effect of tumor growth and resection.

The time course of plasma substrate level alterations in the tumor-bearing (TB) host and their relationship to tumor burden or weight loss has not been well defined. Plasma glucose, lactate, pyruvate, cholesterol, triglycerides, acetoacetate, beta-hydroxy butyrate, and glycerol levels were measured in postabsorptive TB and nontumor-bearing (NTB) male Fisher 344 rats at weekly intervals for 5 weeks. Chronically starved rts (CS) had plasma substrates measured at Week 3 and compared with TB and NTB levels. Tumor burden was 0.5% of body weight at Week 1, 3.5% at Week 2, 12% at Week 3, and 20% at Week 5. Glucose levels were significantly lower in TB versus NTB (P = 0.01) at Week 1 and progressively declined in TB versus NTB through Week 5 (P less than 0.001). At Week 2, plasma triglycerides, cholesterol, acetoacetate, and beta-hydroxy butyrate were significantly elevated in TB versus NTB (P less than 0.001) and increased further to Week 5. The profile of substrate changes in TB and CS, when compared to NTB rats at Week 3, were different, suggesting that the plasma substrate alterations seen in the TB state were not secondary to starvation. In addition, when tumors were excised from TB animals at Week 3 and plasma substrates were determined 1 week later, all substrates that were different between TB and NTB animals, except cholesterol, returned to NTB levels when compared to sham-operated controls.(ABSTRACT TRUNCATED AT 250 WORDS)

3-Hydroxybutyric Acid↗

The antiketogenic effect of alanine in normal man: evidence for an alanine-ketone body cycle.

The effect of alanine on ketone body levels, independent of hormonal changes, in normal man has been investigated. Five normal subjects were given somatostatin infusions (200 micrograms/hour) for 3 hr. After 1 hr alanine or isotonic saline was infused for 2 hr. With saline blood beta-hydroxybutyrate and acetoacetate levels rose steadily to a peak of 0.230 plus or minus 0.053 and 0.112 plus or minus 0.023 mmole/l respectively. With alanine beta-hydroxybutyrate and acetoacetate levels plateaued at 0.099 plus or minus 0.020 and 0.055 plus or minus 0.006 mmole/l respectively. Alanine levels reached nearly 1 mmole/l but a significant effect on ketone body levels was apparent at physiologic levels (less than 0.6 mmole/l). Plasma fatty acid and glycerol levels did not change significantly. Insulin C-peptide and glucagon levels were suppressed to a similar extent in both experiments. These results support the view that alanine suppresses ketogenesis in man by a direct hepatic effect independent of insulin and glucagon. It is suggested that this forms part of a negative feedback substrate cycle between alanine and ketone bodies.

3-Hydroxybutyric Acid↗

Glucose and ketone body turnover in carnitine-palmitoyl-transferase deficiency.

Most of the patients with carnitine-palmitoyl-transferase deficiency (CPT) show reduced levels of blood ketone bodies in the postabsorptive state. In the present study, we have evaluated ketone body and glucose kinetics in patients with CPT deficiency. Intermediate metabolites of carbohydrate and lipid metabolism have also been studied. Ketone body (KB) turnover was measured by means of sequential intravenous bolus injections of 3-14C acetoacetate and 3-14C D(-) 3-hydroxybutyrate in four patients with liver, platelet, and muscle deficiency of CPT system and in eight normal overnight fasting subjects. 6-3H glucose was also injected, along with 3-14C ketone bodies to measure glucose turnover rate. Three out of four CPT deficiency patients had normal KB turnover, despite a marked reduction in liver CPT activity. Only one subject, with severe defect of CPT activity in liver, showed a significantly reduced, but still present rate of de novo synthesis of acetoacetate and 3-hydroxybutyrate (40 and 51 mumol/m-2/min-1 respectively) in comparison with control subjects (103 +/- 14 and 157 +/- 22 mumol/m-2/min-1). Blood concentrations of dicarboxylic adipic and suberic acids were significantly higher in CPT deficiency patients (0.035 +/- 0.007 and 0.021 +/- 0.005, mmol/L respectively) than in control subjects (0.008 +/- 0.008 and 0.006 +/- 0.003 respectively). Basal glucose turnover was increased in CPT deficiency patients (505 +/- 13 mumol/m-2/min-1) in comparison with normal subjects (433 +/- 18 mumol/m-2/min-1; P less than .01) as well as clearance rates (127 +/- 3 mL/m-2/min-1 and 91 +/- 11 mL/m-2/min-1, respectively; P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)

3-Hydroxybutyric Acid↗

The effects of changes in plasma nonesterified fatty acid levels on oxidative metabolism during moderate exercise in patients with non-insulin-dependent diabetes mellitus.

Blood levels of intermediary metabolites were measured and indirect calorimetry was performed in 10 otherwise healthy, non-insulin-dependent diabetic (NIDDM) patients before, during, and after 30 minutes of moderate exercise on three occasions in random order at weekly intervals with (1) heparin treatment to increase preexercise plasma nonesterified fatty acid (NEFA) levels (HEPARIN); (2) acipimox, a nicotinic acid analogue, to reduce preexercise plasma NEFA levels (ACIPIMOX); and (3) no manipulation of preexercise plasma NEFA levels (NIL). With ACIPIMOX, preexercise blood levels were significantly reduced for NEFAs and glycerol (P < .01) and marginally reduced for acetoacetate and 3-hydroxybutyrate (NS) compared with preexercise levels for the other two treatments; these low levels seen with acipimox treatment increased only slightly during exercise and the postexercise period. Plasma NEFA levels increased by approximately 150% (P < .001) with HEPARIN at the same times. The levels of ketone bodies during either NIL or HEPARIN increased rapidly postexercise by approximately 90% to 110% for both acetoacetate and 3-hydroxybutyrate (both P < .01). Plasma insulin levels tended to be lowest (despite similar plasma glucose levels during the three treatments) with ACIPIMOX, while growth hormone (hGH) and, perhaps, noradrenaline levels were highest both during and after exercise. The respiratory quotient (RQ) was highest with ACIPIMOX (P < .05 for exercise and postexercise periods compared with the other two treatments), which, compared with NIL, reduced fat oxidation by 27% and 60% and increased carbohydrate oxidation by 29% and 74% during and after exercise, respectively (all P < .05). These changes in substrate oxidation due to ACIPIMOX were almost opposite to those observed with HEPARIN.(ABSTRACT TRUNCATED AT 250 WORDS)

3-Hydroxybutyric Acid↗

Temporary and selective anosmia in tiger salamanders (Ambystoma tigrinum) caused by chemical treatment of the olfactory epithelium.

Tiger salamanders (Ambystoma tigrinum) were trained to respond to two dissimilar odorants (i.e., cyclohexanone, dimethyl disulfide) but not to a third odorant (n-butanol). When the training criterion was met, the animals were anesthetized and given nasal lavages of saline or odorants in saline. Lavage with cyclohexanone decreased responding to cyclohexanone but not dimethyl disulfide, and vice-versa, in later behavioral tests. Likewise, ethyl acetoacetate lavage produced selective response decrements to cyclohexanone. Lavage with n-butanol had no effect on responding to presentations of either cyclohexanone or dimethyl disulfide. The effects of lavage increased with increasing odorant concentration. Thus, lavage with 0.05 M cyclohexanone or ethyl acetoacetate produced more persistent response decrements than lavage with 0.01 M concentrations of either odorant. Such results are consistent with several testable interpretations, including the possibility the response decrements could reflect the nature of odorant-receptor interactions.

1-Butanol↗

Concentrations of ketone bodies in the blood of the green lizard Ameiva ameiva (Teiidae) in different physiological situations.

1. The concentrations of acetoacetate and 3-hydroxybutyrate have been determined in the blood of the green lizard Ameiva ameiva (Teiidae) in fed animals and in animals starved for periods from one week to about four months. 2. The concentrations of acetoacetate are low and unaltered in fed and starved animals, being in the range from 0.014 to 0.018 mM. 3. The concentrations of 3-hydroxybutyrate are high: 2.67 mM, in fed animals, falling during starvation down to 0.26 mM. 4. The 3-hydroxybutyrate/acetoacetate ratio is high, 151, in fed animals, falling down to 17. 5. The possible importance of ketone bodies in the metabolism of Ameiva ameiva is discussed.

3-Hydroxybutyric Acid↗

The ketogenic effect of glucose in rumen epithelium of ovine (Ovis aries) and bovine (Bos taurus) origin.

In experiments with rumen epithelium incubated in vitro the ratio of 3-hydroxybutyrate: acetoacetate produced was similar to the ratio reported for portal blood, and the ratio ketogenesis: oxidized to CO2 of butyrate was also close to values reported in vivo. Ovine and bovine epithelium incubated with butyrate differed significantly by the values of about 12-17 and 4-7 obtained for the ratio of 3-hydroxybutyrate: acetoacetate. Increasing levels of butyrate in the incubation medium resulted in a decreasing proportion of butyrate oxidized to CO2 and an increasing proportion of ketogenesis. The addition of glucose to butyrate in the incubation medium significantly increased the rate of ketogenesis from butyrate by ovine and bovine tissues. The addition of glucose to butyrate in the incubation medium significantly decreased the rate of butyrate oxidation to CO2 by ovine and bovine tissues. The ketogenic effect of glucose was also apparent in perfused rumen epithelium with butyrate at the mucosal side and glucose at the serosal side.

3-Hydroxybutyric Acid↗

Changes in ketone body utilization by chick liver, duodenal mucosa and kidney during embryonic and postnatal development.

1. Lipid synthesis from acetoacetate and 3-hydroxybutyrate in chick kidney and duodenal mucosa showed a clear decrease between 15 and 19 days of the embryonic phase, followed by an increase at hatching and a new decrease during the first neonatal period. The hepatic synthesis of lipids presented a different profile: a peak at 19-day-old embryo and a new increase during postnatal development. 2. These changes would be related to those in 3-hydroxybutyrate concentration in chick plasma throughout the perinatal period. 3. Phospholipids were the main kind of lipid formed in the three tissues. An appreciable percentage of radioactivity was also recovered as free cholesterol, especially during the embryonic phase. Triglycerides were also formed from acetoacetate in a high proportion in liver from neonatal animals. 4. Chick kidney showed the maximal ability to incorporate both precursors into amino acids. The peak obtained around hatching time would be related to the availability of the substrates. 5. Ketone body oxidation to CO2 was also maximal in kidney. In this tissue, a drastic decrease was observed during the final embryonic period, followed by a strong increase at day 1 after hatching and a new decrease at 4 days.

3-Hydroxybutyric Acid↗

Metabolic abnormalities of mitochondrial redox potential in postburn multiple system organ failure.

Forty-five burn patients underwent sequential assays for plasma acetoacetate and beta-hydroxybutyrate concentrations as well as plasma amino acid levels. Those patients who went on to develop multiple system organ failure were noted to have a decrease in their acetoacetate concentration with time, whereas there was no change in those patients who failed to develop multiple system organ failure. The plasma concentration of beta-hydroxybutyrate was not altered by multiple system organ failure. In addition, the plasma acetoacetate/beta-hydroxybutyrate ratio was found to be directly related to the plasma concentration of branched chain amino acids and inversely related to the concentration of aromatic amino acids.

Acetoacetates↗

Regulation of food intake by hepatic oxidative metabolism.

It has been proposed that the turnover of metabolic fuels affects feeding. In the present study the effects of different metabolites (glycerol, L-malate, D-3-hydroxybutyrate, L-lactate) or their oxidation products (dihydroxyacetone, oxaloacetate, acetoacetate, pyruvate) on food intake were investigated under various feeding conditions. The results of these experiments increase our knowledge about the nature and origin of metabolic signals in the energostatic control of food intake. In the experiments, glycerol, malate, 3-hydroxybutyrate, lactate or pyruvate (4.5-7.7 mmoles/kg metabolic body weight injected subcutaneously reduced food intake in rats, whereas dihydroxyacetone, acetoacetate or oxaloacetate did not. The effect of glycerol disappeared with high levels of dietary protein, whereas the effects of lactate and pyruvate disappeared with high levels of dietary fat. The hypophagic effects of glycerol, malate, 3-hydroxybutyrate, lactate and pyruvate were abolished by selective hepatic vagotomy. The results suggest that vagally mediated signals originating from hepatic oxidation of metabolic fuels contribute to the regulation of food intake.

3-Hydroxybutyric Acid↗

Influence of early chronic phenobarbital treatment on cerebral arteriovenous differences of glucose and ketone bodies in the developing rat.

The influence of an early chronic phenobarbital treatment on cerebral arteriovenous differences of glucose, lactate, pyruvate, beta-hydroxybutyrate and acetoacetate was studied in suckling rats. The animals were treated from day 2 to 21 after birth by a daily injection of 50 mg/kg phenobarbital or by saline and were studied at 10, 14 and 21 days. Phenobarbital treatment induced a decrease in cerebral arteriovenous difference of glucose at P14 and no change at P10 and P21. The barbiturate did not have any influence on cerebral arteriovenous difference of lactate and pyruvate at the three stages studied. Cerebral uptake of beta-hydroxybutyrate was unchanged at P10 and increased by two-fold at P14 and by threefold at P21 by phenobarbital. Cerebral arteriovenous difference of acetoacetate was low and did not change with the pharmacological treatment. At P14 and P21, the calculated amount of oxygen used by the brain for the oxidation of ketone bodies was twice as high in barbiturate- as in saline-treated rats and reached values of 47 and 16% respectively in phenobarbital-exposed animals. In addition, the barbiturate seemed to affect the carrier process of beta-hydroxybutyrate from blood to brain. The results of the present study are in good agreement with previous data from our laboratory showing that an early chronic phenobarbital treatment is able to induce a shift in the cerebral energy metabolism balance in favor of ketone bodies.

3-Hydroxybutyric Acid↗

Phosphorylation activity of the response regulator of the two-component signal transduction system AtoS-AtoC in E. coli.

Antizyme, long known to be a non-competitive inhibitor of ornithine decarboxylase, is encoded by the atoC gene in Escherichia coli. The present study reveals another role for AtoC, that of a response regulator of the AtoS-AtoC two component system regulating the expression of the atoDAEB operon upon acetoacetate induction. This operon encodes enzymes involved in short-chain fatty acid catabolism in E. coli. Evidence is presented to show that AtoS is a sensor kinase that together with AtoC constitutes a two-component signal transduction system. AtoS is a membrane protein which can autophosphorylate and then transfer that phosphoryl group to AtoC. This process can also be reproduced in vitro. AtoC contains in its amino acid sequence a conserved aspartic acid (D55), which is the putative phosphorylation site, as well as an unexpected "H box" consensus sequence (SHETRTPV), common to histidine kinases, with the histidine contained therein (H73) being a second potential target for phosphorylation. Substitution of either D55 or H73 in His10-AtoC diminished but did not abrogate AtoC phosphorylation suggesting that either both residues can be phosphorylated independently or that the phosphate group can be transferred between them. However, the D55 mutation in comparison to H73 had a more pronounced effect in vivo, on the activation of atoDAEB promoter after acetoacetate induction, although it was the presence of both mutations that rendered AtoC totally unresponsive to induction. These data provide evidence that the gene products of atoS and atoC constitute a two-component signal transduction system, with some unusual properties, involved in the regulation of the atoDAEB operon.

Acetoacetates↗

An acute hyperglycemia or acidosis-induced changes of indolamines level correlates with PKC-alpha expression in rat brain.

Hyperglycemia and ketoacidosis are the two most serious factors in acute metabolic complications of both type 1 and type 2 diabetes. Dysfunction of the central nervous system is a well-documented complication of diabetes. We and others have previously reported that acute or chronic diabetes in animal's results in altered brain neurotransmitter levels. In this study, we investigated the effects of acute (7 days) glucose-induced hyperglycemia and sodium acetoacetate (NaAcAc) or ammonium chloride (NH4Cl) induced acidosis on the level of indolamines (5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA)) as well as PKC-alpha expression/activity in discrete areas of rat brain. Glucose-induced (500 mg/kg, bw) hyperglycemic ( approximately 249 mg%) rats showed significant (p<0.05) increase in 5-HT levels in mid brain (MB), pons medulla (PM) and cerebellum (CB), respectively. 5-HIAA level increased in hippocampus (HC) (p<0.05) as compared to control. The rats treated with sodium acetoacetate (NaAcAc) for 7 days (60 mg/kg, bw) showed significant decrease (p<0.05) of 5-HT level in hypothalamus (HT). Whereas, the 5-HIAA level increased in MB (p<0.05). Similarly, the PKC-alpha expression as well as the enzyme activity showed significant increase in HC, MB, PM and CB under glucose-induced hyperglycemia and that changes correlated the changes of indolamines, suggesting that the hyperglycemia may be the major metabolic disorder in diabetic complications.

Acetoacetates↗

Daily changes in parameters of energy metabolism in brain of rainbow trout: dependence on feeding.

We assessed the daily patterns of parameters involved in energy metabolism in plasma and brain of rainbow trout. Where daily rhythms were found, we analyzed the potential influence of feeding. Immature rainbow trout were randomly distributed in 3 groups: fish fed for 7 days, fish fasted for 7 days, and fish fasted for 7 days and refed for 4 days. On sampling day, fish of fed and refed groups were fed at 11.00 h, and all fish were sampled from each treatment group using the following time schedule: 14.00, 18.00, 21.00, 00.00, 04.00, 07.00, 10.00 and 14.00 h. The results obtained from metabolic parameters assessed in plasma and brain can be grouped into three different categories, such as (i) those displaying no 24 h changes in fed fish such as plasma lactate, protein or acetoacetate levels, as well as brain amino acid and protein levels, and lowKm(glucose) hexokinase, and aspartate aminotransferase activities, (ii) those displaying 24 h changes that were apparently dependent on feeding since they disappeared in fasted fish such as the case of plasma cortisol, glucose and triglyceride levels, as well as brain glycogen, glucose, and lactate levels, and pyruvate kinase and hexokinase IV activities, and (iii) those parameters displaying 24 h changes apparently not dependent on feeding such as plasma amino acids, brain acetoacetate levels as well as several enzyme activities measured in brain such as glucose 6-phosphate dehydrogenase, alpha-glycerophosphate dehydrogenase, glutamate dehydrogenase, and lactate dehydrogenase-oxidase. In general, 24 h changes dependent on feeding indicate an increased use of glucose in brain several hours post-feeding whereas those changes not dependent on feeding were characterized by reduced levels/activity at the night period suggesting a metabolic depression in brain during darkness.

Acetoacetates↗

Derivatization of endogenous and exogenous compounds in plasma for high-performance liquid chromatographic analysis.

Methods are described for the analysis of endogenous (acetoacetic and pyruvic acids) and exogenous (propranolol) compounds in plasma. Acetoacetic and pyruvic acids were converted into 2,4-dinitrophenylhydrazones and identified by chromatography on a normal-phase silica gel column at 360 nm. The propranolol amino group was modified with dansyl chloride and the dansyl derivative, after extraction with a hydrophobic solvent, was analysed on a normal-phase silica gel column using fluorescence detection. The method allows the determination of 0.5 ng/ml of propranolol.

Acetoacetates↗

Effects of noncarbohydrate substrates on protein synthesis in hearts from fed and fasted rats.

An overnight fast reduced RNA content and resulted in lower rates and efficiency of protein synthesis when rat hearts were perfused in vitro and supplied glucose as oxidizable substrate. Decreased efficiency of synthesis was associated with development of a block in peptide chain initiation in hearts of both fed and fasted rats. Provision of pyruvate increased the rate and efficiency of protein synthesis in fasted but not fed tissue, and partially overcame the initiation block in both groups. A mixture of glucose, pyruvate and insulin increased the efficiency of protein synthesis and decreased ribosomal subunit content to similar values in both groups of hearts. Noncarbohydrate substrates, including pyruvate, lactate, acetoacetate and beta-hydroxybutyrate, supported higher rates of protein synthesis than glucose in hearts of fasted, but not fed rats. However, mixtures of glucose and either pyruvate, acetoacetate or beta-hydroxybutyrate increased the synthetic rate in fed tissue. Provision of noncarbohydrate substrates increased energy availability, as indicated by higher creatine-P/creatine ratios in both groups of hearts, but the synthetic rate increased as a function of creatine-P/creatine ratio only in the fasted tissue. Octanoate and leucine accelerated protein synthesis and increased energy availability in the fed tissue. The mixtures of glucose and noncarbohydrate substrates or octanoate elevated glucose-6-P content. These studies indicate that an overnight fast decreased the capacity for protein synthesis and modified the regulation of synthesis by noncarbohydrate substrates.

3-Hydroxybutyric Acid↗