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Histochemical and biochemical investigations concerning the function of larval oenocytes of Tenebrio molitor L. (Coleoptera, Insecta).

Larval oenocytes of Tenebrio molitor were investigated histochemically. In contrast to the lipid droplets of the fat body, they did not stain wit Sudan black. A positive reaction for lipoproteins appeared only after destructive oxidation with sodium hypochlorite. These lipoproteins are the remnants of degenerated membranes, as revealed by ultrastructural analysis. Polyphenols could be identified in th exocuticle of exuvia, and in the newly formed procuticle. Endocuticle, epidermis and oenocytes showed no staining reaction. In oenocytes a great amount of lipase is also present which could be detected with several Tweens as substrates. The significance of these lipases remains unclear, since only few glycerides are synthesized in the cells, as shown below. They may play a role in the extended membrane turnover observed in this cell type. In vitro incubation of oenocytes of the larval generation demonstrated that 14C-labeled acetate was only incorporated into the paraffin fraction. A negligible amount of the label was found in glycerides; wax esters were free of label. Larval epidermis is also capable of paraffin formation, but only to a small degree. Oenocytes of the imaginal generation located between the sternal epidermis cells of pupae and adults do not synthesize paraffins, but other more polar compounds not yet identified. Labeled waxes in cuticular lipids were detected only when 14C-acetate was injected into whole larvae, and the lipids extracted some hours later. Autoradiographs demonstrated that 14C-acetate was intensively incorporated into larval oenocytes, the rate varying in different cells. Incorporation into the epicuticle, probably into the wax layer, was clearly shown. Cuticulin and dense layer do not show an intensive label. The lamellated cuticle also seems to be impregnated with acetate derivatives.

Acetates↗

Estrogen and accessory sex gland lipids.

Administration of estradiol-17 beta elicited differential responses on accessory sex glands of rats. In caput epididymis, the estrogen treatment led to an accumulation of glycerides and phospholipids while in cauda epididymis, there was no significant change. However, in seminal vesicles, depletion of phospholipids was observed. In prostate, the treatment, resulted in an accumulation of glycerides.

Animals↗

Stimulatory effect of glucose upon triglyceride synthesis from acetate, decanoate, and palmitate by mammary gland slices from lactating mice.

Slices prepared from the mammary glands of lactating mice incorporate only small amounts of (1--14C) acetate, (1--14C) decanoate, or (1--14C) palmitate into lipids. However, when glucose is added to the incubation medium, fatty acid incorporation is stimulated-13-fold from acetate, 17-fold from decanoate, and 2-fold from palmitate. Over 90% of the -14C activity in the lipid fraction is in triglycerides. Analysis of fatty acids in the triglycerides showed that almost all of the decanoate and the palmate were incorporated as intact molecules, while acetate yielded acids of varying chain lengths. The glucose stimulation of triglyceride synthesis is not solely due to its effect upon chain elongation but also could involve glyceride-glycerol availability, as well as other unknown factors. However, from measurements of the amounts of glycerol 3-phosphate in the tissue incubated in the presence of palmitate, it would appear that glyceride-glycerol availability is not rate limiting in triglyceride synthesis.

Acetates↗

Cholesterol and fatty acid synthesis in swine.

In incubation studies with swine tissue slices, acetate-1-14C or glucose-U-14C as substrates were incorporated more readily into fatty acids and cholesterol in adipose tissue than other tissues tested. Cholesterol and fatty acid synthesizing acitivity was substantial in the small intestine. When acetate was available, liver, small intestine, and adipose tissue were important sites for cholesterol synthesis. Heart and aortic tissue had marginal levels of cholesterol synthesizing ability. Lipogenesis in adult swine liver, heart, and aortic tissue was extremely low. As in tissue slices, incorporation of acetyl-1-14C CoA into fatty acids by adipose homogenates indicated high lipogenic activity. Subcellular fractionations of heart and aortic tissue indicated that the heart microsomal fraction had the highest lipogenic activity as measured by the incroporation of acetyl-4-14C CoA into fatty acids. In adult swine adipose tissue, the incorporation of glucose-U-14C into fatty acid was higher than its incorporation into glyceride-glycerol. The synthesis of glyceride-glycerol from glucose-U-14C or acetate-1-14C in liver was higher than for fatty acid synthesis. The acitivity of acetyl CoA carboxylase, fatty acid synthetase, citrate cleavage enzyme, nicotinamide adenine dinucleotide phosphat-malate dehydrogenase, glucose-6-phosphate dehydrogenase, and 6-phosphogluconate dehydrogenase was considerably higher in adipose tissue than in other tissues tested, paralleling its high lipogenic capacity.

ATP Citrate (pro-S)-Lyase↗

Pathways of paracetamol absorption from layered excipient suppositories: artificial intelligence approach.

When studying paracetamol availability after rectal administration, the differences between slower and faster release suppositories were discovered. Approach with modelling and simulation of compartment-based models was used to explore the differences. A study of paracetamol from layered excipient suppositories shows that many different mechanisms are involved in the drug pharmacokinetics. There is also a large number of articles, each dealing with only one or with a few of the mechanisms. However, there is little information available on how the mechanisms interact in the organism and thus govern the pharmacokinetics of the drug, which means that systemic view in the expert knowledge is missing. In the case of paracetamol rectal availability the use of partially fuzzyfied model allowed systemic combination of all described mechanisms found in the literature and measured data. In spite of non-identifiability, the model showed that patterns that explained differences in bioavailabilities of the two formulations of suppositories could be found. Results of modelling and simulation show that "in vivo" there is practically no difference in cumulative release profiles between the two formulations. However, due to higher content of mono-di-glycerides in a slower release formulation, the extent of absorption is augmented both by absorption-enhancing effect of mono-di-glycerides and the liver bypass mechanism via diminished viscosity.

Acetaminophen↗

Ester synthesis in an aqueous environment by Streptococcus thermophilus and other dairy lactic acid bacteria.

The ability of Streptococcus thermophilus ST1 and 19 other dairy lactic acid bacteria (LAB) to synthesize esters was investigated in an aqueous environment. These LAB were able to synthesize esters from alcohols and glycerides via a transferase reaction (alcoholysis) in which fatty acyl groups from glycerides were transferred to alcohols. S. thermophilus ST1 was active on tributyrin and on di- or monoglycerides of up to C10 with ethanol as the acyl acceptor. This strain was also active on a diglyceride of C6 and monoglyceride of C8 with 2-phenyl ethanol as the acyl acceptor. Alcoholysis occurred preferentially over hydrolysis. S. thermophilus ST1 had an apparent K(m) value of 250 mM for ethanol and an apparent K(m) value of 1.3 mM for tributyrin, measured against whole cells. Around 80% of both the transferase activity and the esterase activity were detected in the cell-free extract (CFE) of strain ST1. Both activities in the CFEs of five LAB tested were, to a similar degree, enhanced slightly by growth in the presence of ethanol and tributyrin. Using tributyrin and ethanol as substrates, the transferase activities ranged over 0.006-1.37 units/mg cell dry weight among the LAB tested and were both species- and strain-dependent.

Alcohols↗

Retention of acyl groups in LM cell fibroblasts with altered phospholipid composition.

Incorporation and loss of membrane lipids were measured in murine fibroblasts grown in suspension culture with [2-14C]acetate in a chemically defined medium devoid of serum or lipid. Greater than 98 and 94% of the 14C present in the phospholipids and neutral glycerides respectively was found in the fatty acyl groups. During a 3-day growth period LM fibroblasts lost 65% of their 14C-labelled acyl groups from the phospholipids and 36% from the neutral lipids. Selective retention of 14C in individual isolated phospholipid and neutral lipid species occurred only in phosphatidylethanolamine. Retention of acyl groups in the phospholipids or neutral glycerides was not a property of LM suspension cultured fibroblasts. Supplementation of the LM cells with choline analogues, such as N,N'-dimethylethanolamine, N-monomethylethanolamine, or ethanolamine instead of choline, decreased the loss of 14C-labelled fatty acids from phospholipid and neutral lipid. Ethanolamine supplementation resulted in increased loss of 14C from desmesterol as well as decreased incorporation of 14C into desmosterol. Thus, polar head group manipulation affected acyl group composition, acyl group retention, and sterol metabolism in suspension cultured LM cells.

Acetates↗

Specificity of lipoprotein lipase and hepatic lipase toward monoacylglycerols varying in the acyl composition.

We report here that both the hepatic lipase and lipoprotein lipase demonstrate specificity towards the acyl group present on monoacylglycerols. We found that unsaturated glycerides are more readily degraded than saturated glycerides. However, the basis for this specificity appears to be different for each enzyme. The activity of the hepatic lipase, but not the lipoprotein lipase, could be stimulated by Triton X-100 and phosphoglycerides. We interpret these results to show that while both the lipoprotein lipase and hepatic lipase are sensitive to the physical state of the substrate (as shown by fluorescence depolarization), the lipoprotein lipase also has a low affinity for monoacylglycerols that contain a saturated acyl group. In the course of this study we also obtained evidence that some type of phase separation occurs when mixtures of saturated and unsaturated monoacylglycerols are prepared.

Animals↗

Decrease in lipogenesis and glucose oxidation of rat adipose tissue after chronic ethanol feeding.

This investigation was performed to determine whether chronic ethanol feeding affects adipose tissue lipogenesis and glucose metabolism. Female Wistar rats were pair-fed nutritionally adequate liquid diets containing ethanol as 36% of energy or an isocaloric amount of carbohydrate for 3 weeks. Chronic ethanol feeding resulted in a depression of adipose tissue lipogenesis as assessed by labeled glucose incorporation into glyceride glycerol and glyceride fatty acids. Glucose oxidation was also impaired after chronic ethanol feeding. Such changes may contribute to the postprandial hypertriacyglyceridemia observed in alcoholics.

Adipose Tissue↗

The major outer membrane lipoprotein and new lipoproteins share a common signal peptidase that exists in the cytoplasmic membrane of Escherichia coli.

The cell envelope of Escherichia coli possesses several lipoproteins including the major outer membrane lipoprotein. These lipoproteins are synthesized as a signal peptide-carrying precursor that is subsequently modified with glyceride. In this work, lipoprotein signal peptidase that processes the precursor of the major lipoprotein was partially purified from cells harboring a plasmid that carries the gene for this enzyme (1spA). The enzyme was also active against the glyceride-containing precursors of the peptidoglycan-associated lipoprotein and many additional membrane lipoproteins. The unmodified precursor of the major lipoprotein was not attacked by the enzyme. The enzyme was exclusively localized in the cytoplasmic membrane.

Bacterial Proteins↗

High resolution proton NMR studies of perfused rat hearts.

High resolution 1H NMR spectra of perfused rat hearts have been obtained under normoxic, ischemic and hypoxic conditions. Several myocardial metabolites including taurine, carnitine, lactate and tissue glycerides are detected in the 1H NMR spectra. Changes in oxygen availability induce perturbations in the levels of some metabolites, in particular, lactate. Experiments with fasted rats and with substrate-free perfusion suggest that the glycerides detected in 1H spectra are metabolically mobilizable but have a slow rate of turnover. These results demonstrate that utility of 1H NMR in monitoring myocardial metabolism.

Animals↗

Biosynthesis of lung lipids from acetoacetate and glucose in the developing rat in vivo.

1. [3-14C]Acetoacetate and [U-14C]glucose administered to rats were readily utilized for synthesis of phospholipids, cholesterol and glycerides in the lung during suckling period. 2. Among all lipids synthesized, phosphatidylcholine accounted for 31-48% from [14C]acetoacetate and 30-46% from [14C]glucose. The synthesized phosphatidylcholine contained predominantly [14C]-labeled palmitate. 3. Glycerol lipids represented 83-92% and 56-68% of total lipids synthesized from [14C]glucose and [14C]acetoacetate, respectively. 4. Glucose is the preferred substrate for glyceride-glycerol, while acetoacetate appears better for fatty acid and cholesterol synthesis.

Acetoacetates↗

Liver glucose, glycogen and lipid synthesis in fed and 24-hour fasted rats soon after a pulse of [3-14C]pyruvate through the portal vein.

1. A pulse of [3-14C]pyruvate was given to rats through the portal vein and blood was collected at brief intervals from the inferior cava vein at the level of the suprahepatic veins. 2. In 24 hr fasted rats, the appearance of [14C]glucose in blood and blood glucose specific radioactivity were higher than in fed animals from the first minute after delivery of the tracer. At this time total radioactivity did not differ between the two groups. 3. After 5 and 20 min. liver radioactivity present in glycogen and glyceride glycerol was enhanced while in fatty acids it was reduced in fasted as compared with fed animals. 4. It is proposed that, in the fasted state, both glycogen and glyceride glycerol synthesis are predominantly gluconeogenic processes.

Animals↗

Variations of glucose metabolism by fat cells from three adipose depots of the rat.

Fat cells isolated from the epididymal (E) and dorsal subcutaneous (S) depots from 150g male Wistar rats were similar in size, but differed markedly in glucose metabolism. Retroperitoneal (RP) fat cells were slightly larger but were metabolically similar to epididymal fat cells. Basal incorporation of [1-14C] glucose into fatty acids was lower in S than E and RP, CO2 production and glyceride-glycerol synthesis were similar in all three; and lactate production was increased in RP and S compared to E. S adipocytes exhibited a blunted respond to both submaximally and maximally-stimulating concentrations of insulin compared to RP and E adipocytes in glucose incorporation into fatty acids, CO2 and lactate production, but not glyceride-glycerol. Maximally insulin-stimulated fatty acid synthesis by S fat cells was 19% and 29% of the values in E and RP fat cells respectively. Basal and maximally insulin-stimulated glucose transport (2-deoxy [14C] glucose uptake) was depressed by 30%-40% in S cells compared to E and RP. Thus, the decreased basal glucose utilization of S could be attributed primarily to a decreased glucose transport capacity. The markedly lower insulin-stimulated glucose metabolism in S fat cells, however, may be explained by alterations of the capacities for both transport and intracellular metabolism. Subcutaneous fat cells were also somewhat less sensitive to submaximal doses of insulin and this was reflected in rightward shift in the dose-response curves for 2-deoxyglucose uptake and fatty acid synthesis. The decreases in insulin stimulated glucose oxidation and fatty acid synthesis were paralleled by decreases in the major lipogenic enzymes. Although the reason for these variations in the capacity for glucose metabolism among depots is unknown, they are important in assessing the metabolic function of the whole adipose organ.

Adipose Tissue↗

Brown adipose tissue metabolism in hypothalamic-obese rats.

Studies were performed to evaluate the metabolic changes of brown adipose tissue (BAT) in rats with hypothalamic obesity (VMNL). In vitro 14C-palmitate oxidation and incorporation into triglycerides were similar in VMNL and control rats. However, protein and fatty acid content and incorporation of 14C-palmitate into phospholipid were significantly less in both hyperphagic and normophagic VMNL rats. In order to assess in vivo BAT lipogenesis, rats were injected with 3H2O. Plasma H2O incorporation into BAT lipids was significantly greater in VMNL rats. Likewise, BAT lipid content was higher in obese rats. In another experiment BAT was incubated with U-14C-glucose to evaluate glucose utilization by BAT. 14C-glucose was oxidized and incorporated into both lipids and glycogen more rapidly by obese than by normal rat BAT. Glycogen content was greater in VMNL rats. Tissues were also incubated with 1-14C-pyruvate and 2-14C pyruvate. Pyruvate incorporation into glyceride glycerol and oxidation of 2-14C pyruvate through the Krebs cycle were similar in both obese and control rats. However, the incorporation of pyruvate into glyceride fatty acids was increased in VMNL rats. The results indicate that both fatty acid and lipid synthesis are increased in BAT of obese rats whereas lactate production is decreased and Krebs cycle activity is normal. Some of these changes appear to be independent of the level of food intake.

Adipose Tissue, Brown↗

Hypothalamic regulation of lipid metabolism in the rat: effect of hypothalamic stimulation on lipogenesis.

The rates of fatty acid synthesis and syntheses of other lipids were measured in vivo by determining 3H-incorporation from tritiated water or 14C-incorporation from [U-14C]glucose into white adipose tissues (WAT), brown adipose tissue (BAT) and liver, during electrical stimulation of the ventromedial hypothalamic nucleus (VMH) and the lateral hypothalamic nucleus (LH) of female rats. Electrical stimulation of the VMH markedly increased the rate of fatty acid synthesis and the conversion of glucose to total lipids, glyceride glycerol and phospholipids in interscapular BAT, but not in parametrial or retroperitoneal WAT. Conversely, on VMH stimulation, the syntheses of total lipids, glyceride glycerol and phospholipids from glucose in the liver decreased slightly, though not significantly. Electrical stimulation of the LH had no such effect. Administration of insulin increased the rate of fatty acid synthesis in both brown and white adipose tissues and in the liver. The rate of disappearance of [14C]glucose and the concentration of insulin in the blood were not changed significantly by stimulation of the VMH. These data indicate that electrical stimulation of the VMH, but not the LH, enhances lipogenesis in BAT preferentially through a mechanism not involving insulin, but probably through activation of sympathetic innervation of BAT. The physiological significance of the findings is discussed in relation to hypothalamic function in controlling non-shivering thermogenesis and obesity.

Adipose Tissue↗

Interaction between insulin and thyroid hormones on the control of carbohydrate and lipid metabolism in rat adipose tissue.

Adipose tissue segments excised from normal rats and from rats rendered experimentally hypothyroid and hyperthyroid retained insulin responsiveness when studied in vitro. Basal rates of glucose oxidation to CO2, conversion into glyceride-glycerol, fatty acids, and total lipids, and the activities of pyruvate dehydrogenase and fatty acid synthetase were enhanced in fat pads from hyperthyroid rats when compared with values seen with tissue from euthyroid animals. The response of each of these parameters was further enhanced by treating tissue from hyperthyroid rats in vitro with insulin. Basal rates of glucose oxidation and the activities of pyruvate dehydrogenase and fatty acid synthetase were depressed as a result of hypothyroidism. However, all of these values could be restored to levels approaching the values seen in the basal state for euthyroid rats when tissue segments from hypothyroid rats were incubated in vitro with insulin. The basal rates for glucose conversion into glyceride-glycerol, fatty acids, and total lipids were not changed by hypothyroidism but retained insulin responsiveness. These data suggest that the insulin-effector system in adipose tissue is not altered by thyroid status and that thyroid hormones may act independently with insulin to regulate glucose and lipid metabolism in this tissue at multiple intracellular metabolic sites.

Adipose Tissue↗

Liquid filled nanoparticles as a drug delivery tool for protein therapeutics.

In the present study, an attempt was made to study the feasibility of nanoparticulate adsorbents in the presence of an absorption enhancer, as a drug delivery tool for the administration of erythropoietin (EPO) to the small intestine. Liquid filled nano- and micro-particles (LFNPS/LFMPS) were prepared using solid adsorbents such as porous silicon dioxide (Sylysia 550), carbon nanotubes (CNTs), carbon nanohorns, fullerene, charcoal and bamboo charcoal. Surfactants such as a saturated polyglycolysed C8-C18 glyceride (Gelucire 44/14), PEG-8 capryl/caprylic acid glycerides (Labrasol) and polyoxyethylene hydrogenated castor oil derivative (HCO-60) were used as an absorption enhancer at 50mg/kg along with casein/lactoferrin as enzyme inhibitors. The absorption of EPO was studied by measuring serum EPO levels by an ELISA method after small intestinal administration of EPO-LFNPS preparation to rats at the EPO dose level of 100 IU/kg. Among the adsorbents studied, CNTs showed the highest serum EPO level of 62.7 +/- 11.6 mIU/ml. In addition, with the use of casein, EPO absorption was improved, C(max) 143.1 +/- 15.2 mIU/ml. Labrasol showed the highest absorption enhancing effect after intra-jejunum administration than Gelucire 44/14 and HCO-60, 25.6 +/- 3.2 and 22.2 +/- 3.6 mIU/ml, respectively. Jejunum was found to be the best absorption site for the absorption of EPO from LFNPS. The use of CNTs as LFNPS, improved the bioavailability of EPO to 11.5% following intra-small intestinal administration.

Absorption↗