Effects of Liposome and Cardiolipin on Folding and Function of Mitochondrial Erv1

Erv1 (EC number 1.8.3.2) is an essential mitochondrial enzyme catalyzing protein import and oxidative folding in the mitochondrial intermembrane space. Erv1 has both oxidase and cytochrome c reductase activities. While both Erv1 and cytochrome c were reported to be membrane associated in mitochondria, it is unknown how the mitochondrial membrane environment may affect the function of Erv1. Here, in this study, we used liposomes to mimic the mitochondrial membrane and investigated the effect of liposomes and cardiolipin on the folding and function of yeast Erv1. Enzyme kinetics of both the oxidase and cytochrome c reductase activity of Erv1 were studied using oxygen consumption analysis and spectroscopic methods. Our results showed that the presence of liposomes has mild impacts on Erv1 oxidase activity, but significantly inhibited the catalytic efficiency of Erv1 cytochrome c reductase activity in a cardiolipin-dependent manner. Taken together, the results of this study provide important insights into the function of Erv1 in the mitochondria, suggesting that molecular oxygen is a better substrate than cytochrome c for Erv1 in the yeast mitochondria.     

Dietary lipid,fatty acid oxidation and incorporation of carbon into cholesterol

Female rats (200 g) were fed a nutritionally adequate diet containing 1% by weight of corn oil (low-fat, LF), 21% of corn oil (CO) or 20% of beef tallow plus 1% of corn oil (BT) for two weeks. Food was removed for 8–12 hr, then each rat was refed for 1 hr. Each rat was injected ip with Na-³H-acetate and U-¹⁴C-Na-palmitate, (P),-oleate (O) or-linoleate (L). Expired CO₂ was collected for 2 hr. Liver, heart and serum were obtained for analysis of total lipid¹⁴C and³H and cholesterol¹⁴C and³H. Oxidation of L was twice as great as O or P when the LF diet was fed. CO and BT diets doubled oxidation of O to equal L, and increased oxidation of P, 50%. In liver and serum P was retained to a greater extent than O or L on BT and CO diets. Incorporation of acetate into total lipid was highest on LF diet and reduced by feeding either CO or BT. Incorporation of acetate into cholesterol was greater when BT or CO was fed than for LF.¹⁴C was incorporated into cholesterol in such small amounts that it was barely detectable and could not be counted accurately. Conclusions are that (a) dietary fat affects rate of oxidation of uniformly labeled palmitate and oleate, but not linoleate, (b) acetate is a more ready precursor to cholesterol than is fatty acid carbon, and (c) the acetate incorporated into cholesterol when polyunsaturated fat is fed is not derived directly from fatty acid carbon. The failure of incorporation of fatty acid carbon into cholesterol within 2 hr of administration opens the question of compartmentation of acetate as to its metabolic source. Colorado Agricultural Experiment Station Scientific Series Paper No. 1510.     

Effects of dietary heated fats on rat liver enzyme activity

The objective of this study was to evaluate the effects of dietary heated fats from a commercial deep-fat frying operation on rat liver enzyme activity. The fats, partially hydrogenated soybean oil (PHSBO) used for four days and for 7 days (7-DH) for frying foodstuffs in a commercial restaurant, were fed to rats in either free access to food or by pair-feeding graded doses. All diets were isocaloric and contained 15 g/100 g of diet. Experiments were conducted with control rats fed nonheated (NH) PHSBO diet. Animals fed 7-DH diet in each set of experiments had larger amounts of cytochromes P₄₅₀ and b₅ and greater activity of NADPH-cytochrome P₄₅₀ reductase when compared to controls. The activities of carnitine palmitoyltransferase-I and isocitrate dehydrogenase were significantly lower in rats fed test diets in comparison to controls. A significantly depressed activity of glucose 6-phosphate dehydrogenase was also noticed for these animals when compared to those fed NH. In addition, liver and microsomal protein concentrations were significantly greater in rats fed the used oils in comparison to controls, and liver glycogen was significantly lower.     

Inhibition of cytochrome c oxidase and hemolysis caused by lysosphingolipids

Galactosylsphingosine, glucosylsphingosine and sphingosine all inhibited cytochrome c oxidase activity in mitochondria from rat liver; more than 50% inhibition was caused by 5 μM lipid (0.1 μmol/mg mitochondrial protein). However, these lysosphingolipids did not suppress the activity of purified cytochrome c oxidase. When the enzyme was “reconstituted” with phosphatidylcholine, the lysosphingolipids clearly inhibited the activity. On the other hand, galactosylsphingosine, glucosylsphingosine and sphingosine all hemolyzed erythrocytes, indicating that lysosphingolipids can disrupt the membrane. Thus, it appears that the inhibition of cytochrome c oxidase, a membrane-bound enzyme in mitochondria, is due to perturbation of the environment of the enzyme and that the primary attacking site of the lysosphingolipids is the membrane. Because the potency to inhibit cytochrome c oxidase and to hemolyze erythrocytes did not differ among these lysosphingolipids and because galactosylceramide caused neither inhibition of cytochrome c oxidase nor hemolysis, the free amino group in the lysosphingolipids seems to be essential to give the effects. In addition, both inhibition of cytochrome c oxidase and hemolysis caused by lysosphingolipids were completely abolished by albumin, suggesting that toxic effects of lysosphingolipids may not be apparent in blood.     

The influence of linoleic acid intake on membrane-bound respiratory activities

The fatty acid composition of subcellular membranes, like that of depot fats, can be altered by dietary manipulation. Most attention has been directed toward the effects of feeding an essentialfatty-acid-free diet. We chose to examine some responses generated by the feeding of a dietary fat containing a disproportionately high level of an essential fatty acid. Rats were fed diets formulated with beef tallow (BT) to provide 4% (P/S, 0.2) or safflower oil (SO) to provide 24% (P/S, 7.6) of total energy as linoleic acid. Lipids isolated from hepatic mitochondria of rats fed the SO diet contained, in relative terms, 85% more unsaturated bonds. Mitochondria isolated from livers of rats fed either diet were tightly coupled. When all aspects of oxidative metabolism examined in this report are considered, mitochondria of SO group origin exhibited greater oxidative activities but lower ADP/O ratios than did BT mitochondria. Our hypothesis is that the perturbed state of the membrane-bound phospholipids initiates a remodeling-response through which an intramitochondria source of ADP is generated to support state-3 respiratory activity.     

Modified electrodes for probing the metal-reductase activity of metalloproteins: the reduction of iron(III) catalyzed by Desulfovibrio vulgaris Hildenborough cytochrome c3

Cytochromes c₃ are polyheme c-type cytochromes characterized by low redox potentials, that have been shown to develop metal-reductase activity. In this paper, different strategies are explored to immobilize one of them, Desulfovibrio vulgaris Hildenborough cytochrome c₃, a highly basic tetraheme cytochrome, including adsorption, covalent bonding, imprisonment in a layer-by-layer assembly, and entrapment within cast films or a dialysis membrane. The performance and efficiency of modified (carbon or gold) electrodes have been evaluated using electrochemical (cyclic and square-wave voltammetry, current-time curves) techniques in the presence of a soluble Fe(III) complex, ammonium Fe(III) citrate acting as the soluble substrate, and chosen as a model system. The advantages and drawbacks of each strategy are discussed with the view of further extension of environmental interest to more toxic metal contaminants.     

A new cyclometalated rhenium(I) complex

Reaction of Re(CO)₅Cl with 2-phenylbenzothiazole (HΦBT) afforded the cyclometalated Re(CO)₄(ΦBT) complex with the molecular structure of the obtained product confirmed by X-ray investigations. Re(CO)₄(ΦBT) shows intense, hardly dependent on environment, luminescence originating from the π–π* ΦBT intraligand electronic transition. The investigated Re(CO)₄(ΦBT) complex can be considered as a potentially applicable “triplet emitter” for OLED devices.     

The influence of linoleic acid intake on the kinetics of adenine nucleotide translocase

Dietary enrichment of mitochondrial polyenoic fatty acid content was associated with increased respiratory activity (Abuirmeileh and Elson). The influence of the membrane lipid composition on the adenine nucleotide translocase (AdNT) was studied in rats that were fed diets formulated with beef tallow (BT) to provide low or safflower oil (SO) to provide high contents of linoleic acid. The phosphatidylcholine/phosphatidylethanolamine ratio was 40% higher in the SO mitochondria due primarily to an increase in phosphatidylcholine. SO mitochondria exhibited 25% higher state 3 respiration, 50% higher state 4 respiration and 13% higher net ADP-dependent respiration than did the BT mitochondria. The relative RCR and ADP/O values of the SO mitochondria were slightly but significantly (p<0.01) lower. The kinetics of the AdNT were determined by the back exchange method (Pfaff and Klingenberg). At all assay temperatures, SO mitochondria exhibited a higher K_m for ADP. However, addition of 5 mM carnitine to the assay mixture increased the affinity of the SO-AdNT giving K_m values similar to that of the BT-AdNT. Washing the mitochondria with fat-free BSA had a similar, but lesser, effect. At 25 C and 37 C, the V_max of the SO-AdNT were increased by 11–15% (p<0.05) which was independent of either BSA-wash or the presence of carnitine. According to Dixon plot studies, the SO-AdNT had a comparable K_i for atractylate but a slightly lower K_i for palmitoyl-CoA inhibition. The accumulation of acyl-CoA esters in the SO mitochondria was not ruled out. The overall results suggest that changes in mitochondrial membrane lipids accommodated an increased V_max of the SO-AdNT, which in turn accounted for that part of the increased state 3 respiration dependent on ADP translocation.     

Membrane physical properties do not explain increased cyclic AMP production in hepatocytes from rats fed menhaden oil

To study the effect of altering plasma membrane fatty acid composition on the glucagon signal transduction pathway, cAMP accumulation was measured in hepatocytes from rats fed diets containing either menhaden oil (MO) or coconut oil (CO). Hepatocytes from MO-fed animals produced significantly more cAMP in response to glucagon and forskolin compared to CO-fed animals. Glucagon receptor number and affinity were similar in MO- and CO-fed rats. Liver plasma membranes from MO-fed animals were enriched in long-chain n-3 fatty acids and contained significantly lower amounts of saturated C₁₀−C₁₆ and 18∶1n−9 than CO-fed animals. Membrane physical properties were examined using both Fourier transform infrared spectroscopy (FTIR) and the fluorescent probe 1,6-diphenyl-1,3,5-hexatriene (DPH). FTIR analysis revealed that below 34°C, CO membranes were more ordered than MO membranes. However, as assay temperature approached 37°C, MO and CO membranes became similarly ordered. DPH polarization values indicated no differences in membrane order at 37°C, whereas membrane order was decreased in CO-fed animals at 25°C. These data indicate the importance of assay temperature in assessing the influence of membrane physical properties on the activity of signal transduction pathways. Whereas increased signal transduction activity has been correlated to reduced membrane order in MO-fed animals, these data indicate that at physiological temperatures membrane order did not vary between groups. Enhanced cAMP accumulation in response to forskolin indicates that adenylate cyclase activity or content may be elevated in MO-vs. CO-fed rats. Enhanced adenylate cyclase activity may result, in part, from changes in specific fatty acids in hepatocyte plasma membranes without demonstrable changes in membrane physical properties.     

Effect of Dimboa, a Hydroxamic Acid from Cereals, on Peroxisomal and Mitochondrial Enzymes from Aphids: Evidence for the Presence of Peroxisomes in Aphids

2,4-Dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), a hydroxamic acid involved in the resistance of cereals to aphids, was administered to adult individuals of the aphid Sitobion avenae in artificial diets. Effects on the cellular metabolism were inferred from the evaluation of several organelle marker enzymes. Catalase from peroxisomes and cytochrome c oxidase from mitochondria increased their activities about twofold when aphids were fed with 2 mM DIMBOA. The role of these enzymes in the metabolizing of xenobiotics by aphids is discussed. Biochemical and cytochemical evidences for the presence of peroxisomes in aphids are reported here for the first time.     

5c, 11c, 14c-Eicosatrienoic acid and 5c, 11c, 14c, 17c-eicosatetraenoic acid ofBiota orientalis seed oil affect lipid metabolism in the rat

The effects of 5c, 11c, 14c-eicosatrienoic acid (20∶3BSO) and 5c, 11c, 14c, 17c-eicosatetraenoic acid (20∶4BSO), polyunsaturated fatty acids (PUFA) contained inBiota orientalis seed oil (BSO), on lipid metabolism in rats were compared to the effects of fats rich in linoleic acid (LA) or α-linolenic acid (ALA) under similar conditions. The potential effect of ethyl 20∶4BSO as an essential fatty acid also was examined in comparison with the ethyl esters of LA. ALA and γ-linolenic acid (GLA). BSO- and ALA-rich fat decreased the concentration of plasma total cholesterol, high density lipoprotein cholesterol, triglyceride and phospholipid as compared to LA-rich fat. BSO was more effective in reducing plasma cholesterol concentrations than was the ALA-rich fat. Dietary BSO markedly decreased the hepatic triglyceride concentration as compared to the LA-rich or ALA-rich fats. Aortic production of prostaglandin I₂ tended to decrease in rats fed BSO or ALA-rich fat compared to those fed the LA-rich fat. Adenosine diphosphate-induced platelet aggregation was similar in the three groups. The proportion of arachidonic acid (AA) in liver phosphatidylcholine (PC) of rats fed BSO was lowest compared to that of rats fed ALA-rich or LA-rich fats. Administration of 20∶4BSO, ALA or GLA to essential fatty acid-deficient rats decreased the ratio of 20∶3n−9 to AA in liver PC to the same extent; administration of LA was more effective. The results indicate that the effects of specific PUFA contained in BSO on lipid metabolism are different from those of LA and ALA. It is also suggested that 20∶4BSO may exhibit some essential fatty acid effects.     

Cyclic fatty acid monomers from dietary heated fats affect rat liver enzyme activity

This study was conducted to investigate the effects of dietary cyclic fatty acid monomers (CFAM), contained in heated fat from a commercial deep-fat frying operation, on rat liver enzyme activity. A partially hydrogenated soybean oil (PHSBO) used 7 d (7-DH) for frying foodstuffs, or 0.15% methylated CFAM diets was fed to male weanling rats in comparison to a control group fed a nonheated PHSBO (NH) diet in a 10-wk experiment. All diets were isocaloric with 15% fat. Animals fed either CFAM or 7-DH diets showed increased hepatic content of cytochrome (cyt.) b₅ and P₄₅₀ and increased activity of (E.C. 1.6.2.4) NADPH-cyt. P₄₅₀ reductase in comparison to the control rats. In addition, the activities of (E.C. 2.3.1.21) carnitine palmitoyltransferase-l and (E.C. 1.1.1.42) isocitrate dehydrogenase were significantly decreased when compared to that of rats fed the NH diet. A significantly depressed activity of (E.C. 1.1.1.49) glucose 6-phosphate dehydrogenase was also observed for these animals compared to the control rats fed NH diet. Moreover, liver and microsomal proteins were significantly increased when CFAM or 7-DH diets were fed to animals in comparison to controls while liver glycogen was decreased significantly in experimental groups of rats. The results obtained in this study indicate that the CFAM in the diet from either synthetic sources or used fats increase the activity of liver enzyme systems that detoxify them.     

Eicosanoid synthesis in 7,12-dimethylbenz(a)anthracene-induced mammary carcinomas in Sprague-Dawley rats fed primrose oil,menhaden oil or corn oil diet

The comparative effects of high-fat diets (20%, w/w) on eicosanoid synthesis during mammary tumor promotion in 7,12-dimethylbenz(a)anthracene (DMBA)-induced rats were studied using diets containing 20% primrose oil (PO), 20% menhaden oil (MO) or 20% corn oil (CO). Sprague-Dawley rats fed the PO or MO diet had 21% or 24% fewer adenocarcinomas, respectively, than rats fed the CO diet. Histologically (i.e., mitotic figures, inflammatory cell infiltration and necrosis), the CO-fed rats exhibited the highest frequency of changes within tumors. Plasma fatty acid composition was significantly altered by diet, reflecting the composition of the oils which were being fed. Only the plasma of PO-fed rats contained detectable levels of gamma-linolenic acid (GLA). Arachidonic acid (AA) levels were significantly higher (p<0.05) in PO-fed than in CO- or MO-fed rats. MO-fed rats had significantly higher levels of plasma palmitic acid, while palmitoleic, eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids were detected only in MO-fed rats. As expected, linoleic acid (LA) and AA levels were lower (p<0.05) in the MO-fed rats than in PO- or CO-fed groups. The plasma of the CO-fed rats contained significantly higher levels of oleic acid. Eicosanoid synthesis in mammary carcinomas of rats fed the 20%-fat diets was 2–10 times higher than in mammary fat pads of control rats. The synthesis of PGE₁ and LTB₄ was significantly (p<0.05) higher in PO-fed rats than in CO-fed or MO-fed rats, although PGE values were significantly (p<0.05) higher in CO-fed rats than in Mo or PO groups. The synthesis of eicosanoids in both mammary fat pads and mammary carcinomas of MO-fed rats was lower (p<0.05) than in tissues of rats fed either CO or PO diets due to less AA precursor being fed and/or to competition between n−6 and n−3 fatty acids for cyclooxygenase and lipoxygenase. The ratios of monoenoic to dienoic eicosanoids in both mammary fat pads and mammary carcinomas were higher in the PO group than in the MO or CO groups. These results suggest that inclusion of GLA (PO feeding) or EPA and DHA (MO feeding) in the diet may decrease malignancy by altering eicosanoid profiles.     

The up-regulation of hepatic acyl-coA oxidase and cytochrome P<Subscript>450</Subscript> 4A1 mRNA expression by dietary oxidized frying oil is comparable between male and female rats

We previously demonstrated that oxidized frying oil (OFO) activates peroxisome proliferator-activated receptor α (PPARα) and up-regulates hepatic acyl-CoA oxidase (ACO) and cytochrome P₄₅₀ 4A1 (CYP4A1) genes in male rats. As female rats were shown to be less responsive to some peroxisome proliferators (PP), this study compared the expression of a few PPARα target genes in male and female rats fed diets containing OFO. Male and female rats were fed a diet containing 20 g/100 g OFO (O diet) or fresh soybean oil (F diet) for 6 wk. Both male and female rats fed the O diet showed significantly higher liver weight, hepatic ACO and catalase activities, CYP4A protein, and expression of ACO and CYP4A1 mRNA (P<0.05) compared with their control groups. The mRNA expression of two other PPARα target genes, FA-binding protein and HMG-CoA synthase, were marginally increased by dietary OFO (P=0.0669 and 0,0521, respectively). Female rats fed the O diet had significantly lower CYP4A protein than male rats fed the same diet. The remaining OFO-induced effects were not significantly different between male and female rats fed the O diet. These results indicate that dietary OFO, unlike clofibrate or other PP, had minimal sexual dimorphic effect on the induction of hepatic PPARα target gene expression.     

Growth hormone and liver mitochondria: Effects on phospholipid composition and fatty acyl distribution

Effects of growth hormone on phospholipid composition and fatty acyl distribution were studied in liver mitochondria of hypophysectomized rats. After hypophysectomy, only cardiolipin showed a 25% decrease. Its fatty acyl distribution, which consisted mainly of linoleic acid (55–60%) and oleic acid (20%), was unchanged. In phosphatidylcholine and phosphatidylethanolamine fractions the contents of docosahexaenoic and arachidonic acids were decreased with a concomitant increase in linoleic acid content. These changes could be accounted for by small but significant decreases in the activities of Δ⁹-desaturase (sucrose-induced), Δ⁵-desaturase and mitochondrial elongation enzymes. The activities of Δ⁶-desaturase NADH cytochrome b₅ ferri-reductase, cytochrome b₅, NADH cytochrome c reductase and microsomal elongation enzymes remained virtually unchanged. Injection of bovine growth hormone daily for seven days restored cardiolipin and fatty acyl distribution and the enzyme activities. From these and other results, we conclude that growth hormone-dependent increase of respiratory activity of liver mitochondria may be partly mediated by the hormonal effects on membrane lipid distribution.     

Mitochondrial bioelectrocatalysis for biofuel cell applications

Mitochondria modified electrodes have been developed and characterized that utilize whole mitochondria isolated from tubers and immobilized within a quaternary ammonium modified Nafion membrane on a carbon electrode that can oxidize pyruvate and fatty acids. Detailed characterization of the performance of these mitochondria modified electrodes has been accomplished by coupling the mitochondria-based bioanode with a commercial air breathing cathode in a complete pyruvate/air biofuel cell. The studies included the effect of fuel (pyruvate) concentration, mitochondria lysing, temperature and pH on the performance of the mitochondria catalyzed, pyruvate/air biofuel cell. Effect of oxygen and cytochrome c oxidase inhibitors on biofuel cell performance has allowed us to further understand the mechanism of electron transfer with the carbon electrode.     

Direct electrochemistry of dinuclear CuA fragment from cytochrome c oxidase of Thermus thermophilus at surfactant modified glassy carbon electrode

Cytochrome c oxidase is ubiquitous enzyme involved in the terminal step of respiratory electron transfer process. The unique binuclear copper center containing bis-dithiolato bridges form a valance delocalized [Cu^1.5+-Cu^1.5+] state of the metal center located at the subunit II of cytochrome c oxidase. This metal center acts as the electron entry site of the enzyme and accepts electrons from cytochrome c. Direct electrochemistry of this binuclear copper center containing the water soluble protein obtained by genetically truncating the membrane bound part of the subunit II from Thermus thermophilus was achieved by favorable orientation of the protein on glassy carbon electrode surface promoting efficient electron transfer in the presence of various surfactants. Very reproducible, Nernstian responses are obtained with Cu_A. The redox potential and the electrochemical response were enhanced prominently in case of cationic surfactant CTAB indicating that the nature of the surfactant has a significant effect on the microenvironment of the protein-electrode interface. The results have been used to understand the mechanism of electron transfer from cytochrome c to the copper center during the enzymatic reaction.     

Electrical communication of cytochrome enriched Escherichia coli JM109 cells with graphite electrodes

In the present study three different strains of Escherichia coli (JM109 - a native “wild type” strain, JM109/pBSD 1300 - a strain overproducing the membrane anchor domain of Bacillus subtilis succinate-quinone reductase, SQR, a protein that contains two transmembraneously arranged heme groups and JM109/pLUV 1900 - a strain overproducing cytochrome c₅₅₀ from B. subtilis, a protein where the cytochrome domain is anchored to the membrane with a transmembrane helix) were immobilised on the surface of a spectrographic graphite electrode and tested for electrical communication using mediators. Such compounds as ferricyanide, 2,6-dichlorophenolindophenol (DCPIP) and ubiquinone (Q₀) were used as soluble mediators and two flexible osmium redox polymers; poly(1-vinylimidazole)₁₂-[Os-(4,4′-dimethyl-2,2′-di’pyridyl)₂Cl₂]^2+/+ (osmium redox polymer I) and poly(vinylpyridine)-[Os-(N,N′-methylated-2,2′-biimidazole)₃]^2+/3+ (osmium redox polymer II) were co-immobilised with the bacterial cells onto the electrode surface. The effects of applied potential, buffer pH and different substrates were compared for the different combinations bacterial strains - mediators. Through the introduction of the cytochromes in the bacterial membrane it was established that it had great effect on the ability of the bacterial cells to effectively communicate with artificial mediators. The introduction of the transmembraneously arranged heme groups of B. subtilis made it possible for this strain to communicate with the Os-polymers, whereas the introduction of the cytochrome c₅₅₀ had an effect especially increasing ability of Q₀ to act as an efficient e⁻ acceptor.     

Effect of dietary di-2-ethylhexyl phthalate on oxidation of14C-palmitoyl CoA by mitochondria from mammalian heart and liver

Oxidation of [1-¹⁴C] palmitoyl CoA by heart and liver mitochondria from rats fed dietary di-2-ethylhexyl phthalate (DEHP) was investigated in vitro. Oxidation of¹⁴C-palmitoyl CoA to¹⁴CO₂ increased two- to threefold in hepatic mitochondria from rats fed 0.1% DEHP for 2 to 3 days; this increase appeared to be a maximum response since similar data were obtained using hepatic mitochondria from rats receiving 0.5% or 1.0% DEHP in the diet. The response of hepatic mitochondria to DEHP was found to continue throughout the duration of 35-day trials in which 1.0% DEHP was fed. In contrast to hepatic mitochondria, the oxidation of¹⁴C-palmitoyl CoA by heart mitochondria decreased ca. 40% upon addition of 0.1% or 0.5% DEHP to the diet; this effect of DEHP on heart mitochondria was not sustained beyond ca. 8 days of DEHP feeding. Limited studies were also performed in rabbits and pigs. Oxidation of¹⁴C-palmitoyl CoA was increased ca. twofold in hepatic mitochondria from rabbits fed 1% dietary DEHP for 12 days and in hepatic mitochondria from pigs that received 5 doses of DEHP (0.8g/kg) at 12-hr intervals; the oxidation¹⁴C-palmitoyl CoA by heart mitochondria from these same animals was unchanged in the rabbit but increased an average of 37% in the pig. DEHP feeding to rats was associated with increased yields of hepatic mitochondrial protein; standardized preparations of heart mitochondria were not similarly affected.     

Effects of conjugated linoleic acid isomers on lipid-metabolizing enzymes in male rats

Male weanling Wistar rats (n=15), weighing 200–220 g, were allocated for 6 wk to diets containing 1% (by weight) of conjugated linoleic acid (CLA), either as the 9c,11t-isomer, the 10t,12c-isomer, or as a mixture containing 45% of each of these isomers. The five rats of the control group received 1% of oleic acid instead. Selected enzyme activities were determined in different tissues after cellular subfractionation. None of the CLA-diet induced a hepatic peroxisome-proliferation response, as evidenced by a lack of change in the activity of some characteristic enzymes [i.e., acyl-CoA oxidase, CYP4A1, but also carnitine palmitoyltransferase-I (CPT-I)] or enzyme affected by peroxisome-proliferators (glutathione S-transferase). In addition to the liver, the activity of the rate-limiting β-oxidation enzyme in mitochondria, CPT-I, did not change either in skeletal muscle or in heart. Conversely, its activity increased more than 30% in the control value in epididymal adipose tissue of the animals fed the CLA-diets containing the 10t,12c-isomer. Conversely, the activity of phosphatidate phosphohydrolase, a rate-limiting enzyme in glycerolipid neosynthesis, remained unchanged in adipose tissue. Kinetic studies conducted on hepatic CPT-I and peroxisomal acyl-CoA oxidase with CoA derivatives predicted a different channeling of CLA isomers through the mitochondrial or the peroxisomal oxidation pathways. In conclusion, the 10t,12c-CLA isomer seems to be more efficiently utilized by the cells than its 9c,11t homolog, though the Wistar rat species appeared to be poorly responsive to CLA diets for the effects measured.