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PubMed1998

Flow injection determination of free and total cholesterol in animal greases using enzymes in non-aqueous media.

A non-covalently coimmobilized bienzymic reactor of horseradish peroxidase (HRP) and cholesterol oxidase (COD), operating in a continuous organic flowing stream of 1X10-3 M p-anisidine in buffer-saturated (pH 7.0) toluene, has been employed for cholesterol determination in animal greases, such as pig, beef, and chicken fat, and codfish liver oil. The method provides a good linear relationship up to 1.8 X 10-3 M cholesterol and average recoveries of 99.5%, a high sensitivity, with a detection limit of 1 X 10-6 M of cholesterol and a good precision (an interday RSD of 1.8% for the determination of total cholesterol in a codfish oil sample). The method permits the direct spectrophotometric determination of free cholesterol present in animal grease samples without any pre-treatment, and the total cholesterol determination after a microwave-assisted saponification. The bienzymic reactor exhibits a good stability in the water-restricted environment, being possible to perform more than 180 analyses during a period of 10d with 1mg of HRP, and 1 mg of COD, equivalent to 220 purpurogallin units and 23 U, respectively.

Animals↗
PubMedDate not supplied

DNA adduct levels in fish from pristine areas are not detectable or low when analysed using the nuclease P1 version of the 32P-postlabelling technique.

In order to understand and apply DNA adduct formation in fish liver as a biomarker for aquatic pollution, information concerning the natural background levels in noncontaminated organisms, caused by endogenous compounds, is of fundamental importance. In this study, DNA adducts were analysed in liver of 11 fish species from arctic and sub-arctic areas in the northern Atlantic using the nuclease P1 version of the 32P-postlabelling technique. The collected fish were assumed not to have been influenced by anthropogenic pollution apart from possible long-range transported pollutants. As polycyclic aromatic hydrocarbons (PAHs) are thought to be fundamental in forming the type of DNA adducts detected by the method used, biliary PAH metabolite levels were measured in a selection of the investigated species. In all investigated individuals, the levels of PAH metabolites were undetectable. Controlled on-site exposure experiments with benzo[a]pyrene (polar cod) and laboratory experiments with crude oil (polar cod and Atlantic cod) were conducted. DNA adducts were formed in both these species. The field-sampled fish showed undetectable levels of DNA adducts or levels just above the detection limit. The present study supports the assumption that when DNA adducts are detected by the nuclease P1 version of the 32P-postlabelling method in fish liver, it can be interpreted as DNA damage caused by pollutants.

Animals↗
PubMed2003

Biomarkers in fish from Prince William Sound and the Gulf of Alaska: 1999-2000.

To test the hypothesis that biomarker levels in fish collected at Prince William Sound (PWS) sites impacted by the 1989 Exxon Valdez oil spill were higher than those collected at unimpacted sites, a 1999-2000 study collected five fish species and associated benthic sediments from 21 sites in PWS and the eastern Gulf of Alaska (GOA). PWS sites were divided in three oiling categories based upon 1989 shoreline assessments: nonspill path (NSP), spill path oiled (SPO), and spill path not oiled (SPNO). Rockfish (N = 177), rock sole (N = 30), and kelp greenling (N = 49) were collected at near-shore locations (approximately 50-500 m from shore); Pacific halibut (N = 131) and Pacific cod (N = 81) were collected further offshore (approximately 500-7000 m). Fish were assayed for bile fluorescent aromatic contaminants (FAC) and cytochrome P4501A (CYP1A) levels measured as liver ethoxyresorufin O-deethylase (EROD) activity and by immunohistochemistry (IHC) of various tissues. For all species studied at all sites, bile FAC concentrations and CYP1A levels were low and in the same range for fish collected at PWS SPO and SPNO sites relative to NSP sites in PWS and the GOA. Consequently, the hypothesis is rejected for the species studied. The bile FAC results further indicate a pervasive exposure of fish at all sites, including those in the GOA far removed from the effects of the spill, to low levels of polycyclic aromatic hydrocarbons. Analysis of the benthic sediments indicates that the probable sources of this exposure are petrogenic hydrocarbons derived from natural oil seeps and eroding sedimentary rocks in the eastern GOA.

Accidents↗
PubMed2003

Cod protein lowers the hepatic triglyceride secretion rate in rats.

The objective of the present study was to determine the combined effects of cod protein and fish oil on the modulation of triglyceride metabolism in rats, and to evaluate their potential mechanisms of action. Plasma and hepatic lipid concentrations, triglyceride (TG) secretion rates and postheparin plasma lipoprotein lipase (LPL) activity were determined in rats fed for 28 d diets varying in both protein (200 g/kg) and lipid (140 g/kg) sources: 1) casein-menhaden oil, 2) casein-beef tallow, 3) cod protein-menhaden oil or 4) cod protein-beef tallow. Menhaden oil feeding diminished hepatic TG concentrations (P = 0.02), hepatic TG secretion rates (P = 0.003) and triglyceridemia (P = 0.02) compared with beef tallow. Hepatic TG concentrations (P = 0.05) and TG secretion rates (P = 0.04) were reduced in rats fed cod protein compared with those fed casein. The protein source did not exert an independent effect on triglyceridemia, whereas the combination of cod protein and menhaden oil resulted in 50% lower plasma TG compared with the casein-beef tallow mixture, whereas the combination of menhaden oil and casein did not significantly decrease triglyceridemia compared with casein-beef tallow. Menhaden oil (P = 0.005) and cod protein (P = 0.03) also lowered plasma cholesterol concentrations in comparison with beef tallow and casein, respectively. This was associated with a reduction in hepatic cholesterol concentrations when rats fed cod protein were compared with those fed casein (P = 0.006). No diet effect was observed on postheparin plasma LPL activity, but the activity of hepatic triglyceride lipase was reduced in rats fed menhaden oil compared with those fed beef tallow. These findings show that both cod protein and menhaden oil exert independent and beneficial effects on lipid metabolism in rats.

Animal Feed↗
PubMed1988

Distribution and elimination of [14C]octachlorostyrene in cod (Gadus morhua), rainbow trout (Salmo gairdneri, and blue mussel (Mytilus edulis).

Cod (Gadus morhua) and rainbow trout (Salmo gairdneri) were given a single oral dose of 100 microCi/kg b.w. of [14C]octachlorostyrene [( 14C]OCS) in peanut oil. Blue mussel (Mytilus edulis) was exposed to [14C]OCS in water. The distribution and elimination of the compound was studied by liquid scintillation counting and whole-body autoradiography. The highest degree of radioactivity in the cod and rainbow trout was measured in the liver and the visceral fat, respectively. The degree of radioactivity in the brain of cod exceeded that of the rainbow trout by a factor between 2 and 4 at all survival times. In addition to bile excretion of [14C]OCS-derived radioactivity, a possible excretion over the intestinal mucosa was suggested. The rate of elimination was slow in both species, and substantial amounts of radioactivity remained in the tissues 90 d after administration. In the blue mussel, the highest degree of radioactivity was found in the hepatopancreas. Substantial amounts of radioactivity were present in the mussel tissues 60 d after administration.

Animals↗
PubMed1983

Long term exposure of marine fish to crude petroleum-I. Studies on liver lipids and fatty acids in cod (Gadus morhua) and winter flounder (Pseudopleuronectes americanus).

1. Total lipids, total phospholipids, lipid class and fatty acids have been studied in the livers of cod and winter flounder from the Northwest Atlantic following long-term exposure to crude petroleum. 2. The levels of free fatty acids and total phospholipids were elevated and triglycerides depleted significantly in the males and females of both species in the experimental groups. 3. The essential fatty acids of C20.5, C22.5 and C22.6 series were significantly altered in the oil-exposed animals. 4. These findings indicate that marine fish, following long-term exposure to crude petroleum, tend to (a) use up their stored energy of neutral lipids and (b) develop changes in membrane structure and function.

Animals↗
PubMed2000

PAH metabolites in bile, cytochrome P4501A and DNA adducts as environmental risk parameters for chronic oil exposure: a laboratory experiment with Atlantic cod.

In order to perform environmental risk assessments with regard to oil contamination in the sea, it is important to obtain knowledge about threshold levels for possible adverse effects in marine organisms. With this objective in mind, selected biomarkers were studied in Atlantic cod (Gadus morhua) chronically exposed to mechanically dispersed crude oil. The fish were exposed for 30 days in a continuous flow system to nominal concentrations of 0.06, 0.25 and 1 ppm North Sea crude oil. Fish were sampled five times during the exposure period. In addition, the 1 ppm group and the control group were sampled 1 week after the end of exposure. Polyaromatic hydrocarbon (PAH) concentrations in the seawater were analysed regularly by direct fluorescence and, at one occasion, by gas chromatography with mass spectrographic detection (GC/MS) measurements. Liver samples were analysed for parent PAH levels by means of GC/MS measurements, and PAH metabolites in bile were analysed by means of fixed wavelength fluorescence. Cytochrome P450 induction in liver was estimated by ethoxyresorufin-O-deethylase (EROD) activity, and hepatic DNA adducts were analysed by the 32P-postlabelling assay. The parent PAH concentrations in liver showed peak levels 3 days after the start of exposure, followed by a reduction towards the end of the experiment. In contrast, the PAH metabolites in bile and EROD activity showed generally increasing levels throughout the whole exposure period, indicating an increased biotransformation efficiency. The level of DNA adducts in the 1 ppm group showed a stable increase during the entire exposure period. Only a slight, non-significant decrease in DNA adduct levels was observed after 7 days of recovery in clean water. Exposure-dependent responses were observed for all three biomarkers. The lowest nominal concentration of dispersed oil in water, 0.06 ppm, corresponded to a measured total PAH concentration in the water of 0.3 ppb. Atlantic cod exposed to this concentration showed increased levels of PAH metabolites in bile and a slight induction of CYP1A, as well as formation of DNA adducts when compared with control fish. Particularly noteworthy is the detection of DNA adducts at such a low exposure concentration of oil in water, which, to our knowledge, is a novel finding. These dose-response data may serve as useful contributions when assessing environmental risk with regard to marine oil pollution.

Animals↗
PubMed2004

Differential effects of fish oil and folic acid supplementation during pregnancy in rats on cognitive performance and serum glucose in their offspring.

OBJECTIVE: We studied the effect of folic acid and polyunsaturated fatty acid supplementation during pregnancy in Wistar albino rats on cognitive performance and serum glucose concentrations in their pups. METHODS: Pregnant female rats from four groups (n = 6/group) were fed casein diets with 18% protein and 2 mg of folic acid/kg of diet (group I), 12% protein and no folic acid (group II), 12% protein and 8 mg of folic acid/kg of diet (group III), or 12% protein and 70 g of cod liver oil/kg of diet (group IV). All pups were weaned on standard control diet with 18% protein. Cognitive performance, brain fatty acid profile, and serum glucose concentrations were studied in offspring at age 6 mo. RESULTS: There was no significant difference in length of gestation or litter size, but the litter weight for group IV was lower (P = 0.047) than that for group I. After weaning, males in group II had lower (P < 0.05) body weights, but those in group III had weights comparable to those in group I for both sexes. In group IV, body weights were lower beyond 15 wk (P < 0.05). Relative brain weight and cognitive performance were significantly higher (P < 0.05) in group IV males and showed higher levels of brain gamma-linolenic acid. Further, these animals had serum glucose levels comparable to those of control animals at age 6 mo, whereas serum glucose levels were higher in males from groups II (P = 0.01) and III (P = 0.01). CONCLUSION: Fish oil supplementation during pregnancy improved cognitive performance and maintained glucose levels into adulthood, unlike folic acid supplementation, which supported only fetal growth and did not maintain glucose levels.

Animals↗
PubMed1999

Genotype and diet effects in lean and obese Zucker rats fed either safflower or coconut oil diets.

Previously we reported that suckling lean heterozygous (FA/fa) Zucker rats had a number of adipose tissue measurements intermediate between those of homozygous lean (FA/FA) and obese (fa/fa) rats. However, in young adult male rats maintained on a low-fat diet, these differences were no longer apparent (i.e., values for the two lean genotypes were similar). In the present study we determined whether the heterozygous effect of the "fa" gene was dependent on the consumption of a high-fat diet. Mother rats were fed high-fat diets containing either safflower (SOD) or coconut (COD) oil throughout mating and lactation. Homozygous lean male and female rats were bred, as well as obese male and lean heterozygous female rats. Suckling rats were studied at 17 days of age. Additional male rats were maintained on the same diet as their mothers until 11-12 weeks of age. Obese suckling rats had higher body weights than lean pups. Inguinal fat pad weights and pad-to-body weight ratios followed the pattern of obese greater than lean (FA/fa) pups that were greater than lean (FA/FA) pups. A similar relationship was found for adipose tissue lipogenic enzyme activities. At 11-12 weeks of age, measurements followed the general pattern of obese rats having greater values than lean rats (i.e., FA/fa = FA/FA). SOD-fa/fa rats had higher hepatic lipogenic enzyme activities than COD-fa/fa rats. In addition, SOD rats had higher fat cell numbers than COD rats. These results suggest that specific fatty acids can alter adipocyte proliferation and/or differentiation in vivo. In addition, there appears to be a defect of fatty acid regulation in livers of genetically obese rats. The heterozygous effect of the "fa" gene in suckling Zucker rats was confirmed. However, high-fat feeding did not result in a heterozygous effect in young adult lean male rats. We will next evaluate the role of sex on this effect.

Adipose Tissue↗
PubMed1987

Species characteristics of the hepatic xenobiotic and steroid biotransformation systems of two teleost fish, Atlantic cod (Gadus morhua) and rainbow trout (Salmo gairdneri).

Groups of Atlantic cod and rainbow trout were treated (ip) with beta-naphthoflavone (BNF), phenobarbital, or peanut oil (controls), and properties of the hepatic xenobiotic and steroid metabolizing enzyme systems were evaluated. In both species, BNF treatment resulted in significant induction of microsomal 7-ethoxycoumarin O-deethylase, 7-ethoxyresorufin O-deethylase, biphenyl 4-hydroxylase, and phenanthrene oxidation, especially at the 1,2-position. Immunochemical studies with rabbit IgG prepared against the major BNF-inducible cytochrome P-450 in cod, P-450c, revealed increased amounts of immunoreactive protein in liver slices from both species after BNF treatment. The molecular weight of the induced protein was approximately 58,000 Da, as shown by Western blotting. When titrating biphenyl 4-hydroxylation, however, the antibodies distinguished between the two species, inhibiting the activity of BNF-induced cod 90% and that of rainbow trout 40% at 10 mg IgG/nmol P-450. Furthermore, cytochrome b5 content and UDP-glucuronyltransferase activity were significantly induced only in rainbow trout, whereas the specific content of cytochrome P-450 was significantly increased only in cod. Differences between the two species were observed in the levels of constitutive activities, the amount of induction, and in the regioselectivity of phenanthrene oxidation and androstenedione metabolism. Treatment with phenobarbital showed no effect on any of the parameters investigated in either species. The results show that although there are many common features of the hepatic xenobiotic and steroid biotransformation systems of the two teleosts, certain species characteristics exist in constitutive properties and induction responses.

7-Alkoxycoumarin O-Dealkylase↗
PubMed2006

Effects of North Sea oil and alkylphenols on biomarker responses in juvenile Atlantic cod (Gadus morhua).

A consequence of oil drilling at sea is the release of produced water contaminated with e.g. polycyclic aromatic hydrocarbons (PAH) and alkylphenols. In the present study, juvenile Atlantic cod were exposed to North Sea oil, nonylphenol and a combination of the North Sea oil and an alkylphenol mixture in a flow-through system. A suite of hepatic biomarkers were analysed. Exposure to North Sea oil resulted in strong induction of CYP1A protein levels and EROD activities. Exposure to nonylphenol, on the other hand, resulted in decreased CYP1A levels and EROD activities. Thus, nonylphenol appears to down-regulate CYP1A expression in Atlantic cod. Combined exposure to North Sea oil with an alkylphenol mixture resulted in lower EROD induction, compared to that in fish exposed to North Sea oil alone. This difference was not statistically significant, but still we believe that the alkylphenols have inhibited CYP1A activities in the fish which may have compromised CYP1A mediated metabolism of other xenobiotics, including PAH. CYP3A protein levels were lower, compared to controls, in fish exposed to nonylphenol and the combination of North Sea oil and alkylphenol mixture. In contrast, the oil alone had no effect on CYP3A protein content. North Sea oil exposure, alone or in combination with alkylphenols, caused oxidative stress observed as elevated levels of GSSG content and GR and CAT activities. Interestingly, exposure to nonylphenol resulted in a marked depletion of total glutathione levels. This apparent depletion may be a consequence of increased conjugation of glutathione to nonylphenol followed by excretion. An increase in conjugation enzyme GST activity was observed in the nonylphenol exposed group, although the difference was not significant. No sign of oxidative damage, measured as lipid peroxidation, was observed in any of the exposures experiments. This study suggests that North Sea oil may lead to oxidative stress and altered CYP1A and CYP3A expression. Alkylphenols, present in produced water, resulted in decreased CYP1A and CYP3A protein expression in Atlantic cod.

Animals↗
PubMed2003

Effects of polyphenolic fraction of silymarin on lipoprotein profile in rats fed cholesterol-rich diets.

To study the influence of polymerised polyphenolics (PP), a fraction of silymarin (SM), on lipids and oxidant status, rats were fed high-cholesterol (1%), high-fat (10%) diets containing either lard fat (LFD) rich in saturated/monounsaturated fatty acids, or currant oil (COD) rich in polyunsaturated fatty acids. PP and SM were administered as dietary supplements (0.1-0.5-1.0%) for 3 weeks. PP (1%) decreased cholesterol (C) in VLDL (from 0.72+/-0.08 mmol l(-1) in LFD control to 0.35+/-0.07 mmol l(-1), P<0.01, and from 0.33+/-0.05 mmol l(-1) in COD control to 0.09+/-0.02 mmol l(-1), P<0.001), and increased HDL-C/VLDL-C ratio, however, without effect on the total plasma C and LDL-C. Liver C content (LFD 19.32+/-1.50 micromol g(-1), COD 18.64+/-2.13 micromol g(-1), N.S.) decreased after PP (1%) to 12.24+/-0.76 micromol g(-1), P<0.01, and 8.78+/-0.95 micromol g(-1), P<0.001, respectively. Triacylglycerols (TAG) in plasma and VLDL decreased after PP in the LFD group only, which displayed higher TAG levels than the COD group. Likewise, LFD caused a higher liver TAG content than did COD (31.16+/-3.00 micromol g(-1) versus 17.31+/-1.48 micromol g(-1), P<0.01), and PP (1%) decreased liver TAG only in rats fed LFD (19.55+/-2.43 micromol g(-1), P<0.02). Blood glutathione (GSH) increased after PP (1%) in the LFD group from 0.97+/-0.11 to 1.54+/-0.19 mmol l(-1) (P<0.05) and in the COD group from 0.58+/-0.15 to 1.23+/-0.10 mmol l(-1) (P<0.01), while liver GSH and plasma TBARS did not change. On principle, effects of PP were dose-dependent and parallel to SM. These results suggest that the polyphenolic fraction of SM positively modifies lipoprotein profile, counteracts the development of fatty liver and ameliorates an antioxidant status in circulation.

Animals↗
PubMed1991

Application of a cytochrome P-450 IA1-ELISA in environmental monitoring and toxicological testing of fish.

1. An enzyme-linked immunosorbent assay (ELISA) based on polyclonal rabbit anti-cod cytochrome P-450 IA1 IgG has been developed in our laboratory. 2. The antibodies employed in the ELISA demonstrate good cross-reactivity with the homologous protein in a number of other fish species, giving cross-reacting protein bands o 54-59 kDa in liver microsomes, as determined by Western blotting. 3. The ELISA technique has been used in numerous experiments with both field collected and laboratory exposed fish of different species, showing good correlation with contaminant exposure. 4. In some instances of PCB exposure where the classical P-450 IA1 monooxygenase assay 7-ethoxyresorufin O-deethylase (EROD) failed to reveal any induction, the ELISA technique demonstrated increased levels of P-450 IA1 protein, indicating inhibiting effects of the PCBs on EROD measurement. 5. In tissues like gill filaments, and in whole cod larvae, where EROD activity is barely detectable, if at all, the ELISA technique showed induction after exposure to a water soluble fraction (WSF) of North Sea crude oil. 6. The results reviewed indicate the usefulness of the ELISA technique to allow rapid screening of a large number of samples, and especially when catalytic measurement is difficult or impossible due to (a) small sample or tissue size, (b) loss of activities in bad storage conditions, or (c) presence of compounds inhibiting activity.

Animals↗
PubMed2002

Diatom cultivation and biotechnologically relevant products. Part II: current and putative products.

While diatoms are widely present in terms of diversity and abundance in nature, few species are currently used for biotechnologically applications. Most studies have focussed on intracellularly synthesised eicosapentaenoic acid (EPA), a polyunsaturated fatty acid (PUFA) used for pharmaceutical applications. Applications for other intracellular molecules, such as total lipids for biodiesel, amino acids for cosmetic, antibiotics and antiproliferative agents, are at the early stage of development. In addition, the active principle component must be identified amongst the many compounds of biotechnological interest. Biomass from diatom culture may be applied to: (1). aquaculture diets, due to the lipid- and amino-acid-rich cell contents of these microorganisms, and (2). the treatment of water contaminated by phosphorus and nitrogen in aquaculture effluent, or heavy metal (bioremediation). The most original application of microalgal biomass, and specifically diatoms, is the use of silicon derived from frustules in nanotechnology. The competitiveness of biotechnologically relevant products from diatoms will depend on their cost of production. Apart from EPA, which is less expensive when obtained from Phaeodactylum tricornutum than from cod liver, comparative economic studies of other diatom-derived products as well as optimisation of culture conditions are needed. Extraction of intracellular metabolites should be also optimised to reduce production costs, as has already been shown for EPA. Using cell immobilisation techniques, benthic diatoms can be cultivated more efficiently allowing new, biotechnologically relevant products to be investigated.

Amino Acids↗
PubMed2001

Immunohistochemical distribution of cytochrome P4501A in larvae and fingerlings of the Siberian sturgeon, Acipenser baeri.

In this paper we investigate by means of immunohistochemistry, the tissue distribution of constitutive cytochrome P4501A (CYPIA), from hatching until 30 days posthatching in developing Siberian sturgeon, Acipenser baeri. For this purpose, a polyclonal (BN-1) antiserum developed against a conservative sequence of piscine CYP1A and a monoclonal (C10-7) antiserum directed against cod CYP1A were used on paraffin-embedded samples. From hatching onwards, distinct CYP1A immunoreactivity was distinctly observed in the following tissues and cells: envelope of oil droplets, matrix and syncytium of the yolk-sac, sinusoids, biliary epithelial cells and hepatocytes. In the digestive tract, buccopharyngeal, oesophageal, gastric and intestinal epithelia, as well as the cytoplasm and brush border of enterocytes were CYP1A-positive. Interestingly, gastric glands and melanin-plug present within lumen of the digestive system were strongly immunoreactive. Kidney (epithelia of renal tubules), gills (pillar and endothelial cells), skin (epithelial cells), muscle fibres of heart and eye (retina) were positive. In brain, we observed a strong CYP1A staining in the developing telencephalon and especially in olfactory system, as well as in those nerve fibres running ventrally toward the posterior brain. A strong CYP1A staining was observed in vascular endothelia of all organs/tissues, especially in the liver. In general, the intensity of CYP1A immunostaining increased during larval development, suggesting besides its known metabolic function (endogenous and/or exogenous), a possible participation of this heme-protein in control of cell division, regulation of growth and differentiation.

Animals↗
PubMed2004

Effects of alkylphenols on CYP1A and CYP3A expression in first spawning Atlantic cod (Gadus morhua).

Alkylphenols are continuously released into the ocean as a result of offshore oil production. Alkylphenols, including 4-tert-butylphenol (C4), 4n-pentylphenol (C5), 4n-hexylphenol (C6), and 4n-heptylphenol (C7), up to 237 ppb concentrations, have been detected in produced water from oil platforms. Previous studies have shown that alkylphenols induce vitellogenesis in fish. Atlantic cod (Gadus morhua) of both sexes were force-fed with various doses ranging between 0.02 and 80 ppm of a mixture of alkylphenols (C4:C5:C6:C7 ratio 1:1:1:1) or 5 ppm 17 beta-estradiol. We investigated effects on hepatic CYP1A and CYP3A protein expression in protein blots, using antibodies against scup (Stenotomus chrysops) CYP1A1 and rainbow trout (Oncorhynchus mykiss) CYP3A. There was a sexually dimorphic expression of CYP1A and CYP3A protein levels, with females expressing higher levels than males. Treatment of male Atlantic cod with 17 beta-estradiol resulted in increased CYP1A and CYP3A protein levels. Exposure to alkylphenols resulted in a dose-dependent increase of CYP1A and CYP3A protein expression in males, but not in females. However, this increase of CYP1A protein levels was not reflected on the CYP1A-mediated ethoxyresorufin-O-deethylase (EROD) activity, implying that alkylphenols inhibited the CYP1A enzyme activity in vivo. In vitro inhibition studies with pooled liver microsomes from Atlantic cod confirmed that the alkylphenols mixture efficiently inhibited the CYP1A activity (IC50=10 microM), although the inhibitory effect of each individual alkylphenol varied. The IC50 values for each individual alkylphenol on the CYP1A activity were, in a descending order of magnitude: [C7>C6>C5>>C4], ranging from 12 to 300 microM with decreased length of the 4-alkyl chain. The effect of alkylphenols on the CYP3A activity in vitro in liver microsomes also was investigated, using the fluorescent 7-benzyloxy-4-[trifluoromethyl]-coumarin (BFC) as a diagnostic CYP3A substrate. The alkylphenol mixture inhibited CYP3A activity with IC50 value at 100 microM. The IC50 values for each individual alkylphenol on CYP3A activity were, in a descending order of magnitude: [C5>C6>C7>C4], ranging between 60 and 250 microM. Taken together, our results show that the alkylphenol mixture and 17 beta-estradiol resulted in elevated hepatic CYP1A and CYP3A expression in male Atlantic cod. The alkylphenol mixture strongly inhibited CYP1A activities, whereas it weakly inhibited CYP3A activity in Atlantic cod liver microsomes in vitro. In addition, 17 beta-estradiol was a weak inhibitor of CYP3A activity (IC50=75 microM) and did not notably inhibit the CYP1A activity in vitro.

Animals↗
PubMed1998

Dietary proteins modulate the effects of fish oil on triglyceridemia in the rat.

Sprague-Dawley rats were fed purified diets varying in both protein (20%) and lipid (11%) content for 28 d to verify the independent and interactive effects of dietary proteins and lipids on serum and hepatic lipids, and on tissue lipoprotein lipase (LPL) activity in both fasted and postprandial states. These diets consisted of either casein-menhaden oil, casein-coconut oil, soy protein-menhaden oil (SPMO), soy protein-coconut oil, cod protein-menhaden oil, or cod protein-coconut oil. A randomized 3 x 2 factorial design was used. A significant protein-lipid interaction was seen on serum triglyceride levels: menhaden oil, compared with coconut oil, induced a decrease in serum triglyceride levels when combined with soy protein but not when combined with cod protein and casein. The lower serum triglyceride concentrations observed in the SPMO-fed rats could be the result of decreased hepatic triglycerides when soy protein was compared with casein and when menhaden oil was compared with coconut oil. Total LPL activity in the heart was higher in menhaden oil-fed rats than in coconut oil-fed rats in the postprandial state. The higher LPL activity in the heart could, however, explain only 10% of the reduction of serum triglycerides, contributing slightly to the lowering effects of SPMO diet on serum triglycerides. Therefore, the present results indicate that dietary proteins can modulate the effects of fish oil on triglyceridemia in the rat, and that could be mainly related to specific alterations in hepatic lipid concentrations.

Adipose Tissue↗