To improve the interpretation and utilisation of blood lipids, ketones and acylcarnitine concentrations as biomarkers in clinical assessments, more information is needed on their dynamic alterations in response to dietary intake and fasting. The aim of this intervention study was to characterise the changes in serum lipid, ketone and acylcarnitine concentrations 24 h after a standardised breakfast meal. Thirty-four healthy subjects (eighteen males and sixteen females) aged 20–30 years were served a breakfast meal (∼500 kcal, 36 E% fat, 46 E% carbohydrates, 16 E% protein, 2E% fibre), after which they consumed only water for 24 h. Blood samples were drawn before and at thirteen standardised timepoints after the meal. Metabolite concentrations were plotted as a function of time since the completion of the breakfast meal. Results demonstrated that concentrations of HDL-cholesterol and LDL-cholesterol decreased until ∼2 h (–4 % for both), while TAG concentrations peaked at 3 h (+27 %). Acetoacetate and β-hydroxybutyrate were highest 24 h after the meal (+433 and +633 %, respectively). Acetylcarnitine, butyrylcarnitine, hexanoylcarnitine, octanoylcarnitine, decanoylcarnitine and dodecanoylcarnitine reached the lowest values at 60 min (decreases ranging from –47 to –70 %), before increasing and peaking at 24 h after the meal (increases ranging from +86 to +120 %). Our findings suggest that distinguishing between fasting and non-fasting blood samples falls short of capturing the dynamics in lipid, ketone, carnitine and acylcarnitine concentrations. To enhance the utility of serum acylcarnitine analyses, we strongly recommend accounting for the specific time since the last meal at the time of blood sampling.

Aflatoxins in contaminated corn do not degrade in corn fermentation solution (CFS) during biofuel production; rather, they are enriched in the co-product, dried distillers grain. Aflatoxin B1 (AfB1) is the most toxic form of all aflatoxins. Removing AfB1 from CFS is desirable to minimize its toxicity to animals. Smectites can adsorb AfB1 from aqueous solutions and, therefore, inactivate the toxin, but proteins in CFS inhibit the adsorption of AfB1 by smectites. The current study aimed to minimize the interference by CFS in adsorption of AfB1 on smectite by modifying a calcium-smectite (Ca-3MS) with a small nutritive organic compound, e.g. carnitine, choline, arginine, histidine, or tryptophan. The organo-smectites were characterized by X-ray diffraction (XRD) and Fourier-transform infrared (FTIR) spectroscopy, and adsorption of AfB1 in CFS by these composites was examined. Various degrees of intercalation of the organic nutrients into the smectites were observed with XRD and FTIR. After immersing the smectite and organo-smectites in the CFS, the d001 values of Ca-3MS expanded to ~1.82 nm due to protein interaction, but the organo-smectites were confined to ~1.39 nm, which indicated that the protein had limited access to the organo-smectite interlayers. The IR bands at ~1652, 1544, 1538, and 1454 cm–1 from the organo-smectites revealed, however, that complete protein inhibition was not achieved. The organo-smectites were capable of adsorbing AfB1 in simple aqueous solution with maximal adsorption capacity up to 0.55 mol kg–1. Significantly greater (p ≤ 0.05) AfB1 adsorption was achieved by choline- and carnitine-modified smectites compared with the original Ca-3MS in the presence of competing protein (pepsin) in simple aqueous solution. In real CFS, both AfB1 adsorption capacities (Qmax) and affinities (K) by all organo-smectites were greater (Qmax = up to 0.45 mol kg–1 and K = up to 0.165 μM–1) than those by Ca-3MS (Qmax = 0.22 mol kg–1 and K = 0.031 μM–1). The study suggested that using smectites modified with an organic nutritive compound could be an effective, economical, and safe strategy for removing mycotoxins, including aflatoxins, during biofuel production.

We critically review potential involvement of trimethylamine N-oxide (TMAO) as a link between diet, the gut microbiota and CVD. Generated primarily from dietary choline and carnitine by gut bacteria and hepatic flavin-containing mono-oxygenase (FMO) activity, TMAO could promote cardiometabolic disease when chronically elevated. However, control of circulating TMAO is poorly understood, and diet, age, body mass, sex hormones, renal clearance, FMO3 expression and genetic background may explain as little as 25 % of TMAO variance. The basis of elevations with obesity, diabetes, atherosclerosis or CHD is similarly ill-defined, although gut microbiota profiles/remodelling appear critical. Elevated TMAO could promote CVD via inflammation, oxidative stress, scavenger receptor up-regulation, reverse cholesterol transport (RCT) inhibition, and cardiovascular dysfunction. However, concentrations influencing inflammation, scavenger receptors and RCT (≥100 µm) are only achieved in advanced heart failure or chronic kidney disease (CKD), and greatly exceed pathogenicity of <1–5 µm levels implied in some TMAO–CVD associations. There is also evidence that CVD risk is insensitive to TMAO variance beyond these levels in omnivores and vegetarians, and that major TMAO sources are cardioprotective. Assessing available evidence suggests that modest elevations in TMAO (≤10 µm) are a non-pathogenic consequence of diverse risk factors (ageing, obesity, dyslipidaemia, insulin resistance/diabetes, renal dysfunction), indirectly reflecting CVD risk without participating mechanistically. Nonetheless, TMAO may surpass a pathogenic threshold as a consequence of CVD/CKD, secondarily promoting disease progression. TMAO might thus reflect early CVD risk while providing a prognostic biomarker or secondary target in established disease, although mechanistic contributions to CVD await confirmation.

Supplementation of carnitine has been shown to improve performance characteristics such as protein accretion in growing pigs. The molecular mechanisms underlying this phenomenon are largely unknown. Based on recent results from DNA microchip analysis, we hypothesized that carnitine supplementation leads to a downregulation of genes of the ubiquitin proteasome system (UPS). The UPS is the most important system for protein breakdown in tissues, which in turn could be an explanation for increased protein accretion. To test this hypothesis, we fed sixteen male, four-week-old piglets either a control diet or the same diet supplemented with carnitine and determined the expression of several genes involved in the UPS in the liver and skeletal muscle. To further determine whether the effects of carnitine on the expression of genes of the UPS are mediated directly or indirectly, we also investigated the effect of carnitine on the expression of genes of the UPS in cultured C2C12 myotubes and HepG2 liver cells. In the liver of piglets fed the carnitine-supplemented diet, the relative mRNA levels of atrogin-1, E214k and Psma1 were lower than in those of the control piglets (P < 0.05). In skeletal muscle, the relative mRNA levels of atrogin-1, MuRF1, E214k, Psma1 and ubiquitin were lower in piglets fed the carnitine-supplemented diet than that in control piglets (P < 0.05). Incubating C2C12 myotubes and HepG2 liver cells with increasing concentrations of carnitine had no effect on basal and/or hydrocortisone-stimulated mRNA levels of genes of the UPS. In conclusion, this study shows that dietary carnitine decreases the transcript levels of several genes involved in the UPS in skeletal muscle and liver of piglets, whereas carnitine has no effect on the transcript levels of these genes in cultivated HepG2 liver cells and C2C12 myotubes. These data suggest that the inhibitory effect of carnitine on the expression of genes of the UPS is mediated indirectly, probably via modulating the release of inhibitors of the UPS such as IGF-1. The inhibitory effect of carnitine on the expression of genes of the UPS might explain, at least partially, the increased protein accretion in piglets supplemented with carnitine.

This study was performed to investigate whether dietary lysine concentration influences the carnitine status of pigs. Therefore, an experiment with twenty young pigs with an average body weight of 21 kg was performed which were fed either a control diet (9·7 g lysine/kg) or a diet with a moderate excess of lysine (16·8 g lysine/kg). Concentrations of all the other amino acids did not differ between the diets. Pigs fed the high-lysine diet had lower concentrations of free and total carnitine in plasma, liver, kidney and skeletal muscle than control pigs (P < 0·05). Pigs fed the high-lysine diet moreover had an increased concentration of trimethyllysine (TML), a reduced mRNA abundance of TML dioxygenase and reduced concentrations of γ-butyrobetaine (BB) in muscle, indicating that the conversion of TML into BB in muscle was impaired. Concentrations of BB, the metabolic precursor of carnitine, in plasma, liver and kidney were also reduced in pigs fed the high-lysine diet while the activity of BB dioxygenase in kidney was not different and that in liver was even increased compared to control pigs (P < 0·05). In conclusion, this study shows that a moderate dietary excess of lysine lowers plasma and tissue carnitine concentrations in pigs. Reduced concentrations of BB in liver and kidney suggest that the depressed carnitine status was likely caused by a decreased rate of carnitine synthesis due to a diminished availability of carnitine precursor, probably mainly as a result of an impaired BB formation in muscle.

It has been shown that treatment of rats with clofibrate, a synthetic agonist of PPARα, increases mRNA concentration of organic cation transporters (OCTN)-1 and -2 and concentration of carnitine in the liver. Since oxidised fats have been demonstrated in rats to activate hepatic PPARα, we tested the hypothesis that they also up regulate OCTN. Eighteen rats were orally administered either sunflower-seed oil (control group) or an oxidised fat prepared by heating sunflower-seed oil, for 6 d. Rats administered the oxidised fat had higher mRNA concentrations of typical PPARα target genes such as acyl-CoA oxidase, cytochrome P450 4A1 and carnitine palmitoyltransferases-1A and -2 in liver and small intestine than control rats (P < 0·05). Furthermore, rats treated with oxidised fat had higher hepatic mRNA concentrations of OCTN1 (1·5-fold) and OCTN2 (3·1-fold), a higher carnitine concentration in the liver and lower carnitine concentrations in plasma, gastrocnemius and heart muscle than control rats (P < 0·05). Moreover, rats administered oxidised fat had a higher mRNA concentration of OCTN2 in small intestine (2·4-fold; P < 0·05) than control rats. In conclusion, the present study shows that an oxidised fat causes an up regulation of OCTN in the liver and small intestine. An increased hepatic carnitine concentration in rats treated with the oxidised fat is probably at least in part due to an increased uptake of carnitine into the liver which in turn leads to reduced plasma and muscle carnitine concentrations. The present study supports the hypothesis that nutrients acting as PPARα agonists influence whole-body carnitine homeostasis.

This study evaluated carnitine and lipid status of fifty Korean newborns. Each subject was assigned to two groups: one according to body weight at birth and the other according to gestational age. Serum total, HDL- and LDL-cholesterol were significantly lower and triacylglycerols were significantly higher, by 14 %, in the low birth weight infant (LBWI, 1310–2490 g) group compared with the normal birth weight infant (NBWI, 2570–4420 g) group. Neither birth weight nor gestational age affected serum total carnitine concentrations. However, serum ASAC (acid-soluble acylcarnitine) concentrations were 43 % higher (P < 0·001) in the LBWI group compared with the NBWI group, and approximately twice as high (P < 0·05) in the 28–32 gestational age group compared with the other gestational age groups. NEC (non-esterified acyl carnitine) fractions were significantly higher in the NBWI and 28–32 week groups (P < 0·001 and P < 0·05); consequently serum acyl/NEC carnitine ratios were four times higher in the LBWI group compared with the NBWI group and 2–3 times higher in the 25–32 week age group compared with the more advanced gestational age groups. Urinary carnitine excretion, including the NEC fraction and total carnitine, was significantly higher (P < 0·001) for LBWI than for NBWI. By gestational age, NEC excretion of the 28–32 week group was significantly (P < 0·05) higher than that of the other two groups, but total carnitine excretion was not different among the groups. This study demonstrated that Korean immature and preterm newborns have higher serum triacylglycerol concentrations but lower carnitine status than NBWI. Therefore, the lower carnitine status and moderately higher triacylglycerols may suggest that LBWI in Korea might be at risk for poor carnitine status and decreased capacity to utilise fatty acids for energy.

This study compared plasma and urinary carnitine concentrations in pregnant and non-pregnant Korean women. The subjects were fifty pregnant women and thirty non-pregnant women aged 24–28 years. During the first trimester, dietary carnitine intakes in the pregnant women were much lower than in non-pregnant women (70·00 (sd 29·22) μmol/d), but over the course of pregnancy carnitine intake increased from 44·64 (sd 24·84) μmol/d during the first trimester to 96·11 (sd 36·56) μmol/d during the third trimester. Pregnant women had a significantly lower plasma carnitine concentration than non-pregnant women. Plasma concentrations of non-esterified carnitine, acid-soluble acylcarnitine and total carnitine were significantly lower during the second and third trimesters than the first. Plasma acid-insoluble acylcarnitine levels, which tended to be higher in the non-pregnant women compared with the pregnant women, increased significantly as gestation proceeded. The urinary excretion of non-esterified carnitine, acid-soluble acylcarnitine and total carnitine was significantly higher in the pregnant women during the first and second trimesters than in non-pregnant women and decreased significantly as gestation proceeded. We found that there was a significant decrease in plasma carnitine level even though dietary carnitine intake increased as gestation proceeded. The low urinary excretion of carnitine in late pregnancy may be caused by an increased demand during pregnancy.

A study was undertaken to examine the effect of different dietary carnitine (200 and 1000 mg/kg diet) and fat (90 and 190 g/kg diet) supplementation on growth and fatty acid concentrations of fish fed either with a low- (13 g/kg) or a high-lysine (21 g/kg) diet. African catfish (22·7 g/fish), Clarias gariepinus Burchell, juveniles were stocked (sixteen aquaria, twenty-five fish per aquarium) and fed for a maximum of 74 d. Dietary lysine had a clear effect on growth performance and feed conversion ratios, but dietary carnitine supplements had no effect. High-carnitine supplements increased total carnitine content (P<0·0004) and reduced tissue free carnitine: acyl-carnitine ratio (P<0·05) compared with low-carnitine supplements. High-fat supplements decreased liver carnitine concentrations. Clear effects on liver fatty acid concentrations were observed in high-carnitine-fed fish compared with low-carnitine-fed fish. The primary liver fatty acids affected were 18:2n-6 (linoleic acid), 20:5n-3 (eicosapentanoic acid) and 22:6n-3 (docosahexanoic acid). The whole-body fatty acid balance suggested that 20:5n-3 disappeared (apparently by β-oxidation) more readily than 18:2n-6 and/or 22:6n-3. From 774 mg 20:5n-3 eaten by high-lysine–high-fat–low-carnitine fish, 58 % was not assimilated into body tissues. High-carnitine-fed fish showed an increase in 20:5n-3 oxidation by 7 % compared with low-carnitine fish. Although dietary carnitine did not improve body growth, these results support the hypothesis that carnitine can enhance the mobilisation of long-chain fatty acids towards oxidation.