Phytyl Fatty Acid Esters in the Pulp of Bell Pepper (Capsicum annuum).

Phytyl fatty acid esters (PFAE) are esters of fatty acids with the isoprenoid alcohol phytol (3,7R,11R,15-tetramethylhexadec-2E-enol). In this study, PFAE were identified and quantified in bell pepper using gas chromatography with mass spectrometry (GC-MS). All red (n = 14) and yellow (n = 6) samples contained six or seven PFAE at 0.9-11.2 mg/100 g fresh weight. By contrast, PFAE were not detected in green bell pepper samples (n = 3). PFAE might eventually be a source for bioavailable phytol, which can be transformed into phytanic acid by humans. Phytanic acid cannot be properly degraded by patients who suffer from Refsum's disease (tolerable daily intake (TDI) ≤ 10 mg of phytanic acid). The phytol moiety of the PFAE (0.4-5.4 mg/100 g fresh weight) would contribute up to ∼50% to the TDI with the consumption of only one portion of bell pepper fruit pulp.

Chemical offense by means of toxicysts in the freshwater ciliate, Coleps hirtus.

Coleps hirtus is a small common freshwater ciliate belonging to the protostomatid group, its body covered by calcified plates assembled to form an armor. Coleps feeds on bacteria, algae, flagellates, living and dead ciliates, animal and plant tissues. To assist its carnivorous feeding the ciliate is equipped with offensive extrusomes (toxicysts), clustering mainly in and around its oral aperture. In this study, we isolated the discharge of the toxicysts from living cells, evaluating its cytotoxic effects against various ciliate species, and demonstrating that it is essential for the effectiveness of Coleps' predatory behavior. The analysis of the toxicyst discharge performed by liquid chromatography-electrospray-mass spectrometry and gas chromatography-mass spectrometry, revealed the presence of a mixture of 19 saturated, monounsaturated and polyunsaturated free fatty acids with the addition of a minor amount of a diterpenoid (phytanic acid).

Toxicological issues associated with production and processing of meat.

Meat is a very complex and continuously changing ex vivo system of various high- and low-molecular substances that can be used for satisfying needs of the human organism for metabolic energy, building material and fulfilling of the other vital functions. A great majority of these substances are useful and safe for the consumer. Yet, meat and meat products may always contain substances exerting detrimental effects to the consumer's organism. The present paper is a literature review of the most important potentially toxic substances found in meat and meat products; their classification, ways of getting into the meat or formation during meat processing, undesirable physiological outcomes and biochemical mechanisms of their toxic effects, and methods for reduction of these responses.

Novel aspects of health promoting compounds in meat.

Meat is an integral part of the human diet. Besides essential amino acids and nutritive factors of high quality and availability, meat provides often overlooked components of importance for human health. These are amino acids and bioactive compounds that may be very important in i) preventing muscle wasting diseases, such as in sarcopenia, ii) reducing food and caloric intake to prevent metabolic syndrome, iii) blood pressure homeostasis via ACE-inhibitory components from connective tissue, and iv) maintaining functional gut environment through meat-derived nucleotides and nucleosides. In addition, meat could be an important source of phytanic acid, conjugated linoleic acids and antioxidants. Further, it becomes increasingly apparent that design of in vitro meat will be possible, and that this development may lead to improved health benefits from commercially viable and sustainable meat products.

Characterization of two Agrostis-Festuca alpine pastures and their influence on cheese composition.

Recently, there has been a renewed interest in mountain farming, and several studies have been carried out on milk and cheese obtained in the unique environmental conditions of the Alps, a 1300 km mountain chain, located in the north of Italy. In this paper, the influence, on some cheese constituents, of two very similar mountain grasslands, both dominated by Festuca - Agrostis , was investigated. The two pastures were located in the same area in the southeastern Italian alpine region and differed in sunshine orientation and exposure. Milk obtained from cows grazing on these pastures was used to produce a semi-hard traditional cheese. The differences observed between the cheeses of the two areas for both some hydrocarbons (1-phytene and 2-phytene) and trans-fatty acids can be explained by a different rumen environment created by the botanical composition of the two pastures. The multidisciplinary approach can be considered a successful strategy, suitable for studying markers of authenticity.

Phytanic acid and the risk of non-Hodgkin lymphoma.

Greater consumption of red meat, processed meat and dairy products has been associated with an increased risk of non-Hodgkin lymphoma (NHL) in several previous reports. Phytanic acid, a saturated fatty acid obtained primarily through the consumption of ruminant meat and dairy products, may offer a potential underlying mechanism for these associations. In a population-based case-control study of 336 cases and 460 controls conducted in Nebraska during 1999-2002, we examined whether phytanic acid-containing foods or total phytanic acid intake, estimated from a food frequency questionnaire and the published phytanic acid values of 151 food items, were associated with increased NHL risk. Unconditional logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals for overall NHL and the common NHL histologic subtypes. In multivariable models, higher intakes of density-adjusted beef [OR(T3 vs. T1) = 1.5 (1.1-2.2); P(trend) = 0.02], total dairy products [OR = 1.5 (1.1-2.2); P(trend) = 0.02) and milk [OR = 1.6 (1.1-2.3); P(trend) = 0.01] were associated with an increased risk of NHL. Intake of total phytanic acid was positively associated with NHL risk [OR = 1.5 (1.0-2.1); P(trend) = 0.04]. In analyses stratified by NHL subtype, greater consumption of beef was associated with an increased risk of diffuse large B-cell lymphoma, and greater consumption of milk was associated with an increased risk of follicular lymphoma (FL). Total phytanic acid intake was associated with an increased risk of FL and small lymphocytic lymphoma/chronic lymphocytic leukemia. Our results provide support that total phytanic acid and phytanic acid-containing foods may increase NHL risk.

Content and distribution of phytanic acid diastereomers in organic milk as affected by feed composition.

Phytanic acid (PA) is a bioactive compound found in milk that is derived from the phytol chain of chlorophyll, and the content of PA in milk fat depends on the availability of phytol from feed. In this study, the content of PA diastereomers was analyzed in milk sampled from five organic herds twice during the grazing season (May and September). The total content of PA was higher in September compared to May, but was not affected by breed (Danish Holstein or Danish Jersey). Total PA could not be directly related to intake of green feed items. The distribution between diastereomers was closely related to the amount of grazed clovers, where a higher intake resulted in a higher share of the RRR isomer.

Polyunsaturated fats attenuate the dietary phytol-induced increase in hepatic fatty acid oxidation in mice.

The effects of dietary phytol and the type of dietary fat on hepatic fatty acid oxidation were examined in male ICR mice. Mice were fed diets containing 0 or 5 g/kg phytol and 100 g/kg palm, safflower, or fish oil for 21 d. Among the groups fed phytol-free diets, the activities and mRNA abundance of various enzymes involved in fatty acid oxidation were greater in mice fed fish oil than in those fed palm or safflower oil. Dietary phytol profoundly increased the activities and mRNA abundance of hepatic fatty acid oxidation enzymes in mice fed palm oil. However, safflower and fish oils, especially the latter, greatly attenuated the phytol-dependent increase in hepatic fatty acid oxidation. The hepatic concentration of phytanic acid, a metabolite of phytol that is the ligand and activator of retinoid X receptors and peroxisome proliferator-activated receptors, was higher in mice fed fish oil than safflower or palm oil, and in those administered safflower oil than palm oil. The hepatic mRNA abundance of sterol carrier protein-2, a lipid-binding protein involved in phytol metabolism, was inversely correlated with the hepatic concentration of phytanic acid. We demonstrated that polyunsaturated fats attenuate the enhancing effect of dietary phytol on hepatic fatty acid oxidation. Dietary fat-dependent changes in the hepatic phytanic acid concentration cannot account for this phenomenon.

A study on the causes for the elevated n-3 fatty acids in cows' milk of alpine origin.

The influence of grass-only diets either from rye-grass-dominated lowland pastures (400 m above sea level) or botanically diverse alpine pastures (2000 m) on the FA profile of milk was investigated using three groups of six Brown Swiss cows each. Two groups were fed grass-only on pasture (P) or freshly harvested in barn (B), both for two experimental periods in the lowlands and, consecutively, two periods on the alp. Group C served as the control, receiving a silage-concentrate diet and permanently staying in the lowlands. Effects of vegetation stage or pasture vs. barn feeding on milk fat composition were negligible. Compared with the control, alpha-linolenic acid (18:3n-3) consumption was elevated in groups P and B (79%; P< 0.001) during the lowland periods but decreased on the alp to the level of C owing to feed intake depression and lower 18:3n-3 concentration in the alpine forage. Average 18:3n-3 contents of milk fat were higher in groups P and B than in C by 33% (P< 0.01) at low and by 96% (P < 0.001) at high altitude, indicating that 18:3n-3 levels in milk were to some extent independent of 18:3n-3 consumption. The cis-9, trans-11 CLA content in milk of grass-fed cows was higher compared with C but lower for the alpine vs. lowland periods whereas the trans-11, cis-13 isomer further increased with altitude. Long-chain n-3 FA and phytanic acid increased while arachidonic acid decreased with grass-only feeding, but none of them responded to altitude. Grass-only feeding increased milk alpha-tocopherol concentration by 86 and 134% at low and high altitude (P < 0.001), respectively. Changes in the ruminal ecosystem due to energy shortage or specific secondary plant metabolites are discussed as possible causes for the high 18:3n-3 concentrations in alpine milk.

Identification of fatty aldehyde dehydrogenase in the breakdown of phytol to phytanic acid.

Phytol is a branched chain fatty alcohol, which is abundantly present in nature as part of the chlorophyll molecule. In its free form, phytol is metabolized to phytanic acid, which accumulates in patients suffering from a variety of peroxisomal disorders, including Refsum disease. The breakdown of phytol to phytanic acid takes place in three steps, in which first, the alcohol is converted to the aldehyde, second the aldehyde is converted to phytenic acid, and finally the double bond is reduced to yield phytanic acid. By culturing fibroblasts in the presence of phytol, increases in the levels of phytenic and phytanic acid were detected. Interestingly, fibroblasts derived from patients affected by Sjögren Larsson syndrome (SLS), known to be deficient in microsomal fatty aldehyde dehydrogenase (FALDH) were found to be deficient in this. In addition, fibroblast homogenates of these patients, incubated with phytol in the presence of NAD+ did not produce any phytenic acid. This indicates that FALDH is involved in the breakdown of phytol.

Analysis of intact tetraether lipids in archaeal cell material and sediments by high performance liquid chromatography/atmospheric pressure chemical ionization mass spectrometry.

A method combining normal phase high performance liquid chromatography (HPLC) with positive ion atmospheric pressure chemical ionization mass spectrometry (APCI-MS) was developed for the analysis of intact glycerol dialkyl glycerol tetraethers (GDGTs) in archaeal cell material and sediments. All GDGTs previously reported to occur in the thermophilic archaeon Sulfolobus solfataricus could be identified based on their mass spectra and retention time. Positive ion mass spectra consisted of abundant protonated molecules and fragment ions corresponding to loss of water and the glycerol moiety. In addition, two novel GDGTs representing alternative combinations of biphytanyl moieties were observed. Using this method, the tetraethers present in the thermophilic archaeon Metallosphaera sedula and two sediment samples were characterized. This rapid method will greatly contribute to the establishment of the sedimentary record of these compounds and increase our understanding of archaea and their occurrence in widely different environments.

Pre- and postnatal diagnosis of peroxisomal disorders using stable-isotope dilution gas chromatography--mass spectrometry.

Quantitative analysis of the following peroxisomal metabolites is reported: very long-chain fatty acids (VLCFA), pipecolic acid, bile acid intermediates, phytanic and pristanic acid, in plasma, urine, cerebrospinal fluid (CSF), blood spots collected at neonatal screening and amniotic fluid. An overview is given of the concentrations of these metabolites in body fluids from control subjects and all patients investigated so far in this laboratory. The method of choice is gas chromatography -- mass spectrometry (GC-MS) with electron capture detection, combined with the use of stable-isotope-labelled internal standards.

Measurement of very long-chain fatty acids, phytanic and pristanic acid in plasma and cultured fibroblasts by gas chromatography.

Two methods are described, both currently used in our laboratory, for the quantitative analysis of very long-chain fatty acids, phytanic acid and pristanic acid in plasma and cultured fibroblasts by gas-liquid chromatography. The first method is based on the procedure developed by Moser and Moser (1991) and the second is based on the method of Onkenhout and colleagues (1989), which is an application of the original method of Lepage and Roy for plasma and fibroblasts. A survey is given of the concentrations of very long-chain fatty acids, pristanic and phytanic acid in plasma and fibroblasts from control subjects and all patients investigated so far in our laboratory.

Isolation and characterization of novel 2-hydroxy fatty acids from the phospholipids of the sponge Smenospongia aurea.

The Caribbean sponge Smenospongia aurea revealed the presence of six novel branched alpha-hydroxy fatty acids: 2-hydroxy-17-methyloctadecanoic acid, 2-hydroxy-21-methyldocosanoic acid, 2-hydroxy-22-methyltricosanoic acid, and 2-hydroxy-22-methyltetracosanoic acid, 2-hydroxy-24-methylpentacosanoic acid, and 2-hydroxy-23-methylpentacosanoic acid. These novel alpha-hydroxy fatty acids were associated with phosphatidylethanolamine. The sponges Aplysina lacunosa and Aplysina fistularis also contained considerable amounts of alpha-hydroxy fatty acids, the very long-chain 5,9,23-tricontatrienoic acid (30:3), and phytanic acid. The sterol composition of the three sponges was also studied. It indicated that A. lacunosa and A. fistularis contained large amounts of aplysterol and verongulasterol, while S. aurea did not show any of these sterols. The results are discussed in terms of the taxonomy of the species.

Phytanic acid and very long chain fatty acids in genetic peroxisomal disorders.

1. Phytanic acid, phytanyl-triacylglycerols, and very long chain fatty acids were analysed by gas chromatography or thin-layer chromatography in blood and tissues of patients with different genetic peroxisomal disorders (Refsum's disease, X-linked adrenoleukodystrophy, neonatal adrenoleukodystrophy, Zellweger syndrome). 2. We evaluated these analyses in the detection of patients with Refsum's disease, X-linked adrenoleukodystrophy, neonatal adrenoleukodystrophy, and Zellweger syndrome, and of carriers of X-linked adrenoleukodystrophy. In particular, the analysis of phytanyl-triacylglycerols by thin-layer chromatography proved to be a rapid and reliable method for the detection of patients and the monitoring of their dietary treatment in Refsum's disease. In X-linked adrenoleukodystrophy, carrier detection may depend on very long chain fatty acid analysis in more than one material (e.g. plasma and fibroblasts). 3. Analysis of phytanic acid showed that in patients with multiple impairments of peroxisomal functions (Zellweger syndrome, neonatal adrenoleukodystrophy) phytanic acid levels may be increased not only in serum, but also in the tissue (e.g. brain, adrenals, kidney). 4. Analysis of very long chain fatty acids in cholesterol esters from the brain, adrenals, kidney, and liver of patients with peroxisomal disorders revealed four different types of very long chain fatty acid patterns according to the behaviour of C 26:0 and of other saturated and monounsaturated very long chain fatty acids.(ABSTRACT TRUNCATED AT 250 WORDS)

[Refsum's disease. Hereditary atactic polyneuritis]. / Refsum's sygdom. Heredopathia atactica polyneuritiformis.

As it is possible to prevent progression of the disease by early treatment all patients with otherwise unexplained retinitis pigmentosa should be screened for phytanic acid accumulation in the plasma.

The enzymatic and mass spectrometric identification of 2-oxophytanic acid, a product of the peroxisomal oxidation of l-2-hydroxyphytanic acid.

A previously unreported metabolite of the mammalian phytanic acid breakdown pathway, 2-oxophytanic acid, was isolated and analysed by mass spectrometry. The metabolic origin of the 2-oxoacid is the oxidation by a rat kidney peroxisomal H2O2-generating oxidase of L-2-hydroxyphytanic acid, a well-established intermediate in phytanic acid alpha-oxidation.

Tissue distribution of phytanic acid and its analogues in a kinship with Refsum's disease.

Three of six kin were identified, by high performance thin layer chromatography, capillary gas chromatography and mass spectrometry, as having phytanic acid storage disease. Phytanic acid was found in triacylglycerol and, to a lesser degree, in phosphatidylcholine and free fatty acids. An unsaturated analogue of phytanic acid was additionally identified in plasma and erythrocyte triacylglycerols. In plasma, branched chain fatty acids were primarily localized in the low density lipoprotein fraction. The concentration of plasma major fatty acids was not affected by the presence of these branched chain fatty acids. In contrast to plasma, only small amounts of phytanic acid were found in cerebrospinal fluid and biopsied sural nerve. The nerve phytanate was mainly associated with triacylglycerol in epineurial and perineurial tissues. Lack of phytanate accumulation in sural endoneurium, even in cases with severe fiber degeneration, suggests that demyelination in Refsum's disease may not be due to myelin instability resulting from the incorporation of branched chain fatty acids into peripheral nerve membrane.