In vitro determination of the allergenic potential of technologically altered hen's egg.

Hen's egg allergy represents one of the most common and severe IgE-mediated reactions to food in infants and young children. It persists, however, in many cases also lifelong. Therefore, the aim of this study was the detailed analysis of a technological process used to reduce the allergenic potential of hen's egg. The investigation focused on the pasteurized egg as starting material, intermediate, and final products of a nine-step manufacturing process performed for use of eggs in convenience products appropriate for allergic individuals. The steps consisted of a combination of various heat treatments and enzymatic hydrolyses. The alterations were controlled by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), immunoblotting, enzyme allergosorbent test (EAST) inhibition, and mass spectrometry. Thereby it could be demonstrated that the allergenic potential of the raw material was reduced from step to step, and despite the known stability against heat and proteolysis of certain egg proteins, the total allergenic potential was finally below 1/100 that of the starting material without a significant change in texture and flavor as evaluated in various products.

Formation of conjugated linoleic acid metabolites in human vascular endothelial cells.

Conjugated linoleic acids (CLAs) are bioactive lipid compounds showing anti-atherogenic actions in cell culture experiments and animal models of atherosclerosis without exact knowledge about the underlying mechanisms. CLAs were recently reported to be further metabolized to bioactive conjugated metabolites indicating that these metabolites are possibly involved in mediating the anti-atherogenic actions of CLA. Regarding the lack of information with respect to the formation of CLA metabolites in the vascular endothelium, which is strongly involved in the process of atherosclerosis, the present study aimed to explore the potential formation of CLA metabolites in vascular endothelial cells. The results from the present study show for the first time that the CLA isomers cis-9, trans-11 CLA and trans-10, cis-12 CLA are metabolized within endothelial cells to beta-oxidation products such as CD16:2c7t9 and CD16:2t8c10 and elongation products such as CD20:2c11t13, CD20:2t12c14 as well as CD22:2c13t15 and CD22:2t14c16. Different CD16:2/CLA ratios observed between cells treated with different CLA isomers indicate that the metabolism of CLAs depends on the configuration of the conjugated double bonds. In conclusion, regarding the biological activity reported for CD20:2t12c14 and other metabolites of CLA, the present results indicate that metabolites of CLA are possibly also involved in mediating the anti-atherogenic actions of CLA.

CLA isomers inhibit TNFalpha-induced eicosanoid release from human vascular smooth muscle cells via a PPARgamma ligand-like action.

Conjugated linoleic acids (CLAs) were reported to have anti-atherogenic properties in animal feeding experiments. In an attempt to elucidate the molecular mechanisms of these anti-atherogenic effects, the modulatory potential of CLA on cytokine-induced eicosanoid production from smooth muscle cells (SMCs), which contributes to the chronic inflammatory response associated with atherosclerosis, has been investigated in the present study. cis-9, trans-11 CLA and trans-10, cis-12 CLA were shown to reduce proportions of the eicosanoid precursor arachidonic acid in SMC total lipids and to inhibit cytokine-induced NF-kappaB DNA-binding activity, mRNA levels of inducible enzymes involved in eicosanoid formation (cPLA2, COX-2, mPGES), and the production of the prostaglandins PGE2 and PGI2 by TNFalpha-stimulated SMCs in a dose-dependent manner. The effect of 50 micromol/L of either CLA isomer was as effective as 10 micromol/L of the PPARgamma agonist troglitazone in terms of inhibiting the TNFalpha-stimulated eicosanoid production by SMCs. PPARgamma DNA-binding activity was increased by both CLA isomers compared to control cells. Moreover, it was shown that the PPARgamma antagonist T0070907 partially abrogated the inhibitory action of CLA isomers on cytokine-induced eicosanoid production and NF-kappaB DNA-binding activity by vascular SMCs suggesting that PPARgamma signalling is at least partially involved in the action of CLA in human vascular SMCs. With respect to the effects of CLA on experimental atherosclerosis, our findings suggest that the anti-inflammatory effect of CLA is at least partially responsible for the anti-atherogenic effects of CLA observed in vivo.

Concentrations of conjugated linoleic acids in neonatal blood in relationship to those in maternal blood.

In this study, the proportions of conjugated linoleic acids (CLA) in total lipids of plasma, lipoproteins and erythrocytes from maternal blood and from venous cord blood of 20 pregnant women consuming conventional western diets after delivery were determined. cis-9, trans-11 CLA was the only isomer detected, and its proportions in maternal blood lipids were relatively low. Mean proportions in plasma, lipoproteins and erythrocytes of mothers were between 0.20 and 0.25 mol/100 mol of total fatty acids. Proportions in cord blood lipids were even lower than those of maternal lipids (values in mol/100 mol: plasma, 0.19+/-0.04; VLDL, 0.20+/-0.06; LDL, 0.15+/-0.03; HDL, 0.14+/-0.06; erythrocytes, 0.12+/-0.05). There was some significant (P<0.05) linear relationship between CLA in maternal lipids and neonatal lipids. The data of this study suggest that CLA proportions in fetal blood lipids are low if mothers are consuming conventional western diets. It is moreover concluded that CLA concentrations in fetal blood lipids are related with maternal CLA intake.

Model studies of lignified fiber fermentation by human fecal microbiota and its impact on heterocyclic aromatic amine adsorption.

This study examined how shifts in pH and fiber fermentation may alter the adsorption of mutagenic heterocyclic aromatic amines (HAAs) 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and 2-amino-9H-pyrido[2,3-b]indole (AalphaC) to dietary fiber in the human small intestine and colon. Nonlignified and artificially lignified maize cell walls were fermented in vitro with human fecal microbiota for 0, 8, or 24h. We then assessed the adsorption of HAAs to unfermented fiber at pH 6.5 and to unfermented and fermented fibers at pH 7.4 to mimic conditions in the small intestine and colon, respectively. HAAs were effectively adsorbed to lignified fiber by up to 74% at pH 6.5 and by up to 68% at pH 7.4. Increasing the lignin content of unfermented fiber from 0.4% to about 14% increased HAA adsorption by two- to three-fold. This increase in lignification reduced microbial fiber degradation from 51% to minimum 8% after 24h of fermentation, whereas variations in the guaiacyl and syringyl makeup of lignin had smaller but significant impacts on fiber degradation. A 24h fermentation decreased the AalphaC adsorption to lignified fiber at pH 7.4 by up to one-third, while PhIP adsorption was not affected. Our results indicate that lignification increases the adsorption of hydrophobic HAAs to fiber but shifts in pH and fermentation may somewhat diminish adsorption of some HAAs as fiber passes from the small intestine into and through the colon.

Investigation of the allergenic potential of wines fined with various proteinogenic fining agents by ELISA.

Hidden allergens are a common problem in food safety that has been known for many years. This is why the European Parliament adopted Directive 2003/89/EC amending 2000/13/EC. In addition to specific ingredients, Directive 2003/89/EC also requests the declaration of specific products that were used in the production and could be a risk for allergic individuals. This also includes the declaration of fining agents and lysozyme used in wines. In fact, it could be assumed that fining agents would be almost completely removed during the manufacturing process; however, until now there has been no necessity to analyze wine for these fining agents. By applying enzyme-linked immunosorbent assay (ELISA), residuals of fining agent proteins and the stabilizer lysozyme were investigated in various German wines. The results showed no detectable amounts of fining agents in wines, except for dried egg white and lysozyme, both derived from hen's egg white. For those products, adverse reactions against treated wines could not be excluded.

Dietary fiber from coffee beverage: degradation by human fecal microbiota.

Arabinogalactans and galactomannans from coffee beverages are part of the dietary fiber complex. Chemical structures and fermentability of soluble dietary fiber obtained from a standard filter coffee beverage (Coffea arabica, origin Colombia, medium roasted) by human intestinal bacteria were investigated. One cup (150 mL) of filter coffee contained approximately 0.5 g of soluble dietary fiber (enzymatic-gravimetric methodology), 62% of which were polysaccharides. The remainder was composed of Maillard reaction products and other nonidentified substances. Galactomannans and type II arabinogalactans were present in almost equal proportions. Coffee dietary fiber was readily fermented by human fecal slurries, resulting in the production of short-chain fatty acids (SCFA). After 24 h of fermentation, 85% of total carbohydrates were degraded. In general, arabinosyl units from the polysaccharide fraction were degraded at a slower rate than mannosyl and galactosyl units. In the process of depolymerization arabinogalactans were debranched and the ratio of (1-->3)-linked to (1-->6)-linked galactosyl residues decreased. Structural units composed of (1-->5)-linked arabinosyl residues were least degradable, whereas terminally linked arabinosyl residues were easily utilized. The impact of coffee fiber on numerically dominant population groups of the intestinal microbiota was investigated by fluorescence in situ hybridization combined with flow cytometry (FISH-FC). After 24 h of fermentation, an increase of about 60% of species belonging to the Bacteroides-Prevotella group was observed. The growth of bifidobacteria and lactobacilli was not stimulated.

Moderate ferulate and diferulate levels do not impede maize cell wall degradation by human intestinal microbiota.

The degradation of plant fiber by human gut microbiota could be restricted by xylan substitution and cross-linking by ferulate and diferulates, for example, by hindering the association of enzymes such as xylanases with their substrates. To test the influence of feruloylation on cell wall degradability by human intestinal microbiota, nonlignified primary cell walls from maize cell suspensions, containing various degrees of ferulate substitution and diferulate cross-linking, were incubated in nylon bags in vitro with human fecal microbiota. Degradation rates were determined gravimetrically, and the cell walls were analyzed for carbohydrates, ferulate monomers, dehydrodiferulates, dehydrotriferulates, and other minor phenolic constituents. Shifting cell wall concentrations of total ferulates from 1.5 to 15.8 mg/g and those of diferulates from 0.8 to 2.6 mg/g did not alter the release of carbohydrates or the overall degradation of cell walls. After 24 h of fermentation, the degradation of xylans and pectins exceeded 90%, whereas cellulose remained undegraded. The results indicate that low to moderate levels of ferulates and diferulates do not interfere with hydrolysis of nonlignified cell walls by human gut microbiota.

Coffee dietary fiber contents and structural characteristics as influenced by coffee type and technological and brewing procedures.

Coffee brews contain considerable amounts of soluble dietary fiber, mainly low substituted galactomannans and type II arabinogalactans. Factors possibly influencing the content and structures of dietary fiber in coffee brews, such as type of coffee, roasting and grinding degree, and brewing procedure, were studied. In addition, several commercial samples such as instant espresso, instant coffee, instant cappuccino, decaffeinated coffees, and coffee pads were analyzed. The dietary fiber contents of the coffee brews ranged from 0.14 to 0.65 g/100 mL (enzymatic-gravimetric methodology), proving an influence of the factors investigated. For example, the drip brew of an arabica coffee contained significantly more soluble dietary fiber than the drip brew of a comparable robusta coffee, and depending on the brewing procedure, the soluble dietary fiber content of beverages obtained from the same coffee sample ranged from 0.26 to 0.38 g/100 mL. Dietary fiber contents of coffee brews were enhanced only up to a certain degree of roast. Drip brews of decaffeinated arabica coffees (commercial samples) contained significantly less dietary fiber than any non-decaffeinated drip brew investigated in this study. The observed differences in the dietary fiber contents were accompanied by changes in the structural characteristics of fiber polysaccharides, such as galactomannan/arabinogalactan ratio, galactose substitution degree of mannans, or galactose/arabinose ratio of arabinogalactans as analyzed by methylation analysis.

Detection of conjugated dienoic fatty acids in human vascular smooth muscle cells treated with conjugated linoleic acid.

Conjugated linoleic acids (CLA) have attracted scientific interest due to their potential beneficial effects on atherosclerosis. Recent studies demonstrated that conjugated metabolites of CLA are found in tissues of CLA-fed animals and cultured cells treated with CLA. This observation has gained in importance since it has recently been shown that these metabolites of CLA exert specific biological activities. Therefore, the present study aimed to explore the potential formation of metabolites of cis-9, trans-11 CLA, trans-10, cis-12 CLA and trans-9, trans-11 CLA in cells of the vascular wall, which has not yet been shown. Examination of fatty acid composition of total cell lipids using Ag+-HPLC, GC-FID and GC-MS analysis revealed a significant isomer-specific formation of conjugated metabolites of CLA such as CD16:2, CD20:2 and CD22:2 in human coronary artery smooth muscle cells treated with various CLA isomers. Different CD16:2/CLA ratios between various CLA isomers as observed in the present study indicate that fatty acid metabolism is differently affected by the configuration of the double bonds. In conclusion, the observation from the present study suggests that the effects of CLA in vascular cells might not only be mediated by CLA itself but also by its conjugated metabolites. Future studies using highly purified conjugated metabolites of CLA are necessary to study their role in mediating biological effects of CLA in cell culture systems.

Isolation and structural identification of complex feruloylated heteroxylan side-chains from maize bran.

Three complex heteroxylan side-chains acylated with ferulate and one arabinosyl ester of p-coumaric acid have been isolated from maize bran insoluble fibre after acidic hydrolysis and fractionation by gel permeation chromatography and semi-preparative RP-HPLC. The complete structural elucidation of all isolated compounds was achieved by 1D/2D NMR spectroscopy and HPLC-MS in combination with methylation analysis. The absolute configuration of the carbohydrate constituents was determined by chiral GC after acidic hydrolysis and trifluoroacetylation. The identified feruloylated tetrasaccharides alpha-d-xylopyranosyl-(1-->3)-alpha-l-galactopyranosyl-(1-->2)-beta-d-xylopyranosyl-(1-->2)-5-O-trans-feruloyl-l-arabinofuranose (FAXGX) and alpha-d-galactopyranosyl-(1-->3)-alpha-l-galactopyranosyl-(1-->2)-beta-d-xylopyranosyl-(1-->2)-5-O-trans-feruloyl-l-arabinofuranose (FAXGG) are the most complex heteroxylan side-chains from maize bran that have been isolated to date. The isolated trisaccharide alpha-l-galactopyranosyl-(1-->2)-beta-d-xylopyranosyl-(1-->2)-5-O-trans-feruloyl-l-arabinofuranose (FAXG) contributes to the complexity of heteroxylan side-chains from maize bran and 5-O-trans-p-coumaroyl-l-arabinofuranose represents the first p-coumaroylated heteroxylan side-chain isolated from cereal grains. Complex feruloylated heteroxylan side-chains are possibly, like ferulate cross-linking of the heteroxylans and binding of heteroxylans to lignin, a factor contributing to limited enzymatic degradation of fibre.

Identification of conjugated linoleic acid elongation and beta-oxidation products by coupled silver-ion HPLC APPI-MS.

Atmospheric pressure photoionisation (APPI) was used in combination with silver-ion (Ag(+))-HPLC for detection of (conjugated) fatty acid methyl esters (FAME) by tandem-mass spectrometry. APPI-MS of methyl esters of conjugated linoleic acid showed an increase in signal-to-noise ratio by a factor of 40 compared to atmospheric pressure chemical ionization in the positive mode. It was possible to identify double bond position, configuration and chain length of FAME based on chromatographic separation and mass detection. The developed LC-MS method is useful for the analysis of CLA elongation and beta-oxidation products, especially with trans,trans-configuration, which are difficult to analyze by conventional GC-MS techniques.

Structural identification of dehydrotriferulic and dehydrotetraferulic acids isolated from insoluble maize bran fiber.

Two new dehydrotriferulic acids and two dehydrotetraferulic acids were isolated from saponified maize bran insoluble fiber using size exclusion chromatography on Bio-Beads S-X3 followed by Sephadex LH-20 chromatography and semipreparative phenyl-hexyl reversed phase high-performance liquid chromatography. On the basis of UV spectroscopy, mass spectrometry, and one- and two-dimensional NMR experiments, the structures were identified as 8-5(noncyclic)/5-5-dehydrotriferulic acid, 8-8(tetrahydrofuran)/5-5-dehydrotriferulic acid, and 4-O-8/5-5/8-O-4-dehydrotetraferulic acid. The second tetramer was tentatively identified as 4-O-8/5-5/8-5(noncyclic)-dehydrotetraferulic acid. Compounds containing an 8-5(noncyclic)-coupled dimeric unit probably do not exist in planta but are formed from their phenylcoumaran precursors containing an 8-5(cyclic)-coupled dimeric unit during saponification. The presented dehydrotrimers are the first dehydrotriferulates that do not contain an 8-O-4-coupled dimeric unit. The ferulate dehydrotetramers that are reported for the first time are presumed, like the dimers and trimers, to cross-link polysaccharides in the plant. Because both tetramers contain a 5-5/8-O-4-dehydrotriferulate moiety, the predominant dehydrotrimer in maize bran, it is not possible to deduce whether tetramers are formed by coupling of a fourth unit to a preformed dehydrotriferulate or by 5-5-coupling of preformed 8-O-4- and 8-5-dehydrodiferulates. Nevertheless, such compounds document expanded roles for ferulates in cross-linking polysaccharides in plant cell walls.

Influence of lignification and feruloylation of maize cell walls on the adsorption of heterocyclic aromatic amines.

Both epidemiological and experimental data indicate that a diet rich in fiber may reduce cancer risk. One possible mechanism is by adsorbing carcinogens and transporting them out of the body without metabolic activation. We investigated the role of fiber lignification and feruloylation on the adsorption of four of the most relevant heterocyclic aromatic amines in food: 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), and 2-amino-9H-pyrido[2,3-b]indole (AalphaC). Adsorption experiments, under conditions mimicking the small intestine, were carried out using nonlignified and artificially lignified primary maize walls with defined lignin and ferulate/diferulate concentrations and defined lignin compositions. Lignin concentration and composition both influenced the adsorption of heterocyclic aromatic amines, especially the more hydrophobic types. Heterocyclic aromatic amine adsorption increased with lignin concentration. 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine and 2-amino-9H-pyrido[2,3-b]indole were better adsorbed by guaiacyl-rich lignins, 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline by syringyl-rich lignins, whereas the adsorption of 2-amino-3-methylimidazo[4,5-f]quinoline was not clearly influenced by lignin composition. Nonlignified cell walls adsorbed lesser amounts of heterocyclic aromatic amines. Variations in cell wall feruloylation had no effect on heterocyclic aromatic amine adsorption.

Use of biotinylated 17beta-estradiol in enzyme-immunoassay development: spacer length and chemical structure of the bridge are the main determinants in simultaneous streptavidin-antibody binding.

17beta-estradiol (E2) concentrations are in the low pg/ml range in plasma. To develop a sensitive enzyme immunoassay (EIA) for E2-determination a highly specific antibody raised against a 6-carboxymethyl (CMO)-E2-bovine serum albumine conjugate was used. Based on 6-CMO-E2 and 6-amino-E2, four biotinylated tracers with two different spacer lengths between E2 and biotin were synthesized using biotinylation reagents in one step reactions. All amino-based tracers were unsuitable for assay development because the antibody binding was too weak compared to the analyte E2. For 6-CMO-based tracers the simultaneous binding of the tracer to the antibody and streptavidin seems to be the determining step in the procedure depending on incubation temperature and spacer lengths. While a short spacer of 9 carbon atoms was susceptible to room temperature, a longer spacer of 16 carbon atoms showed nearly the same results for incubation at 4 degrees C or at room temperature. The absolute detection limit of this system was 0.63 pg/well. For sample clean-up, porcine plasma was solvent-extracted and depending on the initial plasma volume further purified by solvent partition. Determination of reproducibility resulted in intraassay coefficients of variation of 13% and 5.3% for samples with E2-levels of 15 pg/ml and 236 pg/ml, respectively. Measurement of E2-spiked blood plasma revealed recoveries of 83% up to 100% for E2 concentrations between 50 pg/ml and 1000 pg/ml. Only for the lowest concentration (20 pg/ml) a recovery of 58% was observed. Correlation of the EIA with an established radio immunoassay resulted in r=0.991 using the same antibody.

Isolation and structural characterisation of 8-O-4/8-O-4- and 8-8/8-O-4-coupled dehydrotriferulic acids from maize bran.

Two new dehydrotriferulic acids were isolated from saponified maize bran insoluble fiber using Sephadex LH-20 chromatography followed by semi-preparative RP-HPLC. Based on UV-spectroscopy, mass spectroscopy and one- and two-dimensional NMR experiments, the structures were identified as 8-O-4,8-O-4-dehydrotriferulic acid and 8-8(cyclic),8-O-4-dehydrotriferulic acid. Which of the possible phenols in the initially formed 8-8-dehydrodiferulate was etherified by 4-O-8-coupling with ferulate has been unambiguously elucidated. The ferulate dehydrotrimers which give rise to these dehydrotriferulic acids following saponification are presumed, like the dehydrodiferulates, to cross-link polysaccharides. Neither dehydrotriferulic acid described here involves a 5-5-dehydrodiferulic acid unit; only the 5-5-dehydrodimer may be formed intramolecularly. However, whether dehydrotriferulates are capable of cross-linking more than two polysaccharide chains remains open. Although the levels of the isolated ferulate dehydrotrimers are lower than those of the ferulate dehydrodimers, the isolation now of three different dehydrotriferulates indicates that trimers contribute to a strong network cross-linking plant cell wall polysaccharides.

Isolation and structural identification of diarabinosyl 8-O-4-dehydrodiferulate from maize bran insoluble fibre.

The first saccharide ester of a dehydrodiferulic acid (DFA) other than 5-5-DFA has been isolated from maize bran insoluble fibre after acidic hydrolysis and fractionation by gel chromatography and semi-preparative RP-HPLC. HPLC-MS along with 1D, 2D and 3D NMR spectra provided the requisite structural evidence that it is the di-5-O-l-arabinosyl ester of 8-O-4-DFA. Although a range of DFAs have been well authenticated as components released from the cell walls of grasses, the only structural evidence for a DFA attached to polysaccharides had been from 5-5-DFA. The isolation of the 8-O-4-ester demonstrates that polysaccharides in maize cell walls, and presumably in all grasses, are cross-linked through dehydrodiferulates other than 5-5-dehydrodiferulate.

Association of non-starch polysaccharides and ferulic acid in grain amaranth (Amaranthus caudatus L.) dietary fiber.

The association of ferulic acid, an alkali-extractable phenolic acid in amaranth (Amaranthus caudatus L., Amaranthaceae) insoluble fiber (trans-ferulic acid: 620 microg.g-1, cis-ferulic acid: 203 microg.g-1), and non-starch polysaccharides was investigated. Enzymatic hydrolysis of insoluble amaranth fiber released several feruloylated oligosaccharides that were separated using Sephadex LH-20-chromatography and reversed phase-high performance liquid chromatography (RP-HPLC). Three compounds were unambiguously identified: O-(6-O-trans-feruloyl-beta-D-galactopyranosyl)-(1-->4)-D-galactopyranose, O-(2-O-trans-feruloyl-alpha-L-arabinofuranosyl)-(1-->5)-L-arabinofuranose, and O-alpha-L-arabinofuranosyl-(1-->3)-O-(2-O-trans-feruloyl-alpha-L-arabinofuranosyl)-(1-->5)-L-arabinofuranose. These feruloylated oligosaccharides show that ferulic acid is predominantly bound to pectic arabinans and galactans in amaranth insoluble fiber. 5-O-trans-Feruloyl-L-arabinofuranose was the only compound isolated in pure form from an acid hydrolyzate. This compound may have its origin from pectic arabinans but also from arabinoxylans.

Distribution of conjugated linoleic acid in total and subcellular fractions from normal and cancerous parts of human testes.

The objective of the present study was to examine differences in the fatty acid composition of subcellular fractions from normal and cancerous parts of human testes. The conjugated linoleic acid (CLA) content was significantly higher in total testicular carcinoma (TC), but significantly lower in the mitochondrial fraction of TC in comparison to normal testicular tissue. The subcellular distribution pattern of CLA was similar to that of monounsaturated fatty acids, but different to that of 18:2n-6 (linoleic acid), underlining the different physiological properties of CLA and 18 : 2n-6. Because polyunsaturated fatty acids (PUFAs) have been suggested to have an effect on cancer risk and previous research has found that CLA inhibits the metabolism of 18 : 2n-6 into 20 : 4n-6, the contents of n-6 and n-3 PUFAs were determined. Significant differences were observed for 18 : 2n-6, 18 : 3n-3, 20 : 5n-3, and 22 : 6n-3, with 18 : 2n-6, 18 : 3n-3, and 20 : 5n-3 contents being higher and 22 : 6n-3 content being lower in TC than in normal testicular tissue. These results indicate a changed availability of substrates for the cyclooxygenase (COX) or lipooxygenase (LOX) pathways generating eicosanoids. Although not statistically significant, the reduced content of 20 : 4n-6 shown in this study might be due to an increased metabolism of this fatty acid into eicosanoids.