Effect of diet and dietary fatty acids on the transformation and incorporation of C18 fatty acids in double-muscled Belgian Blue young bulls

Three groups of double-muscled Belgian Blue young bulls were fed during different stages of production diets differing in the proportions of linolenic and linoleic acid by including linseed in the concentrate or giving grass silage as main linolenic acid suppliers. Samples of rumen and abomasal contents and of the longissimus thoracis, subcutaneous fat, and liver were taken to analyze the fatty acid pattern with emphasis on the individual trans (t) C18:1 fatty acids and cis-9,trans-11 conjugated linoleic acid (c9t11CLA). Trans C18:1 isomers represented up to 20 g/100 g of total fatty acids in rumen and abomasal contents, whereas the accumulation of c9t11CLA was limited. Total trans C18:1 content in subcutaneous fat and intramuscular fat of the longissimus thoracis comprised 8.4 and 5.2 g/100 g of total fatty acids, respectively, with t11C18:1 being the most abundant one. Compared to rumen contents, subcutaneous and intramuscular fat were enriched in c9t11CLA and contained fewer tC18:1 isomers, resulting in a higher c9t11CLA/t11C18:1 ratio (0.04, 0.22, and 0.22, respectively). This result suggests that the endogenous synthesis of c9t11CLA in adipose tissue by the Delta(9)-desaturase was more important than its ruminal production.     

Fatty acid composition of yak (Bos grunniens) cheese including conjugated linoleic acid and trans-18:1 fatty acids

The esterified fatty acid composition of cheese (YC) from yak ( Bos grunniens), reared in the highlands of the Nepalese Himalayas, was studied using capillary gas-liquid chromatography and compared with that of dairy cow Cheddar cheese (DC) purchased in a local market. The YC was collected from Dolakha, Nepal. The YC had a lower (P<0.001) myristic acid (C14:0; 6.7 vs 10.3%, YC vs DC, respectively) and palmitic acid content (C16:0; 23.3 vs 29.2%, YC vs DC, respectively) compared to DC. The YC had a lower (P<0.01) total medium-chain saturated fatty acids (C10:0-C16:0) content compared to DC (36.7 vs 47.3%, YC vs DC, respectively). On the other hand, the YC had a 24.8% higher (P<0.01) level of total long-chain saturated fatty acids (C17:0-C26:0) and a 3.2 times higher (P<0.001) content of total n-3 PUFA than DC. The ratio of n-3 PUFA to n-6 PUFA in YC was 0.87 compared to 0.20 in DC. YC had a 2.8 times higher (P<0.001) total trans-18:1 (9.18 vs 3.31%, YC vs DC, respectively) content. The percentage of vaccenic acid ( trans-11-C18:1) in YC was 4.6 times higher (6.23 vs 1.35% of total fatty acids, YC vs DC, respectively) than in DC. Vaccenic acid constituted 67.9% of total trans-C18:1 in YC. The Delta9-desaturase index for YC was lower than that of DC. The total conjugated linoleic acid (CLA) content in YC was 2.3% of total fatty acids compared to 0.57% in DC. The cis-9, trans-11 CLA isomer in YC constituted 88.5% of the total CLA. The results suggest that cheese from yak, grazed on Himalayan alpine pastures, may have a more healthful fatty acid composition compared to cheese manufactured from dairy cattle fed grain-based diets.     

Effect of dietary conjugated linoleic acid and monounsaturated fatty acid content on pig muscle and adipose tissue lipase and esterase activity

Three levels (0%, 1%, and 2%) of conjugated linoleic acid (CLA) were combined with two levels (low and high) of monounsaturated fatty acids (MUFA) for pig feeding. The activity of neutral lipase (NL), acid lipase (AL), phospholipase (PL), neutral esterase (NE), and acid esterase (AE) was measured in extracts from muscle and subcutaneous adipose tissues. The addition of CLA in the diet only affected the lipolytic activity in muscle, whereas differences in MUFA content of pig diets were mainly responsible for the lipolytic enzyme modifications observed in adipose tissue. Nevertheless, a significant effect of the interaction CLA x MUFA on the activity of several lipolytic enzymes was observed in both tissues. The effect of either linoleic acid (LA) or CLA on the activity of muscle and adipose lipolytic enzymes was determined by in vitro assays. Remarkable inhibitory or activation effects were detected depending on the enzyme and kind of tissue.     

Extensive analysis of long-chain polyunsaturated fatty acids, CLA, trans-18:1 isomers, and plasmalogenic lipids in different retail beef types

The objective of this investigation was to provide a comprehensive analysis of the total lipid composition of present-day retail beef meat available at the consumer level and to evaluate the total lipid composition with special emphasis on the nutritional value. For this purpose, 40 beef cuts were obtained from four cattle farms based on either a natural grazing system (NGS) or an intensive production system (IPS). The total lipid composition was analyzed using complementary chemical and chromatographic procedures. The content of n-3 LC-PUFA, CLA, total trans-18:1, and branched-chain fatty acids was significantly higher in NGS beef than in IPS beef. The trans-18:1 and CLA profiles were affected by the different production systems, whereby they can be utilized empirically to differentiate between feeding regimen and production management. Fatty acid ratios that have health implications ( n-6/ n-3, LA/alphaLNA, and AA/EPA) were remarkably beneficial for NGS beef compared with IPS beef. In conclusion, from the human health perspective, beef raised on NGS is clearly superior with regard to a more favorable fatty acid profile in comparison to IPS beef.     

Comparison of different methylation methods for the analysis of conjugated linoleic acid isomers by silver ion HPLC in beef lipids

Four different methods for the methylation of conjugated linoleic acid isomers (CLA) in ruminant lipids were compared by silver ion (Ag+) HPLC. The combination of base-catalyzed methods followed by an acid-catalyzed method with BF3/MeOH was tested under different temperatures (room temperature and at 60 degrees C), along with based-catalyzed methylation with NaOCH3 and methylation with BF3/MeOH after saponification with NaOH. The comparison among these four methods was done on muscle and adipose tissue samples from bulls. The repeatability theta of the combined base- and acid-catalyzed methylation (NaOCH3/BF3) at ambient temperature for 20 min and at 60 degrees C for 10 min was most suitable for the quantitative Ag+-HPLC analysis of CLA isomers. At 60 degrees C the combined methods supplied the highest concentrations of most CLA isomers. The base-catalyzed methylation and the saponification followed by BF3/MeOH methylation for 5 min generated significantly lower concentrations for most CLA isomers compared to the combined methods.     

Changes in the fatty acid profile of the subcutaneous fat of swine throughout fattening as affected by dietary conjugated linoleic acid and monounsaturated fatty acids

The fatty acid profile of the subcutaneous fat of pigs and its evolution throughout fattening as affected by dietary conjugated linoleic acid (CLA), monounsaturated fatty acids (MUFA), and their interaction (CLAxMUFA) were studied. Three levels (0, 1, and 2%) of an enriched CLA oil (28% cis-9, trans-11 and 28% trans-10, cis-12 CLA) were combined with two levels of MUFA (low, 19% average; and high, 39% average) for pig feeding (288 gilts). Subcutaneous shot-biopsies were taken from 48 animals at the beginning of the trial (S1, 70 kg), 14 days later (S2, 80 kg), and at slaughter (S3, 107 kg). Inclusion of CLA in the diet caused an increase during fattening in cis-9, trans-11 CLA, trans-10, cis-12 CLA, and saturated fatty acids (SFA) contents of pig backfat and a decrease in MUFA and polyunsaturated fatty acids (PUFA). MUFA supplementation also led to a MUFA enrichment of backfat. The interaction CLAxMUFA affected the SFA content. The rates of accumulation of CLA isomers, SFA, and MUFA throughout the trial did not follow a linear behavior, such rates being higher from S1 to S2 than from S2 to S3. These rates were also influenced by dietary CLA and MUFA levels. The increase in the ratio of saturated to unsaturated fatty acids of backfat caused by dietary CLA might be balanced by supplementation of pig diets with MUFA.     

Effect of the degree of processing on soy oil conjugated linoleic acid yields

Photoirradiation processing can be used to rapidly synthesize conjugated linoleic acid (CLA) in large quantities in soy oil. The objective of this study was to evaluate the effect of the level of refining of soy oil on CLA yields and oxidative properties after photoirradiation. Crude, alkali-refined, alkali-refined bleached, and alkali-refined bleached and deodorized (RBD) soy oils were photoirradiated in a pilot-plant processing system for 12 h with 0.35% iodine catalyst at 47 degrees C. RBD soy oil gave the highest total CLA yield of 16.3% of total oil with 4.3% cis, trans- and trans, cis-CLA isomers. Oxidative stability as measured by weight gain during incubation at 64 degrees C showed that iodine decreased the induction time of soy oil samples by 2-4 days. Photoirradiation processing further decreased the induction time by 2 days as a result of loss of total tocopherols. Iodine addition increased the titratable acidity in all the samples of soy oil. However, the level of refining affects this increase of titratable acidity, and RBD soy oil was found to be the most stable. The study indicates that RBD soy oil was the most suitable candidate for photoirradiation processing. Thus, soy oil should be alkali-refined, bleached, deodorized, and then photoprocessed followed by a secondary adsorption step to remove the iodine catalyst to obtain a RBD CLA-rich soy oil.     

Preparation of a t,t conjugated linoleic acid methylester (CLA-Me) isomers mixture from synthetic CLA by methylation with BF3/methanol

Mixtures of t,t conjugated linoleic acid methylester (t,t CLA-Me) isomers were prepared from synthetic CLA, consisting of 47.8% t10,c12 CLA; 45.5% c9,t11 CLA; 2.0% t,t CLA; and 4.7% others, by methylation with BF(3)/methanol (designated TT-TC/CT) in conjunction with purification at -68 degrees C for 24 h. The amount or composition of the TT-TC/CT was greatly affected by the concentration of BF(3) in methanol and the duration of methylation. The methylation of 50 mg of synthetic CLA for 30 min with 1 mL of 7.0% BF(3)/methanol produced a TT-TC/CT (21.54 mg) with the composition of 1.3% t12,t14; 5.9% t11,t13; 42.7% t10,t12; 44.0% t9,t11; 5.0% t8,t10; and 1.1% t 7,t9 CLA, whereas the methylation for 60 min with 14.0% BF(3)/methanol produced a TT-TC/CT (28.62 mg) with the composition of t,t CLA isomers different from that of TT-TC/CT by methylation for 30 min with 7.0% BF(3)/methanol. A large quantity of TT-TC/CT (14.15 g) with the composition similar to that of TT-TC/CT prepared from 50 mg of synthetic CLA was also prepared from 25 g of synthetic CLA. The purity of TT-TC/CT samples was greater than 98%. These results suggest that TT-TC/CT with a purity greater than 98% was easily prepared from synthetic CLA by BF(3)-catalyzed methylation, and the amount and composition of t,t CLA isomers of TT-TC/CT samples could be controlled by methylation conditions.     

Impact of dietary conjugated linoleic acid on the oxidative stability of rat liver microsomes and skeletal muscle homogenates

Dietary conjugated linoleic acid (CLA; 0-2.0%) increased CLA concentrations in liver microsomes and skeletal muscle homogenates from rats. Dietary CLA decreased oleic and arachadonic acid concentrations in both liver microsomes and skeletal muscle. The presence of CLA in liver microsomes had no impact on linoleic acid, arachadonic acid, and alpha-tocopherol oxidation rates. Dietary CLA (2.0%) also did not alter alpha-tocopherol oxidation rates in liver microsomes or muscle homogenates. Formation of malonaldehyde (MDA) in oxidizing liver microsomes decreased with increasing CLA concentration as determined by measurement of thiobarbituric acid-MDA complexes by HPLC. The ability of CLA to decrease MDA formation without impacting other lipid oxidation markers such as the disappearance of fatty acid and alpha-tocopherol suggests that decreased MDA concentration was the result of CLA's ability to lower polyenoic fatty acids such as arachadonic acid. While CLA does not appear to act as an antioxidant, its ability to decrease polyenoic fatty acid concentrations could decrease the formation of highly cytotoxic lipid oxidation products such as MDA.     

Effect of conjugated linoleic acids on the activity and mRNA expression of 5- and 15-lipoxygenases in human macrophages

Lipoxygenases are a family of non-heme enzyme dioxygenases. The role of lipoxygenases is synthesis of hydroperoxides of fatty acids, which perform signaling functions in the body. Studies on conjugated linoleic acids (CLAs) as fatty acids with a potential anti-atherosclerotic function have recently been initiated. The aim of the study was to test the effect of CLAs and linoleic acid on 5- and 15-lipoxygenase (5-LO, 15-LO-1) enzyme activity, their mRNA expression, and concentration in the cells. It was also desired to determine whether the CLAs are substrates for the enzymes. For the experiments monocytic cell line (THP-1) and monocytes obtained from human venous blood were used. Monocytes were differentiated to macrophages: THP-1 (CD14+) by PMA administration (100 nM for 24 h) and monocytes from blood (CD14+) by 7-day cultivation with the autologous serum (10%). After differentiation, macrophages were cultured with 30 microM CLAs or linoleic acid for 48 h. The 15- and 5-lipoxygenase products were measured by HPLC method. mRNA expression and protein content were analyzed by real-time PCR and Western blot analysis. The in vitro studies proved that both CLA isomers are not substrates for 15-LO-1; in ex vivo studies hydroxydecadienoic acid (HODE) concentration was significantly reduced (p = 0.019). The trans-10,cis-12 CLA isomer reduced HODE concentration by 28% (p = 0.046) and the cis-9,trans-11 CLA isomer by 35% (p = 0.028). In macrophages obtained from THP-1 fatty acids did not change significantly mRNA expression of the majority of the investigated genes. CLAs did not change the content of 5-LO and 15-LO-1 proteins in macrophages obtained from peripheral blood. Linoleic acid induced 15-LO-1 expression (2.6 times, p < 0.05). CLAs may perform the function of an inhibitor of lipoxygenase 15-LO-1 activity in macrophages.     

Physiological difference between free and triglyceride-type conjugated linoleic acid on the immune function of C57BL/6N mice

Previous studies have shown the physiological significance of dietary conjugated linoleic acid (CLA) in various experimental animals and in human beings. One of the important problems to better elucidate is the difference between triglyceride (TG) and free (FFA) dietary CLA. Here, using splenocytes, this study assesses how TG- and FFA-CLA modulate immunoglobulin and various cytokine productions. In this study, C57BL/6N mice were fed an experimental diet containing 0% CLA, 0.1 or 1% FFA-CLA, or 0.1 or 1% TG-CLA for 3 weeks. The production of immunoglobulin tended to be up-regulated by 1% FFA-CLA. As a result of protein array analysis using the supernatant from splenocytes cultured with no CLA, 1% FFA-CLA, and TG-CLA, some cytokine production was shown to be remarkably regulated by dietary FFA- and TG-CLA. A total of 32 cytokines were examined, and 11-14 produced cytokines that were 2-fold up-regulated as compared with control for FFA- or TG-CLA, respectively. Especially, the production of IL-9 and MCP-5 and other cytokines was remarkably up-regulated by both FFA- and TG-CLA. In addition, seven cytokines were 2-fold down-regulated by TG-CLA. These data show that there is a slight but significant difference between the functionalities of FFA- and TG-CLA.     

Validity of protein, retinol, folic acid and n-3 fatty acid intakes estimated from the food-frequency questionnaire used in the Danish National Birth Cohort

OBJECTIVE: To validate intakes of protein, folic acid, retinol and n-3 fatty acids estimated from a food-frequency questionnaire in week 25 of pregnancy (FFQ-25). DESIGN: Validation was done against a 7-day weighed food diary (FD) and biomarkers of the nutrients in gestation week 32-38. SUBJECTS AND SETTING: The FFQ-25 to be validated was used in the Danish National Birth Cohort comprising 101 042 pregnant Danish women, of whom 88 participated in the present validation study. RESULTS: Estimated intakes of protein, retinol and folic acid did not differ significantly between the two dietary methods, but intake of n-3 fatty acids was one third larger when estimated from the FFQ-25. The intakes estimated from the two dietary methods were all significantly correlated, ranging from 0.20 for retinol intake to 0.57 for folic acid intake. Sensitivities of being correctly classified into low and high quintiles were between 0.22 and 0.77, and specificities were between 0.62 and 0.89. Urinary protein content did not correlate significantly with protein estimated from the FFQ (r = 0.17, P > 0.05), but did with intake estimated from the FD (r = 0.56, P < 0.0001). Erythrocyte folate correlated significantly with the estimated total intake from the FFQ (r = 0.55, P < 0.0001) and the FD (r = 0.52, P < 0.0001). No correlations with plasma retinol were found. Erythrocyte eicosapentaenoic acid (C20:5n-3) correlated significantly with n-3 fatty acids intake estimated from both the FFQ-25 (r = 0.37, P < 0.001) and the FD (r = 0.62, P < 0.0001). CONCLUSION: The FFQ-25 gives reasonable valid estimates of protein, retinol and folic acid intakes, but seems to overestimate intake of n-3 fatty acids.     

Effect of the polyunsaturated fatty acid composition of beef muscle on the profile of aroma volatiles

The effect of n-3 polyunsaturated fatty acids (PUFAs) in beef muscle on the composition of the aroma volatiles produced during cooking was measured. The meat was obtained from groups of steers fed different supplementary fats: (i) a palm-oil-based control; (ii) bruised whole linseed, which increased muscle levels of alpha-linolenic (C18:3 n-3) and eicosapentaenoic acid (EPA, C20:5 n-3); (iii) fish oil, which increased EPA and docosahexaenoic acid (C22:6 n-3); (iv) equal quantities of linseed and fish oil. Higher levels of lipid oxidation products were found in the aroma extracts of all of the steaks with increased PUFA content, after cooking. In particular, n-alkanals, 2-alkenals, 1-alkanols, and alkylfurans were increased up to 4-fold. Most of these compounds were derived from the autoxidation of the more abundant mono- and di-unsaturated fatty acids during cooking, and such autoxidation appeared to be promoted by increased levels of PUFAs.     

Effect of acid deposition on forested catchment in the western Tokyo,Japan

Acid deposition and its effect on Andisols were investigated in the forest experimental station, the Rolling Land Laboratory (RLL), located in the Tama Hill region of Central Japan. The annual volume-weighted mean pH value of open bulk precipitation was 4.8 in the period 1990 to 1992. Nitrate deposition at RLL was larger than for sulfate, which was obviously different from results in Japan Environment Agency (JEA) stations or in study areas of Europe and North America. Abundant nitrate deposition was ascribed to the high emissions from a non-point source, mainly cars. Although sulfate concentration in throughfall fluctuated, its concentration in soil solution was kept at a low level. This was attributed to the high sulfate adsorption capacity of the Andisols.     

Effect of acid rain on sandstone: The Royal Palace and the Riddarholm church,Stockholm

The deterioration of two kinds of sandstone is discussed for two 18:th century buildings in central Stockholm: the Royal Palace, and the Royal Carolean Burial Chapel (Karolinska gravkoret) annexed to the mediaeval Riddarholm church. The facades of calcitic Gotland sandstone show many signs of serious decay, such as gypsum formation, pulverized surface, exfoliation, discolouration, and salt efflorescence. The socles are built of the more resistant quartzitic Roslagen sandstone, displaying some discolouration, cracks, and slight exfoliation. In total about 300 samples have been analysed. The surface concentration of sulphur is highest at ground level and at rain-sheltered positions. Chemical and sulphur isotope data indicate that the stone decay to a large part may be attributed to anthropogenic sources like acid deposition and car traffic.     

A comparative study on the chemical composition of humic acids from forest soil,agricultural soil and lignite deposit: Bound lipid,carbohydrate and amino acid distributions

Base- and acid-hydrolysable fractions of humic acids (HAs) isolated from a forest soil, an agricultural soil and a lignite deposit were analysed, and comparisons were made between the base hydrolysable lipid (bound lipid), carbohydrate and amino acid signatures of the different humic acids.Bound lipids differ depending on the humic acid origin. Their composition were rather similar for the two soil humic acids, with three main lipid classes identified: (i) aliphatic components, (ii) aromatic components and (iii) sterols and triterpenols. The aliphatic subfraction was the most abundant and consisted predominantly of cutin- and suberin-derived moieties some of which could be clearly related to the vegetation. A minor bacterial input was indicated by the presence of short chain α- and β-hydroxyalkanoic acids. Aromatic subfraction contributed to a low amount to the total base hydrolysates and consisted mainly of lignin-derived methoxyphenols. Present in trace amounts, sterols and triterpenols are mainly of higher plant origin. The base hydrolysate from lignite humic acid markedly differs. Bound lipids released from lignite HA comprised almost exclusively aliphatic components, largely dominated by long chain alkanoic acids. Lignin-derived moieties, hardly detected, consisted solely of vanillic and 4-hydroxybenzoic acids indicating a much higher degree of lignin alteration in lignite humic acid. Sterols and triterpenols were absent.Although the composition of monosaccharides released upon acid hydrolysis was rather uniform irrespective of the humic acid origin, the distribution changed with the degree of humification of the HAs. Ratios of (Galactose+Mannose) to (Xylose+Arabinose) increased from soil to lignite humic acids. The high values of the ratios indicate that carbohydrates are primarily of microbial origin.In all humic acids neutral and acidic protein amino acids dominated. Non protein amino acids were only minor components consisting mainly of hydroxy proline and ornithine. The amino acid distributions of both soil HAs were similar. The amino acid distribution of lignite HA resembled that of soil HAs except for the following differences: (1) the absence of hydroxy proline and the greater abundance of ornithine suggesting a higher microbial contribution to the amino acids as the degree of humification increases, (2) the higher contribution of polar amino acids suggesting a preferential preservation of these amino acids possibly by interaction with the humic acid surface through hydrogen bonds.