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Background PPARγ is an isoform of peroxisome proliferator-activated receptor (PPAR) belonging to a super family of nuclear receptors. PPARγ receptor is found to play a crucial role in the modulation of lipid and glucose homeostasis. Its commotion has been reported to play a significant role in a broad spectrum of diseases such as type 2 diabetes mellitus, inflammatory diseases, Alzheimer’s disease, and in some cancers. Hence, PPARγ is an important therapeutic target. Polyunsaturated fatty acids (PUFAs) and their metabolites (henceforth referred to as bioactive lipids) are known to function as agonists of PPARγ. However, agonistic binding modes and affinity of these ligands to PPARγ are yet to be deciphered. Methods In this study, we performed a comparative molecular docking, binding free energy calculation and molecular dynamics simulation to infer and rank bioactive lipids based on the binding affinities with the ligand binding domain (LBD) of PPARγ. Results The results inferred affinity in the order of resolvin E1 > neuroprotectin D1 > hydroxy-linoleic acid > docosahexaenoic acid > lipoxin A4 > gamma-linolenic acid, arachidonic acid > alpha-linolenic acid > eicosapentaenoic acid > linoleic acid. Of all the bioactive lipids studied, resolvin E1, neuroprotectin D1 and hydroxy-linoleic acid showed significant affinity comparable to proven PPARγ agonist namely, rosiglitazone, in terms of Glide XP docking score, H-bond formation with the key residues, binding free energy and stable complex formation with LBD favouring co-activator binding, as inferred through Molecular Dynamics trajectory analysis. Conclusion Hence, these three bioactive lipids (resolvin E1, neuroprotectin D1 and hydroxy-linoleic acid) may be favourably considered as ideal drug candidates in therapeutic modulation of clinical conditions such as type 2 DM, Alzheimer’s disease and other instances where PPARγ is a key player.

A putative diol synthase from the fungus Glomerella cingulate was cloned and expressed in Escherichia coli. The putative diol synthase from G. cingulate was purified by His-Trap affinity chromatography with a specific activity of 0.87 U mg⁻¹, an eightfold purification, and a yield of 28 %. One unit of activity was defined as the amount of enzyme required to produce 1 μmol of 7,8-dihydroxy-9,12(Z,Z)-octadecadienoic acid (7,8-DiHODE) per min. The purified enzyme was estimated as a 127-kDa tetramer with a molecular mass of 510 kDa by gel filtration chromatography. The enzyme converted linoleic acid to a product, identified as 7S,8S-DiHODE by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) and nuclear magnetic resonance (NMR) spectroscopy. The specific activity and catalytic efficiency (k cₐₜ/K ₘ) of 7,8-diol synthase from G. cingulate for the conversion of fatty acid to dihydroxy fatty acid followed the order linoleic acid > α-linolenic acid > oleic acid > palmitoleic acid, indicating that the enzyme is a 7,8-linoleate diol synthase (7,8-LDS). The activity of the enzyme for the conversion of 7,8-DiHODE from linoleic acid was maximal at pH 6.5, 40 °C, and 2.5 % (v/v) dimethyl sulfoxide (DMSO). Under these conditions, 7,8-LDS from G. cingulate converted 1.0 mM linoleic acid to 0.62 mM 7,8-DiHODE for 30 min, with a conversion yield of 62 % (mol/mol), via 8-hydroperoxy-9,12(Z,Z)-octadecadienoic acid (8-HPODE) as an intermediate. The accumulation of 8-HPODE was due to a higher 8-dioxygenase activity in the N-terminal domain than hydroperoxide isomerase activity in the C-terminal domain.

To elucidate the role of enzymatic lipid peroxidation in disease pathogenesis and in food deterioration, we recently achieved stereoselective analysis of phosphatidylcholine hydroperoxide (PCOOH) possessing 13 S -hydroperoxy-9 Z ,11 E -octadecadienoic acid (13( S )-9 Z ,11 E -HPODE) using HPLC-MS/MS with a CHIRALPAK OP (+) column. Because enzymatic oxidation progresses concurrently with auto-oxidation, we need to distinguish them further. Here, we attempted such an analysis. First, we used lipoxygenase, linoleic acid, and lysophosphatidylcholine (LPC) to synthesize the enzymatic oxidation product 13( S )-9 Z ,11 E -HPODE PC, and the auto-oxidation products 13( RS )-9 Z ,11 E -HPODE PC and 13( RS )-9 E ,11 E -HPODE PC, which were used as standards to test the ability of various columns to separate the enzymatic oxidation product from auto-oxidation products. Separation was achieved by connecting in series two columns with different properties: CHIRALPAK OP (+) and CHIRALPAK IB-3. The CHIRALPAK OP (+) column separated 13( R )-9 Z ,11 E -HPODE PC and 13( S )-9 Z ,11 E -HPODE PC, whereas CHIRALPAK IB-3 enabled separation of 13( S )-9 Z ,11 E -HPODE PC and 13( RS )-9 E ,11 E -HPODE PC. The results for the analysis of both enzymatically oxidized and auto-oxidized lecithin (an important phospholipid mixture in vivo and in food) indicate that our method would be useful for distinguishing enzymatic oxidation and auto-oxidation reactions. Such information will be invaluable for elucidating the involvement of PCOOH in disease pathogenesis and in food deterioration.

Probiotics with ability to produce conjugated linoleic acid (CLA) is considered as an additive health benefit property for its known role in colon cancer mitigation. The conversion involves the biohydrogenation of the unsaturated fatty acid into conjugated form. Probiotic strain Pediococcus spp . GS4 was efficiently able to biohydrogenate linoleic acid (LA) into its conjugated form within 48 h of incubation. Quantum of CLA produced with a concentration of 121 μg/ml and sustained cell viability of 8.94 log cfu/ml maximally. Moreover, antibacterial effect of LA on the strain ability for biohydrogenation was examined at different concentrations and concluded to have a direct relationship between LA and amount of CLA produced. The efficiency of the strain for CLA production at different pH was also estimated and found maximum at pH 6.0 with 149 μg/ml while this ability was reduced at pH 9.0 to 63 μg/ml. Sesame oil, which is rich in the triacylglycerol form of LA, was also found to act as a substrate for CLA production by Pediococcus spp. GS4 with the aid of lipase-catalyzed triacylglycerol hydrolysis and amount of CLA produced was 31 μg/ml at 0.2 % while 150 μg/ml at 1.0 % of lipolysed oil in skim milk medium. Conjugated form was analyzed using UV scanning, RP-HPLC, and GC-MS. This study also focused on the alternative use of lipolysed sesame oil instead of costly LA for biohydrogenation and could be a potential source for the industrial production of CLA.

The aim of the available literature review was to focus on the role of the proinflammatory mediators of AA and LA derivatives in pathological conditions related to reproduction and pregnancy. Arachidonic (AA) and linoleic acid (LA) derivatives play important roles in human fertility and the course of pathological pregnancies. Recent studies have demonstrated that uncontrolled inflammation has a significant impact on reproduction, spermatogenesis, endometriosis, polycystic ovary syndrome (PCOS) genesis, implantation, pregnancy and labor. In addition, cyclooxygenase-mediated prostaglandins and AA metabolite levels are higher in women’s ovarian tissue when suffering from PCOS. It has been demonstrated that abnormal cyclooxygenase-2 (COX-2) levels are associated with ovulation failure, infertility, and implantation disorders and the increase in 9-HODE/13-HODE was a feature recognized in PCOS patients. Maintaining inflammation without neutrophil participation allows pregnant women to tolerate the fetus, while excessive inflammatory activation may lead to miscarriages and other pathological complications in pregnancies. Additionally AA and LA derivatives play an important role in pregnancy pathologies, e.g., gestational diabetes mellitus, preeclampsia (PE), and fetal growth, among others. The pathogenesis of PE and other pathological states in pregnancy involving eicosanoids have not been fully identified. A significant expression of 15-LOX-1,2 was found in women with PE, leading to an increase in the synthesis of AA and LA derivatives, such as hydroxyeicozatetraenoic acids (HETE) and hydroxyoctadecadiene acids (HODE). Synthesis of the metabolites 5-, 8-, 12-, and 15-HETE increased in the placenta, while 20-HETE increased only in umbilical cord blood in women with preeclampsia compared to normal pregnancies. In obese women with gestational diabetes mellitus (GDM) an increase in epoxygenase products in the cytochrome P450 (CYP) and the level of 20-HETE associated with the occurrence of insulin resistance (IR) were found. In addition, 12- and 20-HETE levels were associated with arterial vasoconstriction and epoxyeicosatrienoic acids (EETs) with arterial vasodilatation and uterine relaxation. Furthermore, higher levels of 5- and 15-HETE were associated with premature labor. By analyzing the influence of free fatty acids (FFA) and their derivatives on male reproduction, it was found that an increase in the AA in semen reduces its amount and the ratio of omega-6 to omega-3 fatty acids showed higher values in infertile men compared to the fertile control group. There are several studies on the role of HETE/HODE in relation to male fertility. 15-Hydroperoxyeicosatetraenoic acid may affect the integrity of the membrane and sperm function. Moreover, the incubation of sperm with physiologically low levels of prostaglandins (PGE2/PGF2α) improves the functionality of human sperm. Undoubtedly, these problems are still insufficiently understood and require further research. However, HETE and HODE could serve as predictive and diagnostic biomarkers for pregnancy pathologies (especially in women with risk factors for overweight and obesity). Such knowledge may be helpful in finding new treatment strategies for infertility and the course of high-risk pregnancies.

The oxidation kinetics of conjugated methyl linoleate was compared with that of non-conjugated methyl linoleate under mild oxidation conditions (30 °C in the dark). Samples of methyl 9-cis,11-trans-linoleate, methyl 10-trans,12-cis linoleate and methyl 9-cis,12-cis linoleate were assayed separately and in mixtures. For comparative purposes, methyl α-linolenate and methyl oleate were also used. Two complementary analytical approaches were selected to monitor the progress of oxidation, (1) the traditional follow-up of residual substrate by gas liquid chromatography, and (2) an analytical procedure by high-performance size-exclusion chromatography (HPSEC) for direct measurement of the oxidation compounds formed. The HPSEC method enabled us to quantitate oxidized monomers, dimers and polymers concomitantly in a rapid and direct analysis. Results showed that conjugated methyl linoleate samples oxidized later than their non-conjugated counterparts, and showed a very different oxidation pattern. Thus, formation of oxidized monomers was negligible and the first and major compounds formed were polymerization products. Also, under the conditions used, non-conjugated and conjugated methyl linoleate samples in 1:1 mixtures led to decreased oxidation rate of non-conjugated methyl linoleate and increased oxidation rate of conjugated methyl linoleate. This study supports the view that oxidation kinetics of conjugated dienes differ substantially from that of methylene-interrupted dienes.

The common practice in North American feedlot industries is to add antibiotics to the diet to prevent disease and improve both BW gain and feed efficiency. In this study, 240 crossbred steer calves were backgrounded on a 54% silage diet for 80 d and fed a finishing diet consisting of 81% barley grain, 10% barley silage, and 7.5% supplement (DM basis) with and without in-feed antibiotics for approximately 120 d. Calves were assigned to 1 of 5 treatments: a control with no antibiotics, 11 mg/kg of chlortetracycline, 44 mg/kg of chlortetracycline, 44 mg/kg of chlortetracycline plus 44 mg/kg of sulfamethazine, and 11 mg/ kg of tylosin phosphate. A combination of GLC and silver-ion HPLC methods was used to analyze the fatty acid composition of brisket adipose tissue, with emphasis on trans-18:1 and CLA isomers. The inclusion of nonionophore antibiotics in the diet had little effect on the fatty acid composition, except that feeding either 44 mg/kg of chlortetracycline or 11 mg/kg of tylosin caused small increases in 9c-14:1 and 16:0 relative to the control (0.26 and 0.9 g/100 g of total fatty acids, respectively). Likewise, profiles of trans-18:1 and CLA isomers were unchanged by antibiotics, but across treatments the predominant trans-18:1 isomer was 10t-18:1 (where t = trans; 3.22%) at 3 times the concentration of the second most abundant isomer (11t-18:1; vaccenic acid, 1.05%). Rumenic acid (9c,11t-18:2, where c = cis) was the major CLA isomer at 61% of total CLA, followed by 7t,9c-18:2 at 9%. Because no other effects on fatty acid composition were evident, data for trans-18:1 and CLA were pooled across treatments to investigate possible relationships among rumen PUFA metabolites. The total trans-18:1 content in brisket adipose tissue was positively correlated with 10t-18:1, but not with 11t-18:1, whereas the total CLA was positively correlated with 9c,11t-18:2, but not with 7t,9c-18:2. The 7t,9c-18:2 was, however, positively correlated with 10t-18:1 and 6t/7t/8t-18:1 but was negatively correlated with rumenic acid. These metabolic interrelationships suggest the presence of bacterial populations with distinct pathways for PUFA biohydrogenation in which either 10t-18:1 or 11t-18:1 predominate. Overall, the nonionophore antibiotics tested did not appreciably change adipose tissue composition and consequently could not be used to improve the trans-18:1 or CLA profile (i.e., increase vaccenic and rumenic acids at the expense of 10t-18:1).

The present study was designed to investigate the capability of mixed rumen protozoa to synthesize conjugated linoleic acid (CLA) from linoleic (LA) and vaccenic acids (VA). Rumen contents were collected from fistulated cows. The protozoal fraction was separated and washed several times with MB9 buffer and then resuspended in autoclaved rumen fluid. The suspensions were anaerobically incubated up to 18 h at 38.5 °C with substrates in the presence (P-AB) or the absence of antibacterial-agents (P-No-AB). Neither P-AB nor P-No-AB suspensions were capable of producing CLA from VA (11t-18:1). Linoleic acid was catabolized by P-No-AB to a greater extent than P-AB. Different isomers of CLA were synthesized by P-AB from LA. The 9c11t-CLA was predominant. Thirty seven percent of the maximum accumulated 9c11t-CLA was found in the P-AB suspension as early as 0.1 h into the incubation period. Accumulation of 10t12c-CLA in P-AB suspension was approximately 10.0 times lower than that of 9c11t-CLA. There were no significant productions of VA, 10t-18:1, and 18:0 in P-AB compared with the control, indicating that rumen protozoa have no ability to biohydrogenate CLA isomers. On the other hand, the concentrations of 10t-18:1, VA, and 18:0 in P-No-AB were greater (P < 0.05) compared with those in P-AB, indicating the role of symbiotic bacteria associated with P-No-AB in biohydrogenating CLA isomers. We concluded that mixed rumen protozoa are capable of synthesizing CLA from LA through isomerization reactions. However, they are incapable of metabolizing CLA further. They are also incapable of vaccenic acid biohydrogenation and/or desaturation.

Objective: To assess the effects of dietary supplementation using two isomeric blends of conjugated linoleic acid (CLA) on immune function in healthy human volunteers. Design: Double-blind, randomised, placebo-controlled intervention trial. Subjects and intervention: A total of 55 healthy volunteers (n=20 males, n=35 females) were randomised into one of three study groups who received 3 g/day of a fatty acid blend containing a 50:50 cis-9, trans-11: trans-10, cis-12 CLA isomer blend (2 g CLA), and 80:20 cis-9, trans-11: trans-10, cis-12 (80:20) CLA isomer blend (1.76 g CLA) or linoleic acid (control, 2 g linoleic acid) for 8 weeks. Results: Supplementation with the 80:20 CLA isomer blend significantly (P

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