Docosahexaenoic Acid is More Stable to Oxidation when Located at the sn-2 Position of Triacylglycerol Compared to sn-1(3)

Regio-isomeric effects on the oxidative stability of triacylglycerols (TAG) containing docosahexaenoic acid (DHA) were investigated using two pairs of regio-isomerically pure TAG, namely 1,3-dihexadecanoyl-2-(4,7,10,13,16,19-docosahexaenoyl)glycerol (PDP)/1,2-dihexadecanoyl-3-(4,7,10,13,16,19-docosahexaenoyl)glycerol (PPD) and 1,3-dioctadecenoyl-2-(4,7,10,13,16,19-docosahexaenoyl)glycerol (ODO)/1,2-dioctadecenoyl-3-(4,7,10,13,16,19-docosahexaenoyl)glycerol (OOD) where P, O, and D represent palmitic acid, oleic acid, and DHA respectively. Each pair of regio-isomers was subjected to accelerated auto-oxidation (at 40 or 50 °C inside a dark oven). In each case, the TAG oxidized more slowly when DHA was located at the sn-2 position (PDP and ODO) compared to the sn-1(3) position (PPD and OOD), as evidenced by slower development of peroxide value, slower depletion of DHA, and slower generation of secondary oxidation products propanal and trans, trans-2,4-heptadienal. The positional effect on auto-oxidation was more pronounced when DHA occurred in combination with oleic acid than with palmitic acid.     

Dietary fatty acids influence the growth and fatty acid composition of the yellow mealworm <Emphasis Type="Italic">Tenebrio molitor</Emphasis> (Coleoptera: Tenebrionidae)

Fat is the second most abundant component of the nutrient composition of the mealworm Tenebrio molitor (Coleoptera: Tenebrionidae) that represents also an interesting source of PUFA, especially n-6 and n-3 fatty acids, involved in prevention of cardiovascular diseases. This study investigated the possibility of modifying the fat content and the FA composition of yellow mealworms through feeding and how this would be influenced by developmental stages, pupal sex, and generation with the future aim of applying this coleopteran as a diet supplement for human health. Growth rate and cumulative mortality percentage on the different feeding substrates were also evaluated to select the optimal conditions for a mass-raising of this insect species. Despite the different fat content in the six different breeding substrates used, T. molitor larvae and pupae contained a constant fat percentage (>34% in larvae and >30% in pupae). A similar total fat content was found comparing larvae and male and female pupae of the second generation to those of the first generation. On the contrary, FA composition differed both in larvae and pupae reared on the different feeding substrates. However, the exemplars reared on the diets based on 100% bread and 100% oat flour showed SFA, PUFA percentages, and an n-6/n-3 ratio more suitable for human consumption; the diet based on beer yeast, wheat flour, and oat flour resulted in a contemporary diet that most satisfied the balance between a fat composition of high quality and favorable growth conditions.     

Fatty Acids: An Insight into the Pathogenesis of Neurodegenerative Diseases and Therapeutic Potential

One of the most common lipids in the human body is palmitic acid (PA), a saturated fatty acid with essential functions in brain cells. PA is used by cells as an energy source, besides being a precursor of signaling molecules and protein tilting across the membrane. Although PA plays physiological functions in the brain, its excessive accumulation leads to detrimental effects on brain cells, causing lipotoxicity. This mechanism involves the activation of toll-like receptors (TLR) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathways, with the consequent release of pro-inflammatory cytokines, increased production of reactive oxygen species (ROS), endoplasmic reticulum (ER) stress, and autophagy impairment. Importantly, some of the cellular changes induced by PA lead to an augmented susceptibility to the development of Alzheimer’s and Parkinson´s diseases. Considering the complexity of the response to PA and the intrinsic differences of the brain, in this review, we provide an overview of the molecular and cellular effects of PA on different brain cells and their possible relationships with neurodegenerative diseases (NDs). Furthermore, we propose the use of other fatty acids, such as oleic acid or linoleic acid, as potential therapeutic approaches against NDs, as these fatty acids can counteract PA’s negative effects on cells.     

Docosahexaenoic acid (22:6n-3) Ameliorated the Onset and Severity of Experimental Autoimmune Encephalomyelitis in Mice

Multiple sclerosis (MS) is a neurologic autoimmune disease, which is the leading cause of nontraumatic neurologic disability in young adults in United States and Europe. n-3 polyunsaturated fatty acids (PUFA) are reported to mitigate severity of this disease. Recent studies suggest that phospholipid (PL) form of dietary n-3 PUFA may lead to their higher tissue accretion than triacylglycerol (TAG) form. We compared efficacy of PL-docosahexaenoic acid (22:6n-3) (DHA) and TAG-DHA on onset and severity of experimental autoimmune encephalomyelitis (EAE) in a mouse model of MS. Female mice were fed low alpha-linolenic acid (18:3n-3) (ALA) diet (control) for 2 weeks and then fed either control, 0.3%, or 1.0% DHA (PL or TAG) for 4 weeks pre-EAE induction and 4 weeks post-EAE induction. The brain and spinal cord n-6:n-3 ratio was significantly lower in all mice fed DHA compared to control. EAE onset was delayed in mice fed both DHA forms and concentrations, except for 1% TAG-DHA. The inverse association between the EAE score and the brain DHA concentration was nonsignificant at the end of the study (p = 0.08). Daily EAE scores of mice fed different DHA diets did not differ from control, however, the score of all DHA groups combined during days 9–16 was lower (p = 0.028) compared to the control. During days 17–22, the EAE score trended lower in 0.3% TAG-DHA and during days 23–28, the EAE score trended lower in both PL-DHA groups than those in all other groups. These findings suggest that TAG-DHA may be more effective than PL-DHA in the early phases of EAE, and in the final outcome, PL-DHA may be more effective than TAG-DHA.     

Combined Application of Aminoglycosides and Ascorbic Acid in the Elimination of Proteus mirabilis Rods Responsible for Causing Catheter-Associated Urinary Tract Infections (CAUTIs)—A Molecular Approach

Proteus mirabilis is a common cause of catheter-associated urinary tract infections (CAUTIs). In this study, we verified the effectiveness of amikacin or gentamicin and ascorbic acid (AA) co-therapy in eliminating uropathogenic cells, as well as searched for the molecular basis of AA activity by applying chromatographic and fluorescent techniques. Under simulated physiological conditions, a combined activity of the antibiotic and AA supported the growth (threefold) of the P. mirabilis C12 strain, but reduced catheter colonization (≤30%) in comparison to the drug monotherapy. Slight modifications in the phospholipid and fatty acid profiles, as well as limited (≤62%) 2’,7’-dichlorofluorescein fluorescence, corresponding to the hydroxyl radical level, allowed for the exclusion of the hypothesis that the anti-biofilm effect of AA was related to membrane perturbations of the C12 strain. However, the reduced (≤20%) fluorescence intensity of propidium iodide, as a result of a decrease in membrane permeability, may be evidence of P. mirabilis cell defense against AA activity. Quantitative analyses of ascorbic acid over time with a simultaneous measurement of the pH values proved that AA can be an effective urine acidifier, provided that it is devoid of the presence of urease-positive cells. Therefore, it could be useful in a prevention of recurrent CAUTIs, rather than in their treatment.     

Forms of n‐3 (ALA,C18:3n‐3 or DHA,C22:6n‐3) Fatty Acids Affect Carcass Yield,Blood Lipids,Muscle n‐3 Fatty Acids and Liver Gene Expression in Lambs

The effects of supplementing diets with n‐3 alpha‐linolenic acid (ALA) and docosahexaenoic acid (DHA) on plasma metabolites, carcass yield, muscle n‐3 fatty acids and liver messenger RNA (mRNA) in lambs were investigated. Lambs (n = 120) were stratified to 12 groups based on body weight (35 ± 3.1 kg), and within groups randomly allocated to four dietary treatments: basal diet (BAS), BAS with 10.7 % flaxseed supplement (Flax), BAS with 1.8 % algae supplement (DHA), BAS with Flax and DHA (FlaxDHA). Lambs were fed for 56 days. Blood samples were collected on day 0 and day 56, and plasma analysed for insulin and lipids. Lambs were slaughtered, and carcass traits measured. At 30 min and 24 h, liver and muscle samples, respectively, were collected for determination of mRNA (FADS1, FADS2, CPT1A, ACOX1) and fatty acid composition. Lambs fed Flax had higher plasma triacylglycerol, body weight, body fat and carcass yield compared with the BAS group (P < 0.001). DHA supplementation increased carcass yield and muscle DHA while lowering plasma insulin compared with the BAS diet (P < 0.01). Flax treatment increased (P < 0.001) muscle ALA concentration, while DHA treatment increased (P < 0.001) muscle DHA concentration. Liver mRNA FADS2 was higher and CPT1A lower in the DHA group (P < 0.05). The FlaxDHA diet had additive effects, including higher FADS1 and ACOX1 mRNA than for the Flax or DHA diet. In summary, supplementation with ALA or DHA modulated plasma metabolites, muscle DHA, body fat and liver gene expression differently.     

Studies of the fatty acid specificity of the lipase fromRhizomucor miehei toward 20:1n-9, 20:5n-3, 22:1n-9 and 22:6n-3

The fatty acid specificity of the lipase fromRhizomucor miehei toward 20:1n-9, 20:5n-3, 22:1n-9 and 22:6n-3 has been determined by comparing the alcoholysis (byn-propanol) of various mixtures of C₂₀ and C₂₂ fatty acids (FFA) or the corresponding ethyl esters (FAEE) inn-heptane. For all the fatty acids examined, the degree of conversion was much higher when using FFA rather than FAEE. When comparing the experiments with either single FAEE or FAEE mixtures, it was found for all four fatty acids that the degree of conversion depended on whether the FAEE was alone or together with other fatty acids in the reaction mixture. The lipase showed a strong specificity toward 20:1n-9, whereas the polyunsaturated fatty acids were much poorer substrates, especially 22:6n-3. The degrees of conversion for the two n-3 fatty acids show a clear preference for 20:5n-3 over 22:6n-3, not only when present alone but also in the different mixtures examined. The results obtained in the present experiments therefore suggest that when using the lipase fromR. miehei for enrichment of fish oils with n-3 fatty acids, it should not only be possible to diminish the content of 20:1 and 22:1 present in the outer positions in the triacylglycerols, but also to incorporate relatively more 20:5n-3 than 22:6n-3 into the triglycerides.     

Octanoic Acid—An Insecticidal Metabolite of Conidiobolus coronatus (Entomopthorales) That Affects Two Majors Antifungal Protection Systems in Galleria mellonella (Lepidoptera): Cuticular Lipids and Hemocytes

The food flavour additive octanoic acid (C8:0) is also a metabolite of the entomopathogenic fungus Conidiobolus coronatus, which efficiently infects and rapidly kills Galleria mellonella. GC-MS analysis confirmed the presence of C8:0 in insecticidal fraction FR3 extracted from C. coronatus filtrate. Topical administration of C8:0 had a dose-dependent effect on survival rates of larvae but not on pupation or adult eclosion times of the survivors. Topically applied C8:0 was more toxic to adults than larvae (LD100 for adults 18.33 ± 2.49 vs. 33.56 ± 2.57 µg/mg of body mass for larvae). The administration of C8:0 on the cuticle of larvae and adults, in amounts corresponding to their LD50 and LD100 doses, had a considerable impact on the two main defense systems engaged in protecting against pathogens, causing serious changes in the developmental-stage-specific profiles of free fatty acids (FFAs) covering the cuticle of larvae and adults and damaging larval hemocytes. In vitro cultures of G. mellonella hemocytes, either directly treated with C8:0 or taken from C8:0 treated larvae, revealed deformation of hemocytes, disordered networking, late apoptosis, and necrosis, as well as caspase 1–9 activation and elevation of 8-OHdG level. C8:0 was also confirmed to have a cytotoxic effect on the SF-9 insect cell line, as determined by WST-1 and LDH tests.     

Estradiol Favors the Formation of Eicosapentaenoic Acid (20:5n-3) and n-3 Docosapentaenoic Acid (22:5n-3) from Alpha-Linolenic Acid (18:3n-3) in SH-SY5Y Neuroblastoma Cells

Whether neurosteroids regulate the synthesis of long chain polyunsaturated fatty acids in brain cells is unknown. We examined the influence of 17-β-estradiol (E2) on the capacity of SH-SY5Y cells supplemented with α-linolenic acid (ALA), to produce eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA). Cells were incubated for 24 or 72 h with ALA added alone or in combination with E2 (ALA + E2). Fatty acids were analyzed by gas chromatography of ethanolamine glycerophospholipids (EtnGpl) and phosphatidylcholine (PtdCho). Incubation for 24 h with ALA alone increased EPA and DPA in EtnGpl, by 330 and 430% compared to controls (P < 0.001) and DHA by only 10% (P < 0.05). Although DHA increased by 30% (P < 0.001) in ALA + E2-treated cells, the difference between the ALA and ALA + E2 treatments were not significant after 24 h (Anova-1, Fisher’s test). After 72 h, EPA, DPA and DHA further increased in EtnGpl and PtdCho of cells supplemented with ALA or ALA + E2. Incubation for 72 h with ALA + E2 specifically increased EPA (+34% in EtnGpl, P < 0.001) and DPA (+15%, P < 0.001) compared to ALA alone. Thus, SH-SY5Y cells produced membrane EPA, DPA and DHA from supplemental ALA. The formation of DHA was limited, even in the presence of E2. E2 significantly favored EPA and DPA production in cells grown for 72 h. Enhanced synthesis of ALA-elongation products in neuroblastoma cells treated with E2 supports the hypothesis that neurosteroids could modulate the metabolism of PUFA.     

The metabolism of 18∶0 and 18∶2(n−6) by the ovine placenta at 120 and 150 days of gestation

The effects of the approach of parturition in the, sheep on the incorporation of 18∶0 and 18∶2(n−6) into the placental lipids and on the activities of the Δ9- and Δ6-desaturase enzymes of placental tissue have been studied in vitro. The incorporation of 18∶0 into the esterified lipids of placental tissue between the 120th and 150th days of gestation declined markedly; the high level of incorporation of 18∶2(n−6) into the esterified lipids of the placenta (some 2-fold higher than18∶0) remained constant over the gestational period. While placental Δ9-desaturase activity was the same at 150 days as at 120 days of gestation, the activity of the Δ6-desaturase enzyme increased significantly. These results are discussed in relation to the fetal demand for fatty acids near term and the differences that exist between the mechanisms of maternal-fetal transfer of 18∶0 and 18∶2(n−6) in the sheep.     

Analysis of C18:1 cis and trans fatty acid isomers by the combination of gas-liquid chromatography of 4,4-dimethyloxazoline derivatives and methyl esters

Trans fatty acids in foods are usually analyzed by gas-liquid chromatography (GLC) of fatty acid methyl esters (FAME). However, this method may produce erroneously low values because of insufficient separation between cis and trans isomers. Separation can be optimized by preceding silver-ion thin-layer chromatography (Ag-TLC), but this is laborious. We have developed an efficient method for the separation of 18-carbon trans fatty acid isomers by combining GLC of FAME with GLC of fatty acid 4,4-dimethyloxazoline (DMOX) derivatives. We validated this method against conventional GLC of FAME, with and without preceding Ag-TLC. Fatty acid isomers were identified by comparison with standards, based on retention times and mass spectrometry. Analysis of DMOX derivatives allowed the 13t, 14t, and 15t isomers to be separated from the cis isomers. The combination of the GLC analyses of FAME and DMOX derivatives gave results comparable with those obtained by GLC of FAME after preceding Ag-TLC, while saving about 100 h of manpower per 25 samples. It allowed the identification and quantitation of 11 trans and 8 cis isomers and resulted in 25% higher values for total C_18:1 trans, compared with the analysis of FAME alone. The combination of DMOX and FAME analyses, as applied to the analysis of 14 foods that contained ruminant fat and partially hydrogenated vegetable and fish oils, indicated that the most common isomers were 11t in ruminant fats, 9t in partially hydrogenated fish fats, and either 9t or 10t in partially hydrogenated vegetable fats. The combination of GLC analyses of FAME and DMOX derivatives of fatty acids improves the quantitation of 18-carbon fatty acid isomers and may replace the laborious and time-consuming Ag-TLC.     

Susceptibility of Female Mice to the Dietary Omega-3/Omega-6 Fatty-Acid Ratio: Effects on Adult Hippocampal Neurogenesis and Glia

Maternal intake of omega-3 (n-3 PUFAs) and omega-6 (n-6 PUFAs) polyunsaturated fatty acids impacts hippocampal neurogenesis during development, an effect that may extend to adulthood by altering adult hippocampal neurogenesis (AHN). The n-3 PUFAs and n-6 PUFAs are precursors of inflammatory regulators that potentially affect AHN and glia. Additionally, n-3 PUFA dietary supplementation may present a sexually dimorphic action in the brain. Therefore, we postulated that dietary n-6/n-3 PUFA balance shapes the adult DG in a sex-dependent manner influencing AHN and glia. We test our hypothesis by feeding adult female and male mice with n-3 PUFA balanced or deficient diets. To analyze the immunomodulatory potential of the diets, we injected mice with the bacterial endotoxin lipopolysaccharide (LPS). LPS reduced neuroblast number, and its effect was exacerbated by the n-3 PUFA-deficient diet. The n-3 PUFA-deficient diet reduced the DG volume, AHN, microglia number, and surveilled volume. The diet effect on most mature neuroblasts was exclusively significant in female mice. Colocalization and multivariate analysis revealed an association between microglia and AHN, as well as the sexual dimorphic effect of diet. Our study reveals that female mice are more susceptible than males to the effect of dietary n-6/n-3 PUFA ratio on AHN and microglia.     

Fatty Acids of Abyssal Echinodermata,the Sea Star Eremicaster vicinus and the Sea Urchin Kamptosoma abyssale: A New Polyunsaturated Fatty Acid Detected, 22:6(n-2)

Fatty acids (FA) of two species of abyssal Echinodermata—the sea star (Asteroidea) Eremicaster vicinus and the sea urchin (Echinoidea) Kamptosoma abyssale—from the Kuril–Kamchatka Trench, collected at depths of 5210 and 6183 m, were analyzed. Lipids of these deposit-feeding animals showed similar FA compositions: 20.05–16.08% saturated, 42.20–39.50% monoenoic, and 37.75–44.42% polyunsaturated FA, respectively. The contents of odd- and branched-chain FA were 17.35% and 8.80%, respectively. A significant part of FA was represented by uncommon FA such as 21:4(n-7), 22:4(n-8), 22:5(n-5), and 23:4(n-9), earlier discovered in deep-sea foraminifera. Also, the newly found acid, related to the ω2 family, Δ5,8,11,14,17,20–22:6(n-2), amounted 1.60% and 0.33% of total FA, respectively, for the two species. Such unusual FA composition of these abyssal species can be explained by the transfer and modification of FA from consumed foraminifera, which in turn, feed on bacteria.     

Synthesis and activity of a trinuclear platinum complex: [{trans-PtCl(NH3)2}2mu-{trans-Pt(3-hydroxypyridine)2(H2N(CH2)6NH2)2}]Cl4 in ovarian cancer cell lines

This paper describes the synthesis, characterisation, and cytotoxicity of a novel trinuclear platinum complex code named TH1. In addition to its activity against human ovarian cancer cell lines: A2780, A2780(cisR), and A2780(ZD0473R), cell uptake, DNA-binding, and the nature of the compound interaction with pBR322 plasmid DNA have been determined. TH1 is found to be significantly more cytotoxic than cisplatin - two times more active than cisplatin against the parent cell line A2780, thirteen times more active against the cisplatin-resistant cell line A2780(cisR) and 11.5 times more active against the cell line A2780(ZD0473R). Whereas the resistance factors for cisplatin as applied to the cell lines A2780 and A2780(cisR), and A2780 and A2780(ZD0473R) are 12.9 and 3.0 respectively, the corresponding values for TH1 are 1.98 and 0.5. The results suggest that TH1 has been able to significantly overcome resistance in A2780(cisR) and A2780(ZD0473R) cell lines. Whereas cisplatin binds with DNA forming mainly intrastrand GG adduct that causes local bending of a DNA strand, TH1 should bind with DNA forming mainly interstrand GG adducts that would cause more of a global change in DNA conformation. Provided it has favourable toxicity profile, TH1 has the potential to be developed into a highly active anticancer drug with a wider spectrum of activity than cisplatin.     

Occurrence of Hexacosapolyenoic Acids 26:7(n‐3), 26:6(n‐3), 26:6(n‐6) and 26:5(n‐3) in Deep‐Sea Brittle Stars from Near the Kuril Islands

Significant amounts of polyunsaturated fatty acids (PUFA) with a chain length of 26 carbon atoms were detected in lipids of five deep water species of Ophiuroidea besides common fatty acids with chain lengths between 14 and 24. By means of hydrogenation, GC–MS of the methyl esters, and 4,4‐dimethyloxazoline (DMOX) derivatives of these C26 acids were characterized as 5,8,11,14,17,20,23‐hexacosaheptaenoic [26:7(n‐3)]; 8,11,14,17,20,23‐hexacosahexaenoic [26:6(n‐3)]; 5,8,11,14,17,20‐ hexacosahexaenoic [26:6(n‐6)]; and 11,14,17,20,23‐hexacosapentaenoic [26:5(n‐3)]. Concentrations of these acids varied from 0.3 to 4.5 mol% of the total FA. In all the samples investigated, the main component of C26PUFA was hexacosaheptaenoic acid 26:7(n‐3). These C26PUFA are localized mainly in polar lipids. The presence of the possible biosynthesis precursors suggests that the C26PUFA are produced by the brittle stars, and are not accumulated from food sources. This finding can also explain the presence of small amounts of the 26:7(n‐3) acid detected earlier in flesh lipids of the roughscale sole Clidoderma asperrimum, which feeds on deep water brittle stars. We suggest a possible scheme of the biosynthesis of C26 PUFA.