BACKGROUND: To investigate the effect of manganese on lipid peroxidation and compositional changes of fatty acids in hippocampus of rat brain. METHODS: Seven rats in experimental group were given with MnCl2 intraperitoneally for 4 weeks (4 mg/kg once daily, 5 days per week). Twenty four hours after the last injection, rats were decapitated and, hippocampus were separated from the rat brain. RESULT: In Mn-treated group, manganese concentrations increased significantly in the hippocampus by 222% compared with control group (P<0.01). MDA concentrations increased significantly by 149% compared with control group (P<0.05). Among fatty acids, total n-6 polyunsaturated fatty acids (PUFAs) increased significantly by 237% compared with control group (P<0.05). Linoleic acid (LA) and arachidonic acids (AA) increased by 213%, 238% (P<0.05, P<0.01, respectively). Among n-3 PUFAs except linolenic acids, eicosapentanoic acid(EPA) and docosahexanoic acids (DHA) decreased significantly by 70%, 50% respectively compared with control group (both P<0.01). CONCLUSION: Our results suggest that manganese may cause compositional changes of fatty acids in hippocampus of rat brain. Characteristics of fatty acids compositional changes by manganese were the decrease of EPAs and DHAs (n-3 PUFAs), and increase of AA and LA (n-6 PUFAs). These changes with the increase of MDA, suggest that manganese neurotoxicity is caused by lipid peroxidation.
alpha-Linolenic Acid ; Animals ; Arachidonic Acid ; Arachidonic Acids ; Brain* ; Fatty Acids* ; Fatty Acids, Omega-3 ; Fatty Acids, Unsaturated ; Hippocampus* ; Linoleic Acid ; Linolenic Acids ; Lipid Peroxidation* ; Malondialdehyde ; Manganese* ; Rats*

Three long-chain polyunsaturated fatty acids, docosahexaenoic acid (DHA, 22:6n-3), eicosapentaenoic acid (EPA, 20:5n-3) and arachidonic acid (ARA, 20:4n-6), are the most biologically active polyunsaturated fatty acids in the body. They are important in developing and maintaining the brain function, and in preventing and treating many diseases such as cardiovascular disease, inflammation and cancer. Although mammals can biosynthesize these long-chain polyunsaturated fatty acids, the efficiency is very low and dietary intake is needed to meet the requirement. In this study, a multiple-genes expression vector carrying mammalian A6/A5 fatty acid desaturases and multiple-genes expression vector carrying mammalian Δ6/Δ5 fatty acid desaturases and Δ6/Δ5 fatty acid elongases coding genes was used to transfect HEK293T cells, then the overexpression of the target genes was detected. GC-MS analysis shows that the biosynthesis efficiency and level of DHA, EPA and ARA were significantly increased in cells transfected with the multiple-genes expression vector. Particularly, DHA level in these cells was 2.5 times higher than in the control cells. This study indicates mammal possess a certain mechanism for suppression of high level of biosynthesis of long chain polyunsaturated fatty acids, and the overexpression of Δ6/Δ5 fatty acid desaturases and Δ6/Δ5 fatty acid elongases broke this suppression mechanism so that the level of DHA, EPA and ARA was significantly increased. This study also provides a basis for potential applications of this gene construct in transgenic animal to produce high level of these long-chain polyunsaturated fatty acid.
Acetyltransferases ; genetics ; metabolism ; Arachidonic Acid ; biosynthesis ; Docosahexaenoic Acids ; biosynthesis ; Eicosapentaenoic Acid ; biosynthesis ; Fatty Acid Desaturases ; genetics ; metabolism ; Fatty Acid Synthases ; genetics ; metabolism ; Fatty Acids, Unsaturated ; biosynthesis ; Genetic Vectors ; HEK293 Cells ; Humans ; Transfection

BACKGROUND/OBJECTIVES: Adequate dietary fatty acid intake is important for toddlers between 12–24 months of age, as this is a period of dietary transition in conjunction with rapid growth and development; however, actual fatty acid intake during this period seldom has been explored. This study was conducted to assess the intake status of n-3 and n-6 polyunsaturated fatty acids by toddlers during the 12–24-month period using 2010–2015 Korea National Health and Nutrition Examination Survey data. SUBJECTS/METHODS: Twenty-four-hour dietary recall data of 12–24-month-old toddlers (n = 544) was used to estimate the intakes of α-linolenic acid (ALA; 18:3n-3), eicosapentaenoic acid (EPA; 20:5n-3), docosahexaenoic acid (DHA; 22:6n-3), linoleic acid (LA; 18:2n-6), and arachidonic acid (AA; 20:4n-6), as well as the major dietary sources of each. The results were compared with the expected intake for exclusively breastfed infants in the first 6 months of life and available dietary recommendations. RESULTS: Mean daily intakes of ALA, EPA, DHA, LA, and AA were 529.9, 22.4, 37.0, 3907.6, and 20.0 mg/day, respectively. Dietary intakes of these fatty acids fell below the expected intake for 0–5-month-old exclusively breastfed infants. In particular, DHA and AA intakes were 4 to 5 times lower. The dietary assessment indicated that the mean intake of essential fatty acids ALA and LA was below the European and the FAO/WHO dietary recommendations, particularly for DHA, which was approximately 30% and 14–16% lower, respectively. The key sources of the essential fatty acids, DHA, and AA were soy (28.2%), fish (97.3%), and animals (53.7%), respectively. CONCLUSIONS: Considering the prevailing view of DHA and AA requirements on early brain development, there remains considerable room for improvement in their intakes in the diets of Korean toddlers. Further studies are warranted to explore how increasing dietary intakes of DHA and AA could benefit brain development during infancy and early childhood.
Animals ; Arachidonic Acid ; Brain ; Diet ; Eicosapentaenoic Acid ; Fatty Acids ; Fatty Acids, Essential ; Fatty Acids, Unsaturated ; Growth and Development ; Humans ; Infant ; Korea ; Linoleic Acid ; Nutrition Surveys

DHA, one of w-3 fatty acids, modulates cell growth or death though the changes of apoptotic signaling in human endothelial ECV304 cells. We investigated the effects of DHA on the changes of apoptotic signaling in human vascular endothelial ECV304 cells using lipid peroxidation (LPO) metabolites. LPO could be originated by dietary polyunsaturated fatty acids such as linoleic acid (LA), arachidonic acid (AA) and docosahexaenoic acid (DHA). DHA caused cell death of ECV304 cells compared to LA, AA or control as evidenced by changes in cell morphology and MTT assay. LPO levels was significantly elevated by 10 fold in DHA-treated ECV 304 cells and caspase-3 activity was increased by DHA corresponding to increasing incubation times compared to control. One of reasons of the cell death in DHA-treated ECV304 cells could be expected that caspase activity, marker for mitochondrial damages, might be triggered by the increasing LPO levels. Our results strongly indicated that DHA induced LPO production has an important role on apoptotic signaling pathway in ECV304 cells. LPO production in endothelial cells which was metabolized by oxidation of dietary PUFA, might be one of risk factors in the initial progression of atherosclerosis.
Apoptosis* ; Arachidonic Acid ; Atherosclerosis ; Caspase 3 ; Cell Death ; Endothelial Cells ; Fatty Acids ; Fatty Acids, Unsaturated ; Humans* ; Linoleic Acid ; Lipid Peroxidation ; Risk Factors

Studies have reported different changes in the fatty acid composition of red blood cell (RBC) total lipids in patients with various types of cancer. It has been indicated that n-3/n-6 ratio plays a key role in the general consequence of skin photocarcinogenesis. However, to our knowledge there was no study examining the unsaturated fatty acid profile in basal cell carcinoma (BCC) patients. So, we explore the fatty acid composition of RBCs in newly diagnosed BCC patients in a hospital-based case-control study. This study has been conducted on new case BCC patients in Razi Hospital, Tehran, Iran. Fatty acid concentration in erythrocyte membranes defined as relative values after extraction, purification and preparation, by gas chromatography.Analysis revealed that heptadecenoic acid (p = 0.010) and oleic acid (p < 0.001) was significantly higher in BCC patients in comparison with control group. Among polyunsaturated fatty acids (PUFAs), linoleic acid (LA), and arachidonic acid (AA) were significantly higher in BCC patients (p < 0.001). It has been indicated that n-3 was significantly lower (p = 0.040) and n-6 was significantly higher (p = 0.002) in BCC patients. In addition, total PUFA (p < 0.001) and n-6 PUFAs/n-3 PUFAs (p = 0.002) were significantly higher in BCC patients compared to the control group. Here we indicated that new case BCC patient had significantly higher n-6 PUFA and lower n-3 along with other differences in unsaturated fatty acid in comparison with healthy subjects. Our study provides evidence that lipids are important in BCC development.
Arachidonic Acid ; Carcinoma, Basal Cell ; Case-Control Studies ; Erythrocyte Membrane ; Erythrocytes ; Fatty Acids ; Fatty Acids, Unsaturated ; Healthy Volunteers ; Humans ; Iran ; Linoleic Acid ; Oleic Acid ; Skin

The aim of this study was to examine the effects of perilla oil as well as several vegetable oils, including flaxseed oil, canola oil, and rice bran oil on plasma levels of cardioprotective (n-3) polyunsaturated fatty acids in mice by feeding each vegetable oil for a period of eight weeks. Concentrations of docosapentaenoic acid (DHA) and eicosapentaenoic acid (EPA), fish-based (n-3) polyunsaturated fatty acids, showed an increase in the plasma of mice fed perilla and flaxseed oils compared to those of mice in the control group (P < 0.05), whereas rice bran and canola oils did not alter plasma DPA and EPA concentrations. Arachidonic acid concentration was increased by feeding rice bran oil (P < 0.05), but not canola, flaxseed, or perilla oil. In addition, oleic acid, linoleic acid, and docosahexaenoic acid concentrations were altered by feeding dietary rice bran, canola, perilla, and flaxseed oils. Findings of this study showed that perilla oil, similar to flaxseed oil, is cardioprotective and could be used as an alternative to fish oil or even flaxseed oil in animal models.
alpha-Linolenic Acid ; Animals ; Arachidonic Acid ; Eicosapentaenoic Acid ; Fatty Acids ; Fatty Acids, Monounsaturated ; Fatty Acids, Unsaturated ; Flax ; Linoleic Acid ; Linseed Oil ; Mice ; Models, Animal ; Oils ; Oleic Acid ; Perilla ; Plant Oils ; Plasma ; Vegetables

Adequate supply of long-chain polyunsaturated fatty acids (LCPUFAs) is of great importance for neonates, especially preterm infants. In particular,
Fatty Acids ; Fatty Acids, Omega-3 ; Fatty Acids, Unsaturated ; Humans ; Infant ; Infant, Newborn ; Infant, Premature

OBJECTIVES: This study was undertaken to identify the effect of oxidative stress on the pathology of manganese intoxication through an analysis of manganese concentrations, superoxide dismutase (SOD) activities, malondialdehyde (MDA) concentrations, and the compositional changes of fatty acids from the corpus striatum of the rat brain. METHODS: Ten Sprague-Dawley rats were equally divided into two groups. Five rats in the experimental group were administered MnCl2 intraperitoneally for 4 weeks (4 mg/kg once daily, 5 days per week) and another five rats from the control group were given normal saline. Twenty-four hours after the last injection, the rats were decapitated and, the corpus striatum was isolated from the brain. RESULTS: In the corpus striatums of the experimental group, manganese concentrations increased significantly by 139 % (p<0.01). The SOD activities decreased significantly by 81 % (p<0.01) and the MDA concentrations increased significantly by 138 % (p<0.01) as compared to the control group. Among fatty acids, total n-6 polyunsaturated fatty acids (PUFAs) increased significantly by 325 % (p<0.01) as compared with the control group. Arachidonic acids (AA) increased by 341 % (p<0.01), and these increases were composed mostly of n-6 polyunsaturated fatty acids (PUFA). Among n-3 PUFAs, with the exception of linolenic acids, eicosapentanoic acid (EPA) decreased significantly by 72 % (p<0.05) and docosahexanoic acids (DHA) decreased by 67 % (p<0.05) as compared with the control group. CONCLUSIONS: Our results suggest that the oxygen free radicals produced by manganese may cause compositional changes of fatty acids in the corpus striatum of the rat brain. The characteristics of the fatty acids'compositional changes by manganese were a decrease of EPAs and DHAs (n-3 PUFAs), and an increase of AAs (n-6 PUFAs). These changes coupled with the decrease of SOD activity and the increase of MDA, suggest that manganese neurotoxicity is caused by lipid peroxidation mediated with oxygen free radicals, particularly superoxide radicals.
alpha-Linolenic Acid ; Animals ; Arachidonic Acid ; Arachidonic Acids ; Brain* ; Corpus Striatum* ; Eicosapentaenoic Acid ; Fatty Acids ; Fatty Acids, Omega-3 ; Fatty Acids, Unsaturated ; Free Radicals ; Linolenic Acids ; Lipid Peroxidation ; Malondialdehyde ; Manganese ; Oxidative Stress* ; Oxygen ; Pathology ; Rats* ; Rats, Sprague-Dawley ; Superoxide Dismutase ; Superoxides

Cytochrome P450 4A11 (CYP4A11) is a fatty acid hydroxylase enzyme expressed in human liver. It catalyzes not only the hydroxylation of saturated and unsaturated fatty acids, but the conversion of arachidonic acid to 20-hydroxyeicosatetraenoic acid (20-HETE), a regulator of blood pressure. In this study, we performed a directed evolution analysis of CYP4A11 using the luminogenic assay system. A random mutant library of CYP4A11, in which mutations were made throughout the entire coding region, was screened with luciferase activity to detect the demethylation of luciferin-4A (2-[6-methoxyquinolin-2-yl]-4,5-dihydrothiazole-4-carboxylic acid) of CYP4A11 mutants in Escherichia coli. Consecutive rounds of random mutagenesis and screening yielded three improved CYP4A11 mutants, CP2600 (A24T/T263A), CP2601 (T263A), and CP2616 (A24T/T263A/V430E) with ~3-fold increase in whole cells and >10-fold increase in purified proteins on the luminescence assay. However, the steady state kinetic analysis for lauric acid hydroxylation showed the significant reductions in enzymatic activities in all three mutants. A mutant, CP2600, showed a 51% decrease in catalytic efficiency (k cat/K m) for lauric acid hydroxylation mainly due to an increase in K m. CP2601 and CP2616 showed much greater reductions (>75%) in the catalytic efficiency due to both a decrease in k cat and an increase in K m. These decreased catalytic activities of CP2601 and CP2616 can be partially attributed to the changes in substrate affinities. These results suggest that the enzymatic activities of CYP4A11 mutants selected from directed evolution using a luminogenic P450 substrate may not demonstrate a direct correlation with the hydroxylation activities of lauric acid.
Animals ; Arachidonic Acid ; Blood Pressure ; Cats ; Clinical Coding ; Cytochrome P-450 Enzyme System ; Escherichia coli ; Fatty Acids, Unsaturated ; Humans* ; Hydroxylation ; Liver ; Luciferases ; Luminescence ; Mass Screening ; Mutagenesis

Two-pore domain K+ (K2P) channels are recently described. They have properties of background K+ channels, and play a crucial role in setting the resting membrane potential and regulating cell excitability. Mammalian family of K2P channel proteins are encoded by 17 KCNK genes and subdivided into 6 subfamilies on the basis of sequence similarities: TWIK, TREK, TASK, TALK, THIK, and TRESK (TWIK related spinal cord K+ channel). TWIK is weakly inward-rectifying, and TASK (acid sensitive) is a background outward-rectifiers whose activity is inhibited by low pH. TREK and TRAAK (TWIK related arachidonic acid-stimulated K+ channel) are both outward-rectifiers that are activated by polyunsaturated fatty acids, including arachidonic acid. The alkaline-activated K2P channel TALK-1 is primarily expressed in the pancreas. THIK is halothane-inhibited K+ channel. TRESK is a TWIK related spinal cord K+ channel. Many studies with volatile general anesthetics were performed using the patch-clamp technique and they showed the activation of K+ currents and they suggested that the K2P channels might be an another plausible group of targets for general anesthetics. An appreciation of how general anesthetics modulate the activity of K2P channels at the molecular level will be enhanced by future studies using site-directed mutagenesis, high-resolution structural approaches, and molecular dynamics simulations.
Anesthetics, General ; Arachidonic Acid ; Fatty Acids, Unsaturated ; Humans ; Hydrogen-Ion Concentration ; Membrane Potentials ; Mutagenesis, Site-Directed ; Pancreas ; Patch-Clamp Techniques ; Proteins ; Spinal Cord