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Applied Mechanics and Materials ISSN: 1662-7482, Vol. 310, pp 162-165 doi:10.4028/www.scientific.net/AMM.310.162 © 2013 Trans Tech Publications, Switzerland Online: 2013-02-27 Evaluation of using Eicosapentaenoic acid in Plant Tissue Culture Media Kamran Safavi Technology Incubator Center, Khorasgan (Isfahan) Branch, Islamic Azad University, Isfahan, Iran. Email: [email protected] Keywords: Omega-3, Essential fatty acids, Eicosapentaenoic acid, Plant tissue culture Abstract. Biotechnology is name given to the methods and techniques that involve the use of living organisms like bacteria, yeast, plant cells or their parts or products as tools. Plant tissue culture techniques are essential to many types of academic inquiry, as well as to many applied aspects of plant science. Plant tissue culture techniques are also central to innovative areas of applied plant science, including plant biotechnology and agriculture. The omega-3 fatty acids are "essential" fatty acids because they are vital for normal metabolism and cannot be synthesized by the human body. Eicosapentaenoic acid is an omega-3 fatty acid. With this research we obtain the strong plants with benefits of Omega-3 fatty acids. Introduction Plant tissue culture techniques are essential to many types of academic inquiry, as well as to many applied aspects of plant science. In the past, plant tissue culture techniques have been used in academic investigations of totipotency and the roles of hormones in cytodifferentiation and organogenesis. Currently, tissue-cultured plants that have been genetically engineered provide insight into plant molecular biology and gene regulation. Plant tissue culture techniques are also central to innovative areas of applied plant science, including plant biotechnology and agriculture [4]. In addition, the management of genetically engineered cells to form transgenic whole plants requires tissue culture procedures; tissue culture methods are also required in the formation of somatic haploid embryos from which homozygous plants can be generated. Thus, tissue culture techniques have been, and still are, prominent in academic and applied plant science [5]. The omega-3 fatty acids docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) are orthomolecular, conditionally essential nutrients that enhance quality of life and lower the risk of premature death. They function exclusively via cell membranes, in which they are anchored by phospholipid molecules. DHA is proven essential to pre- and postnatal brain development, whereas EPA seems more influential on behavior and mood. Both DHA and EPA generate neuroprotective metabolites. In doubleblind, randomized, controlled trials, DHA and EPA combinations have been shown to benefit attention deficit/hyperactivity disorder (AD/HD), autism, dyspraxia, dyslexia, and aggression [7]. For the affective disorders, meta-analyses confirm benefits in major depressive disorder (MDD) and bipolar disorder, with promising results in schizophrenia and initial benefit for borderline personality disorder. Accelerated cognitive decline and mild cognitive impairment (MCI) correlate with lowered tissue levels of DHA/EPA, and supplementation has improved cognitive function. Huntington disease has responded to EPA. Omega-3 phospholipid supplements that combine DHA/EPA and phospholipids into the same molecule have shown marked promise in early clinical trials [2]. Utilizing DHA and EPA together with phospholipids and membrane Mantioxidants to achieve a “triple cell membrane synergy” mayfurther diversify their currently wide range of clinical applications. All rights reserved. No part of contents of this paper may be reproduced or transmitted in any form or by any means without the written permission of Trans Tech Publications, www.ttp.net. (ID: 130.203.136.75, Pennsylvania State University, University Park, USA-05/03/16,15:49:39) Applied Mechanics and Materials Vol. 310 163 Result and Discussion Metabolism and Biological Effects of Essential Fatty Acids Dietary fat is an important source of energy for biological activities in human beings. It encompasses saturated fatty acids (SFAs), which are usually solid at room temperature, and unsaturated fatty acids (UFAs), which are liquid at room temperature. UFAs can be further divided into monounsaturated (MUFAs) and polyunsaturated fatty acids (PUFAs). PUFAs can be classified, on the basis of their chemical structure, into two groups: omega-3 (n-3) fatty acids and omega-6 (n-6) fatty acids. The omega-3 or n-3 notation means that the first double bond in this family of PUFAs is 3 carbons from the methyl end of the molecule. The same principle applies to the omega-6 or n-6 notation [6]. Of all fats found in food, two alpha-linolenic acid (chemical abbreviation: ALA; 18:3 n-3) and linoleic acid (LA; 18:2 n-6)—cannot be synthesized in the human body, yet these are necessary for proper physiological functioning. These two fats are thus called “essential fatty acids” (EFAs). The EFAs can be converted in the liver to long-chain PUFAs (LC PUFAs), which have a higher number of carbon atoms and double bonds. ALA and LA comprise the bulk of the total PUFAs consumed in a typical North American diet. Typically, LA comprises 89 percent of the total PUFAs consumed, while ALA comprises percent. Smaller amounts of other PUFAs make up the remainder.1 Both ALA and LA are present in a variety of foods. Eicosapentaenoic acid (EPA; 20:5 n-3) and docosahexaenoic acid (DHA; 22:6 n-3) can act as competitors for the same metabolic pathways as arachidonic acid (AA; 20:4 n-6). General scientific agreement supports increased consumption ofomega-3 fatty acids and reduced intake of omega-6 fatty acids to promote good health. However, for omega-3 fatty acid intake, the specific quantitative recommendations vary widely among countries not only in terms of different unit’s ratio, grams and total energy intake but also in quantity. Metabolic Pathways of Omega-3 and Omega-6 Fatty Acids Omega-3 and omega-6 fatty acids share the same pools of enzymes and go through the same oxidation pathways while being metabolized. Once ingested, the parent of the omega- 3 fatty acids, ALA, and the parent of the omega-6 fatty acids, LA, can be elongated and desaturated into LC PUFAs. The long-chain omega-6 fatty acid, AA, is the precursor of a group of eicosanoids including series-2 prostaglandins (PG2) and series-4 leukotrienes (LT4). The omega- 3 fatty acid, EPA, is the precursor to a group of eicosanoids including series-3 prostaglandins (PG3) and series-5 leukotrienes (LT5). The series-2 prostaglandins and series-4 leukotrienes Nderived from AA are involved in intense actions (such as accelerating platelet aggregation, and enhancing vasoconstriction and the synthesis of mediators of inflammation) in response to physiological stressors. The series-3 prostaglandins and series-5 leukotrienes derived from EPA are less physiologically potent than those derived from AA. More specifically, the series-3 prostaglandins are formed at a slower rate and work to attenuate excessive series-2 prostaglandins [6]. Thus, adequate production of the series-3 prostaglandins, which are derived from the omega-3 fatty acid, EPA, may protect against heart attack and stroke as well as certain inflammatory diseases like arthritis, lupus and asthma [6]. EPA can also be converted into the longer chain omega-3 form of docosapentaenoic acid (DPA, 22:5 n-3), and then further elongated and oxygenated into DHA. EPA and DHA are frequently referred to as VLN-3FA—very long chain n-3 fatty acids. DHA, which is thought to be important for brain development and functioning, is present in significant amounts in a variety of food products, including fish, fish liver oils, fish eggs, and organ meats. Similarly, AA can convert into an omega-6 form of DPA. Studies have reported that omega-3 fatty acids decrease triglycerides (Tg) and very low density lipoprotein (VLDL) in hypertriglyceridemic subjects, concomitant with an increase in high density lipoprotein (HDL). However, they appear to increase or have no effect on low density lipoprotein (LDL). Omega-3 fatty acids apparently lower Tg by inhibiting VLDL and apolipoprotein B-100 synthesis, and decreasing post-prandial lipemia. Omega-3 fatty acids, in conjunction with transcription factors (small proteins 164 Engineered Technologies in Materials Science, Geotechnics, Environment and Mechanical Engineering that bind to the regulatory domains of genes), target the genes governing cellular Tg production and those activating oxidation of excess fatty acids in the liver. Inhibition of fatty acid synthesis and increased fatty acid catabolism reduce the amount of substrate available for Tg production. As noted earlier, omega-6 fatty acids are consumed in larger quantities (> 10 times) than omega-3 fatty acids. Maintaining a sufficient intake of omega-3 fatty acids is particularly important since many of the body’s physiologic properties depend upon their availability and metabolism [6]. Omega-3s in Childhood Brain Development During the last trimester of fetal life and the first two years of childhood, the brain undergoes a period of rapid growth the “brain growth spurt.”1 Nutrient insufficiency during this period can compromise brain function. DHA is one nutrient absolutely required for the development of the sensory, perceptual, cognitive, and motor neural systems during the brain growth spurt. EPA’s importance for the brain’s development in utero is unclear, but colostrum and breast milk contain EPA, albeit in lesser amounts than DHA [7]. Established Benefits in Affective Disorders Numerous studies have examined the effects of DHA/EPA for affective disorders and have found them to be beneficial for mood management. Mood disorders that apparently respond to DHA/EPA include major depressive disorder, manic depression (bipolar disorder), and possibly also schizophrenia, borderline personality disorder (BPD), and anorexia nervosa [7]. DHA/EPA Support Cell Membranes, the Pacemakers of Metabolism DHA and EPA literally feed cell membranes, the dynamic structures that manage the vast majority of life processes. Within the membrane bilayer, DHA and EPA are attached to the larger phospholipid molecules via ester bonds. Phospholipids with their attached fatty acids are the molecular building blocks of the membrane. DHA and EPA interact with the other fatty acids in the membrane bilayer – saturates, monounsaturates (omega-9s), and polyunsaturates (omega-6s, minor omega-3s) – and membrane fluidity is a net outcome of all the electron densities. This property renders DHA (six double bonds) and EPA (five double bonds) the most highly fluidizing of the major membrane fatty acids. In general, experts agree the advanced human cell is only as efficient as its membrane system [7]. Orthomolecular Synergy of Cell Membrane Nutrients DHA and EPA have an obvious and predictable synergy with other cell membrane nutrients, specifically phospholipids and antioxidants. Depending on the requirements of the tissue in question, the phospholipids phosphatidylserine (PS), phosphatidylethanolamine (PE), and PC can carry substantial amounts of DHA in their “tail” positions, especially tail position [7]. These phospholipid “parent molecules” also anchor EPA within the membrane. Healthy cells have a complement of antioxidants within their membranes that help protect them from destruction by intrinsic oxidants (obligatorily generated during routine metabolism) or extrinsic oxidants imposed by lifestyle or the environment.90 Membrane, antioxidants are structurally intermingled with fatty acids and function as a protective “first line of defense.” In the presence of antioxidants, fatty acids with the most unsaturated bonds, namely DHA and EPA, are protected against oxidative (“free radical”) destruction [7]. Thus, within the dynamic membrane milieu, DHA and EPA exist in homeostatic synergy with both their parent phospholipids and the antioxidants dispersed in the membrane lipid bilayer, providing “triple cell membrane synergy.” Thus, in addition to protective antioxidants, supplements Applied Mechanics and Materials Vol. 310 165 that deliver DHA and EPA bound to phospholipids – such as omega-3s bound to phosphatidylserine and krill oil that contains omega-3 FAs bound to phospholipids – provide the building blocks for healthy cell membranes [7]. The Correct Intake of DHA/EPA for Brain Benefits Technically, humans can synthesize EPA and DHA from the shorter-chain ALA, but the conversion efficiency is low, even in healthy individuals. Thus flaxseed oil as a source of ALA cannot be assumed to substitute for dietary sources of DHA/EPA. Foods high in omega-3 FAs or supplements with preformed DHA and EPA are required. In regard to food sources of DHA/EPA, the standard American diet is unlikely to contribute more than 50-100 mg/day. Various “functional foods” have appeared with DHA/EPA added. Omega-3 eggs, for example, can be a significant source by providing greater than 200 mg of “omega-3” per egg [7]. However, it may be necessary to confirm which omega-3 FAs are in the food (e.g., DHA/EPA or ALA). Further caution is advised to ensure that other ingredients in the food are healthful. For example, one heavily promoted omega-3 spread carries trans-fatty acids a potential toxic counterbalance to the omega-3 benefits. The current knowledge base on DHA/EPA for brain function does not generate a rational daily intake recommendation. From his studies on national seafood intakes and affective disorder incidence, suggested pregnant women may want to consume a minimum 650 mg/day of DHA and EPA (with a minimum 300 mg/day of DHA) to prevent postpartum depression [7]. Using Eicosapentaenoic acid in Plant Tissue Culture Media In this research we understand that the omega-3 such as Eicosapentaenoic has very good benefits and when we add it to plant tissue culture media the plant can absorb the omega-3. Potato is one of the plants that we candidate for this research [1,4]. Therefore the plants have an omega-3 and when people eat them they obtain the benefit of omega-3. References [1]- A. Badoni and J. S. Chauhan. Effect of Growth Regulators on Meristem-tip Development and in vitro Multiplication of Potato Cultivar ‘Kufri Himalini’. Nature and Science. 2009.7(9):31-34. [2]- P. Kidd. Omega-3 DHA and EPA for Cognition, Behavior, and Mood: Clinical Findings and Structural-Functional Synergies with Cell Membrane Phospholipids. Alternative Medicine Review. 2007. Volume 12, Number 3. [3]- M. Khalafalla, K. G. Abd Elaleem and R. S. Modawi. Callus formation and organogenesis of potato (Solanum Tuberosum L.) cultivar almera. Journal of Phytology Tissue Culture. 2010. 2(5): 40–46. [4]- L. Mineo. Plant tissue culture techniques. Pages 151 174, in Tested studies for laboratory teaching. Volume 11. (C. A. Goldman, Editor). Proceedings of the Eleventh Workshop/Conference of the Association for Biology Laboratory Education (ABLE), 1990. 195 pages. [5]- M. Saljooghian Pour, M. Omidi, I. Majidi, D. Davoodi and P. Ahmadian. In-vitro plantlet propagation and microtuberization of meristem culture in some of wild and commercial potato cultivars as affected by NaCl. African Journal of Agricultural Research. 2010. 5(4): 268-274. [6]- H. Schachter. Effects of Omega-3 Fatty Acids on Mental Health . Evidence Report/Technology Assessment. AHRQ Publication No. 05-E022-2. 2005. Number 116. [7]- A. Simopoulos. Omega-3 Fatty Acids and Antioxidants in Edible Wild Plants. Biol Res. 2004. 37: 263-277. [8]- S. Supinski, J. Vanags, L. Callahan. Eicosapentaenoic acid preserves diaphragm force generation following endotoxin administration. Critical Care. 2010. 14:R35 Engineered Technologies in Materials Science, Geotechnics, Environment and Mechanical Engineering 10.4028/www.scientific.net/AMM.310 Evaluation of Using Eicosapentaenoic Acid in Plant Tissue Culture Media 10.4028/www.scientific.net/AMM.310.162