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Документ Взаємодія гліфосату з амінокислотами генетично модифікованої раундапостійкої і не генетично модифікованої сої у водному розчині в умовах in vitro(Українська медична стоматологічна академія, 2018) Кулик, Ярослава Михайлівна; Обертюх, Ю. В.; Виговська, І. О.; Гончар, Л. О.; Кулик, Ярослава Михайловна; Выговская, И. А.; Гончар, Л. А.; Kulyk, Y. M.; Obertiukh, Y. V.; Vyhovska, I. O.; Honchar, L. O.Встановлено, що діюча речовина Roundup – гліфосат в умовах in vitro вступає у взаємодію з ароматичними амінокислотами сої, а з іншими амінокислотами в організмі, як аналог гліцину, утворює пептидні сполуки, що є підтвердженням утворення в організмі людей і тварин неприродних пептидів за наявності в продуктах харчування і кормах гліфосату; Установлено, что действующее вещество Roundup – глифосат в условиях in vitro вступает во взаимодействие с ароматическими аминокислотами сои, а с другими аминокислотами в организме, как аналог глицина, образует пептидные соединения, что является подтверждением образования в организме людей и животных неестественных пептидов при наличии в продуктах питания и кормах глифосата; Research purpose. To study the interaction of glyphosate with amino acids GM Roundup-resistant and non GM soy in an aqueous solution in conditions in vitro on the principle of Pauli reaction with aromatic amino acids and formol titration. Object and research methods. In the first 2 cups placed 10 g of soybeans not GM and in one of them added 66 mg of Roundup (glyphosate) in dry powder form, and in the 3rd and 4th cups, similar to 10 g of GM Roundupresistant soybean, and in one of them was added also 66 mg of Roundup (glyphosate). In all 4 cups, 150 ml of distilled water was added, and in the 5th cup only 66 mg of Roundup (glyphosate) and 150 ml of distilled water. Whenь boiling during 1 hour, the water evaporated, so the volume was adjusted to 100 ml with distilled water. In the process of boiling, there was inactivation of all biologically active substances soy with the transition to aqueous solution of heat-resistant water-soluble proteins, peptides and amino acids, including aromatic in non-GM soybeans and possibly the remaining glyphosate from the Roundup-resistant soy. In the boiling process, glyphosate, as the active ingredient of the Roundup herbicide, interacts with the amino acids, peptides and proteins of aqueous extracts soy of both variants. After boiling broth filtered through a nylon cloth and thus received aqueous extract for subsequent studies. In the obtained aqueous extract of soy and glyphosate aqueous solution was determined by reaction Pauli content of aromatic amino acids, including phenylalanine, tyrosine, histidine and tryptophan. The interaction of glyphosate with aromatic amino acids of aqueous extract of soya in the boiling process should be manifested by the formation of compounds of glyphosate with aromatic amino acids. Because glyphosate is blocking their synthesis in non-Roundup-resistant plants – weeds when crop spraying of GM soy. In water extracts of soya 2nd variants and in a solution only glyphosate without soy was determined by the content of amine nitrogen on the principle of formol titration. Research results and their discussion. Indicators extinction FEC reaction Pauli content of aromatic amino acids in aqueous extract different samples of non-GM soybeans and the same indicators when boiling with glyphosate showed reduction of aromatic amino acids by boiling soybeans with glyphosate. This is evidence that glyphosate (N-phosphonomethylglycine) contained in Roundup-resistant GM soybeans in the gastrointestinal tract, blood and tissue cells, primarily of people, and animals enter into peptide chain as glycine analogue. In this study, glyphosate reacts with aromatic amino acids reducing the color intensity of the reaction Pauli, i.e., the index of extinction FEC reduced. Obviously, the phosphonomethyl group of N-phosphonomethylglycine reacts with an aromatic ring of these amino acids, which prevents the passage of the Pauli reaction. Realization similar studies with Roundup-resistant GM soy showed a further decrease of extinction FEC of soybeans after boiling with glyphosate. If the percentage of reduction the content of aromatic amino acids in non-GM soybean was 10.4 ± 0.88 % in the range of deviations of 14.0-7.0 %, then in GM soy the average value was 18.1 ± 3.09 % with a variation of 36.0 to 9.0%. The analysis of the comparison of the reaction with aromatic amino acids of non-GM soybeans with GM soya and with glyphosate during boiling indicates a more distinct reaction of the aromatic amino acids of Roundupresistant soy in the understanding of the interaction of the phosphonomethyl group of glyphosate with the aromatic ring of these amino acids. Indicators of formol titration on the content of amino acids in aqueous extracts of non-GM soybeans without glyphosate and when boiling with glyphosate convincingly showed the interaction of amino acids with N-phosphonomethylglycine. As a result of such interaction, compounds of peptides with a phosphonomethyl group of glyphosate are formed. Similar results were obtained in studies with Roundup-resistant GM soybeans. Conclusions 1. Based on Pauli the reaction found that glyphosate under conditions in vitro glyphosate interacts, that is, forming compounds with aromatic amino acids of non-GM soybeans and more pronounced with the same aromatic amino acids Roundup-resistant GM soybeans. 2. The method of formol titration confirmed that in vitro glyphosate when it is boiled in an aqueous solution with whole soybean interacts with amino acids as an analogue of glycine to form unnatural peptide compounds. 3. The studies conducted are an argument for the risk of using GM soy in food for people and especially children.Документ Наявність неприродних пептидів у білку бобів генетично модифікованої раундапостійкої сої(Полтавський державний медичний університет, 2018) Кулик, Я. М.; Чорнолата, Л. П.; Kulyk, Y. M.; Chornolata, L. P.Дослідженнями, які включають кип’ятіння у водному розчині подрібненої сої, осадження білків трихлороцтовою кислотою, одержання фільтрату і визначення на спектрофотометрі вмісту ароматичних амінокислот на принципі поглинання ними ультрафіолетових променів при 280 нм, встановлено нижчий вміст ароматичних амінокислот в генетично модифікованій (ГМ) раундапостійкій сої порівняно з не ГМ соєю, що свідчить про наявність в ГМ сої неприродних пептидів, очевидно, сполуки гліфосату з тирозином, триптофаном і фенілаланіном.Документ Небезпека використання трансгенної раундапостійкої сої в продуктах харчування дітей через наявність у ній неідентифікованих факторів(Українська медична стоматологічна академія, 2017) Кулик, Ярослава Михайлівна; Обертюх, Ю. В.; Хіміч, О. В.; Виговська, І. О.; Кулик, Ярослава Михайловна; Обертюх, Ю. В.; Химич, А. В.; Выговская, И. А.; Kulyk, Y. M.; Obertiukh, Y. V.; Khimich, O. V.; Vygovska, I. O.Проведеними дослідженнями за допомогою біологічного тесту виявлено наявність неідентифікованого фактору в бобах трансгенної раундапостійкої сої, в жовтку і білку яєць курей та м’язовій і кістковій тканині щурів, яким згодовували таку сою, обґрунтовується стимулюванням розмноження інфузорій тетрахімена піріформіс у поживному середовищі водної витяжки сої, водному екстракті після кип’ятіння жовтка і білка яєць курей та водному екстракті (бульйоні) м’язової і кісткової тканини щурів. Це є підтвердженням, що в трансген- ній раундапостійкій сої присутній неідентифікований фактор, який викликає небезпеку використання такої сої в продуктах харчування дітей, підлітків і молодих людей; Проведенными исследованиями с помощью биологического теста выявлено наличие неидентифицированного фактора в бобах трансгенной раундапостойкой сои, в желтке и белке яиц кур и мышечной и костной ткани крыс, которым скармливали такую сою, обосновывается стимулированием размножения инфузорий тетрахимены пириформис в питательной среде водной вытяжки сои, водном экстракте после кипячения желтка и белка яиц кур и в водном экстракте (бульоне) мышечной и костной ткани крыс. Это является подтверждением, что в трансгенной раундапостойкой сои присутствует неидентифицированый фактор, который вызывает опасность использования такой сои в продуктах питания детей, подростков и молодых людей; Research purpose. Detecting the presence of unidentified factor transgenic roundup resistant soybeans by method biological tests. Object and research methods. Ciliates — Tetrahymena pyriformis, after boiling water extract of soybeans and whole plant GM soybeans and non GM soy, aqueous extract of yolks and egg whites of chickens were fed GM soy (experiment) and sunflower meal (control), aqueous extract (broth) muscle and bone tissue of rats were fed GM soy (experiment) and sunflower meal (control). To obtain the aqueous extract of GM and non GM soy weighed 50 grams beans both options and covered in heat- resistant glass cups, and then added 300 ml of distilled water, heated to boiling and boil for 30 minutes. In the process of boiling occurred inactivation of urease, antitrypsin and antyhimotrypsynu and other anti-utritional bioactive substances with the transition to an aqueous solution of water-soluble fraction thermostable proteins of low molecular weight substances from non-GM soybeans and plasmid DNA fragments and other low molecular weight compounds Agrobacterium tumefaciens with GM soya beans. After boiling broth filtered through a nylon cloth and thus prepared aqueous extract of GM and non GM soy. Test each aqueous extract of soybean GM and non GM conducted a 3-time repetition. In three vials of antibiotics contributed to 1 cm3 aqueous extract roundup resistant GM soybeans and similarly in the 3-bottle — non-GM soybeans, and then added to each bottle to 0.1 cm3 3 daily culture ciliates Tetrahymena pyriformis. The following biological test based on the determined presence of unidentified factor transgenic roundup resistant soybeans in the eggs of hens based on stimulating vitality ciliates Tetrahymena pyriformis in conditions in vitro when using a nutrient medium — water extract of yolk and egg whites chickens which a part of the diet were fed transgenic roundup resistant soybeans. The presence of unidentified factor roundup resistant transgenic soy in chicken eggs is their ability to stimulate the vitality of ciliates Tetrahymena pyriformis in nutrient medium aqueous extract after boiling egg yolk and whites chicken, which in part of the diet for 2 months were fed roundup resistant transgenic soybeans compared with water extract the yolk and egg whites chicken, which is not were fed GM soy. To obtain the aqueous extract of yolk and egg whites chicken experimental and control groups performed 3 times the selection of eggs for three of the group. Smashed eggs and yolk and whites separated from the transfer of in heat-resistant glass cups, and then added for 50 ml of distilled water for one egg yolk and whites, after their thoroughly homogenized to obtain a homogeneous aqueous solution and heated to boiling, boil for 30 minutes. In the process of boiling occurred inactivation of biologically active substances with the transition to an aqueous solution of water-soluble fraction thermostable proteins, lipoproteins, phospholipids, low molecular weight compounds and fragments of ring structures of DNA and unidentified substances Agrobacterium tumefaciens from eggs of hens of the experimental group. After the boil in every glass added 2 ml of 9 % acetic acid and stirred welded whites and yolk. After filtration method of push ups through a nylon fabric obtained water extract of yolk and whites, which re-filtered through a paper filter. In the extract obtained drops pH value adjusted to 7.2-7.5 0.1 n solution of NaOH. To obtain an aqueous extract of muscle and bone tissue of experimental and control rats carried their slaughter for 3 heads from each group and carcasses, ie, muscle and bone tissue finely crushed and placed in a heat-resistant glass cups, and then added 200 ml of distilled water, heated to boiling and boil for 30 minutes. In the process of boiling occurred inactivation of biologically active substances with the transition to an aqueous solution of water- soluble fraction thermostable proteins of low molecular weight substances from the muscle and bone tissue of rats in the control group and possible fragments of DNA plasmids and unidentified compounds Agrobacterium tumefaciens tissues of animals of the experimental group. After boiling decoction filtered through a nylon cloth and thus obtained aqueous extract of muscle and bone tissue of rats experimental and control group. Test each aqueous extract of muscle and bone tissue carried from 3 rats. In three vials from antibiotics contributed to 1 cm3 of aqueous extract of muscle and bone tissue of rats research and similar in 3-bottle — the control group, and then added to every bottle to 0.1 cm3 3 daily culture ciliates Tetrahymena pyriformis. Research results and their discussion. The performed a study using a biological test revealed the presence of unidentified factor in transgenic beans resistant roundup soybean in egg yolks and egg whites chicken and muscle and bone tissue of rats were fed soy substantiated such stimulation multiplication ciliates Tetrahimena piriformis nutrient medium in aqueous extract of soy, water the extract after boiling (broth) of muscle and bone tissue of rats. Conclusions. The research results are confirmation that the transgenic soybean roundup resistant present unidentified factor that causes the danger of using such soy in food products children, adolescents and young people.