21 research outputs found

    Admittance spectroscopy of charge traps of FET based on nanotubes

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    Investigation of electrical properties of FET based on polymer wrapped nanotubes, namely charge carrier transport and trap appear mechanisms. Model of carrier transport in the device was performed. Local traps states activation energies were obtained

    Evaluation of biotransformation capacity of transplastomic plants and hairy roots of Nicotiana tabacum expressing human cytochrome P450 2D6

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    Cytochrome P450 monooxygenases (CYPs) are important tools for regio- and stereoselective oxidation of target molecules or engineering of metabolic pathways. Functional heterologous expression of eukaryotic CYPs is often problematic due to their dependency on the specific redox partner and the necessity of correct association with the membranes for displaying enzymatic activity. Plant hosts offer advantages of accessibility of reducing partners and a choice of membranes to insert heterologous CYPs. For the evaluation of plant systems for efficient CYP expression, we established transplastomic plants and hairy root cultures of Nicotiana tabacum carrying the gene encoding human CYP2D6 with broad substrate specificity. The levels of CYP2D6 transcript accumulation and enzymatic activity were estimated and compared with the data of CYP2D6 transient expression in N. benthamiana. The relative level of CYP2D6 transcripts in transplastomic plants was 2-3 orders of magnitude higher of that observed after constitutive or transient expression from the nucleus. CYP2D6 expressed in chloroplasts converted exogenous synthetic substrate loratadine without the need for co-expression of the cognate CYP reductase. The loratadine conversion rate in transplastomic plants was comparable to that in N. benthamiana plants transiently expressing a chloroplast targeted CYP2D6 from the nucleus, but was lower than the value reported for transiently expressed CYP2D6 with the native endoplasmic reticulum signal-anchor sequence. Hairy roots showed the lowest substrate conversion rate, but demonstrated the ability to release the product into the culture medium. The obtained results illustrate the potential of plant-based expression systems for exploiting the enzymatic activities of eukaryotic CYPs with broad substrate specificities

    Creation of glyphosate-resistant Brassica napus L. plants expressing DesC desaturase of cyanobacterium Synechococcus vulcanus

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    Aim. Creation of glyphosate-resistant canola plants expressing bifunctional hybrid desC::licBM3 gene. In the hybrid gene the sequence of DesC desaturase of cyanobacterium S. vulcanus without plastid targeting was fused with the sequence of thermostable lichenase reporter LicBM3 gene. Methods. Agrobacterium tumefaciens-mediated transformation, PCR, quantitative and qualitative determination of lichenase activity, genetic analysis. Results. Transgenic canola plants, carring the enolpyruvat shikimat phosphate syntase gene (epsps), conferring on plants resistance to phosphonomethyl glycine herbicides (Roundup), as well as the desC::licBM3 gene, were selected. The presence of transgenes was confimed by multiplex PCR. The epsps gene expression in canola was shown at the transcription level, during in vitro growth and after greenhouse herbicide treatment. Activity of the licBM3 gene product as a part of hybrid protein allowed quantitative and qualitative estimation of the desaturase gene expression. Inheritance of heterologous genes and their expression in the first generation were investigated. Conclusions. Transgenic canola plants were obtained, the presence of trangenes in plant genome was proved and expression of the target genes was detected

    Dynamics of markers of endothelial dysfunction in experimental antiphospholipid syndrome

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    Analysis of the entire set of currently available factors allows us to consider antiphospholipid syndrome (APS) as a unique model of autoimmune platelet vasculopathy, the study of which is essential for determining the relationship between such fundamental pathological processes as atherosclerosis, vasculitis, blood clotting disorders and the immune system. At the same time, the diagnosis and treatment of APS is based on serological tests, and little attention is paid to hemostatic markers and endothelial dysfunction in this pathology. The aim of the study was to study the dynamics of endothelial state indicators in animals with simulated antiphospholipid syndrome and the effectiveness of their correction by introducing warfarin, immunoglobulin and L-arginine. Research results. Disruption of the functional state of the endothelium in animals with simulated antiphospholipid syndrome was found. A pronounced increase in the von Willebrand factor level (p <0.001) in the blood of rats with experimental APS was proved. It was revealed that the development of experimental antiphospholipid syndrome leads to an increase in the level of homocysteine in the blood of rats (p <0.001). The most pronounced results of correction were found in the fifth group, the animals of which received complex therapy with warfarin, immunoglobulin and L-arginine

    Study of indicators of endothelial dysfunction in rats with experimental peritonitis

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    Gutsulyuk V. G., Savytskyi I. V., Tsypoviaz S. V., Znamerovskyi S. G., Sarahan V. M., Gerasymenko T. V. Study of indicators of endothelial dysfunction in rats with experimental peritonitis. Journal of Education, Health and Sport. 2022;12(1):514-522. eISSN 2391-8306. DOI http://dx.doi.org/10.12775/JEHS.2022.12.01.043 https://apcz.umk.pl/JEHS/article/view/41740 https://zenodo.org/record/7512638 The journal has had 40 points in Ministry of Education and Science of Poland parametric evaluation. Annex to the announcement of the Minister of Education and Science of December 1, 2021. No. 32343. Has a Journal's Unique Identifier: 201159. Scientific disciplines assigned: Physical Culture Sciences (Field of Medical sciences and health sciences); Health Sciences (Field of Medical Sciences and Health Sciences). Punkty Ministerialne z 2019 - aktualny rok 40 punktów. Załącznik do komunikatu Ministra Edukacji i Nauki z dnia 1 grudnia 2021 r. Lp. 32343. Posiada Unikatowy Identyfikator Czasopisma: 201159. Przypisane dyscypliny naukowe:Nauki o kulturze fizycznej (Dziedzina nauk medycznych i nauk o zdrowiu); Nauki o zdrowiu (Dziedzina nauk medycznych i nauk o zdrowiu). © The Authors 2022; This article is published with open access at Licensee Open Journal Systems of Nicolaus Copernicus University in Torun, Poland Open Access. This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author (s) and source are credited. This is an open access article licensed under the terms of the Creative Commons Attribution Non commercial license Share alike. (http://creativecommons.org/licenses/by-nc-sa/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. The authors declare that there is no conflict of interests regarding the publication of this paper. Received: 05.01.2022. Revised: 21.01.2022. Accepted: 31.01.2022. UDC 616.381-018.74 Study of indicators of endothelial dysfunction in rats with experimental peritonitis V. G. Gutsulyuk, I. V. Savytskyi, S. V. Tsypoviaz, S. G. Znamerovskyi, V. M. Sarahan, T. V. Gerasymenko International Academy of Ecology and Medicine, Kyiv, Ukraine Correspondence author: Savytskyi Ivan Volodymyrovich, 65039, Odesa, Fountain road4-а/29, tel.+38050-381-21-83, [email protected] Abstract The article presents the results of the study of the activity of endothelial and inducible NO-synthase, the level of Willebrand factor and endothelin-1 in rats with experimental peritonitis. The most likely mechanism that is damaged in the endothelium during peritonitis is the activation of the synthesis of inducible NO-synthase by neutrophils/macrophages in response to infection. It is possible that hyperproduction of nitric oxide (NO), on the one hand, is aimed at destroying microflora and oxidizing toxins, and on the other hand, at suppressing the expression of tissue factor and cell adhesion molecules. platelet aggregation and cascade disorders in the hemostasis system. All this indicates that the hyperproduction of NO not only reflects the processes that occur in the focus of damage to the vascular endothelium, but also affects the severity of the inflammatory process and the outcome of the disease. In animals with experimental peritonitis on the background of OS, an increase in the number of circulating desquamated endothelial cells in the blood, which is a highly specific marker of endothelial dysfunction, was noted. The level of the Willebrand factor also increased, which can serve as a marker of increased risk of thrombus formation and indicate the pathogenetic dependence of the factors that damage the vascular wall endothelium on the concentration of the Willebrand factor, which contributes to the reduction of vascular permeability by adhesion of platelets to the endothelium. Confirmation of the development of endothelial dysfunction in peritonitis is an increase in the concentration of endothelin-1, which is a regulator of the process of vascular neoangiogenesis in response to endothelial damage. Keywords: peritonitis; endothelial dysfunction; oxidative stress; Willebrand factor; pathogenesis

    Multiplex PCR assay for detection of human interferon alpha2b gene in transgenic plants

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    During the last decade interferons are regarded as potent candidates for generation of plant-based edible vaccines because of broad spectrum of antiviral activities and adjuvant properties. Establishment and certification of numerous interferon producing plant systems requests development of fast and efficient multiplex PCR protocol for the transgene detection in GM plants. Here we represent a protocol for simultaneous amplification in one assay of fragments of hIFN alpha 2b gene and two control genes, namely virD1 of Agrobacterium tumefaciens and conservative region of plant actin gene.В последнее десятилетие интерфероны рассматриваются как перспективные кандидаты для получения из растений в виде съедобных вакцин, поскольку обладают широким спектром антивирусной активности и адъювантными свойствами. Создание и сертификация многочисленных растительных систем, продуцирующих рекомбинантный интерферон, делают актуальной разработку быстрого и эффективного протокола мультиплексной ПЦР для определения данного трансгена в генетически модифицированных растениях. В настоящей публикации мы приводим метод детекции гена человеческого интерферона альфа-2b в трансгенных растениях с помощью совместной амплификации в ходе одной реакции фрагментов гена hINTα2b и двух контрольных генов, virD1 Agrobacterium tumefaciens и консервативного участка гена актина растений.В останнє десятиліття інтерферони розглядаються як перспективні кандидати для отримання з рослин у вигляді їстівних вакцин оскільки, вони мають широкий спектр антивірусної активності й ад’ювантні властивості. Створення і сертифікація численних рослинних систем, які накопичують рекомбінантний інтерферон, роблять актуальною розробку швидкого й ефективного протоколу мультиплексної ПЛР для визначення даного трансгена в генетично модифікованих рослинах. В цій публікації ми наводимо метод детекції гена людського інтерферону альфа-2b у трансгенних рослинах за допомогою сумісної ампліфікації в ході однієї реакції фрагментів гена hINTα2b і двох контрольних генів, virD1 Agrobacterium tumefaciens і консервативної ділянки гена актину рослин

    Creation of transgenic Brassica napus L. Plants expressing human alpha 2b interferon gene

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    Spring rapeseed transgenic lines expressing human interferon alpha 2b were created by Agrobacteriummediated transformation of aseptic plant leaf explants. The maximum antiviral activity of the leaf extracts reached 4500 IU/g fresh weight. It was determined that the antioxidant activity and the activity of an enzyme of plant antioxidant system – superoxide dismutase (SOD) – in the leaf tissues of transgenic plants increased compared to controls. There were no correlations between the interferon and antioxidant activities, as well as between SOD and interferon activities. Using the obtained transgenic rapeseed plants with high interferon and antioxidant activities as a feed additive for animals might have preventive effect on their body, increasing resistance to infections of various origins.Методом агробактериальной трансформации листовых эксплантов асептических растений ярового рапса получены линии, экспрессирующие альфа 2b интерферон человека. Максимальная противо-вирусная активность экстрактов листьев достигала 4500 МЕ/сырого веса. Установлено, что антиоксидантная активность и активность одного из ферментов антиоксидантной системы растений – супероксиддисмутазы (СОД) – в тканях листьев трансгенных растений повышена по сравнению с контрольными. Не выявлено корреляции как между активностью интерферона и антиоксидантной активностью, так и между активностью интерферона и активностью СОД. Использование полученных трансгенных растений рапса с высокой активностью интерферона и повышенной антиоксидантной активностью как добавки к кормам животных могло бы оказывать профилактическое влияние на их организм, повышая устойчивость к различным инфекциям.Методом агробактеріальної трансформації листових експлантів асептичних рослин ярого ріпака отримано лінії, що експресують альфа 2b інтерферон людини. Максимальна противірусна активність екстрактів листків сягала 4500 МО/сирої ваги. Встановлено, що антиоксидантна активність і активність одного з ферментів антиоксидантної системи рослин – супероксиддисмутази (СОД) – в тканинах листків трансгенних рослин підвищена в порівнянні з контрольними. Не виявлено кореляції ні між активністю інтерферона і антиоксидантною активністю, ні між активністю інтерферона і активністю СОД. Використання отриманих трансгенних рослин ріпака з високою активністю інтерферона та підвищеною антиоксидантною активністю як добавки до кормів тварин мало б профілактично впливати на їхній організм, підвищуючи стійкість до інфекцій різного походження
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