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SYNTHESIS OF CALIX[4]ARENES WITH FIXED CONFORMATION AS POTENTIAL INHIBITORS OF FIBRIN POLYMERIZATION 固定构象杯[4]芳烃纤维蛋白聚合抑制剂的合成
Pub Date : 2023-04-28 DOI: 10.15407/biotech16.02.007
D.M. Ahishev
Aim. The purpose of the present study was to develop a method for the fixation of ‘fixed’ conformation for estimation of the impact of calix[4]arene structure on the efficacy of its anticoagulant activity. This was achieved by substitution of the lower rim of C-145 analogue. Methods.Calix[4]arene C-145F (compound 6) was obtained in 4 steps starting with Duff reaction. Calix[4]arene methylene-bis-phosphonic ester 3 was prepared via addition of diisopropylphosphite in presence of metallic sodium to the parent calix[4]arene aldehyde 2. Further steps included Mitsunobu reaction, that afforded dipropoxycalix[4]arene 5 with rather good yields (80%), following the hydrolysis step that resulted in compound 6 in almost quantitative yield. Modeling of 3D-structure of calix[4]arene C-145 and its analogue C-145F was performed in Maestro, Schrodinger software. Results. Using a 2D NMR-NOESY spectroscopy, we can observe a distinct cross-peak between an aromatic singlet with a chemical shift on 7.72 ppm and protons of isopropyl group with a chemical shift on 1.62 ppm, which are moved in the strong field. Conclusions. The easy method of the fixation of conus conformation of calix[4]arene cup will be useful for synthesis of novel functionally active compounds. We believe that further development and study of different calix[4]arenes will help scientists to obtain bioactive molecules that could be prospective anti-thrombotic drugs.
目标本研究的目的是开发一种固定“固定”构象的方法,以评估杯[4]芳烃结构对其抗凝血活性的影响。这是通过取代C-145类似物的下边缘来实现的。方法:从Duff反应开始,分4步合成杯[4]芳烃C-145F(化合物6)。杯[4]芳烃亚甲基双膦酸酯3是通过在金属钠存在下向母体杯[4]芳烃醛2中加入亚磷酸二异丙酯制备的。进一步的步骤包括Mitsunobu反应,该反应以相当好的产率(80%)提供二丙氧基杯[4]芳烃5,随后的水解步骤产生几乎定量产率的化合物6。杯[4]芳烃C-145及其类似物C-145F的三维结构建模是在Maestro,Schrodinger软件中进行的。后果使用2D NMR-NOESY光谱,我们可以观察到在强场中移动的化学位移为7.72ppm的芳香单线态和化学位移为1.62ppm的异丙基质子之间的明显交叉峰。结论。杯[4]芳烃杯圆锥构象的简单固定方法将有助于合成新的功能活性化合物。我们相信,对不同杯[4]芳烃的进一步开发和研究将有助于科学家获得可能成为潜在抗血栓药物的生物活性分子。
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引用次数: 0
RHODIOLA ROSEA AND FERULIC ACID ACTIVATE EXPRESSION OF GENES RELATED TO AUTOPHAGY AND RESISTANCE TO HEAT SHOCK IN MICE OF DIFFERENT AGE 红景天和阿魏酸激活不同年龄小鼠自噬和热休克抵抗相关基因的表达
Pub Date : 2023-04-28 DOI: 10.15407/biotech16.02.015
O. Demianchuk
The aim of our study was testing whether R. rosea extract and ferulic acid activate expression of targets of FoxO, regulators of energy metabolism and autophagy in livers of young and old mice, and to what extent the effects of R. rosea extract and ferulic acid on the genes studied coincide. Methods. . C57BL/6J mice were reared at 22 ± 2 °С, 50-60% humidity, and 12/12 hour light/dark cycle. All groups were reared on a standard chow (4,8% fats, 21,8% protein, and 3,9% fibre). Experimental groups consumed water, supplemented with either sodium ferulate or R. rosea during 12 weeks prior sacrificing. The amounts of ferulate and R. rosea were adjusted to provide 4 mg of phenol-containing substances per 100 g weight, for a mouse, for 24 hours. We tested three-month-old (“young”) and twelve-month-old males (“old”). The levels of messenger ribonucleic acid (mRNA) were assessed using AriaMx real-time polymerase chain reaction (RT-PCR) instrument (Agilent). Ribonucleic acid was purified using the Monarch Miniprep kit (New England BioLabs (NEB), T2010), complementary deoxyribonucleic acid synthesis was performed using the ProtoScript II kit (NEB, E6560), and quantitative RT-PCR (qRT-PCR) was performed using the Luna Universal kit (NEB, E3003). The expression of genes ATG5 (an autophagy marker), HSPB8 (a small heat shock protein, an FoxO target), UCP2 (uncoupling protein 2, a senescence marker), CDKN2 (cell cycle regulator, a senescence marker), PDK2 and PDK4 (pyruvate dehydrogenase kinases 2 and 4, regulators of oxidative metabolism), and TFEB (transcription factor EB, a transcriptional regulator of autophagy) was evaluated. Results. Livers of young mice that consumed food supplemented with either sodium ferulate or R. rosea extract had 3.2-fold and 3.6-fold higher levels of mRNA of the small heat shock protein HspB8 than control mice, respectively. In old mice, the levels of mRNA for this protein were 3.3-fold higher in mice reared on the diet containing R. rosea extract as compared with the control. However, there was no significant difference between control mice and those that consumed ferulate-supplemented food. In young mice, ferulate and R. rosea extract induced synthesis of mRNA of PDK4 by 4.3 and 6.6 times from the control level, respectively. Ferulate and R. rosea extract also affected the levels of mRNA of ATG5 and PDK2 in the livers of old mice. The levels of PDK2 were 1.5-fold higher in the livers of mice that consumed ferulate-supplemented food than in control mice. Conclusions. Both, R. rosea extract and one of its active components – ferulic acid – promote increasing in the levels of mRNA for genes HSPB8 and PDK4, coding for small heat shock protein and pyruvate dehydrogenase kinase 4, respectively. In old mice, R. rosea promote expression of HSPB8, ATG5, PDK2, and PDK4. Thus, ferulic acid and R. rosea exert similar effects on gene expression by supposed activation of heat shock response and autophagy, and concomitant inhibition of mitochond
我们研究的目的是测试玫瑰红提取物和阿魏酸是否激活年轻和老年小鼠肝脏中FoxO靶点、能量代谢和自噬调节因子的表达,以及玫瑰红提取物与阿魏酸对所研究基因的影响在多大程度上一致。方法。C57BL/6J小鼠在22±2°С、50-60%湿度和12/12小时光/暗周期下饲养。所有组均采用标准食物(脂肪含量为4.8%,蛋白质含量为21.8%,纤维含量为3.9%)饲养。实验组在处死前12周饮水,补充阿魏酸钠或玫瑰红。调节阿魏酸盐和玫瑰红的量,以在24小时内为小鼠提供每100g重量4mg含酚物质。我们测试了三个月大(“幼”)和十二个月大的雄性(“老”)。信使核糖核酸(mRNA)的水平使用AriaMx实时聚合酶链式反应(RT-PCR)仪器(安捷伦)进行评估。使用Monarch Miniprep试剂盒(新英格兰生物实验室(NEB),T2010)纯化核糖核酸,使用ProtoScript II试剂盒(NEB,E6560)进行互补脱氧核糖核酸合成,并使用Luna Universal试剂盒(NEX,E3003)进行定量RT-PCR(qRT-PCR)。评估基因ATG5(自噬标记物)、HSPB8(小热休克蛋白,FoxO靶点)、UCP2(解偶联蛋白2,衰老标记物),CDKN2(细胞周期调节因子,衰老标记)、PDK2和PDK4(丙酮酸脱氢酶激酶2和4,氧化代谢调节因子)以及TFEB(转录因子EB,自噬转录调节因子)的表达。后果食用补充有阿魏酸钠或玫瑰红提取物的食物的年轻小鼠的肝脏中,小热休克蛋白HspB8的mRNA水平分别是对照小鼠的3.2倍和3.6倍。在老年小鼠中,与对照组相比,在含有玫瑰红提取物的饮食中饲养的小鼠中,这种蛋白质的mRNA水平高3.3倍。然而,对照小鼠和食用添加了阿魏酸的食物的小鼠之间没有显著差异。在年轻小鼠中,阿魏酸和玫瑰红提取物诱导PDK4 mRNA的合成分别是对照水平的4.3倍和6.6倍。Ferulate和R.rosea提取物也影响老年小鼠肝脏中ATG5和PDK2的mRNA水平。摄入阿魏酸补充食物的小鼠肝脏中PDK2的水平是对照小鼠的1.5倍。结论。玫瑰红提取物及其活性成分之一阿魏酸都能促进HSPB8和PDK4基因mRNA水平的增加,HSPB8基因和PDK4分别编码小热休克蛋白和丙酮酸脱氢酶激酶4。在老年小鼠中,玫瑰红促进HSPB8、ATG5、PDK2和PDK4的表达。因此,阿魏酸和玫瑰红通过激活热休克反应和自噬对基因表达产生类似的影响,并通过促进PDK2和PDK4的表达抑制线粒体代谢。
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引用次数: 0
MICROGLIAL PHAGOCYTOSIS IN RATS WITH DIFFERENT MODELS OF ALZHEIMER'S DISEASE 不同阿尔茨海默病模型大鼠的小胶质细胞吞噬作用
Pub Date : 2023-02-28 DOI: 10.15407/biotech16.01.057
A. Nefedova
Neuroinflammation is a key feature of Alzheimer's disease (AD), a progressive neurodegenerative disorder. Microglia, the resident immune cells of the central nervous system, play a crucial role in the pathogenesis of AD and are active participants in neuroinflammation. Adequate reproduction of neuroinflammation in animal models is one of the main methodological approaches for studying AD pathogenesis and pathophysiology. The aim of the study was to conduct a comparative assessment of the phagocytic activity of microglia in rats with AD induced by intrahippocampal administration of beta-amyloid (Aβ) 1-40 and Aβ25-35. Materials and methods. Wistar male rats were used in the study. Intact and sham-operated animals were used as controls. The development of the disease was confirmed by the assessment of cognitive impairment in the Barnes maze behavioral test, as well as by the level of dopaminergic neurons (DN). The phagocytic activity of microglia, as well as oxidative metabolism and the expression of phenotypic markers CD80 and CD206 were determined by flow cytometry. Results. In animals with Aβ 1-40-induced AD, significant impairment of cognitive activity and loss of DN were registered, microglial cells were characterized by an increase in the proportion of phagocytic cells and an increase in their endocytic activity, augmented oxidative metabolism and overexpression of CD86 and CD206. In animals with Aβ 25-35-induced AD, moderate impairment of cognitive activity was observed, microglial cells were characterized only by an increase in the number of phagocytizing cells without changes in their endocytic activity, oxidative metabolism, and expression of phenotypic markers. Conclusion. Thus, in animals with Aβ1–40-induced AD, the pro-inflammatory functional profile of microglia, which is characteristic for neuroinflammation in the clinical course of the disease, is more adequately reproduced.
神经炎症是阿尔茨海默病(AD)的主要特征,是一种进行性神经退行性疾病。小胶质细胞是中枢神经系统的常驻免疫细胞,在阿尔茨海默病的发病机制中起着至关重要的作用,并且是神经炎症的积极参与者。在动物模型中充分复制神经炎症是研究阿尔茨海默病发病机制和病理生理的主要方法之一。本研究的目的是比较评估海马内给药β -淀粉样蛋白(a β) 1-40和a β25-35诱导AD大鼠小胶质细胞的吞噬活性。材料和方法。本研究采用Wistar雄性大鼠。以完整和假手术的动物作为对照。巴恩斯迷宫行为测试中的认知障碍评估以及多巴胺能神经元(DN)水平证实了疾病的发展。流式细胞术检测小胶质细胞的吞噬活性、氧化代谢及表型标志物CD80、CD206的表达。结果。在Aβ 1-40诱导的AD动物中,认知活动明显受损,DN缺失,小胶质细胞的特征是吞噬细胞比例增加,内吞活性增加,氧化代谢增强,CD86和CD206过表达。在Aβ 25-35诱导的AD动物中,观察到中度认知活动障碍,小胶质细胞仅表现为吞噬细胞数量的增加,而其内吞活性、氧化代谢和表型标记物的表达没有变化。结论。因此,在a - β1 - 40诱导的AD动物中,小胶质细胞的促炎功能谱(这是该疾病临床过程中神经炎症的特征)得到了更充分的再现。
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引用次数: 0
MAIN ASPECTS OF THE MANUFACTURER OF ORGANIC PRODUCTS IN UKRAINE 乌克兰有机产品制造商的主要方面
Pub Date : 2023-02-28 DOI: 10.15407/biotech16.01.040
Nataliia Оhоrоdnyk
The article is devoted to highlighting the state and prospects for the development of organic production in Ukraine. The main requirements for the production, classification and labeling of organic products of animal and plant origin are presented. The current legal norms governing their certification and circulation are emphasized. The key provisions regarding evaluation and regulation in this field of activity are reflected. The stages of improvement of the domestic legislative framework of organic production are shown on the way of adaptation to European standards.Purpose. To highlight the state and prospects for the development of organic production in Ukraine and the improvement of the legislative framework of organic production on the way to adaptation to European standards. Materials and methods. Methodical analysis and abstract-logical method for summarizing the criteria for evaluating the formation, development and integration of domestic organic production into the structure of the world production of safe products of animal husbandry and crop production. Results. The article describes in detail the development of the organic movement, which is a promising lever for food security in Ukraine. Therefore, the work on the legal regulation of the activities of domestic producers of organic products does not stop. The legislation is improving in the direction of introducing effective state support in this area at the regional and national levels. Of course, organic feed production, animal husbandry and crop production will continue to exist in parallel with non-organic production. However, the principles and relationship of these systems will depend significantly on the availability of energy sources, plant protection products, fertilizers, soil fertility, care for the preservation of the natural environment, ensuring welfare population and its needs in healthy nutrition. In addition, for the restoration of agricultural lands, demining and bioremediation with the use of bacterial and phytoremediation of soil and water resources should be applied. For this, after the liberation of our state, a return to the peaceful management of the national economy is necessary. We believe in the victory and restoration of Ukraine with the help of allied states and people of good will.
这篇文章致力于强调乌克兰有机生产发展的现状和前景。介绍了动植物源性有机产品的生产、分类和标识的主要要求。强调了管理其核证和流通的现行法律规范。其中反映了关于这一活动领域的评价和管理的关键规定。在适应欧洲标准的道路上,展示了国内有机生产立法框架的完善阶段。强调乌克兰有机生产的发展现状和前景,以及在适应欧洲标准的道路上有机生产立法框架的完善。材料和方法。用方法分析和抽象逻辑的方法总结了评价国内有机生产的形成、发展和融入世界畜牧业和作物安全产品生产结构的标准。结果。文章详细介绍了有机运动的发展,这是乌克兰粮食安全的一个有希望的杠杆。因此,对国内有机产品生产商的活动进行法律监管的工作并没有停止。立法正在朝着在区域和国家两级对这一领域实行有效的国家支助的方向改进。当然,有机饲料生产、畜牧业和农作物生产将继续与非有机生产并行存在。然而,这些系统的原则和关系将在很大程度上取决于能源、植物保护产品、肥料、土壤肥力、保护自然环境、确保人口福利及其健康营养需求的可得性。此外,为了恢复农业用地,应采用利用细菌和植物修复土壤和水资源的排雷和生物修复。为此,在我国解放以后,恢复对国民经济的和平管理是必要的。我们相信,在盟国和善意人民的帮助下,乌克兰将取得胜利并恢复原状。
{"title":"MAIN ASPECTS OF THE MANUFACTURER OF ORGANIC PRODUCTS IN UKRAINE","authors":"Nataliia Оhоrоdnyk","doi":"10.15407/biotech16.01.040","DOIUrl":"https://doi.org/10.15407/biotech16.01.040","url":null,"abstract":"The article is devoted to highlighting the state and prospects for the development of organic production in Ukraine. The main requirements for the production, classification and labeling of organic products of animal and plant origin are presented. The current legal norms governing their certification and circulation are emphasized. The key provisions regarding evaluation and regulation in this field of activity are reflected. The stages of improvement of the domestic legislative framework of organic production are shown on the way of adaptation to European standards.Purpose. To highlight the state and prospects for the development of organic production in Ukraine and the improvement of the legislative framework of organic production on the way to adaptation to European standards. Materials and methods. Methodical analysis and abstract-logical method for summarizing the criteria for evaluating the formation, development and integration of domestic organic production into the structure of the world production of safe products of animal husbandry and crop production. Results. The article describes in detail the development of the organic movement, which is a promising lever for food security in Ukraine. Therefore, the work on the legal regulation of the activities of domestic producers of organic products does not stop. The legislation is improving in the direction of introducing effective state support in this area at the regional and national levels. Of course, organic feed production, animal husbandry and crop production will continue to exist in parallel with non-organic production. However, the principles and relationship of these systems will depend significantly on the availability of energy sources, plant protection products, fertilizers, soil fertility, care for the preservation of the natural environment, ensuring welfare population and its needs in healthy nutrition. In addition, for the restoration of agricultural lands, demining and bioremediation with the use of bacterial and phytoremediation of soil and water resources should be applied. For this, after the liberation of our state, a return to the peaceful management of the national economy is necessary. We believe in the victory and restoration of Ukraine with the help of allied states and people of good will.","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":"49 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135827611","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
COMBINED NANOCHEMOTHERAPY USING DOXORUBICIN AND CURCUMIN AS AN EXAMPLE 以阿霉素和姜黄素联合纳米化疗为例
Pub Date : 2023-02-28 DOI: 10.15407/biotech16.01.005
M. Kaniuk
The aim of the work was to review literature data on combined nanochemotherapy using the example of two drugs ̶doxorubicin and curcumin. Special attention was paid to the use of substances with synergistic properties in one nanoparticle, capable to penetrate into living cell. The method of combined chemotherapy of nanopreparations improves processing efficiency. The technique of using nanocontainers with synergistic drugs in combination with ligands reduces the side effects of chemotherapy drugs. Results. Literature data indicate that the use of nanopreparations contributes the rapid creation and use of synergistic combinations that were purposefully delivered to target cells, reducing dosage due to precise targeting. A promising direction of nanomedicine is the creation of multifunctional nanomaterials based on several active drugs having synergistic properties, with the simultaneous use of their enhancers and the strategy of active targeting. These structures enabled targeted and controlled penetration of medicinal compounds into the localization of pathological processes, reducing drugs toxicity for normal cells. Conclusions. Combined chemotherapy using polymers and nanoparticles with ligands, in which synergistic drugs are included, ensures to reduce side effects and doses of chemotherapy drugs, and helps to overcome multiple drug resistance as well.
这项工作的目的是回顾文献数据联合纳米化疗使用两种药物的例子——阿霉素和姜黄素。特别注意的是在一个纳米颗粒中使用具有协同特性的物质,能够穿透活细胞。纳米修复剂联合化疗的方法提高了处理效率。将纳米容器与协同药物与配体结合使用的技术减少了化疗药物的副作用。结果。文献数据表明,使用纳米修复有助于快速创建和使用协同组合,有目的地传递到靶细胞,减少剂量,由于精确靶向。以几种具有协同作用的活性药物为基础,同时使用其增强剂和主动靶向策略,制备多功能纳米材料是纳米医学的一个有前途的方向。这些结构使药物化合物能够靶向和受控地渗透到病理过程的局部,降低药物对正常细胞的毒性。结论。利用聚合物和纳米颗粒与配体的联合化疗,包括增效药物,确保减少化疗药物的副作用和剂量,并有助于克服多重耐药。
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引用次数: 0
DETERMINING OF NUTRITIONAL VALUE OF CAVIAR OF SIBERIAN STURGEON IN UKRAINE 乌克兰西伯利亚鲟鱼子酱营养价值的测定
Pub Date : 2023-02-28 DOI: 10.15407/biotech16.01.067
I. P. Palamarchuk
Aim. The purpose of the work was to study the nutritional value of siberian sturgeon caviar Acipenser baerii, grown in aquaculture conditions in the Dnieper reservoirs of Ukraine, and to compare its quality indicators with this product produced abroad. Methods: There are identified organoleptic properties of siberian sturgeon caviar, and its energetic value, composition of amino acids in albumens and content of fatty acids in lipids of the product. The data obtained in the research were compared with the same indices of quality of caviar of sturgeons bred abroad. Results. The organoleptic properties of siberian sturgeon caviar bred in Ukraine (appearance, color, consistency, taste and aroma) conformed to its standardized indices of quality. The caviar contained all eight essential amino acids and belonged to category of products rich by albumen (21.54±2.13%),and fats eicosapentaenoic and docosahexaenoic ω-3 acids (3.46 % and 11.2 %, respectively). (13.20±0.93%). The big content of fat, especially of polyunsaturated fatty acids and 3 acids (the eicosapentaenoic and docosahexaenoic ones — 3.46 % and 11.2 %, respectively) is one more factor, which enables to identify thes siberian sturgeon caviar as the product of high biological value. It was shown that the caviar of siberian sturgeon produced in Ukraine is in close coincidence with those that were produced in other countries. Conclusions. The totality of studied characteristics of caviar of siberian sturgeon produced in Ukraine witnesses its high nutritional value. Therefore this product may be recommended in prophylactics of numerous illnesse and strengthening of state of health.
的目标。这项工作的目的是研究在乌克兰第聂伯河水库水产养殖条件下生长的西伯利亚鲟鱼鱼子酱的营养价值,并将其质量指标与国外生产的该产品进行比较。方法:对西伯利亚鲟鱼子酱的感官特性、能量值、蛋白中氨基酸组成和脂质中脂肪酸含量进行鉴定。并将所得数据与国外鲟鱼鱼子酱的相同品质指标进行了比较。结果。乌克兰养殖的西伯利亚鲟鱼鱼子酱的感官特性(外观、颜色、稠度、味道和香气)符合其标准化的质量指标。鱼子酱含有全部8种必需氨基酸,属于富含蛋白质(21.54±2.13%)和脂肪(分别为3.46%和11.2%)的二十碳五烯和二十碳六烯ω-3酸的产品。(13.20±0.93%)。脂肪含量高,尤其是多不饱和脂肪酸和3种脂肪酸(二十碳五烯酸和二十二碳六烯酸分别为3.46%和11.2%),是确定西伯利亚鲟鱼鱼子酱具有高生物价值的另一个因素。结果表明,乌克兰生产的西伯利亚鲟鱼鱼子酱与其他国家生产的鱼子酱非常吻合。结论。研究了乌克兰生产的西伯利亚鲟鱼鱼子酱的总体特征,证明了其高营养价值。因此,本品可推荐用于预防多种疾病和加强健康状况。
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引用次数: 0
MICROBIAL CO-CULTIVATION: DISCOVERY OF NOVEL SECONDARY METABOLITES WITH DIFFERENT BIOLOGICAL ACTIVITIES 微生物共培养:发现具有不同生物活性的新型次生代谢物
Pub Date : 2023-02-28 DOI: 10.15407/biotech16.01.021
T. Pirog
In recent decades, overuse and misuse of antibiotics as well as social and economic factors have accelerated the spread of antibiotic-resistant bacteria, making them a major problem for humanity. One of the most effective approaches to the discovery of new secondary antimicrobial metabolites is co-cultivation of microorganisms, in which the producer of the target products is grown together with competitive microorganisms ( inductors), in response to the presence of which silent biosynthetic genes of the producer strain are activated and an increase in the biological activity of the synthesized secondary metabolites and/or even the synthesis of new metabolites is observed. The review summarizes the current literature data on the co-cultivation of antimicrobial substances producers with competitive microorganisms, which results in the synthesis of new metabolites with antimicrobial and cytotoxic activity, not typical for monocultures. During the co-cultivation of fungi, bacteria, and fungi with bacteria, the synthesis of new antimicrobial and anticancer metabolites, which are classified as alkaloids, phenylpropanoids, macrolides, polyketides, cyclopeptides, terpenoids, anthraquinones, and steroids, is observed. These data indicate that the mixed fermentation of microorganisms is a simple, cheap, and quite effective way to obtain new metabolites that are promising for use in medicine.
近几十年来,抗生素的过度使用和滥用以及社会和经济因素加速了抗生素耐药细菌的传播,使其成为人类的一个主要问题。发现新的次级抗菌代谢物的最有效方法之一是微生物的共同培养,其中目标产物的生产者与竞争微生物(诱导剂)一起生长,作为对其存在的反应,生产者菌株的沉默生物合成基因被激活,合成的次级代谢物的生物活性增加,甚至可以观察到新的代谢物的合成。本文综述了目前关于抗菌物质生产者与竞争微生物共同培养的文献资料,这种共同培养的结果是合成具有抗菌和细胞毒性活性的新代谢物,而不是单一培养的典型代谢物。在真菌、细菌和真菌与细菌的共培养过程中,观察到新的抗菌和抗癌代谢物的合成,这些代谢物被分类为生物碱、苯丙素、大环内酯、聚酮、环肽、萜类、蒽醌和类固醇。这些数据表明,微生物混合发酵是一种简单、廉价、有效的获取新型代谢物的方法,具有广阔的医学应用前景。
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引用次数: 0
ULTRASOUND-ASSISTED AND ENZYMATIC-BASED METHOD FOR ISOLATION OF β-GLUCANS FROM OAT BRAN 超声辅助酶法从燕麦麸皮中分离β-葡聚糖
Pub Date : 2023-02-28 DOI: 10.15407/biotech16.01.051
V. Korsa
β-Glucans are a group of non-starchy polysaccharides, or (1,3),(1,4)-β-D-glucans, that can be found in the cell walls of several species of bacteria, algae, lichens, fungi, and cereal grains. These carbohydrates are extensively used in food industry, cosmetics, pharmaceuticals and healthcare, therefore optimization of the extraction and isolation of β-glucans from grain sources has an especial importance in various fields of biotechnology, drug design, food science and technology. The aim of the study was to develop an optimized technological scheme for isolation of β-glucans from oat bran based on ultrasonic and enzymatic processing of raw material. Materials and methods. β-Glucans were isolated from grinded oat cereals during multi-stage process, which includes extraction of grain fats, hydrobarothermic processing, ultrasonification, enzymatic hydrolysis of concomitant starch and proteins, precipitation of β-glucan fraction by ethanol, centrifugation, and dry-freezing. Yield of β-glucans from raw material and its concentration in the final product were determined after hydrolysis by sulfuric acid or enzymatic cleavage by endo-1,3(4)-β-glucanase. Results. As shown by acidic hydrolysis of the final product, the yield of β-glucans was 10.8 ± 0.23% and concentration was 79.6 ± 3.89%, while enzymatic hydrolysis gave 8.7 ± 0.82% and 65.1 ± 4.72%, respectively. Thus, the use of hydrobarothermic and ultrasound pre-treatment of raw material in combination with proteolytic digestion of ballast lipids and proteins allowed producing oat β-glucans in amounts comparable with those in case of acid- or alkali-based procedures. Conclusions. The described technological scheme of β-glucan isolation from oat bran based on sequential hydrobarothermic processing, ultrasonification, and enzymatic removing starch and proteins can be widely used for routine β-glucan production for various purposes in food technology, pharmacological industry, and medicine.
β-葡聚糖是一组非淀粉多糖,或称(1,3)、(1,4)-β- d -葡聚糖,存在于多种细菌、藻类、地衣、真菌和谷物的细胞壁中。这些碳水化合物广泛应用于食品工业、化妆品、制药和医疗保健等领域,因此优化从粮食中提取和分离β-葡聚糖在生物技术、药物设计、食品科学和技术等各个领域具有特别重要的意义。本研究的目的是建立一种基于超声波和酶处理的燕麦麸皮β-葡聚糖的优化工艺方案。材料和方法。通过谷物脂肪提取、水气压法、超声法、伴随淀粉和蛋白质的酶解、乙醇沉淀、离心和干冷冻等多阶段工艺从燕麦粉中分离出β-葡聚糖。通过硫酸水解或内切-1,3(4)-β-葡聚糖酶裂解,测定原料中β-葡聚糖的产率和最终产物中β-葡聚糖的浓度。结果。最终产物酸解β-葡聚糖得率为10.8±0.23%,浓度为79.6±3.89%,酶解β-葡聚糖得率为8.7±0.82%,浓度为65.1±4.72%。因此,使用水压温热和超声波预处理原料,结合蛋白质水解消化压舱脂质和蛋白质,可以生产燕麦β-葡聚糖,其数量与酸或碱法生产的葡聚糖相当。结论。本文所述的燕麦麸皮分离β-葡聚糖的工艺方案,基于顺序水热法、超声波法、酶法去除淀粉和蛋白质,可广泛应用于食品工业、药理工业和医药等领域的常规β-葡聚糖生产。
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引用次数: 1
IN VITRO ANTILEUKEMIC ACTIVITY OF EUPHORBIA ECHINUS EXTRACT 针叶树提取物的体外抗白血病活性
Pub Date : 2022-12-30 DOI: 10.15407/biotech15.06.070
Sbayou Houda
Aim. Cancer continues to pose a serious threat to human health. Euphorbia plants are rich in phenolics, aromatic esters, steroids and several bioactive compounds. Studies have shown the presence of a large number of bioactive compounds in E. echinus including flavonoids, phenolics, and proanthocyanins. Method. There it was investigated cytotoxic effects of E. echinus methanolic extract on K562, HL60, Ishikawa, Raji and SH-SY5Y cells. Results. The E. echinus extract was found to be highly cytotoxic against HL60 and K562 (79.78 and 76.44% cytotoxicity, respectively). DNA fragmentation was exclusively observed in K562 cells indicating that the reduction of viable cells following treatment with E. echinus extract is due to apoptosis. Conclusions. Our results suggest that E. echinus extract might have a drug potential against leukemic cells.
的目标。癌症继续对人类健康构成严重威胁。大戟类植物富含酚类物质、芳香酯类、类固醇和几种生物活性化合物。研究表明,棘皮中含有大量的生物活性化合物,包括类黄酮、酚类物质和原花青素。方法。研究了棘猴醇提物对K562、HL60、Ishikawa、Raji和SH-SY5Y细胞的细胞毒作用。结果。结果表明,棘皮提取物对HL60和K562具有较高的细胞毒性(分别为79.78%和76.44%)。DNA片段只在K562细胞中被观察到,这表明棘棘提取物处理后活细胞的减少是由于细胞凋亡。结论。我们的研究结果表明,棘猴提取物可能具有抗白血病细胞的药物潜力。
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引用次数: 0
THE EXPRESSION OF TLR4, IFN-γ, TGF-β AND TNF-αLL LINE OF HUMAN SMALL CELL LUNG CARCINOMA NCI-H69 AND IN CISPLATIN-RESISTANT SUBLINE NCI-H69/CPR tlr4、ifn -γ、tgf -β和tnf -αll在人小细胞肺癌nci-h69及顺铂耐药亚系nci-h69 / cpr中的表达
Pub Date : 2022-12-30 DOI: 10.15407/biotech15.06.055
I. Stupak
Aim: to investigate the effect of teichoic acid Staphylococcus aureus for expression of pro-inflammatory cytokines and of TLR4 in a human small cell lung carcinoma cell line NCI-H69, and cisplatin resistant subline NCI-H69/CPR. Methods. Incubation of cells with teichoic acid (1 ng/m) conducted for 2 days. Expression level of TLR4, TGF-β, INF-γ, TNF-α was evaluated by the real time PCR on 7500 Real-Time PCR System, using specific primers and fluorochrome SYBR Green. The reverse transcription reaction was performed with High-Capacity cDNA Reverse Transcription Kit carried out under the conditions: 25 °C - 10 min, 37 °C ‒ 120 min and 85 °C ‒ 5 min. Results. In cell line culture NCI-H69 addition of teichoic acid increased expression of TLR4 by 1.3 times, and IFN-γ – by 1,1 times. Expression of TGF-β and TNF-α was decreased 2.5 and 4.9 times respectively. In cell line culture NCI-H69/CPR the addition of teichoic acid inhibited the expression of all studied parameters. Expression TLR4 decreased by 4.2 times, IFN-γ – by 1.4 times. Expression TGF-β and TNF-α was depressed 1.6 and 1.2 times. The presented data indicate that teichoic acid of bacterial origin provided the effect of modulating the inflammatory effect in lung cancer cell culture, sensitive and resistant to cisplatin. Conclusions. Teichoic acid as a ligand of TLR4 modulates the expression of pro-inflammatory and anti-inflammatory cytokines in small cell lung cancer cell culture and suppresses the expression of TLR4 and all investigated cytokines in the cisplatin-resistant cell line NCI-H69.
目的:研究磷壁酸金黄色葡萄球菌对人小细胞肺癌细胞株NCI-H69和顺铂耐药亚系NCI-H69/CPR中促炎细胞因子和TLR4表达的影响。方法。用磷壁酸(1ng/m)培养细胞2天。采用特异性引物和荧光染料SYBR-Green在7500实时PCR系统上进行实时PCR检测TLR4、TGF-β、INF-γ、TNF-α的表达水平。使用高容量cDNA逆转录试剂盒在25°C-10分钟、37°C-120分钟和85°C-5分钟的条件下进行逆转录反应。在细胞系培养中,NCI-H69添加磷壁酸使TLR4的表达增加1.3倍,IFN-γ的表达增加1.1倍。TGF-β和TNF-α的表达分别下降2.5倍和4.9倍。在细胞系培养物NCI-H69/CPR中,磷壁酸的加入抑制了所有研究参数的表达。TLR4表达下降4.2倍,IFN-γ表达下降1.4倍。TGF-β和TNF-α的表达分别降低1.6和1.2倍。结果表明,细菌来源的磷壁酸在癌症细胞培养中具有调节炎症效应的作用,对顺铂敏感且耐药。结论。在小细胞肺癌癌症细胞培养中,作为TLR4配体的替酸调节促炎和抗炎细胞因子的表达,并抑制顺铂耐药细胞系NCI-H69中TLR4和所有研究的细胞因子表达。
{"title":"THE EXPRESSION OF TLR4, IFN-γ, TGF-β AND TNF-αLL LINE OF HUMAN SMALL CELL LUNG CARCINOMA NCI-H69 AND IN CISPLATIN-RESISTANT SUBLINE NCI-H69/CPR","authors":"I. Stupak","doi":"10.15407/biotech15.06.055","DOIUrl":"https://doi.org/10.15407/biotech15.06.055","url":null,"abstract":"Aim: to investigate the effect of teichoic acid Staphylococcus aureus for expression of pro-inflammatory cytokines and of TLR4 in a human small cell lung carcinoma cell line NCI-H69, and cisplatin resistant subline NCI-H69/CPR. Methods. Incubation of cells with teichoic acid (1 ng/m) conducted for 2 days. Expression level of TLR4, TGF-β, INF-γ, TNF-α was evaluated by the real time PCR on 7500 Real-Time PCR System, using specific primers and fluorochrome SYBR Green. The reverse transcription reaction was performed with High-Capacity cDNA Reverse Transcription Kit carried out under the conditions: 25 °C - 10 min, 37 °C ‒ 120 min and 85 °C ‒ 5 min. Results. In cell line culture NCI-H69 addition of teichoic acid increased expression of TLR4 by 1.3 times, and IFN-γ – by 1,1 times. Expression of TGF-β and TNF-α was decreased 2.5 and 4.9 times respectively. In cell line culture NCI-H69/CPR the addition of teichoic acid inhibited the expression of all studied parameters. Expression TLR4 decreased by 4.2 times, IFN-γ – by 1.4 times. Expression TGF-β and TNF-α was depressed 1.6 and 1.2 times. The presented data indicate that teichoic acid of bacterial origin provided the effect of modulating the inflammatory effect in lung cancer cell culture, sensitive and resistant to cisplatin. Conclusions. Teichoic acid as a ligand of TLR4 modulates the expression of pro-inflammatory and anti-inflammatory cytokines in small cell lung cancer cell culture and suppresses the expression of TLR4 and all investigated cytokines in the cisplatin-resistant cell line NCI-H69.","PeriodicalId":9267,"journal":{"name":"Biotechnologia Acta","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41561440","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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Biotechnologia Acta
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