Innovative Biosystems and Bioengineering: international scientific e-journal, Vol. 9, No. 2
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Документ Відкритий доступ In silico the Ames mutagenicity predictive modelof environment(Igor Sikorsky Kyiv Polytechnic Institute, 2025) Kislyak, Sergey V.; Duhan, Olexii M.; Yesypenko, Ruslana V.; Starosyla, Darya B.; Yalovenko, Olena I.Background.The classical in vitroand in vivo methods developed and widelyused in the past decades to as-sess the genetic effects of environmental factors are complex in view of their implementation, are expensive, long-lasting, have the problem of reproducibility of the results of experiment in different laboratories and may face ethical problems of using warm-blooded animals in experiments. Objective. Development, optimisation and testing of effective in silicomodels for assessment of Ames muta-genicity of environmental factors.Methods. The genetic assessment of the impact of environmental factors was carried out in accordance with a set of chemical compounds for which information on potential mutagenic activity was obtained experimen-tally, using the in vitroAmes Salmonella/microsome test.Four machine learning models were developed to solve the problem of binary classification to form two classes of xenobiotics (mutagen/non-mutagen).The total sampleis represented by a set of 8,083 xenobiotics. Results. We developed four machine learning models with 85% accuracy, matchingthe reproducibility of Ames test data across laboratories. In addition, we have proposed a binary classifier that subject to dimen-sionality reduction of the input data, taking into account the qualitative composition of molecular descrip-tors, allows us toimprove the accuracy of in silicoprediction of genotoxicity of chemicals. Conclusions.The necessity of updating and expanding the list of effective and more productive methods and approaches for assessing the genotoxic effects of environmental factors is substantiated, which allows avoi-ding the use of warm-blooded animals in the experiment, saving time and reducing the number of false-negative and false-positive results. The possibility of increase the accuracy of predictive machine learning models forassessing the genotoxic potential of environmental factors in conditions of dimensionality reduc-tion of the data set is presented.Документ Відкритий доступ Biosafety aspects of hybridoma technology: nature of risks and approaches to their management(Igor Sikorsky Kyiv Polytechnic Institute, 2025) Shevchuk, Kateryna M.; Baranovska, Anastasia V.; Chernetskyi, Andrii S.; Shchotkina, Nataliia V.; Besarab, Alexander B.This study investigates the biosafety aspects of hybridoma technology, focusing on the identification and management of associated risks. Monoclonal antibodies, essential tools in immunology, biotechnology, and medicine, are primarily produced through hybridoma technology. This process involves fusing B lymphocytes from immunized animals with myeloma cells to create hybridomas, which are then cultured to produce spe-cific antibodies. The research highlights significant contamination risks, particularly from rodent-borne virus-es and other pathogens, during both in vivoand in vitrocultivation. It systematically analyzes existing strate-gies for identifying and mitigating these risks at various stages of monoclonal antibody production, including hybridoma identification, cell fusion, and antibody purification. The study underscores the importance of stringent biosafety protocols and optimized purification methodologies to ensure the production of high-quality, contaminant-free monoclonal antibodies. Additionally, it emphasizes the necessity of comprehensive risk assessments and the implementation of advanced contamination control systems in laboratories. The conclusions drawn from this study provide valuable insights into enhancing the safety and efficacy of monoc-lonal antibody production. By addressing these biosafety concerns, the research supports the widespread ap-plication of monoclonal antibodies in scientific and medical fields, ensuring their reliability and effectiveness in various diagnostic and therapeutic contexts.Документ Відкритий доступ Антиадгезивні властивості 4-(адамантил-1)-1-(1-амінобутил)бензолу щодо staphylococcus aureus(Igor Sikorsky Kyiv Polytechnic Institute, 2025) Гуменюк, Наталія І.Проблематика. Staphylococcusaureusвіднесенідобактерійізвисокимрівнемстійкостідоантимік-робнихпрепаратів, однієюзпричинякоїєздатністьмікроорганізмівформуватибіоплівки. Перспек-тивноюстратегієюантимікробноїтерапіїупацієнтівзінфекційнимизахворюваннями, пов’язанимизбіоплівками, євпливнапершіетапиплівкоутворення.Мета.Встановити здатність 4-(адамантил-1)-1-(1-амінобутил)бензолу впливати на формування біо-плівок S.aureusта експресію генів плівкоутворення.Методика реалізації.Мінімальну інгібуючу концентрацію (МІК) 4-(адамантил-1)-1-(1-амінобутил)бензолу(шифр АМ-166) щодо метицилінрезистентного штаму S.aureus222визначали методом серійних мікро-розведень. Антибіоплівкову активність сполуки АМ-166 досліджували за методикою O’Toole, оцінку інтенсивності утворення біоплівки S.aureusвизначали згідно зіStepanovic.Вплив АМ-166 на експре-сію генів досліджували за допомогою полімеразної ланцюгової реакції (ПЛР) уреальному часі.Результати. МІК сполуки АМ-166 щодо S.aureus222 становить 5мкг/мл. За дії АМ-166 за 0,25та 0,5МІК здатність S.aureusдо плівкоутворення змінюється з середньої (контроль) до слабкої, за 5,0МІКштам втрачає здатність формувати біоплівку. Встановлено, що сполука в концентрації 0,5МІК збіль-шує експресію генаicaRта знижує транскрипційну активність генів icaA, clfB, fib,fnbB, ebpS, eno,якіберуть участь у плівкоутворенні йадгезії.Висновки.Похідне адамантану АМ-166 порушує формування біоплівок MRSA, змінює транскрипційнуактивність генів icaлокусу, пригнічує прикріпленняS.aureusдо біотичної поверхні через вплив на експресію генів.Документ Відкритий доступ Optimization of parameters of saline sodium citrate buffer for stability of colloidal gold nanoparticles usedin dna hybridization biosensor(Igor Sikorsky Kyiv Polytechnic Institute, 2025) Sobolevskyi, Maksym S.; Holubiev, Illia Iu.; Lopatynskyi, Andrii М.; Samoylov, Anton V.; Dorozinsky, Glib V.; Lyapin, Oleksandr M.; Khrystosenko, Roman V.; Chegel, Volodymyr І.; Dzyadevych, Sergiy V.; Soldatkin, Oleksandr O.Background.The use of optical biosensors based on surface plasmon resonance (SPR) spectrometry have long been established as a viable alternative tothe traditional molecular biological methods, such as immu-nostainingor ELISA. Its capacity to perform real-time quantitative measurements is complemented with the possibility for the enhancement of the sensor signal with the use of optically active colloidal nanoparticles. On the other hand, few such nanoparticle-containing DNA biosensors have been developed, owing to poor colloidal stability of nanoparticles in chemical conditions suitable for hybridization of nucleic acid sequences.Objective.This study investigates the possibility of using gold nanoparticles (AuNPs) modified with thiolated oligonucleotides, 6-mercapto-1-hexanol, and lipoic acid as part of a hybridization system for biosensor de-tection of DNA sequences in order to improve its main analytical characteristics.Methods.Astudy of the colloidal stability of nanoparticlesinSSC (saline sodium citrate)media of differentmultiplicitybefore and after modification was carried out in order to select the best environment for the op-eration of the biosensor system.Aggregation of modified AuNPs was facilitated by their centrifugation, after which pelleted nanoparticles were resuspended and investigated by spectrophotometry. The conclusions re-garding the colloidal stability of AuNPs were based on the drop in concentration of colloidal AuNPs after each centrifugation.Results.The possibility of using the studied NPs in biosensor analysis was shown, and0.1SSC buffer solu-tion was determined to bethe optimal mediumfor their operational stability. Approbation of the nanoparticle-containingDNA hybridizationbiosensor based on SPR spectrometry was carried outin 2.0SSC medium.Conclusions.The prospective use ofthestudiedNPs as components of DNA hybridization systems for the detection of genetic markers has been provenДокумент Відкритий доступ Nhibition of cytochrome p450 activities by propoxazepam: safety assessment in context for potential drug interactions(Igor Sikorsky Kyiv Polytechnic Institute, 2025) Larionov, Vitalii B.; Golovenko, Mykola Ya.; Valivodz, Iryna P.; Reder, Anatoliy S.Background.Propoxazepam is a new anelgetic agent of the benzodiazepine group, chemically known as 7-bromo-5-(o-chlorophenyl)-3-propyloxy-1,2-dihydro-3H-1,4-benzodiazepine-2-one. Propoxazepam is con-sidered a possible substrate of the CYP system, so its effect of on the CYP3A4 enzyme activity was investi-gated in vitrousing human liver microsomes.Objective.To evaluate the effects of propoxazepam on CYP3A4 activity in vitrousing testosterone and mida-zolam as markers of metabolic activity for CYP3A4 in human liver microsomes.Methods.Midazolam (1-hydroxylation reaction) and testosterone (6-hydroxylation reaction) were used as markers for CYP3A4-mediated activity. Ketoconazole (0.2 M) was used as a positive control for reversibleinhibition, and troleandomycin (50 M) for metabolism-dependent inhibition. For reversible inhibition, pro-poxazepam was added together with the corresponding substrate and cofactor (NADPH), while for metabo-lism-dependent inhibition, it was incubated with microsomes and cofactor for 30 minutes prior to substrate addition.Results.Propoxazepam at various concentrations (0 to 100 M) consistently inhibited CYP3A4 activities for both substrates, showing a similar "concentration–activity inhibition" dependence, with IC50values of 52.34.9M for midazolam and 46.1 9.2 M for testosterone. For metabolism-dependent inhibition, IC50values were 36.6 8.6 M for midazolam and 28.3 7.4 M for testosterone. Given that the binding of pro-poxazepam to microsomal protein under the experimental conditions, which reflected those in the IC50ex-periments, was low, no microsomal binding correction factor was applied to the reported IC50values.Conclusions.The highest predicted unbound Cmaxplasma concentration of propoxazepam, above which in-teractions can occur, is between 0.462 and 0.524 M, or 462 and 524 nM. This corresponds to concentra-tions of 188 to 214ng/mL (based on the molecular weight of propoxazepam, 414.73g/mol). According to pharmacokinetic data, these concentrations are not achievable after a single oral administration. Furtherstudies are required for multiple-dose administration.Документ Відкритий доступ Biosafety Management: Emphasis on Medicine, Pharmacy, and Biotechnology(Igor Sikorsky Kyiv Polytechnic Institute, 2025) Galkin, Alexander