Biopolym. Cell is an open access peer-reviewed international journal that publishes original contributions in molecular biology and related areas: Structure and function of biopolymers in different cells at different conditions; Genome regulation; Molecular mechanisms of differentiation; Oncogenesis; Cell-virus interaction; Biotechnology; Bioorganic chemistry; Drug design; Biology of peptides, nucleoside derivatives and modified oligonucleotides
Institute of Molecular Biology and Genetics of the NAS of Ukraine.
To the 70th jubilee of Volodymyr Ivanovych Kashuba, Corresponding Member of NAS of Ukraine
Doxorubicin-PLGA nanoparticles are immunomodulational antitumor therapeutics with dual cytotoxic and immune-stimulatory effects
Aim. The aim of this study is to investigate the immunomodulatory and antitumor effects of doxorubicin-loaded poly(lactic-co-glycolic acid) nanoparticles (DOX-PLGA NPs) designed as a dual-function nano-immunotherapeutic system. Methods. DOX-PLGA NPs were synthesized by the emulsion-solvent evaporation method and characterized for particle size, zeta potential, morphology (TEM), drug loading, and release behavior. The cytotoxicity of free DOX and DOX-PLGA NPs was evaluated in MCF-7 and HCT116 cancer cell lines using the MTT assay. «Peripheral blood mononuclear cell (PBMC) — tumor» co-cultures were analyzed for cytokine secretion (IFN-γ, IL-2, TNF-α, and IL-10) and immune cell subsets (CD8+ T-cells, NK cells, and macrophage polarization). The therapeutic efficacy was further examined in CT26 tumor-bearing mice. Results. The DOX-PLGA NPs exhibited a mean size of approximately 145 nm (PDI 0.18, ζ −22 mV), with a drug loading efficiency of 12.5% and encapsulation efficiency of 78%. In comparison to free DOX, the nano-formulation lowered cell viability to a greater extent (p < 0.01) and stimulated greater release of IFN-γ, IL-2, and TNF-α, while IL-10 levels decreased. Flow cytometric analysis shows increases in the populations of CD8+ T and NK cells and a congruent macrophage reversal to the M1 macrophage phenotype. Tumor growth was markedly suppressed, survival was extended, and DOX-PLGA NPs produced insignificant systemic toxicity in the in vivo setting. Conclusions. The immuno-nanoparticles demonstrate controlled release of DOX-PLGA NPs while maintaining powerful immune stimulation due to the dual cytotoxic and immunostimulatory effects they exhibit. Their application as a novel platform for nano-immunotherapy in the treatment of cancer is justified.
Studying the interaction between pentameric oligonucleotides and recombinant signaling proteins and receptors
Aim. This study aimed to synthesize, purify, and investigate the interactions between oligonucleotides (OLN) and recombinant signaling proteins, specifically interferon α2-β, insulin, their receptors, and somatropin, using docking and fluorescence spectroscopy methods. Methods. To analyze these interactions, we employed the Stern-Volmer equation in its general and modified forms, as well as the Gill equation, which enabled us to determine the binding and binding constants. We synthesized oligonucleotides using the solid-phase phosphoamidite method, which offers high efficiency and specificity. The resulting oligonucleotides were then purified by solid-phase extraction, which removed by-products and impurities, as confirmed by spectral analysis. Results. Fluorometric titration revealed that homopolymeric oligonucleotides bind to proteins within the medium-affinity range, forming non-fluorescent complexes. The most significant interactions occurred with shorter oligonucleotides. We observed negative cooperative binding between insulin and the oligoribonucleotide Poly(rG)₅. Notably, all proteins selectively bound only one ligand, the oligonucleotide Poly(dG)₅. Conclusions. Understanding the mechanisms of protein–oligonucleotide interactions may open new possibilities for developing antibiotics, antiviral drugs, and treatments for cancer and genetic diseases.
Intrinsically Disordered Region 2 of G3BP1 is required for RNA recruitment by G3BP1-Caprin1 complex
Aim. The stress granule assembly requires coordinated interactions between RNA-binding proteins and mRNA, yet how the intrinsically disordered regions (IDRs) of stress granule proteins regulate RNA recruitment remains incompletely understood. The aim of this study was to characterize a role of the Intrinsically Disordered Region 2 (IDR2) of Ras GTPase-activating protein-binding protein 1(G3BP1) in mRNA recruitment within the G3BP1-Caprin1 complex. Methods. Protein-protein and protein-RNA interactions in cells were analyzed as colocalization of their fluorescent signals on microtubules (microtubule bench recruitment and mixing assay). Nucleic acid-binding properties of G3BP1, G3BP1-ΔIDR2, a G3BP1 variant with deletion of IDR2, and G3BP1-Caprin1 complex were assessed in vitro by electrophoretic mobility shift assays. Results. Deletion of IDR2 strongly impaired mRNA recruitment by G3BP1 despite the presence of intact RNA-binding domains. G3BP1-ΔIDR2 retained the ability to interact with Caprin1 and exhibited robust heterotypic mixing in the cellular context. However, neither Caprin1 nor other G3BP1 partners were able to compensate for the loss of mRNA recruitment. In vitro assays confirmed reduced nucleic acid binding by G3BP1-ΔIDR2 and showed that Caprin1 did not compensate for the lossof G3BP1-ΔIDR2 nucleic acid binding. These findings indicate that impaired mRNA recruitment is not caused by the altered compartmentalization but reflects an intrinsic defect associated with IDR2 loss. Conclusions. Our results demonstrate that IDR2 plays a critical, indirect role in G3BP1-mediated mRNA recruitment by maintaining an RNA-binding-competent conformation of this protein. This study highlights the importance of intrinsically disordered regions in regulating mRNA engagement through conformational and structural effects rather than direct RNA binding and provides new insight into the molecular principles underlying stress granule assembly.
The anti-Acinetobacter baumannii therapeutic potential of 6-Chloro-4-oxo-4H-chromene-3-carbonitrile
Aim. Acinetobacter baumannii is a Gram-negative opportunistic pathogen responsible for a wide spectrum of hospitalacquired infections. This bacterium has acquired resistance to nearly all existing antibiotics, including aminoglycosides, quinolones, broad-spectrum β-lactams, carbopenems, polymyxins and even colistin which is a last-line antimicrobial agent, providing the stimulus to search for novel antibiotics. The aim of this study is to identify novel anti- Acinetobacter baumannii agents among 4H-chromen-4-one derivatives. Methods. 88 4H-chromen-4-one derivatives were tested by broth microdilution against five bacterial pathogens such as Staphylococcus aureus, Escherichia coli, Klebsiella pneumonia, Acinetobacter baumannii, Pseudomonas aeruginosa and toward two fungal pathogens such as Candida albicans and Cryptococcus neoformans. Results. Among the derivatives we have found two active compounds — 6-Chloro-4-oxo-4H-chromene-3-carbonitrile (1) inhibiting growth of C. albicans (MIC = 0.5 mg/L) and C. neoformans (MIC = 0.25 mg/L) and 6-Chloro-3-(6-hydroxy-3-oxo-3H-benzofuran-2-ylidenemethyl)-chromen- 4-one (2), inhibiting growth of C. neoformans (MIC = 16 mg/L). In addition, compounds 1 and 2, at a concentration of 32 mg/L, inhibited the growth of A. baumannii ATCC 19606 by 24.8% and 23.7%, correspondingly. Compound 1 was revealed to inhibit the growth of multidrug resistant clinical isolate A. baumannii №144 with MIC value of 32 mg/L. Also, it was established that this compound is not cytotoxic toward HEK293 cells. Conclusions. Therefore, 6-Chloro- 4-oxo-4H-chromene-3-carbonitrile can be promising candidate for further research and chemical optimization.
Izatizon — antiviral and immunomodulating substance: over half a century history
Anatolii Potopalsky (1938–2025), a prominent scientist-innovator who worked as the long-time Head of the Laboratory of Modification of Biologically Active Compounds at the Institute of Molecular Biology and Genetics, the NAS of Ukraine and was a founder and director of the Institute of Health Promotion and Rebirth of People of Ukraine.
Molecular dynamics of Trp40 form of B.taurus tyrosyl-tRNA synthetase with replacements of Trp87 and Trp283 by alanine
Aim. Analysis of conformational flexibility of mutant Trp40 form of Bos taurus Tyr-tRNA-synthetase (BtTyrRS) where single Trp was localized near active site. Methods. Computational site directed mutagenesis of Trp87 and 283 residues using the AlphaFold3 server. In silico modeling of 3D structure and molecular dynamics (MD) simulation of mini Trp40 BtTyrRS mutant form. Results. Computational modeling of Trp40 mutant BtTyrRS form has not revealed significant changes at enzyme active site. A model of 3D structure of single-tryptophan form of BtTyrRS presents a compact structure of mutant protein and rigid microenvironment of Trp40 at the active site. Conclusions. The replacement of Trp residues at positions 87 and 283 in the amino acid sequence of mini BtTyrRS with alanine residues does not affect the structure of the enzyme active site. Stable mutant single-tryptophan mini BtTyrRS protein is suitable both for fluorescence studies of structural dynamic and catalytic properties of this enzyme.
Clinically significant molecular alterations of mitochondria in carcinogenesis and age-related diseases: Part 2. Exposome influences, biomarkers and therapeutic strategies
The second part of the review presents an analysis of a number of exposome factors that influence the development of molecular disorders and metabolic shifts in mitochondria in cancer and age-related diseases. A number of mitochondrial biomarkers have been demonstrated to serve as biochemical, genetic, and molecular biological indicators for the diagnosis, prognosis, and treatment of cancer and age-related diseases. Certain areas of therapy and new modern therapeutic approaches to the treatment of cancer and age-related diseases are analyzed in the context of the pathogenetic effect on mitochondrial dysfunction in human cells and organs for the improvement of pathological conditions and the promotion of healthy longevity.
In silico modeling and prediction of antidiabetic potential of bioactive compounds from Galega officinalis L. non-alkaloid extract
Aim. To develop and validate in silico models for predicting the antidiabetic activity of natural compounds and to test their performance on representative medicinal plant components. Materials and Methods. Two machine learning models based on the XGBoost and LightGBM algorithms were constructed and verified using a set of compounds from traditional Chinese medicine (TCM) formulations with experimentally confirmed hypoglycemic activity. The validated models were subsequently applied to analyze components of Galega officinalis L. (non-alkaloid fraction), whose composition was determined by gas and liquid chromatography-mass spectrometry (GC-MS and LC-MS). Results. The constructed models achieved an accuracy of 80-81% and were verified by correctly identifying active compounds among those known from TCM formulations to be effective in type 2 diabetes management. This confirms their ability to accurately classify bioactive natural substances. The models were applied to the components of the NA extract of G. officinalis L., a promising plant for subsequent studies on antidiabetic effects. Conclusions. The developed in silico models enable the prediction of antidiabetic activity of naturally derived compounds. Their verification on reference compound sets and application to G. officinalis L. extract demonstrate the potential of this approach for identifying low-toxicity bioactive substances.
Evaluation of the monomethine cyanine dye FB128 for real-time PCR applications
Aim. To evaluate the suitability of the novel DNA-binding dye fb128 for real-time PCR applications, including amplification efficiency, fluorescence performance, PCR inhibition threshold, and melting curve analysis. Methods. UV-vis absorption and fluorescence spectroscopy, agarose gel electrophoresis, real-time PCR, melt curve analysis. Results. fb128 exhibited low intrinsic fluorescence and strong DNA-specific signal (ΔQ = 160.7). Optimal concentrations (0.2—1.6 μM) provided early Ct values without PCR inhibition, whereas the concentrations of ≥3 μM caused a delayin amplification or complete suppression. PCR efficiency with fb128 was 102.6%, within the optimal range for quantitative applications. Melting peaks with fb128 were stronger and appeared at 74.5 °C, 2.5 °C lower than with SYBR Green I, suggesting weaker dsDNA binding. Conclusions. fb128 demonstrates high amplification efficiency, strong fluorescent signal, broad concentration tolerance, and robust melting analysis performance. These properties establish fb128 as a competitive alternative to SYBR Green I for real-time PCR applications.
Establishment of U-251 MG cell sublines with ectopic overexpression of CHI3L1 mRNA variants as a model system of the isoforms trials in gliomagenesis
Aim. Obtaining glioma derived U-251 MG cell sublines producing the full-length and the short variants of CHI3L1 with subsequent characterization. Methods. Cell lentiviral transduction, Western blot and RT-qPCR analysis. Results. We successfully generated the U-251 MG cell sublines producing the full-length and the short variants of CHI3L1; our results support the suppression of CHI3L1-del8ex secretion and demonstrate differential targeting of microenvironment (LOX and GUSB) and EMT program (TGFB1) modulators by CHI3L1 isoforms. Conclusion. Here we present the cell model for CHI3L1 isoforms trials in gliomagenesis regarding experiments on co-cultivation and/with 3D culturing.
Synthesis and antifungal activity of 3-oxo-2-piperazinylacetamides
Aim. Synthesis of 3-oxo-2-piperazinylacetamides and investigation of their antifungal activity. Methods. Organic synthesis, spectral methods (NMR), pharmaceutical screening. Results. A series of new 3-oxo-2-piperazinylacetamides has been synthesized and the screening of their antifungal activity was performed. N-butyl-3-oxo-2-piperazineacetamide shows antifungal activity against Cryptococcus neoformans comparable with amphotericin B and fluconazole (MIC = 2μg/mL). Conclusions. 3-oxo-2-piperazineacetamide moiety has been identified as a promising template to search for novel antimicrobials against Cryptococcus neoformans.
Assessment of the informativeness of iPBS markers for identifying and differentiating Ukrainian hazelnut varieties
Aim. To assess the informativeness of iPBS markers for the identification and differentiation of Ukrainian hazelnut varieties. Methods. Thirty Corylus spp. specimens (25 varieties and 5 wild plants) were analyzed using PCR amplification with 24 iPBS primers. Informativeness indices were calculated based on PCR banding profiles. Results. Six primers with the highest polymorphism were selected. They produced 95 bands (15.8 per primer on average), only one of which was monomorphic. Mean resolving power (Rp) of these primers was 6.760 and discriminating power (DL) 0.948, while the number of non-differentiated pairs (ND) averaged 3.2 per primer. Primer 2402 distinguished all varieties, and two other primers were unable to differentiate between only one pair. In comparison with SSR markers, iPBS demonstrated higher Rp and DL and significantly lower ND values. Conclusions. iPBS markers are capable of providing an efficient and reliable tool for the differentiation of varieties and can be successfully applied in hazelnut breeding programs.
The analysis for the association of polymorphic variants of the VDR gene(rs2228570) with the risk of herpetic keratitis occurrence after COVID-19
Aim. To investigate the association between polymorphic variants of the VDR gene rs2228570 and clinical features ofherpetic keratitis in adult patients who have had COVID-19. Methods. Peripheral blood samples from patients(N = 50) with a history of herpetic keratitis that developed after COVID-19 were studied. In comparison with (N = 104) unrelated healthy controls. DNA samples were isolated by lymphocyte lysis with proteinase K followed by the use of DNA purification kit (“MBA LABS”). Analysis of the VDR gene (rs2228570) was performed using fluorescently labeled TaqMan probes manufactured by Thermo Fisher Scientific (USA). Statistical analysis was performed using Fisher’s exact test. Results. The results of genotyping followed by comparative statistical analysis revealed significant differences in frequencies of both homozygous genotypes between patients with recurrent HK and controls (p = 0.01636). The GG genotype was significantly more common in the recurrent HK group. Importantly, the frequency of A allelecarriers (AA+GA) was statistically significantly higher in the control group (p < 0.05), OR=0.3381 with 95% CI(0.1433—0.797). Conclusions. It was shown that the G allele of the FokI VDR gene (rs2228570) polymorphism is associated with recurrent herpetic keratitis in patients from Ukraine, indicating it as a marker of hereditary predisposition to the development of recurrent herpetic keratitis after COVID-19.
Alterations in S6K1 isoform expression in MCF-7 cells that induce EMT, affect Akt1–, AMPK–, PKC–, and PKA-related cell signaling
Aim. This study aimed to evaluate how S6K1 isoform expression affects the activities of Akt1, AMPK, PKC, and PKA, as well as their related kinase signaling pathways. Methods. MCF-7 cells and a subline with disrupted p70 and p85 S6K1 isoform expression (MCF7/p60+/p70–/p85–) were cultured under different growth conditions. Western blot analysis was performed on cell lysates using antibodies specific to phosphorylated substrates of Akt1, AMPK, PKC, PKA, and GSK-3β/pSer9. Results. The phosphorylation pattern of kinase substrates showed that changes in S6K1 isoform expression influenced either the activity of Akt1, AMPK, PKC, PKA, or their substrate specificity. Unexpectedly, in the MCF-7 subline exhibiting EMT features, GSK-3β kinase – an Akt1 substrate and EMT inhibitor – was not downregulated by phosphorylation of Ser9. Conclusions. S6K1, a ribosomal protein kinase involved in EMT regulation, can modulate the activity of Akt1, AMPK, PKC, PKA, and GSK-3β, as well as their substrate specificity; therefore, all these kinases may be involved in EMT regulation. GSK-3β could also have a bifunctional role in EMT progression initiated by changes in S6K1 isoform expression that modulate its activity.
Oxidative stress responses of in vitro expanded WJ-MSCs derived from mothers with diabetes mellitus
Aim. To date, there are limited and conflicting data regarding the effects of diabetic microenvironment on perinatal stem cells.Our study aims to assess impact of diabetes mellitus on functional responses of in vitro expanded WJ-MSCs derived from DM-mothers to oxidative stress. Methods. MSCs were obtained by the explant method and cultured according to standard methods. Oxidative stress was induced by hydrogen peroxide (H2O2) added as a pulse. Treated WJ-MSCs were analyzed for metabolic activity by MTT assay. Results. The dose-response effect of different concentration of H2O2 (6,25–440 µM) on WJ-MSCs derived from mothers with DM and healthy mothers was evaluated in vitro. The difference between diabetic and control groups in NOAEL values (no observed adverse effect level) was analyzed. Conclusions. The effect of oxidative stress on the metabolic activity in vitro of both normal and diabetic WJ-MSCs manifested according to the hormetic model of the response to the stressor (dose-response with stimulation by a low dose and inhibition by a high dose) and the threshold model with adaptation to low doses without stimulation. Although diabetic WJ-MSCs were more tolerant to moderate oxidative stress compared to the control cells according to NOAEL, there was no statistically significant difference in this parameter between groups according to the Mann-Whitney U test. Further studies on more amount of samples are needed to use this cell model for diagnostic applications and to develop criteria for donor cells.
Clinically significant molecular alterations of mitochondria in carcinogenesis and age-related diseases: Part 1. Disease susceptibility and diagnosic significance
Mitochondrial dysfunction is associated with the hallmarks of cancer, aging, and age-related diseases. In recent years, numerous molecular mitochondrial alterationss have been identified at the DNA, RNA, and protein levels, as well as metabolic shifts and metabolic reprogramming in various types of cancer and age-related diseases. The review analyzes a number of clinically significant parameters of mitochondrial disorders, including susceptibility to disease development, diagnosis, prognosis, and the course of these pathologies. The manifestations of mitochondrial dysfunction at the molecular level are constantly being refined and revised, so in the future, we can expect the emergence of new clinically significant signs of these disorders in cancer and age-related diseases.
somatic and germline genetic alterations
Spectral-fluorescent response of dioxaborine dyes with varying chromophore charge in the presence of serum albumins
Aim. The aim of the work is the study of a spectral-fluorescent response of a series of dioxaborine dyes in the presence of serum albumins. Methods. UV-Vis absorption and fluorescence spectroscopy. Results. The fluorescent response of eight polymethine dyes (six of them being dioxaborine-based) in the presence of human, bovine, and equine serum albumins was studied and compared to the response in the presence of DNA and ovalbumin. It was shown that dianionic dioxaborine A74 and thiobarbituric-based merocyanine In4tb demonstrated0 the most promising properties as fluorescent probes for human serum albumin detection, with fluorescence intensity increasing by 62- and 56-fold, respectively. Meanwhile, A74 shows the highest selectivity to HSA among other serum albumins: in the presence of ESA and BSA, the signal increased by only 28-fold and 7.9-fold, respectively. Thus, one can suppose that A74 may be sensitive to slight variations in the globular conformation of serum albumins. Besides, dioxaborine merocyanines B33 and B36 demonstrated nearly 10-fold fluorescence intensity increase in the presence of serum albumins. Conclusions. The most promising dyes A74 and In4tb can be further studied for various applications that require fluorescent detection or imaging of HSA, and, in the case of A74, as a probe sensitive to minor changes in the HSA globular structure.
Comparison of EV isolation methods from MSCs
Aim. To evaluate the impact of isolation methods and culture conditions on the purity, yield, and phenotype of extracellular vesicles (EVs) derived from human mesenchymal stromal cells (MSCs). Methods. EVs were isolated using tangential flow filtration (TFF) with membranes of different molecular weight cut-offs (100–300 kDa), combined with ultracentrifugation (UC), under serum-containing and serum-free conditions. Particle concentration and size distribution (40–1000 nm) were analyzed by flow cytometry, total protein content was measured fluorometrically, morphology was assessed by transmission electron microscopy, and expression of tetraspanins (CD9, CD63, CD81) was determined by immunophenotyping. Results. Serum-containing media yielded a higher number of EVs but resulted in substantial protein contamination. The purest isolates (0.09–0.2 mg/mL total protein) were obtained under serum-free conditions using TFF with <100 kDa membranes combined with ultracentrifugation. Under these conditions, CD63⁺ exosomes predominated (up to 98%). Transmission electron microscopy confirmed typical EV morphology with diameters of 30–200 nm. Conclusions. TFF combined with UC enables modulation of EV purity and yield. Serum-free culture significantly improves the biochemical purity and phenotypic specificity of EV isolates, making them more suitable for downstream applications. Optimization of filtration parameters remains critical for balancing vesicle concentration and quality.
To the 85th jubilee of Anna Valentynivna Elska
Altered expression of genes related to inflammation and coagulation in peripheral blood mononuclear cells of patients with severe COVID-19
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the virus that causes coronavirus disease 2019 (COVID-19), the respiratory illness responsible for COVID-19 pandemic. Hyperinflammation and coagulopathy contribute to disease severity and death in patients infected with SARS-CoV-2. Gene expression analysis in peripheral blood mononuclear cells (PBMCs) is valuable to evaluate disease-associated and drug-response related genes. In this study, we aimed to investigate the expression of genes related to inflammation and coagulation in PBMCs of patients with severe COVID-19. The gene expression in PBMCs (52 patients with severe COVID-19 and 59 healthy volunteers) was determined by RT-qPCR. Overexpression of the OAS1, RNASEL, MX1, EIE2AK2, IL8, IL6, IL10, F5 genes and downexpression of CD4 gene were found out in the PBMCs of SARS-CoV-2 infected patients compared to the healthy volunteers. Using the ROC curve, we found genes with an excellent (IL10, EIF2AK2, F5) and with good (FN1, MX1, RNASEL) diagnostic AUCROC. The results of this study indicate the genes in PBMCs that can be potential candidate biomarkers for diagnosis of SARS-CoV-2-induced hyperinflammation, hypercoagulation, and the evaluation of therapy effectiveness for patients with severe COVID-19.
Challenges in PCR detection of Bacillus anthracis
Bacillus anthracis is large Gram-positive bacteria which causes anthrax – one of the deadliest known diseases. Different approaches have been developed to detect it, from more precise, time-consuming culture-based methods to more rapid PCR-based methods. The latter rely on the identification of specific regions in the B. anthracis genome, which is composed of two plasmids, pXO1 and pXO2, and the chromosome. Targeting only plasmid markers may provide unreliable results due to high similarities between B. anthracis and other genetically related species of Bacillus cereus sensu lato group, which necessitates the use of additional chromosomal targets. This article aims to provide a brief non- exhaustive overview on the relevant genetic markers used as targets for PCR-based detection of B. anthracis.
Development and comparison of two alanine aminotransferase-sensitive biosensors based on pyruvate oxidase and glutamate oxidase
Aim. To compare two developed biosensors based on pyruvate oxidase (POx) and glutamate oxidase (GlOx) for measuring alanine aminotransferase activity and to determine their main advantages and disadvantages. Methods. A three-electrode amperometric measurement scheme was used in the study. Platinum disk electrodes were used as working electrodes, and the signal was recorded using a PalmSens potentiostat. Immobilized POx and GlOx were used as recognition biomaterials. Results. The method and the optimal conditions for immobilization of each of the enzymes were selected. The procedure for creating the developed ALT-sensitive biosensors based on POx and GlOx was optimized. The optimal operating parameters for each sensor were chosen and analytical characteristics of both proposed biosensors were compared. POx biosensor has 1.5 times higher sensitivity, wider linear range and 2.5 lower detection limit, and unlike GlOx biosensor, can be used in a multibiosensor system for simultaneous measurement of ALT and AST. Conclusions. Taking into account the biosensor based on POx has more advantages to be used for ALT activity determination in the biological fluids.
ESR1 gene editing and its impact on S6K1 signaling and cell behavior
Aim. The aim of this study was to validate the link between the expression of estrogen receptor alpha (ESR1) isoforms and the regulation of S6K1-dependent signaling using the previously generated MCF-7 sublines with the CRISPR/Cas9 editing of the ESR1 gene. Methods. Western blotting, RT-PCR analysis, resazurine cell proliferation assay, and wound healing assay. Results. The presented study reveals a complex and multifaceted relationship between ESR1 isoforms, the S6K1 signaling pathway, and key aspects of MCF-7 cell behavior. It was demonstrated that complete inactivation of full-length p66 ESR1 leads to a significant decrease in S6K1 expression, which, in turn, negatively impacts cell proliferation and motility. Interestingly, the increased expression of the p46 ESR1 isoform, despite maintaining overall S6K1 levels, disrupts its activation dynamics, suggesting a possible regulatory role for p46 ESR1 in modulating cellular responses to growth factors. Conclusions. These findings are crucial for understanding the mechanisms by which different estrogen receptor isoforms can influence the growth, aggressiveness, and metastatic potential of breast cancer cells, particularly through their effect on S6K1 expression and activity. Further research is needed to elucidate the precise molecular mechanisms by which p46 ESR1 modulates S6K1 activation.
Interleukins and their autoantibodies in health and disease
Anti-interleukin autoantibodies (AIAs) are a unique type of immunoglobulins that target the human own ILs, which are key protein mediators of the immune system. Although the presence of AIAs is mostly associated with several immunodeficiency conditions, recent studies show that their presence in healthy individuals is normal, however in small amounts. In addition, the elevated titters may also be a marker of a positive prognosis or even a decrease in the severity of some diseases. Research into AIAs opens up new perspectives for understanding the pathogenesis of autoimmune and infectious diseases, as well as for the development of new approaches to effective and targeted therapy.
autoimmune polyendocrine syndrome type 1
In silico prediction of neuroprotective properties of natural compounds using Scutellaria baicalensis as an example
Aim. To develop, optimize, and evaluate effective in silico models for predicting the neuroprotective and anxiolytic properties of natural compounds using Scutellaria baicalensis as a case study. Methods. Construction and validation of machine learning models. Results. Three machine learning models, constructed using the Random Forest, XGBoost, and LightGBM algorithms, were developed for in silico prediction of the neuroprotective and anxiolytic activity of natural compounds. The classifiers achieved an accuracy of 75–78%. A binary classification approach was proposed, incorporating molecular descriptors and structural fingerprints, which, after preprocessing and optimization, enabled the identification of compounds with potential neuroprotective activity. The study confirms the effectiveness of these modeling approaches in predicting the neuroprotective, anxiolytic properties of S. baicalensis compounds. Application of the models to known phytochemicals from this plant verified previously reported bioactive substances: of 78 analyzed compounds, 46 were predicted to be potentially active. Conclusions. The in silico prediction of neuroprotective properties of bioactive compounds shows promise for screening and identifying phytocomplexes, particularly for applications in modern medicine such as the prevention and management of PTSD and other neurological disorders.
XIX All-Ukrainian Conference of Young Scientists of the Institute of Molecular Biology and Genetics of the NAS of Ukraine, dedicated to the Day of Science in Ukraine
The XIX All-Ukrainian Conference of Young Scientists of the Institute of Molecular Biology and Genetics of the NAS of Ukraine was held on May 20–21, 2025, to mark the Day of Science in Ukraine (registration certificate issued by UkrISTEI of the Ministry of Education and Science of Ukraine, No. 212, dated February 12, 2025). It was organized by the Council of Young Scientists (Head – PhD Antonenko S.V.). Three types of participation were available for registered scientists: oral presentation, poster presentation, and abstract publication only. The conference included four sections in the field of biological sciences, namely: Molecular Biology and Bioorganic Chemistry; Molecular Physiology and Biophysics; Biotechnology and Microbiology; Molecular Oncogenetics, Genetics, and Epigenetics. More than 60 participants registered for the conference, representing research institutions, universities, and medical establishments from various regions of Ukraine. In addition, young scientists from the Institute of Biosciences, Life Sciences Center, Vilnius University (Lithuania), participated in the event.
Computational prediction of biological activity of 5-(2-oxoindolin-3-ylidene)-substituted of 3-(benzo[d]thiazol-2-ylamino)-2-thioxothiazolidin-4-ones
Aim. To carry out in silico research of potential affinity of 5-isatinylidene derivatives of 3-(benzo[d]thiazol-2-ylamino)-2-thioxothiazolidin-4-one to biotargets and determine their possible belonging to ATC classes. Methods. Web tool SuperPred 3.0. Results. Determined the predominant antitumour activity of the compounds. The potential biological activity of derivatives 5–19 was compared with the in vitro efficacy of the core heterocycle, previously synthesized compounds 1–4 and the known effect of the antineoplastic drug Sutent (Pfizer Inc., USA). The compounds exhibited the hignest group efficiency against two targets: Cathepsin D and Casein kinase II alpha/beta (Model accuracy are 98.95% and 99.23%, respectively). Determined the important role of the isatinylidene moiety on the biological activity of the derivatives. Conclusions. The highest probability of structural similarity to drugs with ATC code L01XE is predicted for compound 2 (38.18%). Compounds 15 and 19 are considered potential multi-hitters. The highest affinity is predicted for compounds: 19 (99.80% to Cath-D) and 1 (93.56% to CK2). These values exceed those predicted for Sutent. The results obtained herein provide a platform for structure-based optimization of these derivatives.
Morphological changes in the somatosensory cortex of a rat brain after acute ischemia-reperfusion modelling followed by administration of mesenchymal stem cells of various origins, rat embryonic fibroblasts and Wharton's jelly MSCs lysate
Aim. To evaluate the effect of mesenchymal stem cells of various origins, rat embryonic fibroblasts and Wharton's jelly lysate on the dynamics of destructive changes in the somatosensory cortex of rats with artificially induced acute cerebrovascular injury. Methods. Modelling of transient bilateral ischemia-reperfusion (IR) in rats by ligation of the internal carotid arteries. Transplantation of mesenchymal stem cells (MSCs) of different origins into the femoral vein of experimental animals. Cytological study on 7th and 14th days after IR with counting the total number of neuronal nuclei in 1 mm2 in the somatosensory cortex of animals and the ratio of the intact neuronal nuclei to the pathologically altered nuclei. Results. Transplantation of MSCs, rat embryonic fibroblasts, MSC lysate from human umbilical cord Wharton's jelly (hUC-WJ) and administration of citicoline led to an increase in the number of intact neurons and nuclei without pathological changes in the somatosensory cortex of rats. The most effective in terms of cytomorphology was the transplantation of hUC-WJ MSCs. Conclusions. The MSC transplantation is effective in the treatment of cerebral ischemia. The most effective prevention of destructive changes was observed in the somatosensory cortex of rats that were transplanted with MSCs from human umbilical cord Wharton's jelly after IR.
Biotechnological production of chitosan: extraction and characterization from Trichoderma sp.
Aim. The study aimed at extraction and physicochemical characterization of fungal chitosan from Trichoderma reesei (MTCC 4876). Methods. Fungal chitosan was isolated using submerged fermentation over 4 days in three different growth media: Potato Dextrose Broth (PDB), Yeast Potato Dextrose Broth (YPDB), and Czapek Dox Broth (CDB). Growth kinetic parameters, including specific growth rate (µ) and doubling time (td), were determined for each medium. Chitosan extraction from the harvested biomass was performed using an alkaline treatment method. Physicochemical characterization of extracted chitosan using FTIR & DSC, UV-Vis spectroscopy for optical absorption properties and Photoluminescence (PL) spectroscopy to assess fluorescence behavior. Results. Among the tested media, PDB demonstrated the highest biomass yield of 116 mg/g. FTIR analysis confirmed the presence of functional groups analogous to commercial chitosan, indicating structural similarity. Thermal analysis via DSC validated the thermal stability of fungal chitosan. UV-Vis spectroscopy revealed significant optical absorption, while PL spectra exhibited notable fluorescence behavior, suggesting potential use in advanced material applications. Conclusions. In this study chitosan was successfully isolated from Trichoderma reesei (MTCC 4876), demonstrating distinct physicochemical properties comparable to the commercial chitosan. These findings highlight the potential of fungal chitosan for diverse industrial and commercial applications, particularly where optical and thermal properties are crucial.