Acta Naturae最新文献

{"title":"A Method for the Production of Recombinant VSVs with Confirmation of Biological Activity.","authors":"V D Moroz, N B Gasanov, A D Egorov, A S Malogolovkin, M O Nagornykh, E N Subcheva, E S Kolosova, A Yu Fizikova, R A Ivanov, A V Karabelsky","doi":"10.32607/actanaturae.27314","DOIUrl":"10.32607/actanaturae.27314","url":null,"abstract":"<p><p>The design of new effective cancer treatment methods is a promising and important research field in translational medicine. Oncolytic viruses can induce immunogenic cell death by activating the body's immune system to recognize tumor cells. This work presents the results for optimizing the production of recombinant vesicular stomatitis viruses (rVSVs). To ensure the assembly of viral particles, we developed the HEK293TN-T7 cell line, which stably expresses DNA-dependent RNA polymerase 7 for viral genome transcription, and obtained helper plasmids encoding viral genes under the control of the CAG promoter. The oncolytic activity of the purified virus preparation was assessed in a murine model of B16F10Red melanoma cells expressing a red fluorescent protein. The presented method makes it possible to obtain purified viral preparations with a high titer and oncolytic activity. The amplification of viral particles in a HEK293 suspension culture allows for rapid scalability. Therefore, the developed approach can be used to obtain other recombinant VSV-based oncolytic viruses for tumor immunotherapy.</p>","PeriodicalId":6989,"journal":{"name":"Acta Naturae","volume":"16 1","pages":"59-66"},"PeriodicalIF":2.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11062106/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140852359","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Post-transcriptional Regulation of Gene Expression via Unproductive Splicing.","authors":"L G Zavileyskiy, D D Pervouchine","doi":"10.32607/actanaturae.27337","DOIUrl":"10.32607/actanaturae.27337","url":null,"abstract":"<p><p>Unproductive splicing is a mechanism of post-transcriptional gene expression control in which premature stop codons are inserted into protein-coding transcripts as a result of regulated alternative splicing, leading to their degradation via the nonsense-mediated decay pathway. This mechanism is especially characteristic of RNA-binding proteins, which regulate each other's expression levels and those of other genes in multiple auto- and cross-regulatory loops. Deregulation of unproductive splicing is a cause of serious human diseases, including cancers, and is increasingly being considered as a prominent therapeutic target. This review discusses the types of unproductive splicing events, the mechanisms of auto- and cross-regulation, nonsense-mediated decay escape, and problems in identifying unproductive splice isoforms. It also provides examples of deregulation of unproductive splicing in human diseases and discusses therapeutic strategies for its correction using antisense oligonucleotides and small molecules.</p>","PeriodicalId":6989,"journal":{"name":"Acta Naturae","volume":"16 1","pages":"4-13"},"PeriodicalIF":2.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11062102/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140848337","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ultrastructural 3D Microscopy for Biomedicine: Principles, Applications, and Perspectives.","authors":"K E Mochalov, D S Korzhov, A V Altunina, O I Agapova, V A Oleinikov","doi":"10.32607/actanaturae.27323","DOIUrl":"10.32607/actanaturae.27323","url":null,"abstract":"<p><p>Modern biomedical research often requires a three-dimensional microscopic analysis of the ultrastructure of biological objects and materials. Conceptual technical and methodological solutions for three-dimensional structure reconstruction are needed to improve the conventional optical, electron, and probe microscopy methods, which to begin with allow one to obtain two-dimensional images and data. This review discusses the principles and potential applications of such techniques as serial section transmission electron microscopy; techniques based on scanning electron microscopy (SEM) (array tomography, focused ion beam SEM, and serial block-face SEM). 3D analysis techniques based on modern super-resolution optical microscopy methods are described (stochastic optical reconstruction microscopy and stimulated emission depletion microscopy), as well as ultrastructural 3D microscopy methods based on scanning probe microscopy and the feasibility of combining them with optical techniques. A comparative analysis of the advantages and shortcomings of the discussed approaches is performed.</p>","PeriodicalId":6989,"journal":{"name":"Acta Naturae","volume":"16 1","pages":"14-29"},"PeriodicalIF":2.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11062107/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140850265","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Studying Signaling Pathway Activation in TRAIL-Resistant Macrophage-Like Acute Myeloid Leukemia Cells.","authors":"Y V Lomovskaya, K S Krasnov, M I Kobyakova, A A Kolotova, A M Ermakov, A S Senotov, I S Fadeeva, E I Fetisova, A I Lomovsky, A I Zvyagina, V S Akatov, R S Fadeev","doi":"10.32607/actanaturae.27317","DOIUrl":"10.32607/actanaturae.27317","url":null,"abstract":"<p><p>Acute myeloid leukemia (AML) is a malignant neoplasm characterized by extremely low curability and survival. The inflammatory microenvironment and maturation (differentiation) of AML cells induced by it contribute to the evasion of these cells from effectors of antitumor immunity. One of the key molecular effectors of immune surveillance, the cytokine TRAIL, is considered a promising platform for developing selective anticancer drugs. Previously, under <i>in vitro</i> conditions of the inflammatory microenvironment (a three-dimensional high-density culture of THP-1 AML cells), we demonstrated the emergence of differentiated macrophage-like THP-1ad clones resistant to TRAIL-induced death. In the present study, constitutive activation of proinflammatory signaling pathways, associated transcription factors, and increased expression of the anti-apoptotic <i>BIRC3</i> gene were observed in TRAIL-resistant macrophage-like THP-1ad AML cells. For the first time, a bioinformatic analysis of the transcriptome revealed the main regulator, the <i>IL1B</i> gene, which triggers proinflammatory activation and induces resistance to TRAIL in THP-1ad macrophage-like cells.</p>","PeriodicalId":6989,"journal":{"name":"Acta Naturae","volume":"16 1","pages":"48-58"},"PeriodicalIF":2.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11062100/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140846363","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dimeric Bis-Benzimidazole-Pyrroles DB2Py(n) - AT-Site-Specific Ligands: Synthesis, Physicochemical Analysis, and Biological Activity.","authors":"O Y Susova, S S Karshieva, A A Kostyukov, N I Moiseeva, E A Zaytseva, K V Kalabina, E Zusinaite, K Gildemann, N M Smirnov, A F Arutyunyan, A L Zhuze","doi":"10.32607/actanaturae.27327","DOIUrl":"10.32607/actanaturae.27327","url":null,"abstract":"<p><p>Its broad spectrum of biological activity makes benzimidazole a fundamental pharmacophore in pharmaceutics. The paper describes newly synthesized AT-specific fluorescent bis-benzimidazole molecules DB2Py(n) that contain a pyrrolcarboxamide fragment of the antibiotic drug netropsin. Physico-chemical methods using absorption, fluorescence, and circular dichroism spectra have shown the ability of bis-benzimidazole- pyrroles to form complexes with DNA. The new DB2Py(n) series have turned out to be more toxic to human tumor lines and less vulnerable to non-tumor cell lines. Bis-benzimidazole-pyrroles penetrated the cell nucleus, affected the cell-cycle synthesis (S) phase, and inhibited eukaryotic topoisomerase I in a cellfree model at low concentrations. A real-time tumor cell proliferation test confirmed the molecule's enhanced toxic properties upon dimerization. Preliminary cytotoxicity data for the bis-benzimidazole-pyrroles tested in a cell model with a MDR phenotype showed that monomeric compounds can overcome MDR, while dimerization weakens this ability to its intermediate values as compared to doxorubicin. In this respect, the newly synthesized cytotoxic structures seem promising for further, in-depth study of their properties and action mechanism in relation to human tumor cells, as well as for designing new AT-specific ligands.</p>","PeriodicalId":6989,"journal":{"name":"Acta Naturae","volume":"16 1","pages":"86-100"},"PeriodicalIF":2.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11062108/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140847081","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Broadly Reactive Nanobody Targeting the H3 Hemagglutinin of the Influenza A Virus.","authors":"D V Shcheblyakov, D V Voronina, I A Favorskaya, I B Esmagambetov, I A Alekseeva, A I Korobkova, E I Ryabova, A A Derkaev, V Yu Kan, A Sh Dzharullaeva, A I Tukhvatulin, A S Bandelyuk, M M Shmarov, D Yu Logunov, A L Gintsburg","doi":"10.32607/actanaturae.27374","DOIUrl":"10.32607/actanaturae.27374","url":null,"abstract":"<p><p>Monoclonal antibodies and recombinant antibody fragments are a very promising therapeutic tool to combat infectious diseases. Due to their unique paratope structure, nanobodies (VHHs) hold several advantages over conventional monoclonal antibodies, especially in relation to viral infections. Influenza A viruses (IAVs) remain a major threat to public health. The hemagglutinin (HA) protein is the main protective and immunodominant antigen of IAVs. In this study, three broadly reactive nanobodies (D9.2, E12.2, and D4.2) to H3N2 influenza strains were isolated and Fc-fusion proteins (VHH-Fcs) were obtained and characterized <i>in vitro</i>. This modification improved the nanobodies' binding activity and allowed for their interaction with a wider range of strains. The D9.2-Fc antibody showed a 100% protection rate against mortality <i>in vivo</i> in a mouse lethal model. Furthermore, we demonstrated that the observed protection has to do with Fc-FcγR interactions. These results indicate that D9.2-Fc can serve as an effective antiviral agent against the H3N2 influenza infection.</p>","PeriodicalId":6989,"journal":{"name":"Acta Naturae","volume":"16 1","pages":"101-110"},"PeriodicalIF":2.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11062109/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140847584","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Création d’une liste régionale catégorisée d’espèces exotiques envahissantes et pistes d’utilisation pour les gestionnaires d’espaces naturels","authors":"Iris LANG, Camille GILLIOT, Olivier SCHER, Laurent PONTCHARRAUD, Daniel MARC","doi":"10.5852/naturae2023a7","DOIUrl":"https://doi.org/10.5852/naturae2023a7","url":null,"abstract":"Les espèces exotiques envahissantes (EEE) constituent une des cinq causes majeures d’érosion de la biodiversité à l’échelle mondiale. Devant leur diversité et leur abondance, il est impossible d’agir sur toutes les EEE. Le règlement UE 1143/2014 et la stratégie nationale relative aux EEE de 2017 offrent un cadre pour la prévention de l’introduction et de la propagation des EEE et leur gestion. Le Conservatoire d’Espaces naturels d’Occitanie a élaboré et anime actuellement la stratégie régionale relative aux EEE Faune d’Occitanie, dont l’objectif est d’orienter et de coordonner les actions menées vis-à-vis de ces espèces. Un état des lieux des connaissances des EEE Faune présentes ou susceptibles d’être introduites en Occitanie a d’abord été réalisé. Il n’existe pas, à l’heure actuelle, de méthode standardisée partagée par tous les acteur·rice·s pour classer ces espèces selon la menace qu’elles représentent. Une méthode intégrative a donc été développée à partir de quatre méthodes standardisées internationales et régionales (EICAT, SEICAT, ISEIA, ISSIA) afin de catégoriser les espèces exotiques en Occitanie selon leur potentiel d’invasion et leurs impacts écologiques, socio-économiques et sanitaires. L’objectif de cet article est de partager notre retour d’expérience sur le développement de la méthode pour l’élaboration de la liste catégorisée, et sur les utilisations possibles de cette dernière. Nous montrons notamment comment les acteur·rice·s régionaux, peuvent s’approprier la liste catégorisée, qui constitue le premier outil de connaissance sur les EEE Faune en Occitanie. Les catégories ainsi définies, associées au regard de l’expert·e selon le contexte, permettent d’identifier les grands types d’actions à mener sur ces espèces (prévention, gestion, amélioration des connaissances, communication). Ce travail, qui peut servir aux autres régions déclinant la stratégie nationale, offre un cadre pour l’articulation des stratégies régionales et l’amélioration des modes d’organisation au niveau national.","PeriodicalId":6989,"journal":{"name":"Acta Naturae","volume":"39 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136210398","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Soft X-ray Microscopy in Cell Biology: Current Status, Contributions and Prospects.","authors":"S A Golyshev, E P Kazakov, I I Kireev, D G Reunov, I V Malyshev","doi":"10.32607/actanaturae.26551","DOIUrl":"10.32607/actanaturae.26551","url":null,"abstract":"<p><p>The recent advances achieved in microscopy technology have led to a significant breakthrough in biological research. Super-resolution fluorescent microscopy now allows us to visualize subcellular structures down to the pin-pointing of the single molecules in them, while modern electron microscopy has opened new possibilities in the study of protein complexes in their native, intracellular environment at near-atomic resolution. Nonetheless, both fluorescent and electron microscopy have remained beset by their principal shortcomings: the reliance on labeling procedures and severe sample volume limitations, respectively. Soft X-ray microscopy is a candidate method that can compensate for the shortcomings of both technologies by making possible observation of the entirety of the cellular interior without chemical fixation and labeling with an isotropic resolution of 40-70 nm. This will thus bridge the resolution gap between light and electron microscopy (although this gap is being narrowed, it still exists) and resolve the issue of compatibility with the former, and possibly in the near future, the latter methods. This review aims to assess the current state of soft X-ray microscopy and its impact on our understanding of the subcellular organization. It also attempts to look into the future of X-ray microscopy, particularly as relates to its seamless integration into the cell biology toolkit.</p>","PeriodicalId":6989,"journal":{"name":"Acta Naturae","volume":"15 4","pages":"32-43"},"PeriodicalIF":2.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10790358/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139486935","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"System for Self-excited Targeted Photodynamic Therapy Based on the Multimodal Protein DARP-NanoLuc-SOPP3.","authors":"E I Shramova, A Yu Frolova, V P Filimonova, S M Deyev, G M Proshkina","doi":"10.32607/actanaturae.27331","DOIUrl":"10.32607/actanaturae.27331","url":null,"abstract":"<p><p>Despite the significant potential of photodynamic therapy (PDT) as a minimally invasive treatment modality, the use of this method in oncology has remained limited due to two serious problems: 1) limited penetration of the excitation light in tissues, which makes it impossible to affect deep-seated tumors and 2) use of chemical photosensitizers that slowly degrade in the body and cause photodermatoses and hyperthermia in patients. To solve these problems, we propose a fully biocompatible targeted system for PDT that does not require an external light source. The proposed system is based on bioluminescent resonance energy transfer (BRET) from the oxidized form of the luciferase substrate to the photosensitizing protein SOPP3. The BRET-activated system is composed of the multimodal protein DARP-NanoLuc-SOPP3, which contains a BRET pair NanoLuc-SOPP3 and a targeting module DARPin. The latter provides the interaction of the multimodal protein with tumors overexpressing tumor-associated antigen HER2 (human epidermal growth factor receptor type II). <i>In vitro</i> experiments in a 2D monolayer cell culture and a 3D spheroid model have confirmed HER2-specific photo-induced cytotoxicity of the system without the use of an external light source; in addition, experiments in animals with subcutaneous HER2-positive tumors have shown selective accumulation of DARP-NanoLuc-SOPP3 on the tumor site. The fully biocompatible system for targeted BRET-induced therapy proposed in this work makes it possible to overcome the following limitations: 1) the need to use an external light source and 2) the side phototoxic effect from aberrant accumulation of chemical photosensitizers. The obtained results demonstrate that the fully protein-based self-excited BRET system has a high potential for targeted PDT.</p>","PeriodicalId":6989,"journal":{"name":"Acta Naturae","volume":"15 4","pages":"100-110"},"PeriodicalIF":2.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10790359/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139484877","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Animal Models of Mitochondrial Diseases Associated with Nuclear Gene Mutations.","authors":"O A Averina, S A Kuznetsova, O A Permyakov, P V Sergiev","doi":"10.32607/actanaturae.25442","DOIUrl":"10.32607/actanaturae.25442","url":null,"abstract":"<p><p>Mitochondrial diseases (MDs) associated with nuclear gene mutations are part of a large group of inherited diseases caused by the suppression of energy metabolism. These diseases are of particular interest, because nuclear genes encode not only most of the structural proteins of the oxidative phosphorylation system (OXPHOS), but also all the proteins involved in the OXPHOS protein import from the cytoplasm and their assembly in mitochondria. Defects in any of these proteins can lead to functional impairment of the respiratory chain, including dysfunction of complex I that plays a central role in cellular respiration and oxidative phosphorylation, which is the most common cause of mitopathologies. Mitochondrial diseases are characterized by an early age of onset and a progressive course and affect primarily energy-consuming tissues and organs. The treatment of MDs should be initiated as soon as possible, but the diagnosis of mitopathologies is extremely difficult because of their heterogeneity and overlapping clinical features. The molecular pathogenesis of mitochondrial diseases is investigated using animal models: i.e. animals carrying mutations causing MD symptoms in humans. The use of mutant animal models opens new opportunities in the study of genes encoding mitochondrial proteins, as well as the molecular mechanisms of mitopathology development, which is necessary for improving diagnosis and developing approaches to drug therapy. In this review, we present the most recent information on mitochondrial diseases associated with nuclear gene mutations and animal models developed to investigate them.</p>","PeriodicalId":6989,"journal":{"name":"Acta Naturae","volume":"15 4","pages":"4-22"},"PeriodicalIF":2.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10790356/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139484918","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}