Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines

The last decade has seen a sharp increase in the number of scientific publications describing physiological and pathological functions of extracellular vesicles (EVs), a collective term covering various subtypes of cell-released, membranous structures, called exosomes, microvesicles, microparticles, ectosomes, oncosomes, apoptotic bodies, and many other names. However, specific issues arise when working with these entities, whose size and amount often make them difficult to obtain as relatively pure preparations, and to characterize properly. The International Society for Extracellular Vesicles (ISEV) proposed Minimal Information for Studies of Extracellular Vesicles ( MISEV ) guidelines for the field in 2014. We now update these MISEV2014 guidelines based on evolution of the collective knowledge in the last four years. An important point to consider is that ascribing a specific function to EVs in general, or to subtypes of EVs, requires reporting of specific information beyond mere description of function in a crude, potentially contaminated, and heterogeneous preparation. For example, claims that exosomes are endowed with exquisite and specific activities remain difficult to support experimentally, given our still limited knowledge of their specific molecular machineries of biogenesis and release, as compared with other biophysically similar EVs. The MISEV2018 guidelines include tables and outlines of suggested protocols and steps to follow to document specific EV-associated functional activities. Finally, a checklist is provided with summaries of key points

Theory of Generations in Russia: on the Problem of Determining the Age Boundaries of Youth

Статья посвящена обзору различных авторских подходов в рамках отечественной школы теории поколений. В качестве предмета исследования выступает анализ российских моделей теории поколений в контексте определения возраста современной молодежи. Объектом исследования стали теории А. С. Сумской и С. А. Свердлова, группы авторов российской школы теории поколений «RuGenerations». В статье представлены результаты анализа отечественных теорий. Автор уточняет на этой основе возрастные границы цифровой молодежи.The article is devoted to the review of various author›s approaches within the framework of the national school of generational theory. The subject of the study is the analysis of the Soviet and Russian models of the theory of generations in the context of determining the age of modern youth. The object of the study were the theories of Yu. Levada, A. S. Sumskaya and S. A. Sverdlov, the group of authors of the Russian 385 school of generational theory “RuGenerations”. The article presents the results of the analysis of domestic theories. The author clarifies on this basis the age limits of digital youth

The Experience of Studying the Media Studies of Tambov Schoolchildren

The article presents the results of the media study related to the behavior of Tambov schoolchildren during the pandemic. The subject of the study was the values of generation Z at the age of 13 to 17 years, the object — the process of changing values under the influence of the pandemic. As research methods, we used general scientific methods-analysis and synthesis, and an empirical method of measuring the audience of representatives of generation Z using questionnaires. The results of the study allow us to predict changes in the moral priorities of young people, which will affect the media behavior of centenials in the post-pandemic period.В статье представлены результаты исследования медиаповедения тамбовских школьников в период пандемии. Предметом исследования стали ценностные установки поколения Z в возрасте от 13 до 17 лет, объектом — процесс изменения ценностей под влиянием пандемии. В качестве методов исследования использовались общенаучные методы — анализ и синтез, и эмпирический метод измерения аудитории представителей поколения Z при помощи анкетирования. Результаты исследования позволяют прогнозировать изменения нравственных приоритетов молодежи, которые отразятся на медиаповедении центениалов в постпандемический период

Media Texts of Modern Youth Media: the Specifics of Building the “Addressee — Addressee” Relationship

Статья поступила в редакцию 10.02.2023 г.Статья посвящена актуальной проблеме формирования у молодежи коммуникативной культуры. С целью выявления специфики построения отношений «адресант — адресат» был проведен анализ медиатекстов, ориентированных на молодых медиапотребителей. Для достижения цели исследования при помощи текстологического метода изучался контент современных молодежных медиа «Маруся», «Янгспейс», NR. В работе представлены результаты глубинного анализа страниц в социальных медиа представителей молодежи — участников проекта Тамбовского государственного университета имени Г. Р. Державина «Школа развития Derzhavinskills», наблюдений среди учеников 4–8-х классов тамбовской школы, а также интервью с работниками образования (школьные учителя и психологи). В заключение приведены выводы о средствах построения отношений «адресант — адресат»: преобладание приема диалогизации, тяготение к игрореализации как форме диалога с аудиторией, усиление воздействия манипулятивных механизмов и нивелирование этических норм в контексте речевого общения.The article is devoted to the actual problem of the formation of a communicative culture among young people. In order to identify the specifics of building the “addressee — addressee” relationship, the study of media texts aimed at young media consumers is carried out. To achieve the research goal, the content of modern youth media “Marusya”, “Yangspace”, NR was studied based on the textual method. The paper presents the results of an in–depth analysis of the social media pages of youth representatives participating in the Tambov State University named after G. R. Derzhavin School of Development Derzhavinskills, the results of observation among students of grades 4–8 of the Tambov school and interviews with educators (school teachers and psychologists). In conclusion, the conclusions about the means of building the “addressee — addressee” relationship are presented: the predominance of the method of dialogization, the attraction to game realization as a form of dialogue with the audience, the strengthening of the impact of manipulative mechanisms and the leveling of ethical norms in the context of speech communication

Diverse lipid conjugates for functional extra-hepatic siRNA delivery in vivo

Small interfering RNA (siRNA)-based therapies are proving to be efficient for treating liver-associated disorders. However, extra-hepatic delivery remains challenging, limiting therapeutic siRNA utility. We synthesized a panel of fifteen lipid-conjugated siRNAs and systematically evaluated the impact of conjugate on siRNA tissue distribution and efficacy. Generally, conjugate hydrophobicity defines the degree of clearance and the liver-to-kidney distribution profile. In addition to primary clearance tissues, several conjugates achieve significant siRNA accumulation in muscle, lung, heart, adrenal glands and fat. Oligonucleotide distribution to extra-hepatic tissues with some conjugates was significantly higher than with cholesterol, a well studied conjugate, suggesting that altering conjugate structure can enhance extra-hepatic delivery. These conjugated siRNAs enable functional gene silencing in lung, muscle, fat, heart and adrenal gland. Required levels for productive silencing vary (5-200 mug/g) per tissue, suggesting that the chemical nature of conjugates impacts tissue-dependent cellular/intracellular trafficking mechanisms. The collection of conjugated siRNA described here enables functional gene modulation in vivo in several extra-hepatic tissues opening these tissues for gene expression modulation. A systemic evaluation of a panel of conjugated siRNA, as reported here, has not previously been investigated and shows that chemical engineering of lipid siRNAs is essential to advance the RNA therapeutic field

Loss of huntingtin function slows synaptic vesicle endocytosis in striatal neurons from the htt(Q140/Q140) mouse model of Huntington\u27s disease

Huntington\u27s disease (HD) is caused by CAG repeat expansion within the HTT gene, with the dysfunction and eventual loss of striatal medium spiny neurons a notable feature. Since medium spiny neurons receive high amounts of synaptic input, we hypothesised that this vulnerability originates from an inability to sustain presynaptic performance during intense neuronal activity. To test this hypothesis, primary cultures of either hippocampal or striatal neurons were prepared from either wild-type mice or a knock-in HD mouse model which contains 140 poly-glutamine repeats in the huntingtin protein (htt(Q140/Q140)). We identified a striatum-specific defect in synaptic vesicle (SV) endocytosis in htt(Q140/Q140) neurons that was only revealed during high frequency stimulation. This dysfunction was also present in neurons that were heterozygous for the mutant HTT allele. Depletion of endogenous huntingtin using hydrophobically-modified siRNA recapitulated this activity-dependent defect in wild-type neurons, whereas depletion of mutant huntingtin did not rescue the effect in htt(Q140/Q140) neurons. Importantly, this SV endocytosis defect was corrected by overexpression of wild-type huntingtin in homozygous htt(Q140/Q140) neurons. Therefore, we have identified an activity-dependent and striatum-specific signature of presynaptic dysfunction in neurons derived from pre-symptomatic HD mice, which is due to loss of wild-type huntingtin function. This presynaptic defect may render this specific neuronal subtype unable to operate efficiently during high frequency activity patterns, potentially resulting in dysfunctional neurotransmission, synapse failure and ultimately degeneration

The Practice of Media Consumption of «Digital Youth» in Social Media

Предметом исследования данной работы является активность «цифровой молодежи» в социальных медиа. Объект исследования – медиапотребление молодежной аудитории в социальных сетях и мессенджерах. К проведению глубинного анализа медиаконтента молодежи привлечена группа студентов различных специальностей, проходящих обучение в проекте «Derzhavinskills» Тамбовского государственного университета имени Г. Р. Державина. В результате исследования авторы статьи формулируют выводы о функционале и степени доверия «цифровой молодежи» популярным социальным медиа. Отмечается проявление в медиапотреблении практики «Digital detox», свидетельствующей об «усталости от публичности» и стремлении «цифровой молодежи» к ассимиляции в небольшие группы.The subject of this work is the activity of “digital youth” in social media. The object of the study is the media consumption of the youth audience in social networks and instant messengers. A group of students of various specialties studying in the Derzhavinskills project of Derzhavin Tambov State University. As a result of the study, the authors of the article formulate conclusions about the functionality and degree of trust of “digital youth” in popular social media. There are manifestations in media consumption of the practice of “Digital detox”, indicating “publicity fatigue” and the desire of “digital youth” to assimilate into small groups

5-Vinylphosphonate improves tissue accumulation and efficacy of conjugated siRNAs in vivo

5-Vinylphosphonate modification of siRNAs protects them from phosphatases, and improves silencing activity. Here, we show that 5-vinylphosphonate confers novel properties to siRNAs. Specifically, 5-vinylphosphonate (i) increases siRNA accumulation in tissues, (ii) extends duration of silencing in multiple organs and (iii) protects siRNAs from 5-to-3 exonucleases. Delivery of conjugated siRNAs requires extensive chemical modifications to achieve stability in vivo. Because chemically modified siRNAs are poor substrates for phosphorylation by kinases, and 5-phosphate is required for loading into RNA-induced silencing complex, the synthetic addition of a 5-phosphate on a fully modified siRNA guide strand is expected to be beneficial. Here, we show that synthetic phosphorylation of fully modified cholesterol-conjugated siRNAs increases their potency and efficacy in vitro, but when delivered systemically to mice, the 5-phosphate is removed within 2 hours. The 5-phosphate mimic 5-(E)-vinylphosphonate stabilizes the 5 end of the guide strand by protecting it from phosphatases and 5-to-3 exonucleases. The improved stability increases guide strand accumulation and retention in tissues, which significantly enhances the efficacy of cholesterol-conjugated siRNAs and the duration of silencing in vivo. Moreover, we show that 5-(E)-vinylphosphonate stabilizes 5 phosphate, thereby enabling systemic delivery to and silencing in kidney and heart

2\u27-O-Methyl at 20-mer Guide Strand 3\u27 Termini May Negatively Affect Target Silencing Activity of Fully Chemically Modified siRNA

Small interfering RNAs (siRNAs) have the potential to treat a broad range of diseases. siRNAs need to be extensively chemically modified to improve their bioavailability, safety, and stability in vivo. However, chemical modifications variably impact target silencing for different siRNA sequences, making the activity of chemically modified siRNA difficult to predict. Here, we systematically evaluated the impact of 3\u27 terminal modifications (2\u27-O-methyl versus 2\u27-fluoro) on guide strands of different length and showed that 3\u27 terminal 2\u27-O-methyl modification negatively impacts activity for \u3e60% of siRNA sequences tested but only in the context of 20- and not 19- or 21-nt-long guide strands. These results indicate that sequence, modification pattern, and structure may cooperatively affect target silencing. Interestingly, the introduction of an extra 2\u27-fluoro modification in the seed region at guide strand position 5, but not 7, may partially compensate for the negative impact of 3\u27 terminal 2\u27-O-methyl modification. Molecular modeling analysis suggests that 2\u27-O-methyl modification may impair guide strand interactions within the PAZ domain of argonaute-2, which may affect target recognition and cleavage, specifically when guide strands are 20-nt long. Our findings emphasize the complex nature of modified RNA-protein interactions and contribute to design principles for chemically modified siRNAs