The composition of low-polarity extracts obtained by sequential extraction of the aerial parts of Rhododendron adamsii Rehd. with hexane and methyl tert-butyl ether (MTBE) was investigated using GC-MS. The hexane extract was dominated by non-polar components: squalene, n-alkanes (nonacosane, hentriacontane), sesquiterpenes (trans-nerolidol, spathulenol, β-farnesene), and β-sitosterol. The subsequent MTBE extract was enriched in more polar lipids, primarily free triterpenic acids (ursolic and oleanolic acids). A critical finding was the complete absence of diterpene grayanotoxins in all tested extracts, confirming the safety of the non-polar extraction approach. In bioactivity assays, the total hexane extract demonstrated potent inhibitory activity against the SARS-CoV-2 main protease (3CLpro) with IC50 values of 0.0125–0.025 mg/mL, only one order of magnitude higher than the reference inhibitor disulfiram. Fractionation revealed that the activity was distributed among free acids, bound acids, and the unsaponifiable residue, indicating a multicomponent mechanism. Importantly, none of the samples inhibited HIV-1 protease (IC50 > 0.1 mg/mL), demonstrating selectivity for the cysteine protease 3CLpro over the aspartyl protease of HIV-1. These results highlight that sequential non-polar extraction of R. adamsii provides a grayanotoxin-free lipophilic complex with selective anti-SARS-CoV-2 protease activity, paving the way for bioactivity-guided identification of individual inhibitors.

The composition of low-polarity extracts obtained by sequential extraction of the aerial parts of Rhododendron adamsii Rehd. with hexane and methyl tert-butyl ether (MTBE) was investigated using GC-MS. The hexane extract was dominated by non-polar components: squalene, n-alkanes (nonacosane, hentriacontane), sesquiterpenes (trans-nerolidol, spathulenol, β-farnesene), and β-sitosterol. The subsequent MTBE extract was enriched in more polar lipids, primarily free triterpenic acids (ursolic and oleanolic acids). A critical finding was the complete absence of diterpene grayanotoxins in all tested extracts, confirming the safety of the non-polar extraction approach. In bioactivity assays, the total hexane extract demonstrated potent inhibitory activity against the SARS-CoV-2 main protease (3CLpro) with IC₅₀ values of 0.0125–0.025 mg/mL, only one order of magnitude higher than the reference inhibitor disulfiram. Fractionation revealed that the activity was distributed among free acids, bound acids, and the unsaponifiable residue, indicating a multicomponent mechanism. Importantly, none of the samples inhibited HIV-1 protease (IC₅₀ > 0.1 mg/mL), demonstrating selectivity for the cysteine protease 3CLpro over the aspartyl protease of HIV-1. These results highlight that sequential non-polar extraction of R. adamsii provides a grayanotoxin-free lipophilic complex with selective anti-SARS-CoV-2 protease activity, paving the way for bioactivity-guided identification of individual inhibitors.

The composition of the lipophilic components of Centaurea scabiosa L. has been studied. The raw material was subjected to extraction with hexane and methyl tert-butyl ether (MTBE) using both exhaustive and sequential schemes for a detailed characterization. The resulting extracts were fractionated into acidic and neutral components via treatment with alkali solutions. The acidic compounds were converted into methyl esters for subsequent gas chromatography–mass spectrometry (GC-MS) analysis, while the neutral unsaponifiable fractions were separated into groups of different polarities using column chromatography on silica gel. This approach enabled the identification of a complex profile of lipophilic substances. In the acidic fractions, aliphatic acids with chain lengths from C10 to C32, including unsaturated variants, were characterized. The neutral fractions revealed over compounds, encompassing n-alkanes, substantial levels of the unsaturated branched hydrocarbon squalene, and a diverse array of oxygenated terpenoids. The latter were mainly represented by highly active triterpene alcohols and ketones belonging to the ursane, oleanane, lupane, and cycloartane types. The sterol composition was dominated by β-sitosterol and accompanied by cholesterol, campesterol, stigmasterol, stigmast-7-en-3-β-ol, fucosterol, and stigmastan-3-β-ol. Bioactivity screening demonstrated that several of the obtained lipophilic extracts, particularly those of lower polarity, exhibited high inhibitory activity against the main protease of SARS-CoV-2, underscoring the potential of C. scabiosa as a valuable source of anti-coronavirus agents.

The purpose of the article is to prove the possibilities and limitations of studying childhood events to justify practice-oriented projects in the field of youth policy and childhood support. The article describes main directions of the evidence-based approach in applied research. Authors argue the need to use sociological data on the scale, specifics of various situations, episodes, debut events in childhood for making management decisions in evidence-based state policy in the field of childhood and youth. The data of panel studies “Eventfulness of Childhood” implemented in 2018 and 2023 are analyzed. From an array of 100 significant childhood events, those that are most relevant for designing various programs for children and youth (deviant childhood events, introduction to subcultural practices, mastering information and communication technologies, traditional practices of mastering culture) were selected to argue the evidence-based approach. It has been revealed which groups of children and adolescents are the target audience for projects on the prevention of deviation and associative subcultural groups, introduction to reading, digital hygiene programs, etc. The prospects for development of an evidence-based approach have been identified: the imperative of using it not only at the initial stage of creating projects, but also upon completion of programs to prove effectiveness, the need to introduce research as a mandatory condition in grant programs, and a change in the priorities for selecting experts.

The annual number of reported human cases of flavivirus infections continues to increase. Measures taken by local healthcare systems and international organizations are not fully successful. In this regard, new approaches to treatment and prevention of flavivirus infections are relevant. One promising approach is to use monoclonal antibody preparations. The mouse mAb 10H10 is capable of interacting with viruses belonging to the genus Orthoflavivirus which are pathogenic to humans. ELISA and molecular modeling data can indicate that mAb 10H10 recognizes the fusion loop region of E protein. The KD of interaction between the mAb 10H10 and recombinant analogs of the E protein of the tick-borne encephalitis (TBEV), Zika (ZIKV) and dengue (DENV) viruses range from 1.5 to 4 nM. The aim of this study was to map the epitope of this antibody using phage display technology. After three rounds of biopanning, 60 individual phage clones were chosen. The amino acid sequences of the selected peptides were conveniently divided into five groups. Based on the selected peptides, bacteriophages were obtained carrying peptides on the surfaces of the pIII and pVIII proteins, which were tested for binding to the antibody in ELISA. Thus, the epitope of the mAb 10H10 is the highly conserved region 98-DRGWGNXXGLFGK-110 of the flavivirus E protein. The structures of the complexes of the identified peptides with the antibody paratope are proposed using the molecular docking and dynamics methods.

Журнал для непрерывного медицинского образования врачей РЕДАКЦИОННАЯ СТАТЬЯ РезюмеЦель работы -сравнительная оценка куриных (IgY) и мышиных моноклональных (IgG) антител к IgG человека в качестве зонда для тест-систем иммуноферментного анализа (ИФА) и дот-иммуноанализа при диагностике клещевого энцефалита.Материал и методы.Проведены эксперименты по сравнению способности зондов на основе мышиных IgG и куриных IgY антител к IgG человека, а также протеинов А и G, связанных с пероксидазой хрена и коллоидным золотом, определять человеческие иммуноглобулины разных классов, а также выявлять в сыворотке крови человека IgG, специфические антитела к вирусу клещевого энцефалита (ВКЭ).В качестве реагента захвата в ИФА и дот-анализе использовали рекомбинантный аналог домена III гликопротеина Е ВКЭ.Исследования выполнены в сравнении с коммерческим набором «ВектоВКЭ-IgG» с применением панелей сывороток крови, содержащих антитела к ВКЭ, желтой лихорадке и денге, а также образцов от здоровых доноров.Результаты и обсуждение.Показано, что сконструированные экспериментальные наборы для ИФА и дот-анализа адекватно определяют положительные и отрицательные образцы и позволяют дифференцировать антитела к ВКЭ от антител к другим флавивирусам.Зонды с пероксидазой и коллоидным золотом обеспечивают одинаковые лимиты обнаружения IgG, а зонды на основе куриных антител демонстрируют более интенсивные оптические сигналы и преимущества в чувствительности по сравнению с конъюгатами мышиных IgG.Заключение.Полученные данные позволяют рассматривать конъюгаты на основе куриных антител в качестве перспективных реагентов для иммуноанализа.Финансирование.Исследование проведено в рамках выполнения государственного задания Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человека.

A recombinant plasmid pBSU-PLC containing the B. thuringiensis phospholipase C gene has been developed. Expression of the target genes in this vector is under the control of the strong synthetic Pgrac promoter. B. mojavensis cells were transformed with the resulting recombinant plasmid. The obtained recombinant strain was used for experimental production of recombinant B. thuringiensis phospholipase C in the pilot cycle of cultivation. As a result of the fermentation cycle, with successive transfer from a 15-liter fermenter to a 250-liter one, an enzyme was obtained with high activity after 13 hours of cultivation.

In December 2019, a new coronavirus, SARS-CoV-2, was found to in Wuhan, China. Cases of infection were subsequently detected in other countries in a short period of time, resulting in the declaration of the COVID-19 pandemic by the World Health Organization (WHO) on 11 March 2020. Questions about the impact of herd immunity of pre-existing immune reactivity to SARS-CoV-2 on COVID-19 severity, associated with the immunity to seasonal manifestation, are still to be resolved and may be useful for understanding some processes that precede the emergence of a pandemic virus. Perhaps this will contribute to understanding some of the processes that precede the emergence of a pandemic virus. We assessed the specificity and virus-neutralizing capacity of antibodies reacting with the nucleocapsid and spike proteins of SARS-CoV-2 in a set of serum samples collected in October and November 2019, before the first COVID-19 cases were documented in this region. Blood serum samples from 799 residents of several regions of Siberia, Russia, (the Altai Territory, Irkutsk, Kemerovo and Novosibirsk regions, the Republic of Altai, Buryatia, and Khakassia) were analyzed. Sera of non-infected donors were collected within a study of seasonal influenza in the Russian Federation. The sample collection sites were located near the flyways and breeding grounds of wild waterfowl. The performance of enzyme-linked immunosorbent assay (ELISA) for the collected sera included the usage of recombinant SARS-CoV-2 protein antigens: full-length nucleocapsid protein (CoVN), receptor binding domain (RBD) of S-protein and infection fragment of the S protein (S5-6). There were 183 (22.9%) sera reactive to the S5-6, 270 (33.8%) sera corresponding to the full-length N protein and 128 (16.2%) sera simultaneously reactive to both these proteins. Only 5 out of 799 sera had IgG antibodies reactive to the RBD. None of the sera exhibited neutralizing activity against the nCoV/Victoria/1/2020 SARS-CoV-2 strain in Vero E6 cell culture. The data obtained in this study suggest that some of the population of the analyzed regions of Russia had cross-reactive humoral immunity against SARS-CoV-2 before the COVID-19 pandemic started. Moreover, among individuals from relatively isolated regions, there were significantly fewer reliably cross-reactive sera. The possible significance of these data and impact of cross-immunity to SARS-CoV-2 on the prevalence and mortality of COVID-19 needs further assessment.

Despite the cancellation of the COVID-19 pandemic status, the question of the dynamics of specific antibodies to the new coronavirus is still relevant. The aim of this study is to assess the level of class M antibodies to the receptor-binding domain of the SARS-CoV-2 spike protein in recovered COVID-19 patients and to search for its relationship with such indicators and characteristics as the positivity coefficient, sex, age, severity and duration of the period after the disease.

Currently, SARS-CoV-2 spike receptor-binding-domain (RBD)-based vaccines are considered one of the most effective weapons against COVID-19. During the first step of assessing vaccine immunogenicity, a mouse model is often used. In this paper, we tested the use of five experimental animals (mice, hamsters, rabbits, ferrets, and chickens) for RBD immunogenicity assessments. The humoral immune response was evaluated by ELISA and virus-neutralization assays. The data obtained show hamsters to be the least suitable candidates for RBD immunogenicity testing and, hence, assessing the protective efficacy of RBD-based vaccines.

Despite the rapid accumulation of facts about the humoral immune response in COVID-19, there are still no evidencebased answers to questions about the factors influencing the level and duration of the detection period of antibodies to SARS-CoV-2 in the blood.Objective: To assess the prevalence, clinical and demographic associations of IgG antibodies to RBD of the SARSCoV-2 spike protein at different times after COVID-19.Materials and methods. Residents of the Altai region of Russia, Caucasians aged 20-93 years, who had COVID-19 from May 2020 to February 2021 (n = 314), took part in a onetime observational study. The level of antibodies in the blood was measured by enzyme-linked immunosorbent assay 1-14 months after the onset of the clinical manifestation of COVID-19.Results. Anti-RBD IgG antibodies of the SARS-CoV-2 spike protein were detected in 86.9% of the study participants. The dependence of the antibody titer on the duration of the period after COVID-19 was not revealed. The antibody titer was positively correlated with the complication of COVID-19 pneumonia and the volume of lung tissue lesions. The presence of pneumonia COVID-19 and the volume of lung tissue lesions are positively associated with age. Age positively correlated with antibody titer regardless of the pneumonia COVID-19 in the anamnesis.Conclusion. IgG antibodies to RBD of the SARS-CoV-2 spike protein are present in most of the COVID-19 patients. The titer of these antibodies in adults depends on age, complications of pneumonia COVID-19, and probably persists up to 14 months after the first symptoms of infection appear.

Identification of factors behind the level and duration of persistence of the SARS-CoV-2 antibodies in the blood is assumed to set the direction for studying humoral immunity mechanisms against COVID-19, optimizing the strategy for vaccine use, antibody-based drugs, and epidemiological control of COVID-19. Objective: This study aimed to study the relationship between clinical and demographic characteristics and the level of IgG antibodies to the RBD of SARS-CoV-2 spike protein after COVID-19 in the long term. Residents of the Altai Region of Western Siberia of Russia, Caucasians, aged from 27 to 93 years (median 53.0 years), who recovered from COVID-19 between May 2020 and February 2021 (n = 44) took part in this prospective observational study. The titer of IgG antibodies to the RBD of SARS-CoV-2 spike protein was measured repeatedly in the blood at 4–13 months from the beginning of the clinical manifestation of COVID-19 via the method of enzyme-linked immunosorbent assay. The antibody titer positively correlated with age (p = 0.013) and COVID-19 pneumonia (p = 0.002) at 20–40 and 20–24 weeks from the onset of COVID-19 symptoms, respectively. Age was positively associated with antibody titer regardless of history of COVID-19 pneumonia (beta regression coefficient p = 0.009). The antibody titer decreased in 15 (34.1%) patients, increased in 10 (22.7%) patients, and did not change in 19 (43.2%) patients from the baseline to 48–49 weeks from the onset of COVID-19 symptoms, with seropositivity persisting in all patients. Age and COVID-19 pneumonia are possibly associated with higher IgG antibodies to the spike protein RBD of SARS-CoV-2 following COVID-19 in the long term. Divergent trends of anti-RBD IgG levels in adults illustrate inter-individual differences at 4–13 months from the onset of COVID-19 symptoms.

Changes in the repertoire of circulating antibodies is a promising biomarker for various diseases. We have developed microarrays containing more than thousands of peptides with random amino acid sequences that can be used to analyze the entire repertoire of circulating antibodies. Using these peptide microarrays, we have shown that in the blood of patients diagnosed with breast cancer (BC) antibodies interacting with certain peptides are found. Moreover, circulating antibodies of patients with different molecular subtypes of BC (luminal A, luminal B and Basal-like) interact with different, but overlapping, panels of peptides. We have selected 634 peptides from 120K peptides presented on microarrays that specifically interact with circulating antibodies of blood plasma from patients with various subtypes of BC. This panel of peptides that specifically interact with the blood plasma immunoglobulins of patients with various molecular BC subtypes can be used for the development of non-invasive diagnostic test systems. Microchips containing thousands of peptides can be used as a tool for molecular screening populations at risk for various diseases. Thus, peptide microarrays can be used to create a new tool for studying the repertoire of circulating antibodies for different diseases evaluation, as well as for studying and evaluating protein-protein interactions.

The article substantiates the expansion of factors of children's mobility and the inclusion of a temporal factor associated with the characteristics of time: age-related debuts of movements, the transformation of individual episodes of activity into group routes, the sequence of development of environmental objects, the influence of the specific historical situation of residence. There are three areаs of movements in independent mobility – intrafamily, extrafamily, and virtual mobility. Based on the author's study in 2018, the temporal characteristics of many events of modern childhood are presented and some forecasts and hypotheses of the relationship between mobility and growing up are described.

The article describes the features of the childhood events of two conditional generations of young people - Y and Z. In methodological terms, the novelty lies in the substantiation of the new concept "event of a child's life" - an event in everyday life that stands out in the stream of life as a necessary means of growing up, and is performed by the child, and not by adults towards him/her. The rationale for the new concept /notion of "an event of child's life" is given - the event of everyday life, which is highlighted in the stream of life as a necessary means of growing up, and is committed by the child, and not by adults. The article reveals the methodological possibilities of studying the eventfulness of growing up with the help of the author's "Event-Generation Z" method. Limited application of the modern theory of XYZ generation is proved: the difficulty of a clear boundary between the generations at the turn of the century. The use of a new Russian conditional generation Z in childhood has been established to use a different format for the practices used in information and communication technologies, bodily practices, social activity. It has been proved that the age of adulthood events is related to the place of residence, the socio-economic situation of the family, and composition of the family. The coincidence of the chronology of events and the average age of the subjective termination of childhood is recorded. Noted are the facts of narrowing the space of children's self-dependence and the "extension of childhood" of separate groups, the paradoxes of the events of childhood as not consistent, not logical from the point of view of the development and upbringing of children.