FLIM reveals alternative EV-mediated cellular up-take pathways of paclitaxel

In response to physiological and artificial stimuli, cells generate nano-scale extracellular vesicles (EVs) by encapsulating biomolecules in plasma membrane-derived phospholipid envelopes. These vesicles are released to bodily fluids, hence acting as powerful endogenous mediators in intercellular signaling. EVs provide a compelling alternative for biomarker discovery and targeted drug delivery, but their kinetics and dynamics while interacting with living cells are poorly understood. Here we introduce a novel method, fluorescence lifetime imaging microscopy (FLIM) to investigate these interaction attributes. By FLIM, we show distinct cellular uptake mechanisms of different EV subtypes, exosomes and microvesicles, loaded with anti-cancer agent, paclitaxel. We demonstrate differences in intracellular behavior and drug release profiles of paclitaxel-containing EVs. Exosomes seem to deliver the drug mostly by endocytosis while microvesicles enter the cells by both endocytosis and fusion with cell membrane. This research offers a new real-time method to investigate EV kinetics with living cells, and it is a potential advancement to complement the existing techniques. The findings of this study improve the current knowledge in exploiting EVs as next-generation targeted drug delivery systems.Peer reviewe

Development of the potato cryocollection preserved in the VIR cryobank

Background. The article presents the results of the studies carried out within the framework of a complex program for the long-term conservation of genotyped accessions of advanced potato cultivars in the cryobank of the N.I. Vavilov All-Russian Institute of Plant Genetic Resources (VIR). These accessions are genetically identical to the nomenclature standards of the corresponding cultivars.Materials and methods. The research material included 24 accessions from the in vitro collection of VIR, corresponding to the nomenclature standards of Russian potato cultivars developed in different breeding centers of the Russian Federation. Cryopreservation was carried out using the droplet vitrification method, several stages of which were modified in the Department of Biotechnology of VIR.Results and discussion. The frequency of post-cryogenic regeneration for the analyzed 24 cultivars in control variants with short-term immersion of explants in liquid nitrogen varied from 20 to 47.3%, and in 9 accessions it exceeded 39% – the current level of reliable long-term preservation of samples in cryobanks. The data of the statistical analysis failed to reveal a significant effect of the genotype or the calendar dates of different experiment replications on the regeneration ability. These 24 accessions were added to the potato cryocollection stored in the VIR cryobank. An analysis of the composition and structure of VIR’s potato cryocollection, which currently includes 140 accessions, is presented

Cryopreservation of raspberry cultivar accessions bred in Russia from the VIR <i>in vitro</i> collection

Cryobanks use plant cryocollections for long-term preservation of crops which cannot be preserved in seed collections. These are vegetatively propagated crops, accessions of species which form either a small amount of seeds, or recalcitrant seeds. Shoot tips (apexes) of in vitro plants are used for cryopreservation for most berry crops, therefore maintenance of in vitro collections is very important. The VIR in vitro collection includes 150 accessions of Rubus L. species, 85 of them are raspberry cultivars, 59 of which were bred in Russia. These cultivars reflect a wide ecogeographic diversity. Among them, there are raspberry cultivars created at the end of the 19th – first half of the 20th centuries, including cultivars bred by I.V. Michurin and by the pioneer of northern horticulture V.V. Spirin. More than half of national raspberry varieties (33) are listed in the State Register for Selection Achievements Admitted for Usage. Raspberry cultivars from Russian breeding programs have a very limited representation in foreign genebanks. The first aim of the present work was cryopreservation of mostly folk and old Russian raspberry cultivars received by VIR from 1925 till 1950 and their transfer into the cryobank. The second aim of the work was to monitor post-cryogenic regeneration of raspberry cultivars transferred to the cryobank earlier. A modified protocol of the droplet vitrification method by “DV-biotech” was used for cryopreservation of shoot tips of in vitro plants of 10 raspberry cultivars (7 of which are folk and old Russian ones) from the VIR in vitro collection. Post-cryogenic regeneration was evaluated for 17 raspberry cultivars preserved in the cryobank from one to five years. Ten raspberry cultivars (900 apexes) with an average mean post-cryoregenic regeneration value of 38.2±3.0% determined in control tests, were placed in the cryobank for long-term storage. A statistically significant effect of the genotype on the viability of explants after cryopreservation was noted, while the post-cryogenic regeneration was genotype insensitive. Additionally, levels of post-cryogenic regeneration were evaluated for 17 raspberry cultivars (296 apexes) preserved in the cryobank from one to five years. Post-cryogenic regeneration within the 20-70% range was displayed by four raspberry cultivars preserved in the cryobank for one year, and for 8 cultivars conserved there from three to five years post-cryogenic regeneration was within the 10-50% range. According to the results of monitoring, regeneration displayed by 12 raspberry cultivars was within the 10-70% range, which can be considered as a reliable rate of apex preservation in liquid nitrogen vapors in the VIR cryobank. Monitoring of the post-cryogenic regeneration of the raspberry accessions preserved in the VIR cryobank and cryopreservation of new raspberry cultivars will be continued

ВПЛИВ ФІНАНСОВОЇ КРИЗИ В УКРАЇНІ 2014—2015 РОКІВ НА РІВЕНЬ ФІНАНСОВОЇ БЕЗПЕКИ БАНКІВ

It has been determined that commercial banks operate in a complex operational environment, which is due to the influence of a number of factors of political, social, economic, which are largely unpredictable and destructive. It is substantiated that external key parameters through which the influence on the process of providing financial security of banks is mediated. The relationship between the size of the rate of mandatory reserve requirements and the level of financial security of the bank is researched. The analysis of the state of assets, liabilities and equity capital by groups of banks during 2012—2016. The role of the central bank in regulating the activities of banks in order to ensure their financial security has been determined. The expediency of imposing sanctions on carrying out of any financial operations of banks with Russian capital in favor of parent structures was substantiated. The key problems that restrain the process of providing financial security of banks in terms of their functional components (capital-resource security, credit-investment, currency and safe level of incomes and expenditures) are determined. The methodical approach to the rating of banks estimation on the level of their financial security is offered on the basis of the analysis of indicators The stimulator / disintegrators of the analyzed banks is based on the rating rating of the bank according to the level of its financial security on the basis of comparison of j-th KB with the «standard» — the bank which The best results for each indicator are the maximum values for stimulators and the minimum for disinfectants. The rating is defined as the arithmetic mean of standardized indicators summed up in the matrix of key indicators. The necessity of grouping the analyzed banks according to their financial security level has been proved. Using the method of grouping of research objects is substantiated as an instrument for streamlining the analyzed banks. It is determined empirically, taking into account qualitative, non-formalized characteristics of four groups of banks for the p vnem their financial security (with a high level of financial security, sufficient, low and critical).Визначено, що комерційні банки функціонують у складному операційному середовищі, а це зумовлено впливом низки факторів політичних, соціальних, економічних, що здебільшого є малопрогнозованими і деструктивними. Обґрунтовано зовнішні ключові параметри, через які опосередковується вплив на процес забезпечення фінансової безпеки банків. Досліджено взаємозв’язок між розміром ставки нормативів обов’язкового резервування і рівнем фінансової безпеки банку. Проаналізовано стан активів, зобов’язань і власного капіталу за групами банків упродовж 2012—2016 рр.. Визначено роль центрального банку в регулюванні діяльності банків щодо забезпечення їхньої фінансової безпеки. Обґрунтовано доцільність запровадження НБУ санкцій на проведення будь-яких фінансових операцій банків із російським капіталом на користь материнських структур. Визначено ключові проблеми, які стримують процес забезпечення фінансової безпеки банків у розрізі і функціональних складових (капітало-ресурсної безпеки, кредитно-інвестиційної, валютної та безпечного рівня доходів і витрат. Запропоновано методичний підхід до рейтингової оцінки банків за рівнем їхньої фінансової безпеки на основі аналізу показників-стимуляторів / дестимуляторів аналізованих банків. Обґрунтовано проведення рейтингової оцінки банку за рівнем його фінансової безпеки на основі порівняння&nbsp;j-го КБ з «еталоном» — банком, який має найкращі результати щодо кожного показника — максимальні значення для стимуляторів і мінімальні — для дестимуляторів. Рейтинг визначено як середнє арифметичне стандартизованих показників, зведених у матрицю ключових показників. Доведено необхідність групування аналізованих банків за рівнем їхньої фінансової безпеки. Обґрунтовано використання методу групування об’єктів дослідження як інструменту упорядкування аналізованих банків. Визначено емпіричним шляхом з обліком якісних, неформалізованих характеристик чотири групи банків за рівнем їхньої фінансової безпеки (з високим рівнем фінансової безпеки, достатнім, низьким і критичним)

Comparative analysis of anxiety-depressive spectrum disorders in patients with rheumatic diseases

Research objective - comparative analysis of incidence and structure of anxiety-depressive spectrum disorders (ADD) in patients with various rheumatic diseases (RD). Materials and methods. 613 patients with RD were enrolled in the study: 180 with a reliable diagnosis of systemic lupus erythematosus (SLE), 128 with rheumatoid arthritis (RA), 110 with systemic sclerosis (SSc), 115 with Behcet's disease (BD), 80 with primary Sjögren's syndrome (pSS). Female prevailed in all groups (95% of patients with pSS, 88,2% - SSc, 87,2% - RA, 85,5% of SLE) except BD patients (70% male). The mean age was 42.3±1.54 years and was lower in patients with BD (33.3±0.98 years) and SLE (34.6±0.93 years) compared to patients with SSc (49.9±2.47 years), RA (47.4±0.99 years) and pSS (46.2±2.3 years). The mean RD duration was 130,0±8,65 months and was more at BD - 148,5±10,4 months, pSS - 141,6±8,92 months, RA - 138,4±10,1months, and less at SLE - 134,9±8,8 months and SSc - 87,0±5,04 months. The mean SLE activity index SLEDAI was 9,13±0,63 points (high), RA (DAS28) - 5,26±0,17 points (high), BD (BDCAF) - 3,79±0,2 points (moderate) and SSc by G. Valentini - 1,1±0,20 points (moderate). Glucocorticoids took 100% of patients with pSS, 91,1% - SLE, 90% - SSc, 87% - BD and 67,2% - RA patients; conventional disease modifying anti-rheumatic drugs (cDMARDs) took 90% of patients with SSc, 84% - BD, 79,6% - RA, 68% - pSS, 40,6% - SLE. Biologic DMARDs took 32% of patients with RA, 17,4% - BD, 7,3% - SSc and 7,2% - SLE. Mental disorders were diagnosed by psychiatrist as a result of screening by the hospital anxiety and depression scale (HADS) and in semi-structured interview in accordance with the ICD-10/ DSM-IV. The severity of depression was evaluated by Montgomery-Asberg Depression Rating Scale (MADRS) and anxiety - by Hamilton Anxiety Rating Scale (HAM-A). Projective psychological methods were used for cognitive impairment detection. Results. Screening of depressive disorders (HADS-D≥8) was positive in 180 (29,4%) patients with RD, including 74 (41%) patients with SLE, 38 (35%) - SSc, 29 (23%) - RA, 23 (20%) - BD and 16 (20%) - pSS; anxiety disorders (HADS-A≥8) - in 272 (44,4%) patients, including 66 (52%) patients with RA, 40 (50%) - pSS, 77 (43%) - SLE, 45 (41%) - SSc and 44 (38%) - BD. In accordance with the ICD-10/ DSM-IV depressive disorders have been identified in 389 (63%) patients, including 94 (73%) patients with RA, 71 (64,5%) - SSc, 69 (60%) - BD, 90 (50%) - SLE and 39 (49%) - pSS; anxiety disorders - in 377 (61,5%) patients, including 20 (25%) patients with pSS, 44 (24,5%) - SLE, 29 (23%) - RA, 20 (17%) - BD and 7 (6,4%) - SSc. Conclusion. Anxiety-depressive spectrum disorders are typical for most patients with RA, SLE, SSc, pSS and BD. ADDs diagnosis in RD patients with the use of the HADS did not reveal a significant proportion. To obtain objective data on the frequency and structure of ADDs, psychopathological and clinical psychological diagnosis is necessary

Global phylogeography and ancient evolution of the widespread human gut virus crAssphage

Microbiomes are vast communities of microorganisms and viruses that populate all natural ecosystems. Viruses have been considered to be the most variable component of microbiomes, as supported by virome surveys and examples of high genomic mosaicism. However, recent evidence suggests that the human gut virome is remarkably stable compared with that of other environments. Here, we investigate the origin, evolution and epidemiology of crAssphage, a widespread human gut virus. Through a global collaboration, we obtained DNA sequences of crAssphage from more than one-third of the world’s countries and showed that the phylogeography of crAssphage is locally clustered within countries, cities and individuals. We also found fully colinear crAssphage-like genomes in both Old-World and New-World primates, suggesting that the association of crAssphage with primates may be millions of years old. Finally, by exploiting a large cohort of more than 1,000 individuals, we tested whether crAssphage is associated with bacterial taxonomic groups of the gut microbiome, diverse human health parameters and a wide range of dietary factors. We identified strong correlations with different clades of bacteria that are related to Bacteroidetes and weak associations with several diet categories, but no significant association with health or disease. We conclude that crAssphage is a benign cosmopolitan virus that may have coevolved with the human lineage and is an integral part of the normal human gut virome

Variants of psychiatric disorders in patients with systemic lupus erythematosus

Objective. To analyze prevalence and structure of psychiatric disorders in pts with systemic lupus erythematosus (SLE) examining in the Institute of rheumatology of RAMS. Material and methods. 115 pts with SLE with median age 34 [24; 45] years and median disease duration 8 [4; 17] years were included. SLE activity was assessed with SLEDAI. Psychiatric disorders were diagnosed by a psychiatrist according to ICD-10 using some psychiatric and psychological scales. Results. Psychiatric disorders were revealed in 76 from 115 (66%) pts. Anxiety-depressive spectrum disorders prevailed (83%): depressive episode (40%), adjustment disorders (24%), generalized anxiety disorder (10%), dysthymia (9%). Severe cognitive dysfunction was revealed in 7% of pts. Pts with and without psychiatric disorders did not significantly differ in age, sex, duration and activity of the disease, duration of treatment and cumulative dose of prednisolone and cytotoxic drugs. Conclusion. Psychiatric disorders are frequent in pts with SLE (66%). Anxiety-depressive disorders prevail among them (83%). Relationship between SLE and psychiatric disorders requires further examination