AAV9-mediated central nervous system-targeted gene delivery via cisterna magna route in mice

Current barriers to the use of adeno-associated virus serotype 9 (AAV9) in clinical trials for treating neurological disorders are its high expression in many off-target tissues such as liver and heart, and lack of cell specificity within the central nervous system (CNS) when using ubiquitous promoters such as human cytomegalovirus (CMV) or chicken-β-actin hybrid (CAG). To enhance targeting the transgene expression in CNS cells, self-complementary (sc) AAV9 vectors, scAAV9-GFP vectors carrying neuronal Hb9 and synapsin 1, and nonspecific CMV and CAG promoters were constructed. We demonstrate that synapsin 1 and Hb9 promoters exclusively targeted neurons in vitro, although their strengths were up to 10-fold lower than that of CMV. In vivo analyses of mouse tissue after scAAV9-GFP vector delivery via the cisterna magna revealed a significant advantage of synapsin 1 promoter over both Hb9 variants in targeting neurons throughout the brain, since Hb9 promoters were driving gene expression mainly within the motor-related areas of the brain stem. In summary, this study demonstrates that cisterna magna administration is a safe alternative to intracranial or intracerebroventricular vector delivery route using scAAV9, and introduces a novel utility of the Hb9 promoter for the targeted gene expression for both in vivo and in vitro applications

Optical Rectification of Phase Modulated Signal Based on Injection Locking:Paper

We experimentally demonstrate feasibility of simultaneous use of Differential Phase Shift Keying (DPSK) and Amplitude Shift Keying (ASK) formats (orthogonal modulation) using injection-locked semiconductor laser. Experimental study shows significant improvement of the bit-error-rate (BER) and doubling of the system capacity

Приближенные симметрии и законы сохранения дробно-дифференциального обобщения уравнения Бюргерса

The classical Burgers equation is well studied and is often used in problems of hydrodynamics and nonlinear acoustics. In recent years, there has been a significantly increasing interest in mathematical models that take into account the power-law memory effect of the medium. Such models are described by equations in which the time derivative is replaced by a fractional derivative.The study object is a generalized Burgers equation with a fractional Riemann-Liouville time derivative. A memory effect of the medium is assumed to be small, so a small parameter, with respect to which the fractional derivative is expanded into a series, is extracted from the fractional-order differentiation. As a result, the initial fractional differential generalization of the Burgers equation is approximated by an equation with a small parameter. The aim of the work is to study the symmetry properties of such a partial differential equation with a small parameter and to construct conservation laws for it. To achieve the goal, methods of modern group analysis are used, as well as widely known methods of integrating systems and partial differential equations of first order.The group classification of the equation under study is carried out according to the function standing at the first derivative with respect to the spatial variable. It is shown that if this function is of arbitrary form, then the admissible approximate group of point transformations is three-parameter. For power-law and linear functions, the admissible approximate group of point transformations extends to five- and seven-parameter groups, respectively. Examples of approximately invariant solutions for some admissible operators are constructed. It is proved that the studied equation with a small parameter is approximately nonlinearly self-adjoint. Based on the principle of nonlinear self-adjointness, conservation laws are constructed for each group generator. It is shown that all conservation laws are either trivial or have the form of the original equation.The results develop the theory of approximate transformation groups for fractional differential equations. The obtained symmetries can be used to construct approximate invariant solutions of the equation in question.Классическое уравнение Бюргерса хорошо исследовано и часто используется в задачах гидродинамики и нелинейной акустики. В последние годы наблюдается значительный рост интереса к математическим моделям, учитывающих эффект степенной памяти среды. Такие модели описываются уравнениями, в которых производная по времени заменена на производную дробного порядка.Объектом исследования является обобщенное уравнения Бюргерса с дробной производной Римана-Лиувилля по времени. Влияние памяти среды предполагается малым, поэтому из порядка дробного дифференцирования выделяется малый параметр, по которому выполняется разложение в ряд дробной производной. В результате исходное дробно-дифференциальное обобщение уравнения Бюргерса приближается уравнением с малым параметром. Целью работы является исследование симметрийных свойств такого дифференциального уравнения в частных производных с малым параметром и построение законов сохранения для него. Для достижения цели используются методы современного группового анализа, а также широко известные методы интегрирования систем и дифференциальных уравнений в частных производных первого порядка.Проведена групповая классификация исследуемого уравнения по функции, стоящей при первой производной по пространственной переменной. Показано, что если эта функция произвольного вида, то допускаемая приближенная группа точечных преобразований является трехпараметрической. Если функция степенная или линейная, то допускаемая приближенная группа точечных преобразований расширяется до пяти- и семипараметрической, соответственно. Построены примеры приближенно инвариантных решений для некоторых допускаемых операторов.Доказано, что исследуемое уравнение с малым параметром является приближенно нелинейно самосопряженным. На основе принципа нелинейной самосопряженности для каждого оператора группы построены законы сохранения. Показано, что все законы сохранения являются либо тривиальными, либо имеют вид исходного уравнения.Результаты развивают теорию приближенных групп преобразований для дробно-дифференциальных уравнений. Найденные симметрии могут быть использованы для построения приближенных инвариантных решений рассматриваемого уравнения

Nonlinear Pulses in Dispersion-Managed Fiber-Optic Systems in Presence of High Losses

A dispersion managed fiber-optic system can support dynamically stable pulses - DM-solitons and two branches of stable pairs of coupled DM-solitons – lower and upper bisolitons. Using semi-analytical model, we have found the dependency of the shape of these pulses on the level of losses and other parameters of the system and have verified the validity of our findings both numerically and experimentally. The obtained data can be used to define whether a dispersion-managed fiber-optic system can be considered as the one with constant dispersion

El impacto del método de proyectos en el desarrollo de habilidades de liderazgo en los estudiantes

The impact of the project method on the development of leadership skills in students / Valentyna O. Tyurina, Hanna Y. Shevchuk, Yuliya D. Kriukova and ot/ // Revista de Investigación Apuntes Universitarios. - 2022. - Vol. 12, Issue 2. – P. 179-197. -DOI: https://doi.org/10.17162/au.v12i2.1045.Досліджено вплив методу проектів на розвиток лідерських якостей у студентів. Проаналізовано розвиток особистості у студентському віці. Використовано комплекс методів дослідження: педагогічний експеримент, тест, спостереження. У дослідженні взяли участь 292 людини. Виявлено переважання середніх показників лідерських якостей у респондентів. Найменш розвиненими виявилися такі компоненти, як мотивація успіху та вольовий самоконтроль. Метод групових проектів довів ефективність для розвитку всіх вивчених навичок, крім вольового самоконтролю. Достовірність усіх виявлених змін знаходиться на рівні p = 0,05, що визначає необхідність подальшої модифікації використовуваного методу. Отримані дані можуть бути використані в практичній діяльності викладачів і психологів вищих навчальних закладів для розвитку лідерських якостей учнів.The article examines the influence of the project method on the development of students' leadership skills. Information on the use of the project method in high school and the details of the leader's personality development at the student's age are analyzed. A set of research methods was used: pedagogical experiment, test, observation, Student's t-test. The study involved 292 people. The predominance of average indicators of leadership skills of the respondents was revealed. Components such as motivation for success and volitional self-control were the least developed. The method of group projects has proven effective for the development of all the skills studied, except volitional self-control. The significance of all the changes detected is at the level of p = 0.05, which determines the need for further modification of the method used. The data obtained can be used in the practice of teachers and psychologists of higher education institutions (HEIs) in the development of students' leadership skills. Other perspectives for scientific research include studying the leadership competence of students and finding the correlation of leadership skills with structural components of personality.Рассматривается влияние метода проектов на развитие лидерских качеств у студентов. Проанализировано развитие личности в студенческом возрасте. Был использован комплекс методов исследования: педагогический эксперимент, тест, наблюдение. В исследовании приняли участие 292 человека. Выявлено преобладание средних показателей лидерских качеств у респондентов. Наименее развиты такие компоненты, как мотивация на успех и волевой самоконтроль. Метод групповых проектов доказал свою эффективность для развития всех изучаемых навыков, кроме волевого самоконтроля. Значимость всех выявленных изменений находится на уровне p = 0,05, что определяет необходимость дальнейшей модификации используемого метода. Полученные данные могут быть использованы в практике преподавателей и психологов высших учебных заведений в развитие лидерских качеств учащихся

SUMO Pathway Dependent Recruitment of Cellular Repressors to Herpes Simplex Virus Type 1 Genomes

Components of promyelocytic leukaemia (PML) nuclear bodies (ND10) are recruited to sites associated with herpes simplex virus type 1 (HSV-1) genomes soon after they enter the nucleus. This cellular response is linked to intrinsic antiviral resistance and is counteracted by viral regulatory protein ICP0. We report that the SUMO interaction motifs of PML, Sp100 and hDaxx are required for recruitment of these repressive proteins to HSV-1 induced foci, which also contain SUMO conjugates and PIAS2β, a SUMO E3 ligase. SUMO modification of PML and elements of its tripartite motif (TRIM) are also required for recruitment in cells lacking endogenous PML. Mutants of PML isoform I and hDaxx that are not recruited to virus induced foci are unable to reproduce the repression of ICP0 null mutant HSV-1 infection mediated by their wild type counterparts. We conclude that recruitment of ND10 components to sites associated with HSV-1 genomes reflects a cellular defence against invading pathogen DNA that is regulated through the SUMO modification pathway

Синтез, антиексудативна та антимікробна активність 6-арилідензаміщених імідазо[2,1-b]тіазолів

Aim. To expand the range of 6-arylidene-2-methyl-2,3-dihydroimidazo[2,1-b]thiazolones as potential objects for studying the antiexudative and antimicrobial activities. Results and discussion. It has been shown that the condensation of synthetically affordable 2-methyl-2,3-dihydroimidazo[2,1-b]thiazolone with aromatic aldehydes can be successfully used for obtaining the corresponding 6-ylidene-functionalized derivatives. The biological screening of the compounds synthesized revealed that they possessed a low or moderate anti-inflammatory activity and inhibited the inflammation process in the range from 3 to 44 %. During the study of the antimicrobial activity of the substances obtained it was determined that their minimum bacteriostatic and minimum fungistatic concentrations ranged from 31.25 to 250 μg/mL. Experimental part. The interaction of 2-methyl-2,3-dihydroimidazo[2,1-b]thiazolone with a series of benzaldehydes and salicylic aldehydes in refluxing acetic acid in the presence of anhydrous sodium acetate leads to new 6-arylidene-2-methyl-2,3-dihydroimidazo[2,1-b]thiazolones. The antiexudative activity screening was performed on the model of carrageenan-induced paw oedema of white outbred male rats. The antimicrobial activity of the compounds was studied using the microtechnique of two-fold serial dilutions in a liquid nutrient medium. Conclusions. It has been found that the Knoevenagel condensation of 2-methyl-2,3-dihydroimidazo[2,1-b]thiazolone with aromatic aldehydes is a convenient way for the structural modification of the position 6 of the heterocyclic system by the arylidene moiety. The arylidene derivatives obtained show a moderate antiexudative activity in the carrageenan-induced rat paw oedema assay, as well as the antimicrobial activity against some gram-positive and gram-negative bacteria and fungi.Мета. Розширити ряд 6-ариліден-2-метил-2,3-дигідроімідазо[2,1-b]тіазолонів як потенційних об’єктів для вивчення антиексудативної та антимікробної активностей. Результати та їх обговорення. Виявлено, що конденсація синтетично доступного 2-метил-2,3-дигідроімідазо[2,1-b]тіазолону з ароматичними альдегідами може бути вдало використана для одержання відповідних 6-іліденофункціоналізованих похідних. Біологічний скринінг синтезованих сполук виявив, що показник інгібування запального процесу лап щурів складає 3 – 44 %. Під час дослідження антимікробної активності отриманих речовин визначено, що їх мінімальна бактеріостатична та мінімальна фунгістатична концентрації коливаються в діапазоні 31,25 – 250 мкг/мл. Експериментальна частина. Взаємодією 2-метил-2,3-дигідроімідазо[2,1-b]тіазолону з низкою бензальдегідів та саліцилових альдегідів у киплячій оцтовій кислоті в присутності безводного натрій ацетату синтезовано нові 6-ариліден-2-метил-2,3-дигідроімідазо[2,1-b]тіазолони. Скринінг антиексудативної активності проводили на моделі індукованого карагеніном набряку лапи білих безпородних щурів-самців. Антимікробну дію вивчали з використанням мікрометоду дворазових серійних розведень у рідкому поживному середовищі. Висновки. Виявлено, що конденсація Кньовенагеля 2-метил-2,3-дигідроімідазо[2,1-b]тіазолону з ароматичними альдегідами є зручним варіантом структурної модифікації положення 6 цієї гетероциклічної системи ариліденовим фрагментом. Одержані ариліденпохідні виявляють помірну антиексудативну активність на моделі карагенін-індукованого набряку лапи щурів, а також антимікробну дію щодо деяких грам-позитивних та грам-негативних бактерій і грибів

EBV Tegument Protein BNRF1 Disrupts DAXX-ATRX to Activate Viral Early Gene Transcription

Productive infection by herpesviruses involve the disabling of host-cell intrinsic defenses by viral encoded tegument proteins. Epstein-Barr Virus (EBV) typically establishes a non-productive, latent infection and it remains unclear how it confronts the host-cell intrinsic defenses that restrict viral gene expression. Here, we show that the EBV major tegument protein BNRF1 targets host-cell intrinsic defense proteins and promotes viral early gene activation. Specifically, we demonstrate that BNRF1 interacts with the host nuclear protein Daxx at PML nuclear bodies (PML-NBs) and disrupts the formation of the Daxx-ATRX chromatin remodeling complex. We mapped the Daxx interaction domain on BNRF1, and show that this domain is important for supporting EBV primary infection. Through reverse transcription PCR and infection assays, we show that BNRF1 supports viral gene expression upon early infection, and that this function is dependent on the Daxx-interaction domain. Lastly, we show that knockdown of Daxx and ATRX induces reactivation of EBV from latently infected lymphoblastoid cell lines (LCLs), suggesting that Daxx and ATRX play a role in the regulation of viral chromatin. Taken together, our data demonstrate an important role of BNRF1 in supporting EBV early infection by interacting with Daxx and ATRX; and suggest that tegument disruption of PML-NB-associated antiviral resistances is a universal requirement for herpesvirus infection in the nucleus

3D Reconstruction of VZV Infected Cell Nuclei and PML Nuclear Cages by Serial Section Array Scanning Electron Microscopy and Electron Tomography

Varicella-zoster virus (VZV) is a human alphaherpesvirus that causes varicella (chickenpox) and herpes zoster (shingles). Like all herpesviruses, the VZV DNA genome is replicated in the nucleus and packaged into nucleocapsids that must egress across the nuclear membrane for incorporation into virus particles in the cytoplasm. Our recent work showed that VZV nucleocapsids are sequestered in nuclear cages formed from promyelocytic leukemia protein (PML) in vitro and in human dorsal root ganglia and skin xenografts in vivo. We sought a method to determine the three-dimensional (3D) distribution of nucleocapsids in the nuclei of herpesvirus-infected cells as well as the 3D shape, volume and ultrastructure of these unique PML subnuclear domains. Here we report the development of a novel 3D imaging and reconstruction strategy that we term Serial Section Array-Scanning Electron Microscopy (SSA-SEM) and its application to the analysis of VZV-infected cells and these nuclear PML cages. We show that SSA-SEM permits large volume imaging and 3D reconstruction at a resolution sufficient to localize, count and distinguish different types of VZV nucleocapsids and to visualize complete PML cages. This method allowed a quantitative determination of how many nucleocapsids can be sequestered within individual PML cages (sequestration capacity), what proportion of nucleocapsids are entrapped in single nuclei (sequestration efficiency) and revealed the ultrastructural detail of the PML cages. More than 98% of all nucleocapsids in reconstructed nuclear volumes were contained in PML cages and single PML cages sequestered up to 2,780 nucleocapsids, which were shown by electron tomography to be embedded and cross-linked by an filamentous electron-dense meshwork within these unique subnuclear domains. This SSA-SEM analysis extends our recent characterization of PML cages and provides a proof of concept for this new strategy to investigate events during virion assembly at the single cell level

The Intrinsic Antiviral Defense to Incoming HSV-1 Genomes Includes Specific DNA Repair Proteins and Is Counteracted by the Viral Protein ICP0

Cellular restriction factors responding to herpesvirus infection include the ND10 components PML, Sp100 and hDaxx. During the initial stages of HSV-1 infection, novel sub-nuclear structures containing these ND10 proteins form in association with incoming viral genomes. We report that several cellular DNA damage response proteins also relocate to sites associated with incoming viral genomes where they contribute to the cellular front line defense. We show that recruitment of DNA repair proteins to these sites is independent of ND10 components, and instead is coordinated by the cellular ubiquitin ligases RNF8 and RNF168. The viral protein ICP0 targets RNF8 and RNF168 for degradation, thereby preventing the deposition of repressive ubiquitin marks and counteracting this repair protein recruitment. This study highlights important parallels between recognition of cellular DNA damage and recognition of viral genomes, and adds RNF8 and RNF168 to the list of factors contributing to the intrinsic antiviral defense against herpesvirus infection