39 research outputs found
Invariant amino acids essential for decoding function of polypeptide release factor eRF1
In eukaryotic ribosome, the N domain of polypeptide release factor eRF1 is involved in decoding stop signals in mRNAs. However, structure of the decoding site remains obscure. Here, we specifically altered the stop codon recognition pattern of human eRF1 by point mutagenesis of the invariant Glu55 and Tyr125 residues in the N domain. The 3D structure of generated eRF1 mutants was not destabilized as demonstrated by calorimetric measurements and calculated free energy perturbations. In mutants, the UAG response was most profoundly and selectively affected. Surprisingly, Glu55Arg mutant completely retained its release activity. Substitution of the aromatic ring in position 125 reduced response toward all stop codons. This result demonstrates the critical importance of Tyr125 for maintenance of the intact structure of the eRF1 decoding site. The results also suggest that Tyr125 is implicated in recognition of the 3d stop codon position and probably forms an H-bond with Glu55. The data point to a pivotal role played by the YxCxxxF motif (positions 125–131) in purine discrimination of the stop codons. We speculate that eRF1 decoding site is formed by a 3D network of amino acids side chains
СОДЕРЖАНИЕ ЭССЕНЦИАЛЬНЫХ И УСЛОВНО ЭССЕНЦИАЛЬНЫХ ХИМИЧЕСКИХ ЭЛЕМЕНТОВ В ИНТРАОПЕРАЦИОННЫХ БИОПТАТАХ ДЕТЕЙ С ВРОЖДЕННЫМИ ПОРОКАМИ СЕРДЦА И АРИТМИЕЙ
Background. Despite recent numerous studies, etiopathogenesis, treatment and rehabilitation of children with heart rhythm disorders haven’t been studied well. We paid attention to the significant impact of mineral nutrients on cardiac activity, while addressing to the viable solutions. Aim. To measure the levels of essential and conditionally essential mineral nutrients and to determine any relationships between their concentrations in hair and in the intraoperative biopsy specimens obtained from children with congenital heart disease and heart rhythm disorders. Methods 55 children (34 boys and 21 girls) aged 6 to 17 years with different heart rhythm disorders and congenital heart disease were included in the study. Levels of 15 essential mineral nutrients (calcium, potassium, magnesium, sodium, phosphorus, sulfur, chromium, copper, iron, iodine, cobalt, manganese, molybdenum, selenium, zinc) and 3 conditionally essential nutrients (boron, silicon, vanadium) were measured in hair and in the intraoperative biopsy specimens obtained from children with congenital heart disease using inductively coupled plasma atomic emission spectrometry and electrothermal atomization atomic absorption spectrometry. The results of the study were processed using variational and alternative statistic methods with the commercially available software “MedStat”. Results The lack of essential nutrients (K, Mn, Se, Cr, P, Co, S, Cu, Na, Mo) and conditionally essential mineral, Si, in the intraoperative biopsy specimens of the heart and great arteries has been found. The direct strong correlation between the levels of essential (K, Mn, Se, Cr, Co) and conditionally essential (Si) minerals in hair and heart tissues of children with congenital heart disease and heart rhythm disorders has been determined. Conclusion The deficit of essential (potassium, manganese, selenium, chromium, phosphorus, cobalt, sulfur, copper, sodium, molybdenum) and conditionally essential (silicon) mineral nutrients, and the direct strong correlation between their levels in hair and heart tissues of children with congenital heart disease and heart rhythm disorders allows using hair as a biosubstrate, which is highly informative for the measurement of nutrients in the human body. Актуальность. Несмотря на многочисленность проводимых исследований, особенно в последнее десятилетие, до настоящего времени ряд аспектов этиопатогенеза, лечения и реабилитации детей с нарушением ритма сердца (НРС) остаются не изученными. В поиске возможных путей решения ряда задач нами было обращено внимание на существенную значимость химических элементов (ХЭ) в обеспечении деятельности сердечно-сосудистой системы. Цель. Оценить содержание и выявить взаимосвязь между концентрацией эссенциальных и условно эссенциальных химических элементов в волосах и в интраоперационных биоптатах детей с ВПС и аритмией. Материалы и методы Обследованы 55 детей (34 мальчика и 21 девочка) в возрасте от 6 до 17 лет с различными видами НРС и ВПС. Определено содержание 15 эссенциальных ХЭ (кальций, калий, магний, натрий, фосфор, сера, хром, медь, железо, йод, кобальт, марганец, молибден, селен, цинк) и 3 условно эссенциальных ХЭ (бор, кремний, ванадий) в волосах и в интраоперационных биоптатах детей с ВПС методами атомно-эмиссионной спектрометрии в индуктивно-связанной плазме и атомно-абсорбционной спектрометрии с электротермической атомизацией. Обработку результатов исследования проводили методами вариационной и альтернативной статистики с использованием лицензионного программного пакета «MedStat». Результаты. По результатам проведенного спектрального многоэлементного анализа интраоперационных биоптатов тканей сердца и магистральных сосудов выявлен дефицит эссенциальных ХЭ (K, Mn, Se, Cr, P, Со, S, Cu, Nа, Mo) и условно эссенциального микроэлемента Si. Доказана прямая сильная корреляционная зависимость между содержанием эссенциальных ХЭ (K, Mn, Se, Cr, Co) и условно эссенциального ХЭ Si в волосах и тканях сердца у детей с ВПС и НРС. Заключение. Выявленный дефицит эссенциальных ХЭ калия, марганца, селена, хрома, фосфора, кобальта, серы, меди, натрия, молибдена и условно эссенциального микроэлемента кремния, доказанная прямая сильная корреляционная зависимость между содержанием калия, марганца, селена, хрома, кобальта и кремния в волосах и тканях сердца у детей с ВПС и НРС подтверждает информативность использования данного биосубстрата для оценки содержания ХЭ в организме
Информационно-образовательная среда как средство освоения компетенций в ординатуре по специальностям «Педиатрия» и «Детская кардиология»
Introduction. At present, in connection with the development of new state educational standards in the system of higher medical education of the Donetsk People»s Republic, including at the postgraduate level, a competency-based approach is being widely introduced. Purpose setting. To familiarize teachers with the peculiarities of mastering competencies in residency in the specialties «Pediatrics» and «Pediatric Cardiology» at the Department of Pediatrics no. 3 of the Faculty of Internship and Postgraduate Education of the Donetsk State Medical University named after M. Gorky using remote technologies. Methodology and methods of the study. At the Department of Pediatrics no. 3, postgraduate training programs have been developed, including for the training of highly qualified personnel in residency in the specialties «Pediatrics» and «Pediatric Cardiology». The purpose of the residency is to train a qualified pediatrician or pediatric cardiologist who has a system of theoretical knowledge and professional competencies, capable and ready for independent professional activity, using modern scientific and technological achievements in the diagnosis and treatment of major diseases in children and adolescents. Results. Teachers of the Department of Pediatrics no. 3 have created selfstudy courses for pediatric residents and pediatric cardiologists on the university»s remote website. Each course contains general information, lectures and materials to prepare for practical exercises. The use of the information and educational environment stimulated the independent purposeful work of a pediatric intern or a pediatric cardiologist intern to form knowledge and further master all the competencies necessary for successful professional activity. Conclusion. The effectiveness of distance learning depends on the methodologically competently structured by the teacher of the educational process and the information and communication capabilities of the higher education institution.Введение. В настоящее время в связи с разработкой новых государственных образовательных стандартов в системе высшего медицинского образования Донецкой Народной Республики, в том числе на последипломном уровне, широко внедряется компетентностный подход. Постановка задачи. Цель – ознакомить преподавателей с особенностями освоения компетенций в ординатуре по специальностям «Педиатрия» и «Детская кардиология» на кафедре педиатрии № 3 факультета интернатуры и последипломного образования ФГБОУ ВО «Донецкий государственный медицинский институт им. М. Горького» с использованием дистанционных технологий. Методика и методология исследования. На кафедре педиатрии № 3 разработаны программы последипломного обучения, в том числе для подготовки кадров высшей квалификации в ординатуре по специальностям «Педиатрия» и «Детская кардиология». Целью ординатуры является подготовка квалифицированного врача-педиатра либо врача-детского кардиолога, обладающего системой теоретических знаний и профессиональных компетенций, способного и готового для самостоятельной профессиональной деятельности, применяющего современные научно-технические достижения диагностики и лечения основных заболеваниях у детей и подростков. Результаты. Преподавателями кафедры педиатрии № 3 на дистанционном сайте университета созданы курсы самостоятельного обучения для ординаторов-педиатров и ординаторов-детских кардиологов. Каждый курс содержит общую информацию, лекции и материалы для подготовки к практическим занятиям. Применение информационно-образовательной среды стимулирует самостоятельную целенаправленную работу ординатора-педиатра либо ординатора-детского кардиолога по формированию знаний и в дальнейшем освоению всех компетенций, необходимых для успешной профессиональной деятельности. Выводы. Эффективность дистанционного обучения зависит от методологически грамотно структурированного преподавателем учебного процесса и информационно-коммуникационных возможностей высшего учебного заведения
Eardrum perforation treatment blood plasma enriched with platelet growth factors
Introduction. The problem of the development of acute post-traumatic otitis media has not lost its relevance. Long-term preservation of the defect of the tympanic membrane contributes to the development of chronic inflammation in the middle ear and a deterioration in the quality of life of patients.Objective. To study the effect of blood plasma enriched with platelet growth factors on the regeneration of the tympanic membrane tissues in patients with acute post-traumatic perforation.Materials and methods. Patients with acute post-traumatic perforation of the tympanic membrane were divided into the main (24 people) and control (19 people) groups. Patients of the main group in the area of perforation of the tympanic membrane were injected once by application with a clot of autoplasma, enriched with platelet growth factors. The control group patients underwent dynamic observation of the processes of natural regeneration of the tympanic membrane tissues. On the 5, 10 and 15th days, the tissue regeneration of the tympanic membranes was assessed subjectively (patient complaints, audiometry) and objectively (otoscopy with the calculation of the dynamics of the relative perforation area, otoacoustic emission).Results. A single application of blood plasma enriched with platelet growth factors to the perforation area was significantly more often accompanied by closure of the tympanic membrane perforation (p ≤ 0.01) and a decrease in the average relative area of the tympanic membrane perforation on days 10 and 15 of the study (p ≤ 0.01). The dynamics of the average relative area of the tympanic membrane perforation reflected a high rate of closure of the tympanic membrane defect in the main group. Better healing of the tympanic membrane in patients of the main group was accompanied by an improvement in the auditory function of the affected ear. The quality of hearing was, on average, significantly better in the patients of the main group than in the patients of the control group, both on the 10th and 15th days of observation (p ≤ 0.01).Conclusion. Clinical use of blood plasma enriched with platelet growth factors has a beneficial effect on the rate, intensity of closure of the tympanic membrane perforation and hearing
GTP-dependent structural rearrangement of the eRF1:eRF3 complex and eRF3 sequence motifs essential for PABP binding
Translation termination in eukaryotes is governed by the concerted action of eRF1 and eRF3 factors. eRF1 recognizes the stop codon in the A site of the ribosome and promotes nascent peptide chain release, and the GTPase eRF3 facilitates this peptide release via its interaction with eRF1. In addition to its role in termination, eRF3 is involved in normal and nonsense-mediated mRNA decay through its association with cytoplasmic poly(A)-binding protein (PABP) via PAM2-1 and PAM2-2 motifs in the N-terminal domain of eRF3. We have studied complex formation between full-length eRF3 and its ligands (GDP, GTP, eRF1 and PABP) using isothermal titration calorimetry, demonstrating formation of the eRF1:eRF3:PABP:GTP complex. Analysis of the temperature dependence of eRF3 interactions with G nucleotides reveals major structural rearrangements accompanying formation of the eRF1:eRF3:GTP complex. This is in contrast to eRF1:eRF3:GDP complex formation, where no such rearrangements were detected. Thus, our results agree with the established active role of GTP in promoting translation termination. Through point mutagenesis of PAM2-1 and PAM2-2 motifs in eRF3, we demonstrate that PAM2-2, but not PAM2-1 is indispensible for eRF3:PABP complex formation
Selectivity of stop codon recognition in translation termination is modulated by multiple conformations of GTS loop in eRF1
Translation termination in eukaryotes is catalyzed by two release factors eRF1 and eRF3 in a cooperative manner. The precise mechanism of stop codon discrimination by eRF1 remains obscure, hindering drug development targeting aberrations at translation termination. By solving the solution structures of the wild-type N-domain of human eRF1 exhibited omnipotent specificity, i.e. recognition of all three stop codons, and its unipotent mutant with UGA-only specificity, we found the conserved GTS loop adopting alternate conformations. We propose that structural variability in the GTS loop may underline the switching between omnipotency and unipotency of eRF1, implying the direct access of the GTS loop to the stop codon. To explore such feasibility, we positioned N-domain in a pre-termination ribosomal complex using the binding interface between N-domain and model RNA oligonucleotides mimicking Helix 44 of 18S rRNA. NMR analysis revealed that those duplex RNA containing 2-nt internal loops interact specifically with helix α1 of N-domain, and displace C-domain from a non-covalent complex of N-domain and C-domain, suggesting domain rearrangement in eRF1 that accompanies N-domain accommodation into the ribosomal A site
Personally oriented approach as a condition for the development of competencies of foreign students during education at the department of therapeutic dentistry
Новая, личностно ориентированная концепция образования предполагает формирование личности с собственными целями, содержанием, технологиями. В современной высшей школе по-прежнему превалирует традиционный тип образования с ориентацией на знания. Компетентности обозначаются, как способности личности осуществлять сложные виды действий. На кафедре терапевтической стоматологии сформировать компетентность у студентов-иностранцев позволяет освоение четырех типов – опыт познавательной деятельности, опыт осуществления известных способов деятельности, опыт творческой деятельности, опыт осуществления эмоционально-ценностных отношений;
Нова, особистісно орієнтована концепція освіти передбачає формування особистості з власними цілями, змістом, технологіями. У сучасній вищій школі як і раніше превалює традиційний тип освіти з орієнтацією на знання. Компетентності позначаються, як здатності особистості здійснювати складні види дій. На кафедрі терапевтичної стоматології сформувати компетентність у студентівіноземців дозволяє освоєння чотирьох типів - досвід пізнавальної діяльності, досвід здійснення відомих способів діяльності, досвід творчої діяльності, досвід здійснення емоційно-ціннісних відносин;
New, student-oriented education concept involves the formation of identity with their own goals, content and technologies. In modern higher education still prevailing traditional type of education with a orientation on knowledge. Competencies identified as the individual's ability to perform out complex types of actions. At the Department of dentistry form competence of international students allows development of four types of experience - the experience of cognitive activity, the experience of the known methods of work, the experience of creativity, experience of emotional and value relations
Modern approaches to assessing the quality of life in children with arrhythmias
The paper presents the special questionnaire «Quality of Life in Children with Arrhythmias» that reflects the subjective assessment of quality of life (QOL) in patients with cardiac arrhythmias and a procedure for the comprehensive assessment of QOL in children with arrhythmias on the basis of subjective and objective data. It gives the results of using the above procedures for assessing QOL in 151 children (71 girls and 80 boys) aged 6–18 years with various cardiac arrhythmias. Complaints were absent in 21,9±3,4% of the children with arrhythmia and were nonspecific, being rarely observed, in 29,8±3,7%, suggesting that there was no reduction in QOL according to their subjective assessments. Comprehensive assessment of QOL showed that its indicators were significantly lower (p<0,05) than those obtained by subjective assessments. Thus, cardiac arrhythmias accompanied by a varying decrease in QOL were seen in 84,8±2,9% of the children, as shown by its comprehensive assessment and in 48,3±4,1% of the patients, as evidenced by their subjective assessments. Patients with organic heart disease and/or signs of chronic heart failure exhibited a decrease in QOL significantly more often (p<0,001). The leading causes of worse QOL were health changes in 56,3±4,0% of the patients, psychoemotional disorders in 77,5±3,4%, autonomic dysfunction in 81,5±3,2%, reduced adjustment in 82,8±3,1%, and pathological changes in cardiovascular function in 84,1±3,0%, as indicated by ECG, 24-hour ECG monitoring, and Doppler echocardiography
МНЕНИЕ ЮРИСТА В СОВРЕМЕННОМ ГРАЖДАНСКОМ ПРАВЕ
Purpose to define value of freedom and fight of opinions, views and lawyer’s positions in development of science of civil law.Methodology theoretical analysis, inductive and deductive methods.Results It is established that fight of opinions, collision of various positions allows to understand more deeply the discussed problem, to come nearer to truth. So, free expression of opinions is the engine of development of civil law.Practical implications introduction in educational process on disciplines of civil jurisprudence, further research of fight of opinions in civil law.Tendencies of the present stage of development of legal system are characterized by aspiration to fix in the Russian legal system of the beginning of private law, where at the head of a corner – people as a legal entity. Opinions of lawyers on various legal problems, and the attitudes towards these opinions are subject to considerable dynamics. The centuries-old history of development of the right showed that fight of opinions, collision of various positions allows to understand more deeply the discussed problem, to come nearer to truth. The modern civil law widely uses a method of comparative jurisprudence, studying experience of the civilized countries which promoted in development of the civil legislation.Цель: определить значение свободы и борьбы мнений, взглядов и позиций юриста в развитии науки гражданского права.Метод или методология проведения работы: теоретический анализ, индуктивные и дедуктивные методы.Результаты: Установлено, что борьба мнений, столкновение различных позиций позволяет глубже понять обсуждаемую проблему, приблизиться к истине. А значит, свободное выражение мнений есть двигатель развития цивилистики.Область применения результатов: внедрение в учебный процесс по дисциплинам гражданского правоведения, дальнейшее исследование борьбы мнений в цивилистике.Тенденции современного этапа развития правовой системы характеризуются стремлением закрепить в российской правовой системе начала частного права, где во главе угла – человек как субъект права. Мнения юристов на различные правовые проблемы, так и отношения к этим мнениям подвержены значительной динамике. Многовековая история развития права показала, что борьба мнений, столкновение различных позиций позволяет глубже понять обсуждаемую проблему, приблизиться к истине. Современная цивилистика широко использует метод сравнительного правоведения, изучая опыт цивилизованных стран, продвинувшихся в развитии гражданского законодательства
Mastocytosis in Children: Literature Review and Own Clinical Observation
The review of literature presents the data on neoplastic disease — mastocytosis. We provide the modern information of the etiopathogenesis, classification, clinical presentation, methods of diagnosis and treatment in children. We presented own experience of 20-month follow-up of female patient with cutaneous mastocytosis