Thermodynamic properties of myo-inositol

© 2017 Elsevier Ltd In the present work, the temperature dependence of heat capacity of vitamin B8 (myo-inositol) has been measured for the first time over the range from 8 K to 340 K by precision adiabatic vacuum calorimetry. Based on the experimental data, the thermodynamic functions of the vitamin B8, namely, the heat capacity, enthalpy H°(T)–H°(0), entropy S°(T)–S°(0) and Gibbs function G°(T)–H°(0) have been determined for the range from T → 0 K to 340 K. The value of the fractal dimension D in the function of multifractal generalization of Debye's theory of the heat capacity of solids was estimated and the character of heterodynamics of structure was detected. The enthalpy of combustion (−2747.0 ± 2.1) kJ·mol−1 of the vitamin B8 was measured for the first time using high-precision combustion calorimeter. The standard molar enthalpy of formation in the crystalline state (−1329.3 ± 2.3) kJ·mol−1 of B8 at 298.15 K was derived from the combustion experiments. Using combination of the adiabatic and combustion calorimetry results the thermodynamic functions of formation of the myo-inositol at T = 298.15 K and p = 0.1 MPa have been calculated. The low-temperature X-ray diffraction was used for the determination of coefficients of thermal expansion

Comprehensive thermodynamic study of methylprednisolone

© 2016 Elsevier LtdIn the present work the temperature dependence of heat capacity for methylprednisolone has been measured for the first time over the temperature range from 6 to 350 K using by precision adiabatic vacuum calorimetry. Based on the experimental data, the thermodynamic functions of the methylprednisolone, namely, the heat capacity, enthalpy H°(T) − H°(0), entropy S°(T) − S°(0) and Gibbs function G°(T) − H°(0) have been evaluated from the experimental values for the range from T → 0 to 350 K. Standard molar enthalpy of combustion (−11898.9 ± 6.7) kJ·mol−1 of the methylprednisolone was measured for the first time using high-precision combustion calorimeter. The standard molar enthalpy of formation in the crystalline state (−1045.8 ± 7.3) kJ·mol−1 of compound at 298.15 K was derived from the combustion experiments. The standard molar enthalpy of sublimation at 298.15 K (194.5 ± 2.2) kJ·mol−1 was measured by using the quartz-crystal microbalance (QCM). Using combination of the adiabatic and combustion calorimetry with the result from QCM, the thermodynamic functions of the methylprednisolone at T = 298.15 K and p = 0.1 MPa have been calculated

Физико-химические и биологические свойства биоподобного и референтного препаратов тканевого активатора плазминогена

Recombinant tissue plasminogen activator (international nonproprietary name — alteplase) which was developed by «GENERIUM» (Russia) and received a marketing authorisation in Russia is completely analogous to Actilyse® which is used to treat medical conditions accompanied by thrombosis, such as acute myocardial infarction, pulmonary embolism, and ischemic stroke. The aim of the study was to carry out a comprehensive comparison of physico-chemical and biological properties of Revelyse® and the reference product Actilyse® in order to assess their biosimilarity. Materials and Methods: comparative peptide mapping and determination of comparability of chromatographic profiles of tryptic hydrolysates was performed using RP-HPLC and massspectrometry; the molecular weight distribution was determined by mass-spectrometry and polyacrylamide gel electrophoresis (Laemmli method). The purity and homogeneity of products as well as the content of related impurities (oligomers and fragments) were determined using gel filtration; N-glycosylation profile was analysed by hydrophilic HPLC, total sialic acid was quantified by the Svennerholm resorcinol method. Protein binding to fibrin and human fibrinogen was assessed by surface plasmon resonance, and the specific activity was compared by fibrin clot lysis. Results: the research demonstrated a complete overlap of the products’ peptide maps, which indicates the identity of аlteplase amino acid sequences in the two medicines being compared. The authors of the study also determined the molecular weight and the content of the intact single-stranded form of the protein, and quantified post-translational modifications, the content of sialic acids and neutral sugars. The analysis of the N-glycosylation profile revealed insignificant differences in the percentage of multiantenna complex glycans. The specificity of alteplase was evaluated by analysing the formation of protein complexes with natural alteplase ligands – fibrin and plasminogen activator inhibitor-1, but no significant differences were found. The comparison of specific activation of plasminogen fibrinolytic activity was performed based on the results of the assay analysing the fibrin clot lysis rate, and it demonstrated comparability of Revelyse® and Actilyse®. Conclusions: comparative experimental studies have shown no differences in the structure, charge distribution heterogeneity, impurities content, and specific activity of alteplase as a component of Revelyse® and the reference product Actilyse®, which leads to the conclusion that they are similar in terms of physicochemical and biological properties.Рекомбинантный тканевой активатор плазминогена (международное непатентованное название — алтеплаза), разработанный и зарегистрированный компанией АО «ГЕНЕРИУМ» (Россия), является полным аналогом лекарственного препарата Актилизе®, используемого для лечения заболеваний, сопровождающихся тромбообразованием, таких как инфаркт миокарда, тромбоэмболия легочной артерии и ишемический инсульт. Цель работы: провести комплексное исследование сопоставимости физико-химических и биологических характеристик лекарственного препарата Ревелиза® и референтного препарата Актилизе® для оценки их биоподобия. Материалы и методы: сравнительное пептидное картирование с определением сопоставимости хроматографических профилей триптических гидролизатов проводилось методами ОФ-ВЭЖХ и масс-спектрометрии, определение молекулярно-массового распределения — методами масс-спектрометрии и электрофореза в полиакриламидном геле по Леммли. Для определения чистоты и гомогенности препаратов, а также содержания родственных примесей (олигомеров, фрагментов) использовали метод гель-фильтрации; исследование профиля N-гликозилирования осуществляли методом гидрофильной ВЭЖХ, определение тотального содержания сиаловых кислот — методом Свеннерхольма. Оценку связывания белка с фибрином и фибриногеном человека проводили методом поверхностного плазмонного резонанса, сравнение специфической активности — методом лизиса фибринового сгустка. Результаты: в ходе исследований было продемонстрировано полное совпадение пептидных карт анализируемых препаратов, что свидетельствует об идентичности аминокислотной последовательности алтеплазы в составе сравниваемых препаратов. Также сравнительно были определены молекулярная масса и содержание интактной одноцепочечной формы белка в лекарственных препаратах, количественно охарактеризованы посттрансляционные модификации, содержание сиаловых кислот и нейтральных сахаров. При изучении профиля N-гликозилирования обнаружены незначительные различия в процентном содержании мультиантенных комплексных гликанов. Специфичность алтеплазы оценивали при образовании белковых комплексов с природными лигандами алтеплазы — фибрином и ингибитором активатора плазминогена первого типа, при этом достоверных различий обнаружено не было. При сравнении специфической активации фибринолитической активности плазминогена в тесте на скорость лизиса фибринового сгустка была продемонстрирована сопоставимость препаратов Ревелиза® и Актилизе®. Выводы: проведенные сравнительные экспериментальные исследования показали отсутствие различий в структуре, гетерогенности распределения зарядов, содержании примесей, а также специфической активности алтеплазы в составе лекарственного препарата Ревелиза® и референтного препарата Актилизе®, что позволяет сделать вывод о сопоставимости препаратов по физико-химическим и биологическим свойствам

Molecular Determinants and Dynamics of Hepatitis C Virus Secretion

The current model of hepatitis C virus (HCV) production involves the assembly of virions on or near the surface of lipid droplets, envelopment at the ER in association with components of VLDL synthesis, and egress via the secretory pathway. However, the cellular requirements for and a mechanistic understanding of HCV secretion are incomplete at best. We combined an RNA interference (RNAi) analysis of host factors for infectious HCV secretion with the development of live cell imaging of HCV core trafficking to gain a detailed understanding of HCV egress. RNAi studies identified multiple components of the secretory pathway, including ER to Golgi trafficking, lipid and protein kinases that regulate budding from the trans-Golgi network (TGN), VAMP1 vesicles and adaptor proteins, and the recycling endosome. Our results support a model wherein HCV is infectious upon envelopment at the ER and exits the cell via the secretory pathway. We next constructed infectious HCV with a tetracysteine (TC) tag insertion in core (TC-core) to monitor the dynamics of HCV core trafficking in association with its cellular cofactors. In order to isolate core protein movements associated with infectious HCV secretion, only trafficking events that required the essential HCV assembly factor NS2 were quantified. TC-core traffics to the cell periphery along microtubules and this movement can be inhibited by nocodazole. Sub-populations of TC-core localize to the Golgi and co-traffic with components of the recycling endosome. Silencing of the recycling endosome component Rab11a results in the accumulation of HCV core at the Golgi. The majority of dynamic core traffics in association with apolipoprotein E (ApoE) and VAMP1 vesicles. This study identifies many new host cofactors of HCV egress, while presenting dynamic studies of HCV core trafficking in infected cells

THE DEVELOPMENT OF A PRODUCTION SCALE TECHNOLOGY OF RECOMBINANT FOLLICLE STIMULATING HORMONE BIOPROCESS IN CHO CELL LINE

The article discusses the development of a production technique for recombinant follicle-stimulating hormone used in the treatment of infertility, in the culture of Chinese hamster ovary cells. As part of the creation of the technological platform receiving hormone the bioprocess in wave-induced motion bioreactor has been optimized. A number of parameters that influence on the bioprocess was studied, as follows: optimal cell density, concentration of metabolites, temperature shift and its duration

Analysis of cholic acid amides by high-performance liquid chromatography

The method of preparative purification of the obtained amides of cholic acid by HPLC was developed. The structure of the compounds was confirmed by 1H NMR spectroscopy and mass spectrometry. Purity of compounds was confi rmed by chromatography

INFORMATION INFRASTRUCTURE OF THE EDUCATIONAL ENVIRONMENT WITH VIRTUAL MACHINE TECHNOLOGY

Subject of research. Information infrastructure for the training environment with application of technology of virtual computers for small pedagogical systems (separate classes, author's courses) is created and investigated. Research technique. The life cycle model of information infrastructure for small pedagogical systems with usage of virtual computers in ARIS methodology is constructed. The technique of information infrastructure formation with virtual computers on the basis of process approach is offered. The model of an event chain in combination with the environment chart is used as the basic model. For each function of the event chain the necessary set of means of information and program support is defined. Technique application is illustrated on the example of information infrastructure design for the educational environment taking into account specific character of small pedagogical systems. Advantages of the designed information infrastructure are: the maximum usage of open or free components; the usage of standard protocols (mainly, HTTP and HTTPS); the maximum portability (application servers can be started up on any of widespread operating systems); uniform interface to management of various virtualization platforms, possibility of inventory of contents of the virtual computer without its start, flexible inventory management of the virtual computer by means of adjusted chains of rules. Approbation. Approbation of obtained results was carried out on the basis of training center "Institute of Informatics and Computer Facilities" (Tallinn, Estonia). Technique application within the course "Computer and Software Usage" gave the possibility to get half as much the number of refusals for components of the information infrastructure demanding intervention of the technical specialist, and also the time for elimination of such malfunctions. Besides, the pupils who have got broader experience with computer and software, showed better results within tests due to the usage of information infrastructure with virtual computers. Practical application. Obtained output can be recommended for introduction into the activity of educational institutions of secondary and higher education, and also for developing the author's and specialized courses focused on collective actions of pupils

PHARMACEUTICAL DEVELOPMENT OF FORMULATION BASED ON BIOPHARMACEUTICAL PREPARATIONS FOR GENE THERAPY

Biopharmaceuticals intended for gene therapy are mostly presented by supercoiled plasmid DNA obtained with bacteria cell lines. The properties of their formulations, including stability under different storage conditions, are barely studied. This article describes the typical pharmaceutical development of freeze-dried gene therapeutics. By means of process analytical technological methods the preparations formulated were controlled during storage. Furthermore, the composition of the formulations was optimized for prolonged storage