Hyperoxia Causes Mitochondrial Fragmentation in Pulmonary Endothelial Cells by Increasing Expression of Pro-Fission Proteins

Objective—We explored mechanisms that alter mitochondrial structure and function in pulmonary endothelial cells (PEC) function after hyperoxia. Approach and Results—Mitochondrial structures of PECs exposed to hyperoxia or normoxia were visualized and mitochondrial fragmentation quantified. Expression of pro-fission or fusion proteins or autophagy-related proteins were assessed by Western blot. Mitochondrial oxidative state was determined using mito-roGFP. Tetramethylrhodamine methyl ester estimated mitochondrial polarization in treatment groups. The role of mitochondrially derived reactive oxygen species in mt-fragmentation was investigated with mito-TEMPOL and mitochondrial DNA (mtDNA) damage studied by using ENDO III (mt-tat-endonuclease III), a protein that repairs mDNA damage. Drp-1 (dynamin-related protein 1) was overexpressed or silenced to test the role of this protein in cell survival or transwell resistance. Hyperoxia increased fragmentation of PEC mitochondria in a time-dependent manner through 48 hours of exposure. Hyperoxic PECs exhibited increased phosphorylation of Drp-1 (serine 616), decreases in Mfn1 (mitofusion protein 1), but increases in OPA-1 (optic atrophy 1). Pro-autophagy proteins p62 (LC3 adapter–binding protein SQSTM1/p62), PINK-1 (PTEN-induced putative kinase 1), and LC3B (microtubule-associated protein 1A/1B-light chain 3) were increased. Returning cells to normoxia for 24 hours reversed the increased mt-fragmentation and changes in expression of pro-fission proteins. Hyperoxia-induced changes in mitochondrial structure or cell survival were mitigated by antioxidants mito-TEMPOL, Drp-1 silencing, or inhibition or protection by the mitochondrial endonuclease ENDO III. Hyperoxia induced oxidation and mitochondrial depolarization and impaired transwell resistance. Decrease in resistance was mitigated by mito-TEMPOL or ENDO III and reproduced by overexpression of Drp-1. Conclusions—Because hyperoxia evoked mt-fragmentation, cell survival and transwell resistance are prevented by ENDO III and mito-TEMPOL and Drp-1 silencing, and these data link hyperoxia-induced mt-DNA damage, Drp-1 expression, mt-fragmentation, and PEC dysfunction

A Methodological Approach to Integral Assessment of the Development Level of Small and Medium-Sized Enterprises in the Ukrainian Economy

The article states that the development of small and medium-sized enterprises (SMEs) is an important prerequisite for the stable functioning of the country’s economy, a source of providing jobs for its citizens, a guarantee of their financial well-being, the basis for the middle class formation. The aim of the article is to develop a methodological approach to integral assessment of the level of SME development in the Ukrainian economy. The article examines the existing methodological approaches to assessing the level of development and significance of SMEs and suggests a methodological approach based on the relevant system of indicators that can be calculated using available official statistical information and form the components of an integral indicator characterizing various aspects of the SME development in Ukraine (the development dynamics and significance for the economy). Using the proposed approach, an integral indicator for assessment of the level of development and significance of SMEs by economic activity (EA) and, in general, in the economy of Ukraine, as well as by its components, is calculated. On the basis of the conducted research, it is established that in 2012-2015 the majority of economic activities of the country had a disproportionate correlation between the dynamics of development and the significance of SMEs, which had a negative impact on the level of social and economic development of Ukraine as a whole

Методичний підхід до оцінки ефективності функціонування малого і середнього підприємництва в Україні

The importance of small and medium-sized enterprises (SMEs) for the socio-economic development of countries indicates the need to assess both the dynamics of their development and their performance. The purpose of the article is to elaborate methodological recommendations for assessing the performance of SMEs in the Ukrainian economy based on the calculation of the integral indicator, which will allow revealing the most influential types of economic activity and elaborating effective recommendations for the selection of priority directions in stimulating the SME development. A methodological approach to assessing performance of SMEs, which implies a comprehensive assessment and analysis of the indicators of return on enterprises’ assets, their liquidity, financial independence, as well as turnover of working capital of SMEs, has been proposed. On the basis of the elaborated approach, the assessment of the performance of SMEs by types of economic activity in whole in the Ukrainian economy has been carried out. A structural study of the dynamics of the integral indicator of SME performance in Ukraine has been conducted, and the types of economic activity that had a positive and negative influence on it have been determined. It has been proved that the performance indicators of SMEs were the most negatively influenced by the indicators of return on assets and financial independence.Важность малого и среднего предпринимательства (МСП) для социально-экономического развития стран указывает на необходимость оценки как динамики его развития, так и эффективности функционирования. Целью статьи является разработка методических рекомендаций по оценке эффективности функционирования МСП в экономике Украины на основе разработки интегрального показателя, что позволит выявить наиболее влиятельные виды экономической деятельности и разработать действенные рекомендации по выбору приоритетных направлений стимулирования развития МСП. Предложен методический подход к оценке эффективности функционирования МСП, который предусматривает комплексную оценку и анализ показателей рентабельности активов предприятий, их ликвидности, финансовой независимости и оборачиваемости оборотных средств предприятий МСП. На основе разработанного подхода осуществлена оценка эффективности функционирования МСП по видам экономической деятельности в целом по экономике Украины. Проведено структурное исследование динамики интегрального показателя эффективности функционирования МСП в Украине и определены виды экономической деятельности, которые осуществляли позитивное и негативное влияние на него. Доказано, что на показатели эффективности функционирования МСП, наиболее негативное влияние оказали показатели рентабельности активов и финансовойнезависимости.Важливість малого і середнього підприємництва (МСП) для соціально- економічного розвитку країн вказує на необхідність оцінки як динаміки його розвитку, так і ефективності функціонування. Метою статті є розробка методичних рекомендацій щодо оцінки ефективності функціонування МСП в економіці України на основі розробки інтегрального показника, що дозволить виявити найбільш впливові види економічної діяльності та розробити дієві рекомендації щодо вибору пріоритетних напрямів стимулювання розвитку МСП. Запропоновано методичний підхід до оцінки ефективності функціонування МСП, що передбачає комплексну оцінку та аналіз показників рентабельності активів підприємств, їх ліквідності, фінансової незалежності та оборотності обігових коштів підприємств МСП. На основі розробленого підходу здійснено оцінку ефективності функціонування МСП за видами економічної діяльності в цілому по економіці України. Проведено структурне дослідження динаміки інтегрального показника ефективності функціонування МСП в Україні та визначено види економічної діяльності, що здійснювали позитивний та негативний вплив на нього. Доведено, що на показники ефективності функціонування МСП, найбільш негативний вплив чинили показники рентабельності активів та фінансової незалежності

The New Methodology for Analyzing the Regulatory Impact of a Legal Act: Pros and Cons

Advantages and problems of the existing methodologies for analyzing the impact and monitoring of the effectiveness of a regulatory act (RIA) have been evaluated. The urgency of developing a methodological approach for evaluating the quality of the implementation of projects of normative legal acts by RIA regulatory authorities has been justified. There have been analyzed RIAs of the projects of normative legal acts developed by the regulatory authorities in March and April 2016 for their compliance with the new methodology. An algorithm of applying the methodological approach has been formed. The integral index has been proved and coefficients of the completeness of qualitative and quantitative evaluations have been determined. To carry out the assessment, the state and local regulatory authorities were selected. The conclusions as to the completeness and quality of the analyzed RIAs, as well as their compliance with the provisions of the new methodology have been made. Proposals to improve existing techniques have been developed

Hyperoxia Causes Mitochondrial Fragmentation in Pulmonary Endothelial Cells by Increasing Expression of Pro-Fission Proteins

Objective— We explored mechanisms that alter mitochondrial structure and function in pulmonary endothelial cells (PEC) function after hyperoxia. Approach and Results— Mitochondrial structures of PECs exposed to hyperoxia or normoxia were visualized and mitochondrial fragmentation quantified. Expression of pro-fission or fusion proteins or autophagy-related proteins were assessed by Western blot. Mitochondrial oxidative state was determined using mito-roGFP. Tetramethylrhodamine methyl ester estimated mitochondrial polarization in treatment groups. The role of mitochondrially derived reactive oxygen species in mt-fragmentation was investigated with mito-TEMPOL and mitochondrial DNA (mtDNA) damage studied by using ENDO III (mt-tat-endonuclease III), a protein that repairs mDNA damage. Drp-1 (dynamin-related protein 1) was overexpressed or silenced to test the role of this protein in cell survival or transwell resistance. Hyperoxia increased fragmentation of PEC mitochondria in a time-dependent manner through 48 hours of exposure. Hyperoxic PECs exhibited increased phosphorylation of Drp-1 (serine 616), decreases in Mfn1 (mitofusion protein 1), but increases in OPA-1 (optic atrophy 1). Pro-autophagy proteins p62 (LC3 adapter–binding protein SQSTM1/p62), PINK-1 (PTEN-induced putative kinase 1), and LC3B (microtubule-associated protein 1A/1B-light chain 3) were increased. Returning cells to normoxia for 24 hours reversed the increased mt-fragmentation and changes in expression of pro-fission proteins. Hyperoxia-induced changes in mitochondrial structure or cell survival were mitigated by antioxidants mito-TEMPOL, Drp-1 silencing, or inhibition or protection by the mitochondrial endonuclease ENDO III. Hyperoxia induced oxidation and mitochondrial depolarization and impaired transwell resistance. Decrease in resistance was mitigated by mito-TEMPOL or ENDO III and reproduced by overexpression of Drp-1. Conclusions— Because hyperoxia evoked mt-fragmentation, cell survival and transwell resistance are prevented by ENDO III and mito-TEMPOL and Drp-1 silencing, and these data link hyperoxia-induced mt-DNA damage, Drp-1 expression, mt-fragmentation, and PEC dysfunction. </jats:sec