Morphogenesis of post-Golgi transport carriers

The trans-Golgi network (TGN) is one of the main, if not the main, sorting stations in the process of intracellular protein trafficking. It is therefore of central importance to understand how the key players in the TGN-based sorting and delivery process, the post-Golgi carriers (PGCs), form and function. Over the last few years, modern morphological approaches have generated new insights into the questions of PGC biogenesis, structure and dynamics. Here, we present a view by which the “lifecycle” of a PGC consists of several distinct stages: the formation of TGN tubular export domains (where different cargoes are segregated from each other and from the Golgi enzymes); the docking of these tubular domains onto molecular motors and their extrusion towards the cell periphery along microtubules; the fission of the forming PGC from the donor membrane; and the delivery of the newly formed PGC to its specific acceptor organelle. It is now important to add the many molecular machineries that have been described as operating at the TGN to this “morphofunctional map” of the TGN export process

AAV-mediated transcription factor EB (TFEB) gene delivery ameliorates muscle pathology and function in the murine model of Pompe Disease

Pompe disease (PD) is a metabolic myopathy due to acid alpha-glucosidase deficiency and characterized by extensive glycogen storage and impaired autophagy. We previously showed that modulation of autophagy and lysosomal exocytosis by overexpression of the transcription factor EB (TFEB) gene was effective in improving muscle pathology in PD mice injected intramuscularly with an AAV-TFEB vector. Here we have evaluated the effects of TFEB systemic delivery on muscle pathology and on functional performance, a primary measure of efficacy in a disorder like PD. We treated 1-month-old PD mice with an AAV2.9-MCK-TFEB vector. An animal cohort was analyzed at 3 months for muscle and heart pathology. A second cohort was followed at different timepoints for functional analysis. In muscles from TFEB-treated mice we observed reduced PAS staining and improved ultrastructure, with reduced number and increased translucency of lysosomes, while total glycogen content remained unchanged. We also observed statistically significant improvements in rotarod performance in treated animals compared to AAV2.9-MCK-eGFP-treated mice at 5 and 8 months. Cardiac echography showed significant reduction in left-ventricular diameters. These results show that TFEB overexpression and modulation of autophagy result in improvements of muscle pathology and of functional performance in the PD murine model, with delayed disease progression

Abnormal cell-clearance and accumulation of autophagic vesicles in lymphocytes from patients affected with Ataxia-Teleangiectasia

Ataxia-Teleangiectasia (A-T) is a neurodegenerative disorder due to mutations in ATM gene. ATM in the nucleus ensures DNA repair, while its role in the cytosol is still poorly clarified. Abnormal autophagy has been documented in other neurodegenerative disorders, thus we evaluated whether alteration in this process may be involved in the pathogenesis of A-T by analyzing the autophagic vesicles and the genes implicated in the different stages of autophagy. Through transmission electron microscopy (TEM) and immunofluorescence analysis we observed an accumulation of APs associated with a LC3 puncta pattern, and a reduced number of ALs. We also documented an increased expression of genes involved in AP and lysosome biogenesis and function, and a decrease of Vps18 expression, involved in their vesicular trafficking and fusion. mTORC1-controlled proteins were hyperphosphorylated in A-T, in keeping with an increased mTOR inhibitory influence of autophagy. Betamethasone is able to promote the degradation of SQSTM1, a biomarker of autophagy. Collectively, our results indicate that in cells from A-T patients, the APs maturation is active, while the fusion between APs and lysosomes is inappropriate, thus implying abnormalities in the cell-clearance process. We also documented a positive effect of Betamethasone on molecules implicated in autophagosome degradation

Information and communication technologies in the development of women’s professional activity

The study describes the role of information and communication technologies (ICT), which have an impact on the development of professional activity of women. It is established that the problem of gender inequality is relevant because of the significant changes in modern society, in particular the rethinking of the roles of different categories of the population, including gender roles and status-role expectations. Based on the time-series analysis female ratio in the ICT organizations was calculated. It allowed to consider the real picture of the development of the analyzed sector with all the changes taking place, as well as to suggest the main priority areas that will promote the active entry of women into the digital economy. The study concluded that, despite the fact that the average number of female employees in ICT organizations is increasing, there is still a significant gender gap among those employed in the sector considered, which indicates that the potential of women in overcoming the acute shortage of IT professionals is not used. It has been established that reducing the digital gender gap in employment is not an end in itself, but the need to make greater use of the labor and intellectual potential of the entire employed population of the country - not only men, but also women. At the same time, the active involvement of women in the digital sector will increase the level of their professional activity and ensure their full participation in the modern labor market

Characterization of the most frequent ATP7B mutation causing Wilson disease in hepatocytes from patient induced pluripotent stem cells

H1069Q substitution represents the most frequent mutation of the copper transporter ATP7B causing Wilson disease in Caucasian population. ATP7B localizes to the Golgi complex in hepatocytes but moves in response to copper overload to the endo-lysosomal compartment to support copper excretion via bile canaliculi. In heterologous or hepatoma-derived cell lines, overexpressed ATP7B-H1069Q is strongly retained in the ER and fails to move to the post-Golgi sites, resulting in toxic copper accumulation. However, this pathogenic mechanism has never been tested in patients' hepatocytes, while animal models recapitulating this form of WD are still lacking. To reach this goal, we have reprogrammed skin fibroblasts of homozygous ATP7B-H1069Q patients into induced pluripotent stem cells and differentiated them into hepatocyte-like cells. Surprisingly, in HLCs we found one third of ATP7B-H1069Q localized in the Golgi complex and able to move to the endo-lysosomal compartment upon copper stimulation. However, despite normal mRNA levels, the expression of the mutant protein was only 20% compared to the control because of endoplasmic reticulum-associated degradation. These results pinpoint rapid degradation as the major cause for loss of ATP7B function in H1069Q patients, and thus as the primary target for designing therapeutic strategies to rescue ATP7B-H1069Q function

DUALISTIC LEGAL REGULATION OF ACQUIRED SUBJECTIVE HUMAN RIGHTS IN THE RUSSIAN FEDERATION

Purpose of the study: This article analyzes dualistic approaches to the determination of legal facts that grant citizens of the Russian Federation subjective rights. It is obvious that the concept of birth is still not specified in the existing Russian legislation. Different norms of the modern Russian legislation associate the legal fact of birth with the criterion of live birth. An embryo (fetus) cannot have a complex of general civil rights until the time of live birth. Methodology: The authors of the article have compared this constitutional directive with existing rules of other branches of Russian law and revealed a completely opposite situation. Main Findings: Of course, the existing discrepancies in certain legal acts that regulate the same or related social relations represent a classical legal conflict. Moreover, partial contradictions in the Russian legal system are caused by the fact that many sectoral norms conflict with the Constitution. Novelty/Originality: Legal certainty is a basic and system-forming principle that dialectically complements and develops other general-legal, intersectoral and sectoral principles, justifies their functional necessity, determines the accuracy and clarity of legal instructions, established order and limits their actions

Development of Human Resources Structure for Health Resort Facilities of the Republic of Crimea

The subject matter of this study focuses on the laws and mechanisms of employment in various sectors of the labor market of the Republic of Crimea. The article is devoted to the research of staffing structure for the development of business activities in the Republic of Crimea, in particular, for the health resort complex, to identify priority areas of regional employment policy.The main hypothesis of the research is as follows: there is a discrepancy between the basic parameters of the Crimean labor market transformation and the educational system that makes their proper interaction impossible, causing inconsistencies between the needs and demands of the labor market and the level of training (in particular, in the health resort complex) and the formal qualification of graduates of educational establishments.The methodological framework of the research is based on a systemic approach in studying the labor market of the Republic of Crimea and the use of statistical methods of analysis of the labor market conjuncture.The study analyzed the dynamics of socio-economic development of the Crimean region. The Crimean regional labor market conjuncture was studied, allowing the identification of the main causes of the imbalances in the development of the labor market. The analysis was made with regard to the qualifications of medical professionals graduating from educational establishments and groups of factors affecting the staffing structure were properly identified. The study of reasons of the Crimean labor market discrepancies showed that the current imbalance was caused by both objective and subjective reasons.Priority directions for employment policies have been suggested for the health resort complex. Their implementation will make the stabilization of the situation related to medical staff possible, including for the health resort complex. Such steps will also help improve the potential of human resources, capable of providing highly professional services to holidaymakers; contribute to the effective solution of problems of development of the Republic of Crimea under new economic conditions

Wilson Disease Protein ATP7B Utilizes Lysosomal Exocytosis to Maintain Copper Homeostasis

SummaryCopper is an essential yet toxic metal and its overload causes Wilson disease, a disorder due to mutations in copper transporter ATP7B. To remove excess copper into the bile, ATP7B traffics toward canalicular area of hepatocytes. However, the trafficking mechanisms of ATP7B remain elusive. Here, we show that, in response to elevated copper, ATP7B moves from the Golgi to lysosomes and imports metal into their lumen. ATP7B enables lysosomes to undergo exocytosis through the interaction with p62 subunit of dynactin that allows lysosome translocation toward the canalicular pole of hepatocytes. Activation of lysosomal exocytosis stimulates copper clearance from the hepatocytes and rescues the most frequent Wilson-disease-causing ATP7B mutant to the appropriate functional site. Our findings indicate that lysosomes serve as an important intermediate in ATP7B trafficking, whereas lysosomal exocytosis operates as an integral process in copper excretion and hence can be targeted for therapeutic approaches to combat Wilson disease