Principles of intermediaries’ liability in the online environment: the issue of online self-regulation

This article examines Internet self-regulation as an adjunct to network regulation. The paper analysed the elements of an integrated approach to be undertaken to enhance regulatory effectiveness. The paper analyses and investigates the aspect of self-regulation as a way to strengthen legal relations in the network, considering the preservation of the balance of interests of the individual, society and the state. The “Manila Principles for Intermediaries”, which is one of the multi-stakeholder approaches to strengthen self-regulatory practices, were analysed in detail. The principles reviewed highlighted a number of advantages that are contained in the current legislation in promoting self-regulation. The results of the analysis highlighted some critical success factors in Internet regulatory issues that are essential for further official recognition

The Translational Regulation of Lipoprotein Lipase by Epinephrine Involves an RNA Binding Complex Including the Catalytic Subunit of Protein Kinase A

The balance of lipid flux in adipocytes is controlled by the opposing actions of lipolysis and lipogenesis, which are controlled primarily by hormone-sensitive lipase and lipoprotein lipase (LPL), respectively. Catecholamines stimulate adipocyte lipolysis through reversible phosphorylation of hormone-sensitive lipase, and simultaneously inhibit LPL activity. However, LPL regulation is complex and previous studies have described translational regulation of LPL in response to catecholamines because of an RNA-binding protein that interacts with the 3′-untranslated region of LPL mRNA. In this study, we identified several protein components of an LPL RNA binding complex. Using an LPL RNA affinity column, we identified two of the RNA-binding proteins as the catalytic (C) subunit of cAMP-dependent protein kinase (PKA), and A kinase anchoring protein (AKAP) 121/149, one of the PKA anchoring proteins, which has known RNA binding activity. To determine whether the C subunit was involved in LPL translation inhibition, the C subunit was depleted from the cytoplasmic extract of epinephrine-stimulated adipocytes by immunoprecipitation. This resulted in the loss of LPL translation inhibition activity of the extract, along with decreased RNA binding activity in a gel shift assay. To demonstrate the importance of the AKAPs, inhibition of PKA-AKAP binding with a peptide competitor (HT31) prevented epinephrine-mediated inhibition of LPL translation. C subunit kinase activity was necessary for LPL RNA binding and translation inhibition, suggesting that the phosphorylation of AKAP121/149 or other proteins was an important part of RNA binding complex formation. The hormonal activation of PKA results in the reversible phosphorylation of hormone-sensitive lipase, which is the primary mediator of adipocyte lipolysis. These studies demonstrate a dual role for PKA to simultaneously inhibit LPL-mediated lipogenesis through inhibition of LPL translation

Alterations in Platelet Secretion Differentially Affect Thrombosis and Hemostasis

We genetically manipulated the major platelet vesicle-associated membrane proteins (VAMP2, VAMP3, and VAMP8) to create mice with varying degrees of disrupted platelet secretion. As previously shown, loss of VAMP8 reduced granule secretion, and this defect was exacerbated by further deletion of VAMP2 and VAMP3. VAMP2Δ3Δ8−/− platelets also had reduced VAMP7. Loss of VAMP2 and VAMP3 (VAMP2Δ3Δ) had a minimal impact on secretion when VAMP7 and VAMP8 were present. Integrin αIIbβ3 activation and aggregation were not affected, although spreading was reduced in VAMP2Δ3Δ8−/− platelets. Using these mice as tools, we asked how much secretion is needed for proper thrombosis and hemostasis in vivo. VAMP2Δ3Δ mice showed no deficiency, whereas VAMP8−/− mice had attenuated formation of occlusive thrombi upon FeCl3-induced arterial injury but no excessive bleeding upon tail transection. VAMP2Δ3Δ8−/− mice bled profusely and failed to form occlusive thrombi. Plasma-coagulation factors were normal in all of the strains, but phosphatidylserine exposure was reduced in VAMP2Δ3Δ and VAMP2Δ3Δ8−/− platelets. From our data, an ∼40% to 50% reduction in platelet secretion in vitro (dense and α granule) correlated with reduced occlusive thrombosis but no compromise in hemostasis. At a \u3e 50% reduction, thrombosis and hemostasis were defective in vivo. Our studies are the first systematic manipulation of platelet exocytic machinery to demonstrate a quantitative linkage between in vitro platelet secretion and hemostasis and thrombosis in vivo. The animals described will be invaluable tools for future investigations into how platelet secretion affects other vascular processes

Composition of Innovative Activity’ Parameters for a System of Analysis and Making Decision in the Innovations

Innovative activity determines the ability of an enterprise, region or country to occupy leading or profitable positions within the global economic system. To understand and interpret the situation of the country in the context of world innovative economic growth, it is necessary to develop a system of criteria and indicators for assessing innovation activity. The definition of the criteria will allow both an assessment of the existing situation and the effectiveness of the programs being conducted, so the research main purpose is to develop a system of indicators of innovation activity and to increase the effectiveness of government programs that are aimed to support and to stimulate innovation activities at the regional and national levels. Given the complexity, diversity and versatility of innovation and of functioning of modern economy, the most effective approach to solve this problem is the use of modern technologies, in particular, artificial intelligence to solve this problem. In this regard, in this article, based on the theoretical analysis of existing approaches to the definition and evaluation of innovation activity, a structured system of characteristics of innovation activity developed for the authors to prepare machine learning and to develop a system of analysis and making decision in the innovations’ sphere is proposed. The necessity to develop the characteristics of innovation activity includes both statistical and economic characteristics, as well as wider possibilities for processing large data with the use of neural network technologies, namely, analysis of weak signals from various sources, taking into account industry specific features and analysis of the dependence of innovation activity on Demand by local, regional or national populations for innovative products and services. This determines the multidimensionality of the set of characteristics of innovation activity, which is effectively solved with the help of neural network technologies, in particular, system of analysis and making decision

The Lipogenic Enzymes DGAT1, FAS, and LPL in Adipose Tissue: Effects of Obesity, Insulin Resistance, and TZD Treatment

Acyl-coenzyme A:diacylglycerol transferase (DGAT), fatty acid synthetase (FAS), and LPL are three enzymes important in adipose tissue triglyceride accumulation. To study the relationship of DGAT1, FAS, and LPL with insulin, we examined adipose mRNA expression of these genes in subjects with a wide range of insulin sensitivity (SI). DGAT1 and FAS (but not LPL) expression were strongly correlated with SI. In addition, the expression of DGAT1 and FAS (but not LPL) were higher in normal glucose-tolerant subjects compared with subjects with impaired glucose tolerance (IGT) (P \u3c 0.005). To study the effects of insulin sensitizers, subjects with IGT were treated with pioglitazone or metformin for 10 weeks, and lipogenic enzymes were measured in adipose tissue. After pioglitazone treatment, DGAT1 expression was increased by 33 ± 10% (P \u3c 0.05) and FAS expression increased by 63 ± 8% (P \u3c 0.05); however, LPL expression was not altered. DGAT1, FAS, and LPL mRNA expression were not significantly changed after metformin treatment. The treatment of mice with rosiglitazone also resulted in an increase in adipose expression of DGAT1 by 2- to 3-fold, as did the treatment of 3T3 F442A adipocytes in vitro with thiazolidinediones. These data support a more global concept suggesting that adipose lipid storage functions to prevent peripheral lipotoxicity

ОСОБЕННОСТИ ТЕМПЕРАМЕНТА СПОРТСМЕНОВ-ЛЕГКОАТЛЕТОВ В ЗАВИСИМОСТИ ОТ СПОРТИВНОЙ СПЕЦИАЛИЗАЦИИ

The purpose of the study was to study the individual typological characteristics of track and field athletes, depending on the specialization and level of sports qualifications. The study involved 30 athletes, a total of 30 elite athletes participated in the study; the average age of the respondents was 23.5 years, among them 21 athletes have had the titles of Honored Master of Sports, Master of Sports of International Degree, Master of Sport. To assess the individual typological characteristics of athletes, the method of   G. Aizenk was used. The results of the study showed that there are significant differences in the individual typological characteristics of track and field athletes in different types of specializations. It was revealed that among the athletes of speed and power athletics (short sprint, long jump, triple jump, high jump and pole vault), athletes of predominantly sanguine and choleric-melancholic temperament types predominate. Among the athletes of the speed–strength endurance group (400 m runners), there are athletes with a bright predominance of a mixed type of sanguine-choleric. In the group of athletes with a specialization in endurance (middle- and long-distance runners), athletes of a sanguine-phlegmatic type of temperament predominate in 75% of cases. The conducted research with the participation of successful and highly qualified athletes makes it possible to identify those individual typological features of athletes that can be considered as models during selecting athletes for various types of specializations.Цель исследования заключалась в изучении индивидуально-типологических особенностей спортсменов-легкоатлетов в зависимости от специализации и уровня спортивной квалификации. В исследовании участвовало 30 легкоатлетов, средний возраст респондентов 23,5 года, среди них  21 спортсмен,  имеющие звания змс, мсмк, мс, 9 человек мсмк, мс. Для оценки индивидуально-типологических особенностей спортсменов-легкоатлетов использовалась методика Г. Айзенка. Результаты исследования показали, что существуют  различия в индивидуально-типологических характеристиках спортсменов-легкоатлетов в разных видах специализаций. Выявлено, что среди спортсменов скоростно-силовых видов легкой атлетики (короткий спринт, прыжки в длину, тройной прыжок, прыжок в высоту и с шестом) преобладают спортсмены преимущественно сангвинистического и холерико-меланхолического типов темперамента. Среди спортсменов группы – скоростно-силовой выносливости (бегуны на 400 м) доминируют спортсмены с преобладанием смешанного типа- сангвинико-холеристического. В группе спортсменов  со специализацией на выносливость  (бегунов на средние и длинные дистанции) преобладают в 75% случаев спортсмены сангвинико-флегматического типа темперамента. Проведенное исследование с участием успешных и высококвалифицированных спортсменов легкоатлетов позволяет выделить те индивидуально-типологические особенности спортсменов, которые могут рассматриваться как модельные при отборе спортсменов на разные виды специализаций

SNARE-Dependent Membrane Fusion Initiates α-Granule Matrix Decondensation in Mouse Platelets

Platelet α-granule cargo release is fundamental to both hemostasis and thrombosis. Granule matrix hydration is a key regulated step in this process, yet its mechanism is poorly understood. In endothelial cells, there is evidence for 2 modes of cargo release: a jack-in-the-box mechanism of hydration-dependent protein phase transitions and an actin-driven granule constriction/extrusion mechanism. The third alternative considered is a prefusion, channel-mediated granule swelling, analogous to the membrane “ballooning” seen in procoagulant platelets. Using thrombin-stimulated platelets from a set of secretion-deficient, soluble N-ethylmaleimide factor attachment protein receptor (SNARE) mutant mice and various ultrastructural approaches, we tested predictions of these mechanisms to distinguish which best explains the α-granule release process. We found that the granule decondensation/hydration required for cargo expulsion was (1) blocked in fusion-protein-deficient platelets; (2) characterized by a fusion-dependent transition in granule size in contrast to a preswollen intermediate; (3) determined spatially with α-granules located close to the plasma membrane (PM) decondensing more readily; (4) propagated from the site of granule fusion; and (5) traced, in 3-dimensional space, to individual granule fusion events at the PM or less commonly at the canalicular system. In sum, the properties of α-granule decondensation/matrix hydration strongly indicate that α-granule cargo expulsion is likely by a jack-in-the-box mechanism rather than by gradual channel-regulated water influx or by a granule-constriction mechanism. These experiments, in providing a structural and mechanistic basis for cargo expulsion, should be informative in understanding the α-granule release reaction in the context of hemostasis and thrombosis

A Rab33b missense mouse model for Smith-McCort dysplasia shows bone resorption defects and altered protein glycosylation

Smith McCort (SMC) dysplasia is a rare, autosomal recessive, osteochondrodysplasia that can be caused by pathogenic variants in either RAB33B or DYM genes. These genes codes for proteins that are located at the Golgi apparatus and have a role in intracellular vesicle trafficking. We generated mice that carry a Rab33b disease-causing variant, c.136A>C (p.Lys46Gln), which is identical to that of members from a consanguineous family diagnosed with SMC. In male mice at 4 months of age, the Rab33b variant caused a mild increase in trabecular bone thickness in the spine and femur and in femoral mid-shaft cortical thickness with a concomitant reduction of the femoral medullary area, suggesting a bone resorption defect. In spite of the increase in trabecular and cortical thickness, bone histomorphometry showed a 4-fold increase in osteoclast parameters in homozygous Rab33b mice suggesting a putative impairment in osteoclast function, while dynamic parameters of bone formation were similar in mutant versus control mice. Femur biomechanical tests showed an increased in yield load and a progressive elevation, from WT to heterozygote to homozygous mutants, of bone intrinsic properties. These findings suggest an overall impact on bone material properties which may be caused by disturbed protein glycosylation in cells contributing to skeletal formation, supported by the altered and variable pattern of lectin staining in murine and human tissue cultured cells and in liver and bone murine tissues. The mouse model only reproduced some of the features of the human disease and was sex-specific, manifesting in male but not female mice. Our data reveal a potential novel role of RAB33B in osteoclast function and protein glycosylation and their dysregulation in SMC and lay the foundation for future studies

Genetic landscape in Russian patients with familial left ventricular noncompaction

BackgroundLeft ventricular noncompaction (LVNC) cardiomyopathy is a disorder that can be complicated by heart failure, arrhythmias, thromboembolism, and sudden cardiac death. The aim of this study is to clarify the genetic landscape of LVNC in a large cohort of well-phenotyped Russian patients with LVNC, including 48 families (n=214).MethodsAll index patients underwent clinical examination and genetic analysis, as well as family members who agreed to participate in the clinical study and/or in the genetic testing. The genetic testing included next generation sequencing and genetic classification according to ACMG guidelines.ResultsA total of 55 alleles of 54 pathogenic and likely pathogenic variants in 24 genes were identified, with the largest number in the MYH7 and TTN genes. A significant proportion of variants −8 of 54 (14.8%) −have not been described earlier in other populations and may be specific to LVNC patients in Russia. In LVNC patients, the presence of each subsequent variant is associated with increased odds of having more severe LVNC subtypes than isolated LVNC with preserved ejection fraction. The corresponding odds ratio is 2.77 (1.37 −7.37; p <0.001) per variant after adjustment for sex, age, and family.ConclusionOverall, the genetic analysis of LVNC patients, accompanied by cardiomyopathy-related family history analysis, resulted in a high diagnostic yield of 89.6%. These results suggest that genetic screening should be applied to the diagnosis and prognosis of LVNC patients

Biological Earth observation with animal sensors

Space-based tracking technology using low-cost miniature tags is now delivering data on fine-scale animal movement at near-global scale. Linked with remotely sensed environmental data, this offers a biological lens on habitat integrity and connectivity for conservation and human health; a global network of animal sentinels of environmen-tal change