SYNCRIP Modulates the Epithelial-Mesenchymal Transition in Hepatocytes and HCC Cells

Heterogeneous nuclear ribonucleoproteins (hnRNPs) control gene expression by acting at multiple levels and are often deregulated in epithelial tumors; however, their roles in the fine regulation of cellular reprogramming, specifically in epithelial–mesenchymal transition (EMT), remain largely unknown. Here, we focused on the hnRNP-Q (also known as SYNCRIP), showing by molecular analysis that in hepatocytes it acts as a “mesenchymal” gene, being induced by TGFβ and modulating the EMT. SYNCRIP silencing limits the induction of the mesenchymal program and maintains the epithelial phenotype. Notably, in HCC invasive cells, SYNCRIP knockdown induces a mesenchymal–epithelial transition (MET), negatively regulating their mesenchymal phenotype and significantly impairing their migratory capacity. In exploring possible molecular mechanisms underlying these observations, we identified a set of miRNAs (i.e., miR-181-a1-3p, miR-181-b1-3p, miR-122-5p, miR-200a-5p, and miR-let7g-5p), previously shown to exert pro- or anti-EMT activities, significantly impacted by SYNCRIP interference during EMT/MET dynamics and gathered insights, suggesting the possible involvement of this RNA binding protein in their transcriptional regulation.

Vitamin D3 Treatment Alters Thyroid Functional Morphology in Orchidectomized Rat Model of Osteoporosis

Vitamin D plays an essential role in prevention and treatment of osteoporosis. Thyroid hormones, in addition to vitamin D, significantly contribute to regulation of bone remodeling cycle and health. There is currently no data about a possible connection between vitamin D treatment and the thyroid in the context of osteoporosis. Middle-aged Wistar rats were divided into: sham operated (SO), orchidectomized (Orx), and cholecalciferol-treated orchidectomized (Orx + Vit. D3; 5 µg/kg b.m./day during three weeks) groups (n = 6/group). Concentration of 25(OH)D in serum of the Orx + Vit. D3 group increased 4 and 3.2 times (p < 0.0001) respectively, compared to Orx and SO group. T4, TSH, and calcitonin in serum remained unaltered. Vit. D3 treatment induced changes in thyroid functional morphology that indicate increased utilization of stored colloid and release of thyroid hormones in comparison with hormone synthesis, to maintain hormonal balance. Increased expression of nuclear VDR (p < 0.05) points to direct, TSH independent action of Vit. D on thyrocytes. Strong CYP24A1 immunostaining in C cells suggests its prominent expression in response to Vit. D in this cell subpopulation in orchidectomized rat model of osteoporosis. The indirect effect of Vit. D on bone, through fine regulation of thyroid function, is small.

Molecular basis of transcriptional repression of anti-CRISPR by anti-CRISPR-associated 2

CRISPR–Cas systems are well known host defense mechanisms that are conserved in bacteria and archaea. To counteract CRISPR–Cas systems, phages and viruses have evolved to possess multiple anti-CRISPR (Acr) proteins that can inhibit the host CRISPR–Cas system via different strategies. The expression of acr genes is controlled by anti-CRISPR-associated (Aca) proteins that bind to an upstream promoter and regulate the expression of acr genes during transcription. Although the role of Aca as a transcriptional repressor has been demonstrated, the mechanism of action of Aca has not been determined. Here, the molecular mechanism underlying the Aca2-mediated transcriptional control of acr genes was elucidated by determining the crystal structure of Aca2 from Oceanimonas smirnovii at a high resolution of 1.92 Å. Aca2 forms a dimer in solution, and dimerization of Aca2 is critical for specific promoter binding. The promoter-binding strategy of dimeric Aca2 was also revealed by performing mutagenesis studies. The atomic structure of the Aca family shown in this study provides insights into the fine regulation of host defense and immune-escape mechanisms and also demonstrates the conserved working mechanism of the Aca family.

A discussion of S.M. Krylov’s book «Neocybernetics» (2008)

A comparative analysis of the “general formal technology (GFT)” by S. M. Krylov is carried out in the context of the published book of the authors “General Theory of Technologies and Microelectronics” (2020) and on the basis of his work of 2008. Despite the abstractness of the algebraic-algorithmic approach, Krylov offers a number of specific constructions that are in demand during the fourth industrial revolution and for the future development of industrial technology in nanoelectronics and biotechnology. Industrial technology is considered as a complex object of management, i.e., it is the object of study of the new discipline «neocybernetics». Although the foundations of this approach were laid in 1930s–1960s within the framework of logical and mathematical research, its expansion is inevitable when using self-organization processes to obtain functional supramolecular structures in technological processes of nanoelectronics (for example, DNA origami engineering). The issues of complexity quantification for a product itself (structure) and its manufacturing technology, or, according to Krylov, the complexity of technological automata, have become even more relevant than before. The theoretical issues of self-organization, the development of artificial life, and the creation of self-replicating technical systems also seem promising for solution. In our opinion, Krylov’s formal technology is an important “block” in the advancement of general theory of technologies (GTT) useful for describing the technology at the levels: operation, route, and process. We would like to encourage a wide range of readers to study the book and form a steady interest in general technological issues. The value of GTT and GFT extends beyond the sphere of technology and, in a narrow sense, factory production, but also into the area of «fine» regulation of physiology in biological objects and pharmacy, as well as into the problem field of cognitive sciences, psychology, and education. when the focus is on the personality structure and heterogeneous constructs «floating in the sea of the unconscious». Both S.M. Krylov and we demonstrate that the issues of industrial technology cannot be considered without abstract formalization and without reference to philosophy.

NAD + Metabolism in Cardiac Health, Aging, and Disease

Nicotinamide adenine dinucleotide (NAD + ) is a central metabolite involved in energy and redox homeostasis as well as in DNA repair and protein deacetylation reactions. Pharmacological or genetic inhibition of NAD + -degrading enzymes, external supplementation of NAD + precursors, and transgenic overexpression of NAD + -generating enzymes have wide positive effects on metabolic health and age-associated diseases. NAD + pools tend to decline with normal aging, obesity, and hypertension, which are all major risk factors for cardiovascular disease, and NAD + replenishment extends healthspan, avoids metabolic syndrome, and reduces blood pressure in preclinical models. In addition, experimental elevation of NAD + improves atherosclerosis, ischemic, diabetic, arrhythmogenic, hypertrophic, or dilated cardiomyopathies, as well as different modalities of heart failure. Here, we critically discuss cardiomyocyte-specific circuitries of NAD + metabolism, comparatively evaluate distinct NAD + precursors for their preclinical efficacy, and raise outstanding questions on the optimal design of clinical trials in which NAD + replenishment or supraphysiological NAD + elevations are assessed for the prevention or treatment of major cardiac diseases. We surmise that patients with hitherto intractable cardiac diseases such as heart failure with preserved ejection fraction may profit from the administration of NAD + precursors. The development of such NAD + -centered treatments will rely on technological and conceptual progress on the fine regulation of NAD + metabolism.

Human TNF-α Affects the Egg-laying Dynamics and Glucose Metabolism of Schistosoma Mansoni Adult Worms in Vitro

Several studies described the effect of human TNF-α on Schistosoma mansoni. It affects the worm’s development, metabolism, egg-laying, changes in the parasite´s gene expression and protein phosphorylation. Data available concerning the influence of hTNF-α on egg-laying are controversial and understanding the mechanism of egg-laying regulation is essential in combating schistosomiasis. We characterized the effects of in vitro treatment of S. mansoni adult worms with different doses of hTNF-α (5, 20 and 40ng/mL) for five days. We explored the effects on the egg-laying rate, glucose, ATP metabolism, mRNA expression levels of lactate dehydrogenase, of glucose transporters and of SmTNFR, the parasite gene for hTNF-α receptor. hTNF-α influenced egg-laying in a time and dose dependent manner: with 40ng/mL, egg-laying increased on day 2 and decreased on days 3 and 4; 20 ng/mL dose, egg-laying decreased on day 3, while at 5ng/mL dose, egg-laying decreased on day 4. The total number of eggs produced was not affected, but the egg-laying dynamic was altered; the median egg-laying time decreased significantly due to treatment. At 5 and 20ng/mL hTNF-alpha, lactate production diminished on days 3 up to 5, while glucose uptake increased on day 5. At 40ng/mL, glucose uptake diminished on days 1 up to 3, while ATP accumulation was detected on day 5. No significant changes in the mRNA expression were detected in all treatments. Crosstalk involving the hTNF-alpha and the parasite signaling play a role in the fine regulation of the worm´s metabolism and physiology and points to new strategies for disease control.

Variety of Non-Coding RNAs in Eukaryotic Genomes

The genomes of large multicellular eukaryotes mainly consist of DNA that encodes not proteins, but RNAs. The unexpected discovery of approximately the same number of protein genes in Homo sapiens and Caenorhabditis elegans led to the understanding that it is not the number of proteins that determines the complexity of the development and functioning of an organism. The phenomenon of pervasive transcription of genomes is finding more and more confirmation. Data are emerging on new types of RNA that work in different cell compartments, are expressed at different stages of development, in different tissues and perform various functions. Their main purpose is fine regulation of the main cellular processes. The presence of a rich arsenal of regulators that can interact with each other and work on the principle of interchangeability determines the physiological complexity of the organism and its ability to adapt to changing environmental conditions. An overview of the currently known functional RNAs expressed in eukaryotic genomes is presented here. There is no doubt that in the near future, using high-tech transcriptome technologies, many new RNAs will be identified and characterized. But it is likely that many of the expressed transcripts do not have a function, but are an evolutionary reserve of organisms.

Calpains are required for efficient microtubule detyrosination

Detyrosination is a major post-translational modification of microtubules (MT), which has significant impact on MT function in cell division, differentiation, growth, migration, polarity, and intracellular trafficking. Detyrosination of α-tubulin occurs via the recently identified complex of vasohibin 1/2 (vash1/2) and small vasohibin binding protein (SVBP). However, there is still remaining detyrosinating activity in the absence of vash1/2/SVBP, and little is known about the regulation of detyrosination. Using cellular and cell-free assays we showed that the calcium-dependent proteases calpains 1 and 2 regulate MT detyrosination. We identified new calpain cleavage sites in the N-terminal disordered region of vash1 using in vitro proteolysis followed by mass spectrometry. However, this cleavage did not affect the detyrosination activity of vasohibin. In conclusion, the regulation of MT detyrosination by calpains occurs via another yet unknown tubulin carboxypeptidase. Importantly, calpains' calcium dependency could allow a fine regulation of MT detyrosination. Thus, identifying the calpain-regulated pathway of MT detyrosination can be of major importance for several basic and clinical research and should be focused on in future studies.