Circular RNA AFF4 modulates osteogenic differentiation in BM-MSCs by activating SMAD1/5 pathway through miR-135a-5p/FNDC5/Irisin axis

Bone marrow-derived mesenchymal stem cells (BM-MSCs), the common progenitor cells of adipocytes and osteoblasts, have been recognized as the key mediator during bone formation. Herein, our study aim to investigate molecular mechanisms underlying circular RNA (circRNA) AFF4 (circ_AFF4)-regulated BM-MSCs osteogenesis. BM-MSCs were characterized by FACS, ARS, and ALP staining. Expression patterns of circ_AFF4, miR-135a-5p, FNDC5/Irisin, SMAD1/5, and osteogenesis markers, including ALP, BMP4, RUNX2, Spp1, and Colla1 were detected by qRT-PCR, western blot, or immunofluorescence staining, respectively. Interactions between circ_AFF4 and miR-135a-5p, FNDC5, and miR-135a-5p were analyzed using web tools including TargetScan, miRanda, and miRDB, and further confirmed by luciferase reporter assay and RNA pull-down. Complex formation between Irisin and Integrin αV was verified by Co-immunoprecipitation. To further verify the functional role of circ_AFF4 in vivo during bone formation, we conducted animal experiments harboring circ_AFF4 knockdown, and born samples were evaluated by immunohistochemistry, hematoxylin and eosin, and Masson staining. Circ_AFF4 was upregulated upon osteogenic differentiation induction in BM-MSCs, and miR-135a-5p expression declined as differentiation proceeds. Circ_AFF4 knockdown significantly inhibited osteogenesis potential in BM-MSCs. Circ_AFF4 stimulated FNDC5/Irisin expression through complementary binding to its downstream target molecule miR-135a-5p. Irisin formed an intermolecular complex with Integrin αV and activated the SMAD1/5 pathway during osteogenic differentiation. Our work revealed that circ_AFF4, acting as a sponge of miR-135a-5p, triggers the promotion of FNDC5/Irisin via activating the SMAD1/5 pathway to induce osteogenic differentiation in BM-MSCs. These findings gained a deeper insight into the circRNA-miRNA regulatory system in the bone marrow microenvironment and may improve our understanding of bone formation-related diseases at physiological and pathological levels.Subject terms: Stem cells, Diseases     

Hydrogen sulfide:A novel component in Arabidopsis peroxisomes which triggers catalase inhibition

Plant peroxisomes have the capacity to generate different reactive oxygen and nitrogen species(ROS and RNS),such as H_2O_2,superoxide radical(O_2~-),nitric oxide and peroxynitrite(ONOO~-).These organelles have an active nitrooxidative metabolism which can be exacerbated by adverse stress conditions.Hydrogen sulfide(H_2S)is a new signaling gasotransmitter which can mediate the posttranslational modification(PTM)persulfidation.We used Arabidopsis thaliana transgenic seedlings expressing cyan fluorescent protein(CFP)fused to a canonical peroxisome targeting signal 1(PTS1)to visualize peroxisomes in living cells,as well as a specific fluorescent probe which showed that peroxisomes contain H_2S.H_2S was also detected in chloroplasts under glyphosate-induced oxidative stress conditions.Peroxisomal enzyme activities,including catalase,photorespiratory H_2O_2-generating glycolate oxidase(GOX)and hydroxypyruvate reductase(HPR),were assayed in vitro with a H_2S donor.In line with the persulfidation of this enzyme,catalase activity declined significantly in the presence of the H_2S donor.To corroborate the inhibitory effect of H_2S on catalase activity,we also assayed pure catalase from bovine liver and pepper fruit-enriched samples,in which catalase activity was inhibited.Taken together,these data provide evidence of the presence of H_2S in plant peroxisomes which appears to regulate catalase activity and,consequently,the peroxisomal H_2O_2 metabolism.     

The influence of colonizing methylobacteria on morphogenesis and resistance of sugar beet and white cabbage plants to <Emphasis Type="Italic">Erwinia carotovora</Emphasis>

The influence of colonization of sugar beet (Beta vulgaris var. saccharifera (Alef) Krass) and white cabbage (Brassica oleracea var. capitata L.) plants by methylotrophic bacteria Methylovorus mays on the growth, rooting, and plant resistance to phytopathogen bacteria Erwinia carotovora was investigated. The colonization by methylobacteria led to their steady association with the plants which had increased growth speed, root formation and photosynthetic activity. The colonized plants had increased resistance to Erwinia carotovora phytopathogen and were better adapted to greenhouse conditions. The obtained results showed the perspectives for the practical implementation of methylobacteria in the ecologically clean microbiology substances used as the plant growth stimulators and for the plant protection from pathogens.     

Influence of mutual arrangement of the electric dipole and the spatial nonuniformity of brain electrical conductivity on the solution of the direct task of electroencephalography using the method of finite elements

The results of comparing the solutions of the direct task of electroencephalography on a spherical model and a spherical model with one nonuniformity are discussed. The nonuniformity was simulated by two parabolas situated on the same axis of symmetry and crossing the boundary of the gray and the white matter. The region between the larger and the smaller parabolas had the physical characteristics of the gray matter, and the region inside the smaller parabola had the characteristics of the cerebrospinal fluid. The task was to find a combination of parameters (distance between the dipole and the nonuniformity, angle of rotation of the dipole relative to the nonuniformity, sizes of the dipole and the nonuniformity, etc.) that provides the maximum effect of the difference of potentials on the outer surface of the scalp in the spherical model with one nonuniformity and the spherical model. The influence of the points of grounding on the value of the effect was analyzed (ground only at the right ear and ground at both ears). The data obtained show that a maximum difference of potentials is reached at the positions of dipoles close to tangential relative to the scalp surface.     

Choice of hydrogen uptake (Hup) status in legume‐rhizobia symbioses

The H₂ is an obligate by-product of N-fixation. Recycling of H₂ through uptake hydrogenase (Hup) inside the root nodules of leguminous plants is often considered an advantage for plants. However, many of the rhizobium-legume symbioses found in nature, especially those used in agriculture are shown to be Hup⁻, with the plants releasing H₂ produced by nitrogenase activity from root nodules into the surrounding rhizosphere. Recent studies have suggested that, H₂ induces plant-growth-promoting rhizobacteria, which may explain the widespread of Hup⁻ symbioses in spite of the low energy efficiency of such associations. Wild legumes grown in Nova Scotia, Canada, were surveyed to determine if any plant-growth characteristics could give an indication of Hup choice in leguminous plants. Out of the plants sampled, two legumes, Securigera varia and Vicia cracca, showed Hup⁺ associations. Securigera varia exhibited robust root structure as compared with the other plants surveyed. Data from the literature and the results from this study suggested that plants with established root systems are more likely to form the energy-efficient Hup⁺ symbiotic relationships with rhizobia. Conversely, Hup⁻ associations could be beneficial to leguminous plants due to H₂-oxidizing plant-growth-promoting rhizobacteria that allow plants to compete successfully, early in the growing season. However, some nodules from V. cracca tested Hup⁺, while others were Hup⁻. This was similar to that observed in Glycine max and Pisum sativum, giving reason to believe that Hup choice might be affected by various internal and environmental factors.     

Editorial: Twin peaks mission

The Y human chromosome has many ancient genes whose fidelity seems to have been preserved by tandem sequences and palindromic ‘hairpins’, compared/repaired by ‘gene conversion’. That a primary function of recombination machinery is DNA repair has been suggested, and rejected, several times; this new evidence is very persuasive. The process, better called gene conservation than gene conversion, could operate in all diploid organisms, accounting for the retention of long gene sequences without ‘informational meltdown’ ('concerted evolution'). It resembles rocket‐science computer‐redundancy error‐checking, comparison of three or four sequences, not just two. If recognition of errors in ‘converted’ sequences can be followed by either repair or rejection, the rejection option can account for the vast wastage of meiotic products. The repair option might be used in Drosophila oocytes and even zygotic nuclei, possibly other oocytes, ancient asexual lineages such as mycorrhizal fungi, perhaps the Y itself. Both evolutionary stasis (conservatism) and development and deployment of complex developmental modules can be understood in these terms so both the evolution of biodiversity and the practice of systematics may have these mechanisms as their bases. The main individual‐fitness and evolutionary advantages of diploidy were not primarily cloaking of recessive al‐leles, or allelic recombination and Mendelism, but conserving long DNA sequences.     

The effects of afterload and stimulation delay on the slow force response in the heterogeneous myocardium

The results of numerical simulations and physiological experiments that assessed the dependence of the intramyocardial slow force response under an afterload in the heterogeneous myocardium are described. Muscle duplexes of our design were used as the simplest experimental and theoretical models of the heterogeneous myocardium. It is shown that the degree of the slow force response increases with an afterload and the time of the stimulation delay between the elements of the duplex. This effect is explainable by an increase in the time of the mechanical interaction between the duplex elements in the isometric phase of contraction. This leads to changes in individual contractility, the configuration of the action potential, and the kinetic characteristics of intracellular calcium in cardiomyocytes of interacting muscles.