MiR-27b promotes sheep skeletal muscle satellite cell proliferation by targeting <Emphasis Type="Italic">myostatin</Emphasis> gene

To investigate the role of miR-27b in sheep skeletal muscle development, here we first cloned the sequence of sheep pre-miR-27b, then further investigated its expression pattern in sheep skeletal muscle in vivo, the relationship of miR-27b expression and sheep skeletal muscle satellite cell proliferation and differentiation in vitro, and then finally confirmed its target gene during this development process. MiR-27b sequence, especially its mature sequence, was conservative among different species. MiR-27b highly expressed in sheep skeletal muscle than other tissues. In skeletal muscle of Suffolk and Bashbay sheep, miR-27b was upregulated during foetal period and downregulated during postnatal period significantly (\(P{<}0.01\)), but it still kept a relatively higher expression level in skeletal muscle of postnatal Suffolk sheep than Bashbay. There is a potential target site of miR-27b on \(3^\prime \)-UTR of sheep myostatin (MSTN) mRNA, and the double luciferase reporter assay proved that miR-27b could successfully bind on this site. When sheep satellite cells were in the proliferation status, miR-27b was upregulated and MSTN was downregulated significantly (\(P{<}0.01\)). When miR-27b mimics was transfected into sheep satellite cells, the cell proliferation was promoted and the protein level of MSTN was significantly downregulated (\(P{<}0.01\)). Moreover, miR-27b regulated its target gene MSTN by translation repression at an early step, and followed by inducing mRNA degradation in sheep satellite cells. Based on these results, we confirm that miR-27b could promote sheep skeletal muscle satellite cell proliferation by targeting MSTN and suppressing its expression.     

Peculiarities of isolated blastomere development of the polyhaete <Emphasis Type="Italic">Alitta virens</Emphasis>

The ability of regulation, including restoration of the missing parts, to normalize the development, is a fundamental characteristic of living systems. However, until recently, representatives of the group Spiralia (animals with spiralian development) have been considered a typical example of determinative development. Meanwhile, studies in recent years give reason to speak about the presence of inductive cell-cell interactions at the early stages of Spiralian embryo development. In this paper, for the first time, we have shown the development of the polychaete Alitta virens blastomeres isolated at different stages. The results indicate the inability of A. virens partial embryos of regulation to form a whole normal larva and a high degree of autonomous determination of trochoblast cells. At the same time, the findings support probable early induction of blastomere D and acquisition of competence by blastomere D to further development of trochoblast ciliated cells.     

<Emphasis Type="Italic">Plasmodium</Emphasis><Emphasis Type="Italic">falciparum</Emphasis> GFP-E-NTPDase expression at the intraerythrocytic stages and its inhibition blocks the development of the human malaria parasite

Plasmodium falciparum is the causative agent of the most dangerous form of malaria in humans. It has been reported that the P. falciparum genome encodes for a single ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase), an enzyme that hydrolyzes extracellular tri- and di-phosphate nucleotides. The E-NTPDases are known for participating in invasion and as a virulence factor in many pathogenic protozoa. Despite its presence in the parasite genome, currently, no information exists about the activity of this predicted protein. Here, we show for the first time that P. falciparum E-NTPDase is relevant for parasite lifecycle as inhibition of this enzyme impairs the development of P. falciparum within red blood cells (RBCs). ATPase activity could be detected in rings, trophozoites, and schizonts, as well as qRT-PCR, confirming that E-NTPDase is expressed throughout the intraerythrocytic cycle. In addition, transfection of a construct which expresses approximately the first 500 bp of an E-NTPDase-GFP chimera shows that E-NTPDase co-localizes with the endoplasmic reticulum (ER) in the early stages and with the digestive vacuole (DV) in the late stages of P. falciparum intraerythrocytic cycle.     

<Emphasis Type="Italic">Xenopus laevis</Emphasis> peroxiredoxins: Gene expression during development and characterization of the enzymes

Reactive oxygen species (ROS) are produced via catabolic and anabolic processes during normal embryonic development, and ROS content in the cell is maintained at a certain level. Peroxiredoxins are a family of selenium-independent peroxidases and play a key role in maintaining redox homeostasis of the cell. In addition to regulating the ROS level, peroxiredoxins are involved in intracellular and intercellular signaling, cell differentiation, and tissue development. The time course of peroxiredoxin gene (prx1–6) expression was studied in Xenopus laevis during early ontogeny (Nieuwkoop and Faber stages 10–63). The highest expression level was observed for prx1 at these developmental stages. The prx1, prx3, and prx4 expression level changed most dramatically in response to oxidative stress artificially induced in X. laevis embryos. In X. laevis adults, prx1–6 were all intensely expressed in all organs examined, the prx1 expression level being the highest. The X. laevis prx1–6 genes were cloned and expressed in Escherichia coli, and physico-chemical characteristics were compared for the recombinant enzymes. The highest peroxidase activity and thermal stability were observed for Prx1 and Prx2. It was assumed that Prx1 plays a leading role in X. laevis early development.     

Interspecific variation in extracellular polysaccharide content and colony formation of <Emphasis Type="Italic">Microcystis</Emphasis> spp. cultured under different light intensities and temperatures

Single cells of five different Microcystis species (M. ichthyoblabe, M. viridis, M. flos-aquae, M. wesenbergii, and M. aeruginosa) were batch-cultured at different temperatures and light intensities: (a) 25 °C and 50 μmol photons m^?2 s^?1 (control culture); (b) 25 °C and 10 μmol photons m^?2 s^?1; and (c) 15 °C and 50 μmol photons m^?2 s^?1. The extracellular polysaccharide content was significantly higher in treatments b and c than in the control treatment. All Microcystis species existed as single cells under the control treatment but formed colonies in treatments b and c. All of the colonies were irregular with indistinct margins. M. ichthyoblabe, M. viridis, M. flos-aquae, and M. wesenbergii formed colonies with similar morphologies and their cells were loosely aggregated. In contrast, M. aeruginosa formed denser colonies with no distinct holes. The colony morphologies differed from the classic morphology of M. ichthyoblabe field-grown colonies but resembled that of small colonies found in Lake Taihu (Yangtze Delta Plain, China) during early spring. This indicates that field- and laboratory-grown colonies are governed by similar formation processes. We suggest that in laboratory and field environments, M. ichthyoblabe (or M. flos-aquae) colonies are representative of small colonies formed from single Microcystis cells, whereas the morphology of older colonies evolves to resemble M. wesenbergii and M. aeruginosa colonies.     

Differentiation of stem cells isolated from rat skeletal muscles towards cardiomyocytes: The effect of an inhibitor of DNA methylation 5-azacytidine

The effects of the inhibitor of DNA methylation 5-azacytidine on stem (satellite) cells isolated from fetal and definitive skeletal muscles of rats lead to the expression of marker genes of cardiomyogenesis (Gata4, Nkx2.5, connexin-43, n-cadherin, as well as Cacna 1 c encoding the cardiac subunit of the L-type Ca²⁺-channel). Through comparative analysis of the dynamics of expression of key markers of cardiomyogenesis, it was established that satellite cells isolated from fetal muscles have a more expressive potency to cardiomyocyte differentiation in vitro (expression of specific marker genes) compared to cells from muscles of adult animals. In the process of induced cardiomyocyte differentiation, the expression of MyoD and m-cadherin marker genes of skeletal muscle differentiation was not detected, suggesting that in the experiments presented the program of myogenic differentiation of skeletal muscles is inhibited.     

Expression of cancer-testis antigens of the Mage family in mouse oocytes and early embryos

Cancer-testis antigens of the Mage family (Melanoma antigens) are expressed predominantly in the spermatogenic and cancer cells, but some genes of this family are expressed ubiquitously. Expression patterns and functional role of Mage family antigens in the regulation of cellular processes in normal embryonic and definitive cells are virtually unknown. Comparative immunofluorescent analysis of Mage expression in mouse oocytes and early embryos identified the expression of Mage antigens at all stages studied. The greatest intensity of the fluorescent staining was detected in the epiblasts and the extraembryonic structures of the egg cylinder at E6.5 stage. At all studied developmental stages of the mouse oocyte and the early embryo, the localization of Mage antigens was found predominantly in the cytoplasm. Quantitative real-time PCR showed that expression levels of most Mage genes in cells of the epiblast and ectoplacental cone were similar, while the gene expression levels of Mage-a10, Mage-b16, and Mage-b18 were higher in cells of the ectoplacental cone than in epiblast cells. Thus, for the first time, our analysis has shown that the Mage family antigens are expressed at the early stages of mouse development and may be involved in the regulation of earliest events of embryogenesis.     

Informative Value of Temperatures in Different Areas of the Human Body for Correcting Body Thermal Imbalance during Extravehicular Activities

Identification of noninvasive and informative sites on the body reflecting the development of body thermal imbalance during extravehicular activities (EVAs) is highly important for enhancing astronaut safety. Temperature changes were evaluated on several areas of the head (the mastoid fossa (T _mf ), the forehead (T _fo ), and the cheek (T _ch )) and on the fingers (T _fing ). Subjects were dressed in a multicompartment liquid cooling/warming garment. Studies I and II consisted of different combinations of hood versus garment cooling or warming imposed across stages; studies III and IV involved sagittally divided cooling or warming regimes with the hood worn (study III) or with the head uncovered (study IV). In studies I and II, T _mf significantly (P < 0.05) differed between stages 2, when the head was cooled and the rest of the body heated, and 3, when the head was heated and the rest of the body cooled. The T _mf changes were consistent with the thermal conditions imposed on the head but not reflective of the developing body heat deficit. In study III, the T _mf at stages 2 and 3 on the right or the left followed the thermal conditions on the ipsilateral side of the body (P < 0.01). In study IV, T _fing showed no significant differences across stages. In studies I–IV, T _fing showed consistent changes across stages (P < 0.05), reflecting the developing body heat deficit. In all studies, there were no significant differences in rectal temperature (T _re ) across stages. T _mf and temperatures at other head skin sites did not respond in accordance with the actual intensity of a heat or cold flux from the garment and were not reflective of the overall development of body thermal imbalance. T _fing was a more adequate indicator of initial thermal destabilization and provided information that would be useful for monitoring the thermal balance and comfort during EVAs.     

Investigation of systemic control of plant cell division and differentiation in the model of tumor growth in radish

The study addresses the control of plant cell division and differentiation using the model of tumor-forming lines of radish. Expression of the genes involved in control of the cell cycle (CycD3), maintenance of meristematic cell activity (STM, WUS, and KNAT1), and primary response to cytokinin (ARR) was studied in inbred radish lines characterized by tumor growth at different stages of development. The influence of exogenic cytokinin on the expression of the genes of interest is analyzed. The possible role of the CycD3, KNAT1, tSTM, WUS, and ARR5 in tumor formation in radish is discussed.