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Changes in cell metabolism accompany the development of a wide spectrum of pathologies including cancer, autoimmune, and inflammatory diseases. Therefore, usage of inhibitors of metabolic enzymes are considered a promising strategy for the development of therapeutic agents. However, the investigation of cellular metabolism is hampered by the significant impact of culture media, which interfere with many cellular processes, thus making cellular models irrelevant. There are numerous reports that show that the results from in vitro systems are not reproduced in in vivo models and patients. Over the last decade a novel approach has emerged, which consists of adaptation of the culture medium composition to that closer to the composition of blood plasma. In 2017‒2019, two plasma-like media were proposed, Plasmax and HPLM. In the review, we have summarized the drawbacks of common media and have analyzed changes in the metabolism of cells cultivated in common and plasma-like media in normal and pathological conditions.

Alternative ORFs in-frame with the known genes are challenging to reveal. Yet they may contribute significantly to proteome diversity. Here we focused on the individual expression of the SERPINA1 gene exon 5 leading to direct translation of alpha1-antitrypsin (AAT) C-terminal peptides. The discovery of alternative ways for their production may expand the current understanding of the serpin gene's functioning. We detected short transcripts expressed primarily in hepatocytes. We identified four variants of hepatocyte-specific SERPINA1 short transcripts and individually probed their potential to be translated in living cells. The long mRNA gave the full-length AAT-eGFP fusion, while in case of short transcripts we deduced four active SERPINA1 in-frame alternative ORFs encoding 10, 21, 153 and 169 amino acids AAT C-terminal oligo- and polypeptides. Unlike secretory AAT-eGFP fusion exhibiting classical AAT behavior, truncated AAT-fusions differ by intracellular retention and nuclear enrichment. Immunofluorescence on the endogenous AAT C-terminal epitope showed its accumulation in the cell nucleoli, indicating that short transcripts may be translated in vivo. FANTOM5 CAGE data on SERPINA1 suggest that short transcripts originate from the post-transcriptional cleavage of the spliced mRNA, initiated mainly from the hepatocyte-specific promoter. CONCLUSION: Short SERPINA1 transcripts may represent a source for the direct synthesis of AAT C-terminal peptides with properties uncommon to AAT.

We studied the effect of neural stem cells (NSC) and mesenchymal stem cells (MSC) from mouse adipose tissue on survival, clonogenic activity, and senescence of NSC after exposure to γ-radiation. It was found that survival and clonogenic activity of NSC irradiated in doses of 1 and 2 Gy was enhanced when irradiated cells were co-cultured with non-irradiated NSC and MSC in permeable Transwell inserts. The proportion of senescent NSC (cells with high β-galactosidase activity) increased with increasing irradiation dose. Co-culturing with non-irradiated NSC in 3 days after irradiation in a dose of 1 Gy led to a decrease in the proportion of senescent cells among irradiated NSC. Factors secreted by NSC and MSC can become the basis for the development of means for prevention and treatment of damage to brain cells resulting from radiation therapy of head and neck cancer.

Progesterone and its synthetic analogues act on cells through different types of receptors, affecting proliferation and apoptosis. These compounds exert their effect through the nuclear receptors and the insufficiently studied membrane progesterone receptors (mPRs) belonging to the progestin and adiponectin Q receptor (PAQR) family. We have identified two selective ligands of mPRs that activate only this type of progesterone receptors - 19-hydroxypregn-4-en-20-one (LS-01) and 19-hydroxy-5β-pregn-3-en-20-one (LS-02). The goal of this work is to study the effect of these compounds on proliferation and death of human pancreatic adenocarcinoma cells BxPC3 and involvement of the two kinases (p38 MAPK and JNK) in signaling pathways activated by progestins through mPRs. It was shown that progesterone and the compound LS-01 significantly (p0.05) inhibited the BxPC3 cell viability, with JNK serving as a mediator. The identified targets of these two steroids are the genes of the proteins Ki67, cyclin D1, PCNA, and p21. Progesterone and the compound LS-01 significantly (p0.05) stimulate DNA fragmentation, enhancing the cell death. The p38 mitogen-activated protein kinase (MAPK) is a key mediator of this process. The BCL2A1 protein gene was identified as a target of both steroids. The compound LS-02 significantly (p0.05) alters membrane permeability and changes the exposure of phosphatidylserine on the outer membrane leaflet, also enhancing the cell death. This compound acts on these processes by activating both kinases, JNK and p38 MAPK. The compound LS-02 targets the genes encoding the proteins HRK, caspase 9, and DAPK.

Leaf pubescence is widespread among higher plants. In bread wheat, a relationship was found between this trait and the efficiency of photosynthetic processes and productivity. In this work, we established the chromosomal localization of the gene for leaf pubescence introgressed from Triticum timopheevii into a bread wheat line 821 and studied its expression in the genetic background of two wheat cultivars differing in genetic control and phenotypic expression of pubescence. To obtain quantitative characteristics of pubescence in cultivars and hybrid populations, the LHDetect2 program was used, which makes it possible to estimate the length and number of trichomes on a leaf fold. A genetic analysis showed the dominant inheritance of the gene. Monosomic analysis F2 was used to establish chromosome localization and investigate the expression of the gene in cultivars Saratovskaya S29 (S29) and Diamant 2 (Dm2). As a result, the gene Hltt, introgressed from T. timopheevii, was identified and localized in the distal region of the long arm of 5A chromosome for the first time. In both F2 populations, the gene reduced the density of trichomes and formed long trichomes, uncharacteristic for the two recipient cultivars S29 and Dm2. A larger number of long trichomes was formed in the genetic background of S29, which carry the bread wheat gene Hl1 and Hl3 for leaf pubescence, than in Dm2. Development of substitution and isogenic lines with the fragment of introgression carrying the gene Hltt will allow determining function and assessing the adaptive significance of the gene more precisely.

The production of new effective hemostatic materials based on a polymer composition of modified chitosan with a three-dimensional cross-linked structure as a polymer matrix and calcium ions coordinated to it is addressed. The optimum concentration of calcium ions in the chitosan-based composition was established as 10 – 20 mM and ensured an effective in vitro blood coagulation time of 40 sec. Compositions of chitosan succinate and chloride in addition to chitosan in solutions with nicotinic acid were shown to have effective hemostatic activity. Statistical differences in the effects of various calcium salts on the blood coagulation time were not found. In vivo tests showed that a chitosan-based composition was effective for stopping bleeding of any degree of severity, both with external and internal injuries to tissues.

The results of biostratigraphic study based on the radiolarian analysis of the rhythmically-layered terrigenous deposits developed in the islands of the Rimsky-Korsakov Archipelago (Peter the Great Bay) are presented. These deposits are most similar to medium-grained turbidiites (according to [21]). In the study region, the development and stratigraphic subdivision of the Upper Triassic and Lower Jurassic boundary deposits, separated by the marking layer, have been validated for the first time. Based on the comparison with the isochronic zonal subdivision of the Pacific and Tethys paleo areas, the layers with Globolaxtorum tozeri (Upper Rhaetian) are distinguished in the Upper Triassic deposits of the studied sections, while the Pantanellium tanuense Zone (Hettangian) is traced and the layers with Parahsuum simplum (Sinemurian–Pliensbachian) are revealed in the Lower Jurassic deposits.

In recent years, studies into the processes of bed agglomeration during the combustion of biomass have become increasingly relevant. The agglomeration processes are most largely influenced by the alkaline components of ash, which form low-melting eutectics by reacting with silicates. Therefore, it is very important to determine the critical concentration, primarily, of potassium that forms the most low-melting eutectics in the bed, at which the particles begin to sinter. The experimental studies are aimed at to investigate the effect of the temperature and the concentrations of alkaline elements on the process of particle agglomeration during the combustion of various types of biomass as well as biomass and coal, studying the presence of bed drain, determining the critical concentration of potassium in the bed, and calculating the estimated time before the start of sintering under conditions of an increase of potassium concentration in the bed. The experimental techniques and characteristics of the four studied types of biomass are described. Data on the fraction of agglomerated particles in the bed at different temperatures and concentrations of alkaline elements are given. Upon replacing a part of the sand by iron oxides, agglomeration is observed only at high temperatures (850°C and more). The obtained data are compared with the results of similar experiments carried out under conditions of the fluidized bed; in these experiments, the effects of the fluidization rate, particle size, and excess air on the agglomeration processes are considered. It is noted that the limiting potassium concentration is lowest in the case of a fixed bed. It is shown that real objects should be used when studying the dependence of the behavior of the potassium concentration in the bed on the operation time. The choice of the fraction of bed drain with the addition of a screened material or fresh sand is of practical interest. The calculation techniques and calculated data obtained with use of them on the behavior of the potassium concentration in the bed under various conditions are presented. The fractions of bed drain during combustion of sunflower husks and bark and wood waste are determined. In addition, it is shown that the joint combustion of husks and coal sharply reduces the probability of bed agglomeration.

Three new organoselenium compounds are synthesized: N-phenyl-2-(2-thienylacetyl)hydrazinecarboselenoamide (I), 4-phenyl-5-(2-thienylmethyl)-2,4-dihydro-3H-1,2,4-triazole-3-selone (II), and 3,3'-di[4-phenyl-5-(2-thienylmethyl)-4H-1,2,4-triazolyl] diselenide (III). Two of them (compounds II and III) are characterized by X-ray diffraction analysis (CIF files CCDC nos. 1956602 (II) and 1956603 (III)). Compound II crystallizes in the monoclinic crystal system (space group P21/n) with two crystallographically independent molecules A and B being different conformers relative to rotation about the NTrz–CTrz–C(H2)–CTph bond, where Trz is triazole and Tph is thiophene (gauche-A (51.4(3)°) and cis-B (4.2(4)°)). In the crystal of compound II, molecules A and B form chains along the crystallographic axis a due to strong hydrogen bonds N–H···Se. Then the chains are bound into a three-dimensional framework via intermolecular nonvalent interactions Se···S (3.3857(11) A). Owing to the anomeric effect, diselenide III is characterized by the typical gauche conformation of the substituents at the Se–Se bond (torsion angle СSeSeС 83.5(4)°) stabilized by a weak intramolecular hydrogen bond С–H···π. In the crystal of compound III, the molecules form chains along the crystallographic acid b due to intermolecular noncovalent interactions Se···π(C–C) (3.404(6) and 3.458(12) A), Se···Se (3.8975(11) A), and S···N (3.250(5) A). Bactericidal and fungicidal activity of the synthesized compounds is studied.

The experimental results obtained in the study on the possibility of sensitizing titanium dioxide (polymorphic anatase phase) to the visible region of the spectrum by doping and co-doping with impurities of non-metals in order to create effective photocatalysts for the decomposition of organic compounds have been analyzed. The presence of impurity atoms appears to result in a change in the electronic structure of the titanium dioxide matrix, in the appearance of “impurity bands” and in the narrowing of the energy gap of titanium dioxide. Such a modification is accompanied by an extension of the spectral range of sensitivity of photoactive solids to the long-wavelength region of the spectrum and, therefore, can be used to improve the catalytic properties of these materials. Spectral manifestations of carbon impurities in titanium dioxide in the form of carbide and carbonate, as well as sulfur in the forms of sulfite, sulfide, and sulfate, have been studied by the density functional theory method. A Ti14H22O39 cluster model was chosen for the titanium dioxide matrix. The calculations were carried out in the framework of the cluster approximation, using functional B3LYP and basis set 6-31G (d, p). Comparison of the results of quantum chemical calculations with the available experimental data shows that the impurity sulfur and carbon atoms in titanium dioxide, which are in different coordination states and different oxidation states, appear in different spectral ranges. This circumstance makes it possible to elucidate the structure of the samples based on the experimental spectra. A change in the coordination and oxidation states of impurity atoms leads to spectral shifts and splitting of peaks, which can reach 1.5 eV (XPS). The presence of admixtures of non-metals leads to a change in color (deepening in the case of sulfide or carbide) of the samples, appearing in the corresponding UV spectra.

Grafted copolymers of gelatin and polymethyl methacrylate are synthesized using the tributylboron–oxygen initiating system in an aqueous dispersion at 60°C. The products are characterized by IR spectroscopy and gel permeation chromatography. The proportion of polymethyl methacrylate in the copolymer is 2–60% and increases with a higher concentration of alkylborane in the initial mixture. The particle size of the aqueous dispersions of the copolymers determined by dynamic light scattering exceeds the particle size of gelatin. The thermogravimetric curves of the copolymers are similar to those of gelatin, with thermal stability tending to increase with a higher concentration of tributylboron. All the copolymers are biodegradable by fungi.

Despite the fact that effects of thyroid hormones were noticed back in the 80s, the underlying molecular mechanisms have not yet been elucidated. Current review summarizes years of investigations upon molecular targets for the action of thyroid hormones, leading to changes in the electrical and mechanical activity of the heart; changes in hemodynamics, affecting the functioning of the heart muscle, are also touched.

Thermal analysis of samples of chitosan–poly(methyl acrylate) block copolymer after exposure to electromagnetic fields, ultraviolet radiation, ultrasound, and biodestructive agents is performed for the first time by means of differential scanning calorimetry in the range of 250–510 K and thermogravimetry in the region of 300–850 K. It is shown that the combined effect of physical factors and biodestructive agents alters certain physicochemical properties: the temperatures of physical transformations, the temperatures of the onset of thermal degradation, and the nature of thermal decomposition.

Expression of prolactin axis genes (prolactin Prl1, prolactin-like hormone Prl2, prolactin receptor “a” PrlRa, and prolactin receptor “b” PrlRb) have been studied in the brains of females and males of the three-spined stickleback Gasterosteus aculeatus upon adaptation to fresh water. The expression of the Prl1 gene in females was significantly higher after 24-h adaptation to the freshwater relative to the control group (seawater) and to that in males at the same time point. No changes in Prl2 gene expression in females and Prl1 in males were detected. In males, after 12 and 18 h of adaptation to fresh water, higher expression of the Prl2 gene was observed relative to the control group. The expression of the PrlRa gene in males in seawater is higher than that in females; upon adaptation to fresh water, it does not change in females, and it decreases in males gradually after 3 h of being transferred to fresh water. No differences in the expression of the PrlRb gene between females and males in seawater were detected; however, after 24 h of freshwater adaptation, this parameter was higher in females than in males. Changes in the expression of prolactin axis elements during freshwater adaptation are gender-dependent, and the contribution of prolactin and prolactin-like hormone to adaptation of females and males of the three-spined stickleback to fresh water is significantly different.

Polyamides based on the 5-hydroxymethylfurfural derivative (2,5-furandicarboxylic acid dichloroanhydride) and diamines of both aromatic and aliphatic series were synthesized. The reactions were monitored by 1H NMR and IR spectroscopy. The thermal stability, glass transition temperature, and intrinsic viscosity of the synthesized polymers were determined.

We analyzed the expression of molecular targets of natriuretic action of prolactin in different layers of the kidney in the rat model of cholestasis of pregnancy. Sodium bicarbonate cotransporter NBCe1 was most sensitive to the conditions of cholestasis and cholestasis of pregnancy: the expression NBCe1 mRNA and protein in the renal outer medulla decreased in comparison with the normal. All forms of cholestasis affected the mRNA expression of sodium-potassium chloride co-transporter NCC, α-subunit of the ENaCα epithelial sodium channel, and Nedd4-2 ubiquitin ligase in different layers of the kidney. The obtained data suggest that prolactin provides fine tuning of various sodium transporters in different parts of the nephron under pathological conditions.

Genetic engineering allows for an extension beyond the limits of a species' natural variability. This enables the production of ornamental plants with novel colors and shapes of flowers, enhanced resistance, and a complex of characteristics gaining aesthetic and economic advantages. As a rule, success in achievement of desirable effects requires changes in the expression patterns of particular genes. A key regulatory element determining the level and tissue and temporal specificity in gene expression is a promoter. Therefore, an appropriate promoter must be chosen in the first place to build the planned structures of genetic constructs for transgene expression in plants. In recent years, many novel constitutive, tissue-specific, and induced promoters of plant origin have been explored. They may broaden advancements for derivation of novel forms and cultivars by means of a strictly directed expression of the transferred genes. The review discusses literary data on promoter application to biotechnology for the cases of ornamental plants.

The spatial structure and electronic structure of oxygen defect and nitrogen impurity centers of anatase and their manifestation in water adsorption, O1s XPS spectra were analyzed. The role of various forms of adsorbed water on anatase surface is considered. The results of experimental studies are compared with the properties of molecular models for the bulk and surface of titanium dioxide studied by the quantum chemical density functional theory method.

High-risk financial markets are considered in which transactions with financial instruments without reservation are conducted. The following risk-free trading strategies with a guaranteed profitability or loss are analyzed: scalping, spoofing, flipping, and scalping with hedging. A mathematical model is constructed for them. Examples of calculations performed by the obtained algorithms in the process of option trading are given.

Progesterone (P4) and its analogues regulate various reproductive processes, such as ovulation, implantation, pregnancy maintenance and delivery. In these processes, an important role is played by the immune cells recruited to the female reproductive organs and tissues, where they are exposed to the action of P4. Progestins regulate cellular processes, acting through nuclear steroid receptors (nSRs), membrane P4 receptors (mPRs), and through the sensors. It remains unclear, what type of receptors is used by P4 and its derivatives to exert their effect on the immune cells and how similar their effects are in different types of these cells. We have previously synthesized new progesterone derivatives, among which two selective mPRs ligands, not interacting with nSRs were identified. The objective of this study was to examine the effects of P4 and new selective mPRs ligands on the expression of pro- and anti-inflammatory cytokines in activated human peripheral blood mononuclear cells (PBMCs), THP-1 monocyte cells, and Jurkat T cells. It was demonstrated that the action of P4 and selective ligands was unidirectional, but in different types of the immune cells, their effects were different, and sometimes even opposite. In PBMCs, exposure to these steroids resulted in the increase of mRNA and secreted protein levels of IL-1β, TNFα, and IL-6 cytokines, as well as in the increase of INFγ mRNA level, decrease of IL-2 mRNA level, increase of TGFβ mRNA level, and decrease of IL-4 mRNA and IL-10 secreted protein levels. In monocytes, similarly to PBMCs, expression of IL-1β and TNFα mRNA was increased, but expression of IL-10 was also increased, and the TGFβ expression statistically significantly remained the same. In Jurkat T cells, expression of IL-2 and TNFα mRNA decreased, while expression of IL-10 increased, and expression of TGFβ did not change. Thus, progestins act on the immune cells through mPRs and have both pro- and anti-inflammatory effects, depending on the phenotypes of these cells. The data obtained are important for understanding the complexity of the immune system regulation by progestins, which depends on the type of the immune cells and individual characteristics of the immune system.

The inhibition of the initiated oxidation of cumene by oxygen under homophase conditions, where acetonitrile or dimethylsulfoxide are used as the solvent, by ascorbic acid is studied. The kinetics of the oxidation process is investigated via a gas volumetric method at a constant temperature of 348 K and a partial pressure of oxygen of 760 mmHg. It is shown during the investigation of the process of radical-chain oxidation of model systems upon varying the concentration of the inhibitor that the replacement of the solvent noticeably affects the value of the induction period. No significant influence of the medium on the rate of the oxidation process after the induction period is found. Concentration dependences of the kinetic parameters of ascorbic acid-inhibited oxidation of cumene in different media are obtained.

The review describes functional and structural features of different isoforms of prolactin receptor, mechanisms of signaling pathway activation, and molecular messengers involved in the transmission and termination of signal from the prolactin receptor isoforms. Changes in the ratio between prolactin receptor isoforms, key mediators of prolactin signal transduction and termination in various organs and tissues, are analyzed. Special attention is given to the role of molecular mediators and the ratio between the isoforms in normal physiological functions and pathologies. Approaches for therapeutic correction of prolactin signaling impairments are discussed.

—Differential scanning calorimetry (DSC) analysis was used to study the dependences of the main parameters of crystallization and melting of small water clusters dispersed in Sephadex resins on the amount of water (30–55%) in the wet polymer. The temperatures and heats of the processes were shown to depend on the frozen water (FW) concentration in Sephadex, with the dependence being considered as a manifestation of the size effect. Hysteresis between FW melting and crystallization was detected in a certain Sephadex humidity range; i.e., Tm was higher than Tcr and Qm higher than Qcr. The two findings reflected the basic properties of low-dimensional systems. Sephadex and amorphous starch, which was examined previously, were compared as two polysaccharide systems with different structural organizations, and a significant difference was observed in FW crystallization in the systems. Nucleation and crystallite growth, which determine water crystallization, were found to occur separately in time in Sephadex with a low humidity under these experimental conditions. Nucleation occurred during cooling, while growth was observed mainly during heating. Both of the processes occurred during cooling and partly overlapped only at a high humidity, far from Tg of Sephadex. In contrast, the two processes always occurred during cooling in amorphous starch regardless of its humidity. The difference in FW crystallization was assumed to arise because water mobility differs between the systems as a result of their difference in molecular mobility of the biopolymer matrix proper near the glass transition region.

Fluorinated esters of 2,5-furandicarboxylic acid have been synthesized starting from 2,5-furan dicarboxylic acid dichloride and fluorine-containing diols. The reaction control was performed by means of 1H, 19F NMR, and IR-spectroscopy. Thermal stability, as well as glass transition temperature, and intrinsic viscosity parameters of the obtained oligomers have been determined.

A new numerical method for finding roots of the characteristic equation in problems of radio wave propagation in tropospheric ducts with a piecewise linear profile of the refractive index is proposed. For the selected model of the vertical profile of the refractive index of an surface duct, dynamics of propagation constants of modes and features of the behavior of the attenuation function of the electromagnetic field as functions of the troposphere parameters and the wavelength are studied. The contribution of the whispering gallery modes is also estimated.

Identification of progesterone selective agonists and antagonists that act through one of the nuclear progesterone receptor isoforms is of particular importance for the development of tissue-specific drugs in gynecology and anticancer therapy. Fourteen pregna-D'6- and pregna-D'3-pentarane progesterone derivatives with 16α,17α-cycloalkane groups and two progesterone 3-deoxyderivatives were examined for their ability to regulate transcriptional activity of human nuclear progesterone receptor isoform B (nPR-B) expressed in Saccharomyces cerevisiae yeast. Transcriptional activity of nPR-B was measured from the expression of the β-galactosidase reporter gene with a hormone-responsible element in the promoter. Among the compounds tested, two were full progesterone agonists, four were partial agonists, one compound possessed both agonistic and antagonistic activity, one compound displayed only partial antagonistic activity, and eight compounds did not show any activity. Modifications of the pentarane structure, precisely, introduction of an additional double bound in the A or B rings and/or modification at the 6th position of progesterone, lead to a switch from the complete agonistic activity to partial agonistic or mixed activities. These modifications enable progestins to act as selective modulators of progesterone receptor. Steroids with reduced A-ring and 3-ketogroups lose their ability to regulate PR-B activity. Both 3-deoxycompounds, being selective ligands of progesterone membrane receptors, do not affect PR-B activity.

The effect of concentrations in the media of nanoparticles of silica, magnetite, and carbon nanotubes, as well as magnetic metallopolymer nanocomposites based on unsaturated dicarboxylates of cobalt (II), nickel (II), and iron (iron itaconate and citraconate and maleates of nickel, cobalt, and iron), on the growth of the soil-forming micromycete Trichoderma asperellum is studied in this work. As a result, the toxicity of a nanocomposite prepared by the thermal decomposition of cobalt maleate is revealed at a concentration of 10–4 parts by weight in the culture media. For the rest of the nanoparticles studied, neither acute toxicity nor the significant inhibition of the micromycete growth are found. For all types of particles studied, it is revealed that the particle concentrations in the media of (1–5) × 10–6 parts by weight nonspecifically stimulate the regrowth of fungi.

The results of studies of the vegetative cover and ontogenetic composition of the cenopopulations of Picea obovata Ledeb. and Larix sibirica Ledeb. in the Gorno-Khadytinskii Forest Reserve, which is the refugium of the northern taiga forests of Western Siberia, have been summarized. According to the Brown–Blanke system, two associations of forests were identified and described, as well as two tundra associations, for comparison. The unique association of the Aconito septentrionalis–Piceetum obovatae taiga forests Zaugolnova et Morozova 2009 were analyzed in detail, and a comparison of these associations was carried out in the Gorno-Khadytinskii and Pechoro-Ilychskii reserves, where these forests are the reference standards of the zonal-type forests. An estimate of the ontogenetic composition of the cenopopulations of P. obovata Ledeb. and L. sibirica Ledeb. is given, as well as the absolute age of the trees of these species.

Transformations of structural defects, the hydrogen state, and electrophysical properties of silicon treated in hydrogen plasma are studied. Treatment in plasma (150°C) produces bands in Raman spectra at 2095 and 2129 cm–1 that are associated with scattering by Si–H vibrations. Subsequent heat treatment (275°C) causes a band for gaseous molecular H2 to appear at 4153 cm–1. A comparison of Raman spectra and scanning probe microscopy results shows that hydrogenation forms defects (platelets) of average size 43 nm and surface density 6.5·109 cm–2 that are due to precipitation of H2 and formation of Si–H bonds. Inclusions of average size 115 nm and surface density 1.7·109 cm–2 that are filled with molecular H2 are observed after heat treatment. The concentration of free charge carriers remains constant after treatment in plasma and subsequent heat treatment.