Hormones, oxidized proteins, and lipids in alcoholism. Duration of remission.

The instability of remission in alcohol dependence (AD) creates a need to search for criteria for predicting its duration. The aim of study was to determine the hormones, oxidized proteins, and lipids in patients with AD, and the possible relations between these parameters and the duration of remission. Blood samples were obtained from 49 male patients with AD after alcohol detoxification (Total group). Two groups of patients were formed: with unstable therapeutic remission up to 6 months (UTR-group); with stable therapeutic remission which lasted 12 months or longer (STR-group). The control group comprised men without AD. The levels of carbonylated proteins (CP), lipid peroxidation (TBA-RS) were determined in the blood plasma. The levels of cortisol, testosterone total, thyroid-stimulating hormone (TSH), triiodothyronine free (fT3), and thyroxine free (fT4) in the blood serum were measured. The Total group of patients showed an increase in CP, TBA-RS, Cortisol, cortisol/testosterone ratio, and a decrease in TSH, fT3, and fT3/fT4 levels. A set of parameters (T, fT3, fT4, cortisol/testosterone ratio) associated with unstable remission was identified.

Green synthesis of nanoparticles with extracellular and intracellular extracts of basidiomycetes.

Au, Ag, Se, and Si nanoparticles were synthesized from aqueous solutions of HAuCl4, AgNO3, Na2SeO3, and Na2SiO3 with extra- and intracellular extracts from the xylotrophic basidiomycetes Pleurotus ostreatus, Lentinus edodes, Ganoderma lucidum, and Grifola frondosa. The shape, size, and aggregation properties of the nanoparticles depended both on the fungal species and on the extract type. The bioreduction of the metal-containing compounds and the formation rate of Au and Ag nanoparticles depended directly on the phenol oxidase activity of the fungal extracts used. The biofabrication of Se and Si nanoparticles did not depend on phenol oxidase activity. When we used mycelial extracts from different fungal morphological structures, we succeeded in obtaining nanoparticles of differing shapes and sizes. The cytotoxicity of the noble metal nanoparticles, which are widely used in biomedicine, was evaluated on the HeLa and Vero cell lines. The cytotoxicity of the Au nanoparticles was negligible in a broad concentration range (1-100 µg/mL), whereas the Ag nanoparticles were nontoxic only when used between 1 and 10 µg/mL.

Antioxidant Properties of the Artist's Conk Medicinal Mushroom, Ganoderma applanatum (Agaricomycetes), upon Cultivation with para-Substituted Phenolic Compounds and Tea Leaf Extracts.

In this study we elucidate antioxidative properties of the mushroom Ganoderma applanatum (Pers.) Pat. enhanced by submerged culture with para-hydroxyphenolic compounds and tea leaf extracts. The tea extract has been shown to increase to different extents the antioxidative efficiency of para-substituted phenolics, with the most profound effect for 2-(4-hydroxyphenyl)ethane-1-ol (tyrosol). Within the range of physiological concentrations, the symbatic correlation of the antioxidative action of the fungal samples with the volume of tea extract in the submerged culture medium was observed. We propose an approach to obtain, through the use of black tea extracts as the nutrient medium component, large amounts of G. applanatum seeding mycelia; the extract exerts a profound positive effect on the level of phenolic-type antioxidants.

Nanoparticles synthesis by Agaricus soil basidiomycetes.

We examined the effect of various concentrations of HAuCl4, AgNO3, Na2SeO3, Na2SiO3, and GeO2 on mycelial growth of the soil basidiomycetes Agaricus bisporus and A. arvensis in submerged and solid media. Fungal mycelial extracts and cell-free culture filtrates were able to reduce ions of Au, Ag, Se, Si, and Ge compounds, forming Au0, Ag0, Se0, Si0/SiO2 and Ge0/GeO2 nanoparticles. The physical characteristics of the mycogenic nanoparticles differed depending on the species of Agaricus and the type of extract. Au nanospheres obtained with cell-free culture filtrates were of 2-5 nm diameter in A. bisporus and of 2-10 nm in A. arvensis. Nanoparticles produced by extracts of mycelia were several times larger and highly heterogenous. Ag nanoparticles produced by cell-free culture filtrates were spherical or irregular-shaped and agglomerated, whereas with extracts of mycelia, small homogenous nanospheres of 1-10 nm were formed. Se nanospheres obtained with cell-free culture filtrates were of 100-250 nm diameter in A. bisporus and of 150-550 nm diameter in A. arvensis. The particles synthesized with extracts of mycelia were of 40-140 nm in A. bisporus and of 100-250 nm in A. arvensis. Incubation of Na2SiO3 with cell-free culture filtrates resulted in porous Si nanoparticles of 30-65 nm in A. bisporus and of 50-200 nm in A. arvensis. Ge nanoparticles synthesized by both Agaricus species were mostly spheres of 50-250 nm diameter.

Brown Mycelial Mat as an Essential Morphological Structure of the Shiitake Medicinal Mushroom Lentinus edodes (Agaricomycetes).

We show here, to our knowledge for the first time, that the brown mycelial mat of the xylotrophic shiitake medicinal mushroom, Lentinus edodes, not only performs a protective function owing to significant changes in the ultrastructure (thickening of the cell wall, increased density, and pigmentation of the fungal hyphae) but also is a metabolically active stage in the development of the mushroom. The cells of this morphological structure exhibit repeated activation of expression of the genes lcc4, tir, exp1, chi, and exg1, coding for laccase, tyrosinase, a specific transcription factor, chitinase, and glucanase, which are required for fungal growth and morphogenesis. This study revealed the maximum activity of functionally important proteins with phenol oxidase and lectin activities, and the emergence of additional laccases, tyrosinases, and lectins, which are typical of only this stage of morphogenesis and have a regulatory function in the development and formation of fruiting bodies.

Role of olfactory reactions, nociception, and immunoendocrine shifts in addictive disorders.

BACKGROUND AND OBJECTIVES: Addictive pathology is associated with nervous, immune, and endocrine shifts. Meanwhile, the nature of intersystemic relationship lying beneath addictive disorders remains unclear. The purpose of the study was to identify neuroimmunoendocrine markers of addictive disorders in male subjects defining the nature of their interaction. METHODS: The study enrolled 69 subjects aged 18-43 years: 59 males and 10 females divided into those with addictive disorders (n = 39) and conditionally healthy subjects (n = 30). EEG testing with olfactory stimulation, olfactometric, and pressure algometric examinations was carried out. Multiplex technique was applied to determine mitogen-induced production of cytokines IL-10, IL-1, IL-1RA, IL-2, IFN-gamma, TNF-alpha. ELISA method was applied to measure serum cortisol and testosterone levels. RESULTS: Olfactory responses to isopropanol with open eyes in addicted patients manifested as increase in alpha-rhythm and beta1-rhythm, with closed eyes presentation of this odorant was accompanied by increase of theta-rhythm in opioid-addicted patients. Male subjects with addictive disorders showed reduced alpha-rhythm in terms of olfactory stimulation with modified emotional evaluation of the odorant, deficient mitogen-induced production of IFN-gamma, and reduced pain sensitivity. Male subjects with opioid addiction had reduced beta1-rhythm in terms of olfactory stimulation, mitogen-induced production of IFN-gamma, and elevated testosterone level. CONCLUSIONS: The findings obtained verify potential involvement of nociception, olfaction, and cytokine production in addiction pathogenesis evidencing their various roles depending on the range of psychoactive substances (PAS) and pathology progression. SCIENTIFIC SIGNIFICANCE: The data obtained may provide background for unification of reward circuit and inhibitory control concepts in regulation of addictive behavior. (Am J Addict 2017;26:640-648).

Lectins from Mycelia of Basidiomycetes.

Lectins are proteins of a nonimmunoglobulin nature that are capable of specific recognition of and reversible binding to the carbohydrate moieties of complex carbohydrates, without altering the covalent structure of any of the recognized glycosyl ligands. They have a broad range of biological activities important for the functioning of the cell and the whole organism and, owing to the high specificity of reversible binding to carbohydrates, are valuable tools used widely in biology and medicine. Lectins can be produced by many living organisms, including basidiomycetes. Whereas lectins from the fruit bodies of basidiomycetes have been studied sufficiently well, mycelial lectins remain relatively unexplored. Here, we review and comparatively analyze what is currently known about lectins isolated from the vegetative mycelium of macrobasidiomycetes, including their localization, properties, and carbohydrate specificities. Particular attention is given to the physiological role of mycelial lectins in fungal growth and development.

Alteration in the ultrastructural morphology of mycelial hyphae and the dynamics of transcriptional activity of lytic enzyme genes during basidiomycete morphogenesis.

The morphogenesis of macromycetes is a complex multilevel process resulting in a set of molecular-genetic, physiological-biochemical, and morphological-ultrastructural changes in the cells. When the xylotrophic basidiomycetes Lentinus edodes, Grifola frondosa, and Ganoderma lucidum were grown on wood waste as the substrate, the ultrastructural morphology of the mycelial hyphal cell walls differed considerably between mycelium and morphostructures. As the macromycetes passed from vegetative to generative development, the expression of the tyr1, tyr2, chi1, chi2, exg1, exg2, and exg3 genes was activated. These genes encode enzymes such as tyrosinase, chitinase, and glucanase, which play essential roles in cell wall growth and morphogenesis.

Biosynthesis of nanoparticles of metals and metalloids by basidiomycetes. Preparation of gold nanoparticles by using purified fungal phenol oxidases.

The work shows the ability of cultured Basidiomycetes of different taxonomic groups-Lentinus edodes, Pleurotus ostreatus, Ganoderma lucidum, and Grifola frondosa-to recover gold, silver, selenium, and silicon, to elemental state with nanoparticles formation. It examines the effect of these metal and metalloid compounds on the parameters of growth and accumulation of biomass; the optimal cultivation conditions and concentrations of the studied ion-containing compounds for recovery of nanoparticles have been identified. Using the techniques of transmission electron microscopy, dynamic light scattering, X-ray fluorescence and X-ray phase analysis, the degrees of oxidation of the bioreduced elements, the ζ-potential of colloidal solutions uniformity, size, shape, and location of the nanoparticles in the culture fluid, as well as on the surface and the inside of filamentous hyphae have been determined. The study has found the part played by homogeneous chromatographically pure fungal phenol-oxidizing enzymes (laccases, tyrosinases, and Mn-peroxidases) in the recovery mechanism with formation of electrostatically stabilized colloidal solutions. A hypothetical mechanism of gold(III) reduction from HAuCl4 to gold(0) by phenol oxidases with gold nanoparticles formation of different shapes and sizes has been introduced.

Antitumor Activity of Extracts from Medicinal Basidiomycetes Mushrooms.

Aqueous extracts from the vegetative submerged mycelia of cultivated Basidiomycetes Ganoderma lucidum, Lentinus edodes, and Grifola frondosa, as well as from the fruiting bodies of G. lucidum, were found to have antitumor activity. The antitumor effect of the mycelial extracts from all 3 fungal species was ascertained in vivo in rats with implanted kidney cancer. Dystrophic changes in tumor cells and tumor necrosis (up to 90%) were noted. In vitro cytotoxicity studies of the A549 human lung adenocarcinoma cell line and HEp-2 human laryngeal epidermoid carcinoma cells showed that the extracts from the G. lucidum fruiting bodies and from the L. edodes vegetative mycelium were the most effective. The animals' immune systems were activated, and the fungal extracts displayed no toxicity when administered orally.

Adjunctive use of interferon γ inducer for treatment of patients with schizophrenia.

OBJECTIVE: The present paper is devoted to evaluation of clinical and immunomodulatory effect of ultra-high dilutions of antibodies to human interferon γ, included in the complex therapy of patients with schizophrenia. Materials and methods The study was carried out at the Mental Health Research Institute, Tomsk, Russian Federation. This double-blind, placebo-controlled randomised in parallel-group study enrolled 40 patients. As a part of complex therapy, patients from the main group (n=20) received anaferon, a drug containing ultra-high dilutions of affinity-purified antibodies to human interferon γ as the active pharmaceutical ingredient; patients from the comparative group (n=20) received placebo. Duration of the therapy was 30±5 days. Assessment of severity of symptoms and changes in them were made using clinical scales: Positive and Negative Syndrome Scale, Clinical Global Impression, Abnormal Involuntary Movements Scale. Spontaneous and phytohemagglutinin-induced production of interferon γ by immunocompetent cells in supernatants of 48 h whole blood culture of patients was measured by enzyme-linked immunosorbent assay (ELISA) method. RESULTS: The reduction of interferon-producing potential by immunocompetent cells in comparison with reference normal value was shown in total group of patients (n=40) before combined therapy. During the treatment, increase of spontaneous interferon γ production and favourable changes in psychopathological symptoms as compared with placebo were shown in subjects receiving anaferon. It was found that favourable changes in clinical symptoms assessed using clinical scales with a high degree of confidence correlated with high level of spontaneous interferon γ production. CONCLUSION: Anaferon as a part of complex therapy of patients with schizophrenia contributes to enhancement of its efficacy acting via mechanism of psychoimmunomodulation.

Reduction of selenite by Azospirillum brasilense with the formation of selenium nanoparticles.

The ability to reduce selenite (SeO(3)(2-)) ions with the formation of selenium nanoparticles was demonstrated in Azospirillum brasilense for the first time. The influence of selenite ions on the growth of A. brasilense Sp7 and Sp245, two widely studied wild-type strains, was investigated. Growth of cultures on both liquid and solid (2 % agar) media in the presence of SeO(3)(2-) was found to be accompanied by the appearance of the typical red colouration. By means of transmission electron microscopy (TEM), electron energy loss spectroscopy (EELS) and X-ray fluorescence analysis (XFA), intracellular accumulation of elementary selenium in the form of nanoparticles (50 to 400 nm in diameter) was demonstrated for both strains. The proposed mechanism of selenite-to-selenium (0) reduction could involve SeO(3)(2-) in the denitrification process, which has been well studied in azospirilla, rather than a selenite detoxification strategy. The results obtained point to the possibility of using Azospirillum strains as endophytic or rhizospheric bacteria to assist phytoremediation of, and cereal cultivation on, selenium-contaminated soils. The ability of A. brasilense to synthesise selenium nanoparticles may be of interest to nanobiotechnology for "green synthesis" of bioavailable amorphous red selenium nanostructures.

Enzymatic formation of gold nanoparticles by submerged culture of the basidiomycete Lentinus edodes.

We report for the first time that the medicinal basidiomycete Lentinus edodes can reduce Au(III) from chloroauric acid (HAuCl4) to elemental Au [Au(0)], forming nanoparticles. Several methods, including transmission electron microscopy, electron energy loss spectroscopy, X-ray fluorescence, and dynamic light scattering, were used to show that when the fungus was grown submerged, colloidal gold accumulated on the surface of and inside the mycelial hyphae as electron-dense particles mostly spherical in shape, with sizes ranging from 5 to 50nm. Homogeneous proteins (the fungal enzymes laccase, tyrosinase, and Mn-peroxidase) were found for the first time to be involved in the reduction of Au(III) to Au(0) from HAuCl4. A possible mechanism forming Au nanoparticles is discussed.

Reduction of organic and inorganic selenium compounds by the edible medicinal basidiomycete Lentinula edodes and the accumulation of elemental selenium nanoparticles in its mycelium.

We report for the first time that the medicinal basidiomycete Lentinula edodes can reduce selenium from inorganic sodium selenite (Se(IV)) and the organoselenium compound 1,5-diphenyl-3-selenopentanedione-1,5 (DAPS-25) to the elemental state, forming spherical nanoparticles. Submerged cultivation of the fungus with sodium selenite or with DAPS-25 produced an intense red coloration of L. edodes mycelial hyphae, indicating accumulation of elemental selenium (Se(0)) in a red modification. Several methods, including transmission electron microscopy (TEM), electron energy loss spectroscopy (EELS), and X-ray fluorescence, were used to show that red Se(0) accumulated intracellularly in the fungal hyphae as electron-dense nanoparticles with a diameter of 180.51±16.82 nm. Under designated cultivation conditions, shiitake did not reduce selenium from sodium selenate (Se(VI)).

Biodegradation of an organoselenium compound to elemental selenium by Lentinula edodes (shiitake) mushroom.

The present paper reports for the first time the transformation of an organic selenium compound into red selenium (Se), which causes the intense red pigmentation of Lentinula edodes (shiitake mushroom) mycelia. The biotransformation of 1,5-diphenyl-3-selenopentanedione-1,5 (diacetophenonyl selenide, preparation DAPS-25) was studied in liquid- and solid-phase cultures of L. edodes. In liquid culture medium, a red color develops in the mycelium at initial DAPS-25 concentrations equal to or higher than 0.1 mmol/l. The intensity and initiation time of coloration is Se concentration-dependent. Semiquantitative data obtained by physicochemical methods on the extent of Se and acetophenone production suggest that L. edodes is able to absorb and/or destruct this organic Se xenobiotic.

Compounds of the 1,5-di(4-R-phenyl)- 3-selenopentanediones-1,5 series interaction with the basidiomycete Lentinula edodes lectins: computations and experiment.

The role of spatial and electron structure, hydrophobic properties and concentration of organoselenium compounds on their interaction with fungal metabolites--extracellular lectins of Lentinula edodes (shiitake mushroom) has been considered. By the hybrid method of density functional theory at the B3LYP/6-31G(d,p) theory level, spatial and electronic structure of the 1,5-diphenyl-3-selenopentanedione-1,5 (preparation DAPS-25), 1,5-di(4-methoxyphenyl)-3-selenopentanedione-1,5 and 1,5-di(4-ethoxyphenyl)-3-selenopentanedione-1,5 molecules has been studied. The above molecules have been stated to be substantially similar to each other by their electronic and spatial characteristics. By means of the QSAR properties evaluation by the atomic-additive schemes, it has been shown that the molecules of the preparation DAPS-25, its dimethoxy- and diethoxy-substituted are close to each other by the hydrophilic-lipophilic balance, whereas di-n-octoxy derivative DAPS-25 is explicitly hydrophobic. The hemagglutinating activity of lectins in the presence of the preparation DAPS-25 and its alkyloxy-substituted increases, therewith the most effective addition is 1,5-di(4-ethoxyphenyl)-3-selenopentanedione-1,5. Apparently, the greater effectiveness of the said substance compared to DAPS -25 is caused by the formation of hydrogen bonds with a participation of unshared electron pairs of oxygen atoms from the ethoxy groups and mobile hydrogen atoms from the OH groups of glycoconjugates on erythrocytes surface. The positive effect of 1,5-di(4-n-octoxyphenyl)-3-selenopentanedione-1,5 is not so prominent, since the enlarged alkyl chain shields the aromatic fragments of organoselenium molecule participating in the binding with lectin.

Laccase and lectin activities of intracellular proteins produced in a submerged culture of the xylotrophic basidiomycete Lentinus edodes.

The white-rot fungus Lentinus edodes produced D-melibiose-specific lectins and two laccase forms in a lignin-containing medium. The maxima of laccase and lectin activities coincided, falling within the period of active mycelial growth. The enzymes and lectins were isolated and purified by gel filtration followed by anion-exchange chromatography. The L. edodes lectins were found to be able to stabilize the activity of the fungus's own laccases. Lectin activity during the formation of lectin-enzyme complexes remained unchanged.

Lectin activity of Lentinus edodes

The hemagglutinating activity of submerged mycelium and culture liquid for four strains of Lentinus edodes (Berk.) Sing [L. edodes (Berk.) Pegler] was studied in the search for lectins. The hemagglutinating activity of culture liquid was substantially higher, compared with mycelium. The carbohydrate-binding capacity of the agglutinins was established, and the lectin activity of extracts from mycelia grown on several agar media was elucidated in relation to fruiting. The lectin activity of L. edodes was examined at different morphogenetic steps: mycelium, brown mycelial film, primordium, and fruiting body. Hemagglutination titers at the brown film step were higher than in the mycelium, whereas activity at the primordial and fruiting bodies steps decreased. Lectins seem to be involved in the formation of hyphal aggregates of brown mycelial film (AU)

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