Exploring Psilocybe spp. mycelium and fruiting body chemistry for potential therapeutic compounds.

Psilocybe mushrooms, otherwise known as "magic" mushrooms, owe their psychedelic effect to psilocin, a serotonin subtype 2A (5-HT2A) receptor agonist and metabolite of psilocybin, the primary indole alkaloid found in Psilocybe species. Metabolomics is an advanced fingerprinting tool that can be utilized to identify the differences among fungal life stages that may otherwise be unaccounted for. In this study, by using targeted and untargeted (metabolomic) multivariate analysis, we demonstrate that the chemical composition of Psilocybe differs among mycelia, grain mycelia, and fruiting bodies. The preferential accumulation of psilocybin, baeocystin, tryptophan, ergothioneine, and phenylethylamine in fruiting bodies differentiated them from mycelia; however, the levels of alpha-glycerylphosphorylcholine (α-GPC), N-acetylglucosamine, and trimethylglycine were found to be proportionally higher in mycelia than in fruiting bodies based on Pareto-scaled data. Considering the wealth of compounds with therapeutic potential that have been isolated from various fungal genera, it would be pertinent to study the compounds found in Psilocybe mycelia as potential naturally derived therapeutic targets.

Reduction of the nematode egg reappearance period in horses after anthelmintic therapy.

Background and Aim: Anthelmintics are used to control equine nematodes. However, helminth resistance to regularly used drugs is a well-known challenge. Among tests to assess effective control and monitor resistance, the most common is the fecal egg count reduction test (FECRT). In the absence of reliable FECRT results, the nematode egg reappearance period (ERP) is taken into account. This study aimed to examine horses from farms around the Moscow Region to assess nematode resistance through ERP after therapy. Materials and Methods: In the first stage, fecal samples from 280 horses were examined by the flotation method with a sodium nitrate solution. The eggs per gram (EPG) in feces were counted using the modified McMaster technique. One hundred and forty out of 280 horses were selected for further work. Five groups were formed: Two groups of horses infected with strongyles (n = 50) and three groups with Parascaris equorum (n = 90). Therapy against strongyles was performed with albendazole and ivermectin. Therapy for parascaridosis was performed with fenbendazole, ivermectin, and aversectin C. Samples from the horses in each group were taken on the 14th day (2 weeks), 28th day (4 weeks), 42nd day (6 weeks), 56th day (8 weeks), and 84th day (12 weeks) after treatment, and the amount of EPG in each sample was determined. Results: Overall, nematodes were found in 65% of the horses examined. P. equorum was most frequently recorded (42.1%) followed by Strongylidae gen. spp. (27.9%). The strongyles ERP after therapy with albendazole and ivermectin was estimated on 42 days (6 weeks). The growth of P. equorum eggs in the feces was observed from the 56th day (8 weeks) after therapy with fenbendazole, from the 42nd day (6 weeks) after therapy with ivermectin, and was observed from the 84th day (12 weeks) after the use of aversectin. Conclusion: Our study shows widespread reductions in nematode ERPs across the Moscow Region after ivermectin therapy in horses, suggesting that additional monitoring of these farms is needed for effective control of anthelmintic resistance.

Increasing physical activity by four legs rather than two: systematic review of dog-facilitated physical activity interventions.

OBJECTIVES: Regular walking is a critical target of physical activity (PA) promotion, and dog walking is a feasible PA intervention for a large segment of the population. The purpose of this paper was to review PA interventions that have involved canine interactions and to evaluate their effectiveness. A secondary aim of this review was to highlight the populations, settings, designs and intervention components that have been applied so as to inform future research. DESIGN: Systematic review. DATA SOURCES: We carried out literature searches to August 2019 using six common databases. ELIGIBILITY CRITERIA: Studies included published papers in peer-reviewed journals and grey literature (theses and dissertations) in the English language that included any PA behaviour change design (ie, randomised controlled trial, quasi-experimental) that focused on canine-related intervention. We grouped findings by population, setting, medium, research design and quality, theory and behaviour change techniques applied. RESULTS: The initial search yielded 25 010 publications which were reduced to 13 independent studies of medium and high risks of bias after screening for eligibility criteria. The approaches to intervene on PA were varied and included loaner dogs, new dog owners and the promotion of walking among established dog owners. Findings were consistent in showing that canine-assisted interventions do increase PA (82% of the studies had changes favouring the canine-facilitated intervention). Exploratory subanalyses showed that specific study characteristics and methods may have moderated the effects. Compared with studies with longer follow-up periods, studies with shorter follow-up favoured behaviour changes of the canine intervention over the control condition. CONCLUSION: Canine-based PA interventions appear effective, but future research should move beyond feasibility and proof of concept studies to increase rigour, quality and generalisability of findings.

Transformation of avobenzone in conditions of aquatic chlorination and UV-irradiation.

Emerging contaminants represent a wide group of the most different compounds. They appear in the environment at trace levels due to human activity. Most of these compounds are not yet regulated. Sunscreen UV-filters play an important role among these emerging contaminants. In the present research the reactions of 4-tert-butyl-4'-methoxydibenzoylmethane (avobenzone), the most common UV filter in the formulation of sunscreens, were studied under the combined influence of active chlorine and UV-irradiation. Twenty five compounds were identified by GC/MS as transformation products of avobenzone in reactions of aquatic UV-irradiation and chlorination with sodium hypochlorite. A complete scheme of transformation of avobenzone covering all the semivolatile products is proposed. The identification of the two primary chlorination products (2-chloro-1-(4-tert-butylphenyl)-3-(4-methoxyphenyl)-1,3-propanedione and 2,2-dichloro-1-(4-tert-butylphenyl)-3-(4-methoxyphenyl)-1,3-propanedione) was confirmed by their synthesis and GC/MS and NMR analysis. Although the toxicities of the majority of these products remain unknown substituted chlorinated phenols and acetophenones are known to be rather toxic. Combined action of active chlorine and UV-irradiation results in the formation of some products (chloroanhydrides, chlorophenols) not forming in conditions of separate application of these disinfection methods. Therefore caring for people «well-being¼ it is of great importance to apply the most appropriate disinfection method. Since the primary transformation products partially resist powerful UV-C irradiation they may be treated as stable and persistent pollutants.

Extraordinary Genetic Diversity in a Wood Decay Mushroom.

Populations of different species vary in the amounts of genetic diversity they possess. Nucleotide diversity π, the fraction of nucleotides that are different between two randomly chosen genotypes, has been known to range in eukaryotes between 0.0001 in Lynx lynx and 0.16 in Caenorhabditis brenneri. Here, we report the results of a comparative analysis of 24 haploid genotypes (12 from the United States and 12 from European Russia) of a split-gill fungus Schizophyllum commune. The diversity at synonymous sites is 0.20 in the American population of S. commune and 0.13 in the Russian population. This exceptionally high level of nucleotide diversity also leads to extreme amino acid diversity of protein-coding genes. Using whole-genome resequencing of 2 parental and 17 offspring haploid genotypes, we estimate that the mutation rate in S. commune is high, at 2.0 × 10(-8) (95% CI: 1.1 × 10(-8) to 4.1 × 10(-8)) per nucleotide per generation. Therefore, the high diversity of S. commune is primarily determined by its elevated mutation rate, although high effective population size likely also plays a role. Small genome size, ease of cultivation and completion of the life cycle in the laboratory, free-living haploid life stages and exceptionally high variability of S. commune make it a promising model organism for population, quantitative, and evolutionary genetics.

Crossing-over in a hypervariable species preferentially occurs in regions of high local similarity.

Recombination between double-stranded DNA molecules is a key genetic process which occurs in a wide variety of organisms. Usually, crossing-over (CO) occurs during meiosis between genotypes with 98.0-99.9% sequence identity, because within-population nucleotide diversity only rarely exceeds 2%. However, some species are hypervariable and it is unclear how CO can occur between genotypes with less than 90% sequence identity. Here, we study CO in Schizophyllum commune, a hypervariable cosmopolitan basidiomycete mushroom, a frequently encountered decayer of woody substrates. We crossed two haploid individuals, from the United States and from Russia, and obtained genome sequences for their 17 offspring. The average genetic distance between the parents was 14%, making it possible to study CO at very high resolution. We found reduced levels of linkage disequilibrium between loci flanking the CO sites indicating that they are mostly confined to hotspots of recombination. Furthermore, CO events preferentially occurred in regions under stronger negative selection, in particular within exons that showed reduced levels of nucleotide diversity. Apparently, in hypervariable species CO must avoid regions of higher divergence between the recombining genomes due to limitations imposed by the mismatch repair system, with regions under strong negative selection providing the opportunity for recombination. These patterns are opposite to those observed in a number of less variable species indicating that population genomics of hypervariable species may reveal novel biological phenomena.

Purifying selection in mammalian mitochondrial protein-coding genes is highly effective and congruent with evolution of nuclear genes.

The mammalian mitochondrial genomes differ from the nuclear genomes by maternal inheritance, absence of recombination, and higher mutation rate. All these differences decrease the effective population size of mitochondrial genome and make it more susceptible to accumulation of slightly deleterious mutations. It was hypothesized that mitochondrial genes, especially in species with low effective population size, irreversibly degrade leading to decrease of organismal fitness and even to extinction of species through the mutational meltdown. To interrogate this hypothesis, we compared the purifying selections acting on the representative set of mitochondrial (potentially degrading) and nuclear (potentially not degrading) protein-coding genes in species with different effective population size. For 21 mammalian species, we calculated the ratios of accumulation of slightly deleterious mutations approximated by Kn/Ks separately for mitochondrial and nuclear genomes. The 75% of variation in Kn/Ks is explained by two independent variables: type of a genome (mitochondrial or nuclear) and effective population size of species approximated by generation time. First, we observed that purifying selection is more effective in mitochondria than in the nucleus that implies strong evolutionary constraints of mitochondrial genome. Mitochondrial de novo nonsynonymous mutations have at least 5-fold more harmful effect when compared with nuclear. Second, Kn/Ks of mitochondrial and nuclear genomes is positively correlated with generation time of species, indicating relaxation of purifying selection with decrease of species-specific effective population size. Most importantly, the linear regression lines of mitochondrial and nuclear Kn/Ks's from generation times of species are parallel, indicating congruent relaxation of purifying selection in both genomes. Thus, our results reveal that the distribution of selection coefficients of de novo nonsynonymous mitochondrial mutations has a similar shape with the distribution of de novo nonsynonymous nuclear mutations, but its mean is five times smaller. The harmful effect of mitochondrial de novo nonsynonymous mutations triggers highly effective purifying selection, which maintains the fitness of the mammalian mitochondrial genome.

Mercury and selenium concentrations in muscle tissue of different species of predatory freshwater fish and correlation between these elements.

Concentrations of total mercury and selenium were determined in 49 and 42 muscle tissue samples, respectively, of six species of predatory freshwater fish, dace (Leuciscus leuciscus), pike perch (Sander lucioperca), pike (Esox lucius), European catfish (Silurus glanis), rainbow trout (Oncorhynchus mykiss) and asp (Aspius aspius). Muscle selenium concentration did not correlate with the corresponding total mercury concentration (R² < 0.198) in all examined predatory fish species. There was an inverse correlation between the ratio Se/Hg content and the total mercury content in the muscle tissues of dace, pike perch, pike, European catfish and asp. The muscle tissue of rainbow trout exhibits a linear correlation between the ratio Se/Hg content and the total mercury content. The total mercury concentration of all examined samples did not exceed the hygienic limit for Hg for predatory fish.

Dynamics in the transient complex of plastocyanin-cytochrome f from Prochlorothrix hollandica.

The nature of transient protein complexes can range from a highly dynamic ensemble of orientations to a single well-defined state. This represents variation in the equilibrium between the encounter and final, functional state. The transient complex between plastocyanin (Pc) and cytochrome f (cytf) of the cyanobacterium Prochlorothrix hollandica was characterized by NMR spectroscopy. Intermolecular pseudocontact shifts and chemical shift perturbations were used as restraints in docking calculations to determine the structure of the wild-type Pc-cytf complex. The orientation of Pc is similar to orientations found in Pc-cytf complexes from other sources. Electrostatics seems to play a modest role in complex formation. A large variability in the ensemble of lowest energy structures indicates a dynamic nature of the complex. Two unusual hydrophobic patch residues in Pc have been mutated to the residues found in other plastocyanins (Y12G/P14L). The binding constants are similar for the complexes of cytf with wild-type Pc and mutant Pc, but the chemical shift perturbations are smaller for the complex with mutant Pc. Docking calculations for the Y12G/P14L Pc-cytf complex did not produce a converged ensemble of structures. Simulations of the dynamics were performed using the observed averaged NMR parameters as input. The results indicate a surprisingly large amplitude of mobility of Y12G/P14L Pc within the complex. It is concluded that the double mutation shifts the complex further from the well-defined toward the encounter state.

Expression of nuclear hormone receptors, their coregulators and type I iodothyronine 5'-deiodinase gene in mammary tissue of nonlactating and postlactating rats.

The aim of the study was to test the hypothesis that expression of retinoid receptors (RARalpha, RARbeta, RARgamma), rexinoid receptors (RXRalpha, RXRbeta), thyroid hormone receptors (TRalpha, TRbeta), estrogen receptors (ERalpha, ERbeta), nuclear receptor coregulators (N-CoR, SRC-1, SMRT), and in addition type I iodothyronine 5'-deiodinase (5'-DI), EGFR and erb-B2/neu would be different in mammary postlactating tissue in comparison with that of nonlactating mammary gland. Using RT-PCR, we have shown that expression of RARalpha, RXRalpha,TRalpha, ERalpha,ERbeta,N-CoR, SRC-1, SMRT and EGFR in rat was significantly increased in postlactating mammary gland when compared to that of nonlactating mammary tissue. Postlactating mammary glands were found to express all RAR and RXR subtypes studied when compared to nonlactating mammary tissues that express exclusively RARalpha and RXRalpha subtypes. Enhanced expression of a number of nuclear hormone receptors, their coregulators in mammary tissue of postlactating rats in comparison with nonlactating animals identify a potential role for retinoid, thyroid and estrogen signalling pathways also after lactation period.

Treatment of 1-methyl-1-nitrosourea-induced mammary tumours with immunostimulatory CpG motifs and 13-cis retinoic acid in female rats: histopathological study.

Histopathological evaluation of the mammary gland tumours of Sprague-Dawley rats induced with 1-methyl-1-nitrosourea (MNU), and treated with either CpG oligodeoxynucleotides (CpG-ODN) and/or 13-cis retinoic acid has been performed in this work. Since, the treatment of animals with CpG-ODN induced a significant decrease of tumour burden and volume in comparison with MNU treated control group (Macejova et al. 2001), it was of high impact to compare histological appearance of tumours in different experimental groups (MNU, CpG-ODN, 13-cis retinoic acid, CpG-ODN plus 13-cis retinoic acid). We have found reduced number of carcinomas with necroses in the CpG motifs treated group when compared to animals treated with MNU only. From the histological point of view the treatment with the CpG-ODN may have some protective effect. Carcinoma patterns proportion in the group treated with CpG-ODN was found to be different in comparison with other experimental groups. Treatment of rats with CpG-ODN had no apparent effect on invasiveness of developed carcinomas.