Polyphenol oxidase from Pectobacterium atrosepticum: identification and cloning of gene and characteristics of the enzyme

© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim In the present study, we attempted to elucidate if the harmful phytopathogenic bacteria of Pectobacterium genus (P. atrosepticum) possess the enzymes for oxidation of phenolic compounds. Polyphenol oxidase (laccase) activity was revealed in P. atrosepticum cell lysates. Using bioinformatic analysis, an ORF encoding a putative copper-containing polyphenol oxidase of 241 amino acids with a predicted molecular mass of 25.9 kDa was found. This protein (named Pal1) shares significant level of identity with laccases of a new type described for several bacterial species. Cloning and expression of the pal1 gene and the analysis of corresponding recombinant protein confirmed that Pal1 possessed laccase activity. The recombinant Pal1 protein was characterized in terms of substrate specificity, kinetic parameters, pH and temperature optimum, sensitivity to inhibitors and metal content. Pal1 demonstrated alkali- and thermo-tolerance. The kinetic parameters K m and kcat for 2,6-dimethoxyphenol were 0.353 ± 0.062 mM and 98.79 ± 4.9 s −1 , respectively. The protein displayed high tolerance to sodium azide, sodium fluoride, NaCl, SDS and cinnamic acid. The transcript level of the pal1 gene in P. atrosepticum was shown to be induced by plant-derived phenolic compound (ferulic acid) and copper sulfate

Algorithm for Physiological Interpretation of Transcriptome Profiling Data for Non-Model Organisms

© 2018, Pleiades Publishing, Inc. Modern techniques of next-generation sequencing (NGS) allow obtaining expression profile of all genes and provide an essential basis for characterizing metabolism in the organism of interest on a broad scale. An important condition for obtaining a demonstrative physiological picture using high throughput sequencing data is the availability of the genome sequence and its sufficient annotation for the target organism. However, a list of species with properly annotated genomes is limited. Transcriptome profiling is often performed in the so-called non-model organisms, which are those with unknown or poorly assembled and/or annotated genome sequences. The transcriptomes of non-model organisms are possible to investigate using algorithms of de novo assembly of the transcripts from sequences obtained as the result of RNA sequencing. A physiological interpretation of the data is difficult in this case because of the absence of annotation of the assembled transcripts and their classification by metabolic pathway and functional category. An algorithm for transcriptome profiling in non-model organisms was developed, and a transcriptome analysis was performed for the basidiomycete Lentinus edodes. The algorithm includes open access software and custom scripts and encompasses a complete analysis pipeline from the selection of cDNA reads to the functional classification of differentially expressed genes and the visualization of the results. Based on this algorithm, a comparative transcriptome analysis of the nonpigmented mycelium and brown mycelial mat was performed in L. edodes. The comparison revealed physiological differences between the two morphogenetic stages, including an induction of cell wall biogenesis, intercellular communication, ion transport, and melanization in the brown mycelial mat

Effects of experimental domestication of silver foxes (Vulpes vulpes) on vocal behaviour

This paper systematizes and generalizes a research cycle devoted to studying the acoustics and vocal behaviour of silver foxes that differ in their tolerance to humans. The research revealed that 50-year selection for tameness toward people resulted in selective use by Tame foxes toward humans of two call types, pant and cackle. At the same time, the selected for aggression toward people Aggressive foxes and the non-selected for behaviour Control foxes, selectively use toward humans cough and snort. Thus, call types representing vocal indicators of friendly and aggressive behaviour of foxes toward humans have been revealed by the research. Nevertheless, experimental domestication did not change vocal behaviour of foxes toward conspecifics; all three strains did not differ by their vocal behaviour toward same-strain silver foxes. Relationship has been investigated between vocal behaviour and degree of tolerance toward people for hybrids between Tame and Aggressive foxes and for backcrosses to Tame and Aggressive foxes. Effect was estimated between fox sex and the degree of human impact on focal fox for variables of fox vocal behaviour. The research revealed the universal for mammals vocal indicators of emotional arousal that are independent of the emotional valence. Characteristics of vocal behaviour that are related with positive and negative emotional valence have been revealed. A simple and effective method for estimating animal discomfort based on ”joint calls” that takes into account the characteristics of all calls irrespective of their acoustic structure has been revealed. The obtained results provide a basis for further comparative studies of the acoustic structure and vocal behaviour for other taxa of the genus Vulpes and the related canid genera (Canis, Cuon, Lycaon)

Kind granddaughters of angry grandmothers: The effect of domestication on vocalization in cross-bred silver foxes

a b s t r a c t The genetic basis of the effects of domestication has previously been examined in relation to morphological, physiological and behavioural traits, but not for vocalizations. According t

Synthesis and Testing of Abscisic Acid with Predominant Replacement of Protium Atoms by Tritium in the Cyclohexene Moiety

© 2018, Pleiades Publishing, Inc. Abstract: A procedure for tritiation of predominantly the cyclohexene moiety of abscisic acid was developed. Tritium was introduced by isotope exchange reaction with 100% tritiated water at 220°C in the presence of diisopropylethylamine. The yield of abscisic acid was 50%, and the specific activity was 30.5 Ci/mmol. The labeled product was tested. It was shown that tritiated abscisic acid synthesized by the proposed method did not differ from the unlabeled precursor and could be used for biological assays

Hypoxia-induced factor-1α and markers of inflammation in patients with ischemic stroke

Ischemic stroke (IS) occurs as a result of local disturbance of hemocirculation and hypoxia in the brain tissue. Hypoxia-inducible factor-1α (HIF-1α), which is involved in the regulation of tissue oxygen levels, plays an important role in the pathophysiology of stroke, including neuronal survival, neuroinflammation, angiogenesis, glucose metabolism, blood-brain barrier permeability, and is important in IS outcomes. The purpose of the study was to determine the relationship between blood levels of HIF-1α and the degree of neurological deficit in the acute period of IS and the outcome of the disease. We examined 58 people with IS aged 73 (67-81) years. Patients were divided into two groups – discharged and dead. The severity of stroke (NIHSS), neurological deficit, comorbidity index, blood levels of HIF-1α, p53 protein, interleukin-6, cystatin C, CRP, creatinine, hematological parameters were determined at admission, on days 3 and 10 of the disease. At admission the blood levels of HIF-1α was lower than in the comparison group and remained reduced until the 10th day. On day 10 the association of HIF-1α with neurological deficit, comorbidity index and disease outcome was determined. We observed a feedback of HIF-1α with the content of erythrocytes, hemoglobin and hematocrit, which can be regarded as a reflection of the hemic component of mixed hypoxia. In dead patients, an increased blood level of cystatin C was detected, which was associated with HIF-1α concentrations. In all periods of observation of IS, a correlation between cystatin C and creatinine and CRP levels was noted. These results may indicate dysfunction of endotheliocytes, inflammation associated with hypoxia in IS. The prognostic significance of the blood level of HIF-1α on the 10th day for the outcome of IS was AUC = 0.900. Blood levels of HIF-1α in the acute period was associated with the severity of IS and the outcome of the disease

Fungicide resistance of <i>Microdochium nivale</i> strains and its interconnection with virulence

The aim of the study was to analyze the resistance of 21 Microdochium nivale strains (fungi that cause pink snow mold in winter crops), living within a single area and a single crop (winter rye), to fungicides that differ in chemical nature and mechanisms of action as well as to test the interconnection between levels of virulence and fungicide resistance of strains. The virulence of M. nivale strains was determined on detached leaves of rye (Ogonek variety) plants as well as on whole plants grown under sterile conditions in vitro. The resistance of strains to fungicides (Provisor (a.s. azoxystrobin) and Ferazim (a.s. carbendazim)) was determined by inhibition of mycelium growth. As a result of the experiments, more strains (13) capable of growing in the presence of Provisor were revealed than Ferazim-resistant strains (2); however, Ferazim-resistant strains had a greater level of resistance (5-16% of growth inhibition) than Provisor-resistant strains (63-94 % of growth inhibition). A negative correlation (Spearman's correlation coefficient -0.604 and -0.532) between the level of virulence of M. nivale strains and the level of their susceptibility to Provisor was found. This evidently means that the acquisition of resistance to Provisor by M. nivale strains is accompanied by an increase in their virulence. No correlations were found between the fungicide resistance of M. nivale strains and their attribution to one or another phylogenetic group, to which the strains were assigned based on the nucleotide sequence of the ITS2 (internal transcribed spacer 2) region. The study shows that for choosing the fungicide application strategy, it is necessary to analyze phytopathogen populations for the presence of strains that are simultaneously characterized by high virulence and resistance to various fungicides and also take into account that the adaptation of fungi to certain fungicides can be accompanied by an increase in their virulence, which will negatively affect the phytopathological state of agrocenosis

Transcriptome profiling helps to identify potential and true molecular switches of stealth to brute force behavior in Pectobacterium atrosepticum during systemic colonization of tobacco plants

© 2018, Koninklijke Nederlandse Planteziektenkundige Vereniging. In the present study, we have monitored the process of systemic plant colonization by the plant pathogenic bacterium Pectobacterium atrosepticum (Pba) using RNA-Seq analysis in order to compare bacterial traits under in planta and in vitro conditions and to reveal potential players that participate in switching from stealth to brute force strategy of the pathogen. Two stages of tobacco plant colonization have been assayed: i) the initial one associated with visually symptomless spread of bacteria throughout the host body via primary xylem vessels where bacterial emboli were formed (stealth strategy), and ii) the advanced stage coupled with an extensive colonization of core parenchyma and manifestation of soft rot symptoms (brute force strategy). Plant-inducible genes in Pba and potential players switching the pathogen’s behavior were revealed. Genes from the cfa locus responsible for the production of coronafacic acid displayed the strongest induction in the asymptomatic zone relative to the symptomatic one and were shown experimentally to act as the true strategy “switchers” of Pba behavior in planta. Surprisingly, cfa genes appeared to be unnecessary for establishment of the asymptomatic stage of plant colonization but were required for the transition to soft-rot-associated symptomatic stage coupled with over-induction of jasmonate-mediated pathway in the plant

EXPERIENCE OF USING WEBINARS IN THE IMPLEMENTATION OF PROGRAMS OF ADDITIONAL PROFESSIONAL EDUCATION IN THE DEPARTMENT OF HYGIENE AND PROFESSIONAL DISEASES WITH A COURSE OF PHYSIOTHERAPY, THERAPEUTIC PHYSICAL TRAINING AND SPORTS MEDICINE

The article is devoted to the experience of conducting webinars for the implementation of programs of additional professional training of doctors at the Department of Hygiene and Occupational Diseases with a course of physiotherapy, exercise therapy and sports medicine of USMU. The main advantages of webinars over traditional forms of teaching are shown, among which the expansion of the geography of the contingent of students is important. The conditions for the successful application of telecommunication technologies in the educational process are given.Рассмотрен опыт проведения вебинаров для реализации программ дополнительного профессионального обучения врачей на кафедре гигиены и профессиональных болезней с курсом физиотерапии, ЛФК и спортивной медицины УГМУ. Показаны основные преимущества вебинаров перед традиционными формами преподавания, в частности расширение географии контингента слушателей. Приведены условия успешного применения телекоммуникационных технологий в образовательном процессе

Silicotuberculosis in the presser of refractory products in modern working conditions

The aim of the study – to study and describe the case of silicotuberculosis in a presser of refractory products in modern working conditions.Цель исследования – изучение и описание случая силикотуберкулеза у прессовщика огнеупорных изделий в современных условиях труда