Novel (p)ppGpp Binding and Metabolizing Proteins of <i>Escherichia coli</i>

The alarmone (p)ppGpp plays pivotal roles in basic bacterial stress responses by increasing tolerance of various nutritional limitations and chemical insults, including antibiotics. Despite intensive studies since (p)ppGpp was discovered over 4 decades ago, (p)ppGpp binding proteins have not been systematically identified in Escherichia coli. We applied DRaCALA (differential radial capillary action of ligand assay) to identify (p)ppGpp-protein interactions. We discovered 12 new (p)ppGpp targets in E. coli that, based on their physiological functions, could be classified into four major groups, involved in (i) purine nucleotide homeostasis (YgdH), (ii) ribosome biogenesis and translation (RsgA, Era, HflX, and LepA), (iii) maturation of dehydrogenases (HypB), and (iv) metabolism of (p)ppGpp (MutT, NudG, TrmE, NadR, PhoA, and UshA). We present a comprehensive and comparative biochemical and physiological characterization of these novel (p)ppGpp targets together with a comparative analysis of relevant, known (p)ppGpp binding proteins. Via this, primary targets of (p)ppGpp in E. coli are identified. The GTP salvage biosynthesis pathway and ribosome biogenesis and translation are confirmed as targets of (p)ppGpp that are highly conserved between E. coli and Firmicutes. In addition, an alternative (p)ppGpp degradative pathway, involving NudG and MutT, was uncovered. This report thus significantly expands the known cohort of (p)ppGpp targets in E. coli

The level of physical activity in the lifestyle of physical education students

Subject: The level of physical activity in the lifestyle of physical education students Introduction: Physical activity is defined as voluntary and spontaneous physical activity practiced during leisure time. Thanks to it, the body can function properly and develop on many levels. It plays an important role in social and physical development, as well as intellectual and emotional development. It finds its place in the fight against civilization diseases, it is also beneficial in resisting many other diseases.    Aim: The aim of the article is to assess the level of physical activity of physical education students of various specializations.   Material and methods: The research was conducted among students of physical education at the University of Rzeszów. 60 people took part in them - 38 women and 22 men aged 19-30. The respondents answered anonymously to 18 closed, single-choice questions.   Results: The majority of students are engaged in physical activity outside a college, and 65% of it is at amateur level activity. The most common reason why respondents take up physical activity is to improve the functional fitness of the body. 58% of the respondents undertook physical activity more than 3 times a week.   Conclusions: Most of the respondents do physical activity outside of their studies. The most common reason for taking up physical activity among the respondents was the improvement of the functional fitness of the body, and the factors influencing the fact that students are physically active were to the greatest extent sports activitiesSubject: The level of physical activity in the lifestyle of physical education students Introduction: Physical activity is defined as voluntary and spontaneous physical activity practiced during leisure time. Thanks to it, the body can function properly and develop on many levels. It plays an important role in social and physical development, as well as intellectual and emotional development. It finds its place in the fight against civilization diseases, it is also beneficial in resisting many other diseases. &nbsp; Aim: The aim of the article is to assess the level of physical activity of physical education students of various specializations. &nbsp; Material and methods: The research was conducted among students of physical education at the University of Rzeszów. 60 people took part in them - 38 women and 22 men aged 19-30. The respondents answered anonymously to 18 closed, single-choice questions. &nbsp; Results: The majority of students are engaged in physical activity outside a college, and 65% of it is at amateur level activity. The most common reason why respondents take up physical activity is to improve the functional fitness of the body. 58% of the respondents undertook physical activity more than 3 times a week. &nbsp; Conclusions: Most of the respondents do physical activity outside of their studies. The most common reason for taking up physical activity among the respondents was the improvement of the functional fitness of the body, and the factors influencing the fact that students are physically active were to the greatest extent sports activities &nbsp; Key words: physical activity; lifestyle; physical education; student

Selected school solutions in the field of a healthy lifestyle among children and adolescents

Introduction and aim: Lifestyle is the daily behaviors, habits and activities adopted by people. It is one of the factors that largely determines the state of human health. It is important to pay attention to what habits and activities we implement in our lives. The source of behaviors are usually attitudes and values acquired in the family and the nearest social environment. The basic categories of pro-health behaviours are related to physical health, psychosocial health, health prevention and avoiding harmful behaviours, preventing an illness and supporting recovery. Anti-health behaviors contribute to health disorders, negative impacts in emotional, physical and psychosocial sphere. The aim of the study was to present selected school solutions in the field of pro-healthy lifestyle among children and adolescents, the influence of pro-healthy lifestyle and assessment of the implementation of health education at schools. Material and method: 85 children and adolescents in the age range of 10-19+ years participated in the study. Twenty-three students in the age range of 10-12 years, 27 students in the age range of 13-16 years and 35 students in the age range of 17-19+ years were surveyed. Twenty-eight males and 57 females took the survey. Results: Respondents most often engage in physical activity 2-3 times per week. The vast majority of respondents actively participate in physical education classes at school as well as undertake extracurricular physical activity. The most popular activities are cycling and team games. The main source of knowledge about pro-healthy lifestyle among children and adolescents is the Internet and the least popular is the press. Conclusions: The majority of students know what a pro-healthy lifestyle is, while less than half of the respondents have no such knowledge. The researched gain knowledge about the way of promoting pro-healthy lifestyle by school, most often through the organization of "Health Day". The majority of children and adolescents participate in extracurricular physical activity, while the rest of the respondents do not undertake such activity

Pyrrolidine nucleoside bisphosphonates as antituberculosis agents targeting hypoxanthine-guanine phosphoribosyltransferase

Therapeutic treatment of tuberculosis (TB) is becoming increasingly problematic due to the emergence of drug resistant Mycobacterium tuberculosis (Mt). Thus, new targets for anti-TB drug discovery need to be identified to combat and eradicate this disease. One such target is hypoxanthine-guanine phosphoribosyltransferase (HGPRT) which synthesises the 6-oxopurine nucleoside monophosphates essential for DNA/RNA production. [3R,4R]-4-Hypoxanthin-9-yl-3-( (S)-2-hydroxy-2-phosphonoethyl)oxy-1-N-(phosphonopropionyl)pyrrolidine and [3R,4R-4-guanin-9-yl-3-((S)-2-hydroxy-2-phosphonoethyl)oxy-1-N-(phosphonopropionyl)pyrrolidine (compound 6) are the most potent inhibitors of MtHGPRT yet discovered having K-i values of 60 nM. The crystal structure of the MtHGPRT.6 complex was obtained and compared with that of human HGPRT in complex with the same inhibitor. These structures provide explanations for the 60-fold difference in the inhibition constants between these two enzymes and a foundation for the design of next generation inhibitors. In addition, crystal structures of MtHGPRT in complex with two pyrrolidine nucleoside phosphosphonate inhibitors plus pyrophosphate provide insights into the final stage of the catalytic reaction. As the first step in ascertaining if such compounds have the potential to be developed as anti-TB therapeutics, the tetra-(ethyl L-phenylalanine) tetraamide prodrug of 6 was tested in cell based assays. This compound arrested the growth of virulent Mt not only in its replicating phase (IC50 of 14 mu M) but also in its latent phase (IC50 of 29 mu M). Furthermore, it arrested the growth of Mt in infected macrophages (MIC50 of 85 mu M) and has a low cytotoxicity in mammalian cells (CC50 of 132 +/- 20 mu M). These inhibitors are therefore viewed as forerunners of new anti-TB chemotherapeutics. (C) 2018 Elsevier Masson SAS. All rights reserved

Role of breast regression protein 39 (BRP-39)/chitinase 3-like-1 in Th2 and IL-13–induced tissue responses and apoptosis

Mouse breast regression protein 39 (BRP-39; Chi3l1) and its human homologue YKL-40 are chitinase-like proteins that lack chitinase activity. Although YKL-40 is expressed in exaggerated quantities and correlates with disease activity in asthma and many other disorders, the biological properties of BRP-39/YKL-40 have only been rudimentarily defined. We describe the generation and characterization of BRP-39−/− mice, YKL-40 transgenic mice, and mice that lack BRP-39 and produce YKL-40 only in their pulmonary epithelium. Studies of these mice demonstrated that BRP-39−/− animals have markedly diminished antigen-induced Th2 responses and that epithelial YKL-40 rescues the Th2 responses in these animals. The ability of interleukin13 to induce tissue inflammation and fibrosis was also markedly diminished in the absence of BRP-39. Mechanistic investigations demonstrated that BRP-39 and YKL-40 play an essential role in antigen sensitization and immunoglobulin E induction, stimulate dendritic cell accumulation and activation, and induce alternative macrophage activation. These proteins also inhibit inflammatory cell apoptosis/cell death while inhibiting Fas expression, activating protein kinase B/AKT, and inducing Faim 3. These studies establish novel regulatory roles for BRP-39/YKL-40 in the initiation and effector phases of Th2 inflammation and remodeling and suggest that these proteins are therapeutic targets in Th2- and macrophage-mediated disorders

Syntéza 4'-N pyrrolidinových nukleosidů, nukleotidů a oligonukleotidů

Synthetic routes to novel pyrrolidine nucleoside, nucleotide, and oligonucleotide analogues, where C-4‘ carbon atom of ribose is replaced with nitrogen atom of pyrrolidine have been developed

Synthesis of PME derivatives on nucleobases with conformation locked via pyrrolidine ring

Among diverse nucleoside phosphonic acids, the acyclic phosphonate derivatives have been earlier shown to involve some exceptionally potent antiviral drugs (e.g., Cidofovir, Adefovir, and Tenofovir). This work presents the synthesis of such acyclic phosphonate nucleotides conformationally locked via pyrrolidine ring