Stimulation of effective ecological and economic interactions in the process of business environment creation

The article substantiates the necessity of introducing a new national framework document that would outline strategic directions for long-term development of Ukraine and facilitate the integration of sustainable development into the national plans, strategies and programs. It provides recommendations concerning the projection of business environment taking into account environmental and economic constraints and green technologies in Ukraine, as well as the use of economic and mathematical modeling. It offers the introduction of ecological and economic projects to enhance control and responsibility for the implementation of the planned measures on all levels, constant examination of the current legislation regarding its compliance with the principles of sustainable development; active use of public examination and independent audit; ensuring continuous dialogue between representatives of business and society

Non-Roundabout design of cancel the intersection signal light on horizontal plane

Traffic congestion is a world problem and an important factor restricting urban development. In order to solve the problem of urban traffic congestion, this paper takes the traffic flow theory and the intersection channel design theory as the research foundation, and conducts in-depth research on the causes of congestion at the intersection and the corresponding solutions, and proposes to cancel the traffic lights at the intersection without any stagnation. This paper proposes a new intersection design scheme, which is like the veins of the flower veins to channel the design intersection, cancel the signal light, and the vehicle can pass through the intersection without stagnation. It proposes a new solution to solve the traffic congestion problem. This new design allows the traffic flow to be spatially separated on the horizontal plane, and due to the cancellation of the signal lights, there is no signal waiting at the intersection, and the vehicle can travel without stopping at the intersection. At the same time, this paper also establishes a plane intersection service capability evaluation system based on simulation and quantitative calculation, which provides an evaluation index and proof basis for the non-stagnation driving channel design of the non-roundabout intersection

Non-Roundabout design of cancel the intersection signal light on horizontal plane

Traffic congestion is a world problem and an important factor restricting urban development. In order to solve the problem of urban traffic congestion, this paper takes the traffic flow theory and the intersection channel design theory as the research foundation, and conducts in-depth research on the causes of congestion at the intersection and the corresponding solutions, and proposes to cancel the traffic lights at the intersection without any stagnation. This paper proposes a new intersection design scheme, which is like the veins of the flower veins to channel the design intersection, cancel the signal light, and the vehicle can pass through the intersection without stagnation. It proposes a new solution to solve the traffic congestion problem. This new design allows the traffic flow to be spatially separated on the horizontal plane, and due to the cancellation of the signal lights, there is no signal waiting at the intersection, and the vehicle can travel without stopping at the intersection. At the same time, this paper also establishes a plane intersection service capability evaluation system based on simulation and quantitative calculation, which provides an evaluation index and proof basis for the non-stagnation driving channel design of the non-roundabout intersection. Document type: Articl

7,8-dihydro-8-oxoadenine, a highly mutagenic adduct, is repaired by Escherichia coli and human mismatch-specific uracil/thymine-DNA glycosylases

Hydroxyl radicals predominantly react with the C8 of purines forming 7,8-dihydro-8-oxoguanine (8oxoG) and 7,8-dihydro-8-oxoadenine (8oxoA) adducts, which are highly mutagenic in mammalian cells. The majority of oxidized DNA bases are removed by DNA glycosylases in the base excision repair pathway. Here, we report for the first time that human thymine-DNA glycosylase (hTDG) and Escherichia coli mismatch-specific uracil-DNA glycosylase (MUG) can remove 8oxoA from 8oxoA•T, 8oxoA•G and 8oxoA•C pairs. Comparison of the kinetic parameters of the reaction indicates that full-length hTDG excises 8oxoA, 3,N4-ethenocytosine (εC) and T with similar efficiency (kmax = 0.35, 0.36 and 0.16 min−1, respectively) and is more proficient as compared with its bacterial homologue MUG. The N-terminal domain of the hTDG protein is essential for 8oxoA-DNA glycosylase activity, but not for εC repair. Interestingly, the TDG status had little or no effect on the proliferation rate of mouse embryonic fibroblasts after exposure to γ-irradiation. Nevertheless, using whole cell-free extracts from the DNA glycosylase-deficient murine embryonic fibroblasts and E. coli, we demonstrate that the excision of 8oxoA from 8oxoA•T and 8oxoA•G has an absolute requirement for TDG and MUG, respectively. The data establish that MUG and TDG can counteract the genotoxic effects of 8oxoA residues in viv

7,8-Dihydro-8-oxoadenine, a highly mutagenic adduct, is repaired by Escherichia coli and human mismatch-specific uracil/thymine-DNA glycosylases

Hydroxyl radicals predominantly react with the C(8) of purines forming 7,8-dihydro-8-oxoguanine (8oxoG) and 7,8-dihydro-8-oxoadenine (8oxoA) adducts, which are highly mutagenic in mammalian cells. The majority of oxidized DNA bases are removed by DNA glycosylases in the base excision repair pathway. Here, we report for the first time that human thymine-DNA glycosylase (hTDG) and Escherichia coli mismatch-specific uracil-DNA glycosylase (MUG) can remove 8oxoA from 8oxoA*T, 8oxoA*G and 8oxoA*C pairs. Comparison of the kinetic parameters of the reaction indicates that full-length hTDG excises 8oxoA, 3,N(4)-ethenocytosine (epsilonC) and T with similar efficiency (k(max) = 0.35, 0.36 and 0.16 min(-1), respectively) and is more proficient as compared with its bacterial homologue MUG. The N-terminal domain of the hTDG protein is essential for 8oxoA-DNA glycosylase activity, but not for epsilonC repair. Interestingly, the TDG status had little or no effect on the proliferation rate of mouse embryonic fibroblasts after exposure to gamma-irradiation. Nevertheless, using whole cell-free extracts from the DNA glycosylase-deficient murine embryonic fibroblasts and E. coli, we demonstrate that the excision of 8oxoA from 8oxoA*T and 8oxoA*G has an absolute requirement for TDG and MUG, respectively. The data establish that MUG and TDG can counteract the genotoxic effects of 8oxoA residues in vivo

Aberrant repair initiated by mismatch-specific thymine-DNA glycosylases provides a mechanism for the mutational bias observed in CpG islands

The human thymine-DNA glycosylase (TDG) initiates the base excision repair (BER) pathway to remove spontaneous and induced DNA base damage. It was first biochemically characterized for its ability to remove T mispaired with G in CpG context. TDG is involved in the epigenetic regulation of gene expressions by protecting CpG-rich promoters from de novo DNA methylation. Here we demonstrate that TDG initiates aberrant repair by excising T when it is paired with a damaged adenine residue in DNA duplex. TDG targets the non-damaged DNA strand and efficiently excises T opposite of hypoxanthine (Hx), 1,N6-ethenoadenine, 7,8-dihydro-8-oxoadenine and abasic site in TpG/CpX context, where X is a modified residue. In vitro reconstitution of BER with duplex DNA containing Hx•T pair and TDG results in incorporation of cytosine across Hx. Furthermore, analysis of the mutation spectra inferred from single nucleotide polymorphisms in human population revealed a highly biased mutation pattern within CpG islands (CGIs), with enhanced mutation rate at CpA and TpG sites. These findings demonstrate that under experimental conditions used TDG catalyzes sequence context-dependent aberrant removal of thymine, which results in TpG, CpA→CpGmutations, thus providing a plausible mechanism for the putative evolutionary origin of the CGIs in mammalian genomes

Genetic and Biochemical Characterization of Human AP Endonuclease 1 Mutants Deficient in Nucleotide Incision Repair Activity

Background: Human apurinic/apyrimidinic endonuclease 1 (APE1) is a key DNA repair enzyme involved in both base excision repair (BER) and nucleotide incision repair (NIR) pathways. In the BER pathway, APE1 cleaves DNA at AP sites and 39-blocking moieties generated by DNA glycosylases. In the NIR pathway, APE1 incises DNA 59 to a number of oxidatively damaged bases. At present, physiological relevance of the NIR pathway is fairly well established in E. coli, but has yet to be elucidated in human cells. Methodology/Principal Finding: We identified amino acid residues in the APE1 protein that affect its function in either the BER or NIR pathway. Biochemical characterization of APE1 carrying single K98A, R185A, D308A and double K98A/R185A amino acid substitutions revealed that all mutants exhibited greatly reduced NIR and 39R59 exonuclease activities, but were capable of performing BER functions to some extent. Expression of the APE1 mutants deficient in the NIR and exonuclease activities reduced the sensitivity of AP endonuclease-deficient E. coli xth nfo strain to an alkylating agent, methylmethanesulfonate, suggesting that our APE1 mutants are able to repair AP sites. Finally, the human NIR pathway was fully reconstituted in vitro using the purified APE1, human flap endonuclease 1, DNA polymerase b and DNA ligase I proteins, thus establishing the minimal set of proteins required for a functional NIR pathway in human cells. Conclusion/Significance: Taken together, these data further substantiate the role of NIR as a distinct and separable functio

The development of the solution search method based on the improved bee colony algorithm

Active digitization of people's daily life leads to the use of the decision-making support systems (DMSS). DMSS is actively used in data processing, forecasting the course of various processes, providing informational support for the decision-making process by decision makers. However, a number of problems arise while evaluating monitoring objects, namely: a large number of destabilizing factors affecting the efficiency of the processes of information collection, processing and transmission; high dynamism of changes in the state and composition of heterogeneous monitoring objects during the conduct of hostilities (operations); high dynamism of conducting hostilities (operations); the uncertainty of the initial situation and the noise of the initial data. In this article, a method of finding solutions based on an improved bee colony algorithm was developed. The efficiency of information processing is achieved by learning the architecture of artificial neural networks; taking into account the type of uncertainty of the information to be evaluated; the use of an improved algorithm of the bee colony, the use of an unordered linguistic scale of measurements with adjustment coefficients for the degree of awareness and the degree of noise of the initial data. An approbation of the use of the proposed method was carried out on the example of assessing the state of the operational grouping of troops (forces). The method is proposed to be used in the development of software for automated systems of control of troops and weapons, namely, in the modernization of existing and development of new automated systems of control of troops and weapons. The evaluation of the effectiveness of the proposed method showed an increase in the efficiency of the evaluation at the level of 21–28 % in terms of the efficiency of information processin

Lys98 Substitution in Human AP Endonuclease 1 Affects the Kinetic Mechanism of Enzyme Action in Base Excision and Nucleotide Incision Repair Pathways

Human apurinic/apyrimidinic endonuclease 1 (APE1) is a key enzyme in the base excision repair (BER) and nucleotide incision repair (NIR) pathways. We recently analyzed the conformational dynamics and kinetic mechanism of wild-type (wt) protein, in a stopped-flow fluorescence study. In this study, we investigated the mutant enzyme APE1K98A using the same approach. Lys98 was known to hydrogen bond to the carboxyl group of Asp70, a residue implicated in binding the divalent metal ion. Our data suggested that the conformational selection and induced fit occur during the enzyme action. We expanded upon the evidence that APE1 can pre-exist in two conformations. The isomerization of an enzyme-product complex in the BER process and the additional isomerization stage of enzyme-substrate complex in the NIR process were established for APE1K98A. These stages had not been registered for the wtAPE1. We found that the K98A substitution resulted in a 12-fold reduction of catalytic constant of 5′-phosphodiester bond hydrolysis in (3-hydroxytetrahydrofuran-2-yl)methyl phosphate (F, tetrahydrofuran) containing substrate, and in 200-fold reduction in 5,6-dihydrouridine (DHU) containing substrate. Thus, the K98A substitution influenced NIR more than BER. We demonstrated that the K98A mutation influenced the formation of primary unspecific enzyme-substrate complex in a complicated manner, depending on the Mg2+ concentration and pH. This mutation obstructed the induced fit of enzyme in the complex with undamaged DNA and F-containing DNA and appreciably decreased the stability of primary complex upon interaction of enzyme with DNA, containing the natural apurinic/apyrimidinic (AP) site. Furthermore, it significantly delayed the activation of the less active form of enzyme during NIR and slowed down the conformational conversion of the complex of enzyme with the cleavage product of DHU-substrate. Our data revealed that APE1 uses the same active site to catalyze the cleavage of DHU- and AP-substrates

Structural and functional investigation of flavin binding center of the NqrC subunit of sodium-translocating NADH:Quinone oxidoreductase from Vibrio harveyi

Na+-translocating NADH:quinone oxidoreductase (NQR) is a redox-driven sodium pump operating in the respiratory chain of various bacteria, including pathogenic species. The enzyme has a unique set of redox active prosthetic groups, which includes two covalently bound flavin mononucleotide (FMN) residues attached to threonine residues in subunits NqrB and NqrC. The reason of FMN covalent bonding in the subunits has not been established yet. In the current work, binding of free FMN to the apo-form of NqrC from Vibrio harveyi was studied showing very low affinity of NqrC to FMN in the absence of its covalent bonding. To study structural aspects of flavin binding in NqrC, its holo-form was crystallized and its 3D structure was solved at 1.56 Å resolution. It was found that the isoalloxazine moiety of the FMN residue is buried in a hydrophobic cavity and that its pyrimidine ring is squeezed between hydrophobic amino acid residues while its benzene ring is extended from the protein surroundings. This structure of the flavin-binding pocket appears to provide flexibility of the benzene ring, which can help the FMN residue to take the bended conformation and thus to stabilize the one-electron reduced form of the prosthetic group. These properties may also lead to relatively weak noncovalent binding of the flavin. This fact along with periplasmic location of the FMN-binding domains in the vast majority of NqrC-like proteins may explain the necessity of the covalent bonding of this prosthetic group to prevent its loss to the external medium