Cloning and sequence analysis of benzo-a-pyreneinducible cytochrome P450 1A in Nile tilapia (Oreochromis niloticus)

Polycyclic aromatic hydrocarbons (PAHs), dioxins, dibenzofurans and polychlorinated biphenyls (PCBs) present in polluted environment induce cytochrome P4501A (CYP1A) isozyme in fish which inturn results in a marked increased production of carcinogenic metabolites. The induction of hepatic CYP1A in fish by certain classes of chemicals has been suggested as an early warning system, a “mostsensitive biological response” for assessing environmental contamination conditions. This has implications for human fish consumption as well as for the health status of aquatic organisms.Considering the importance of Oreochromis niloticus fish as a laboratory animal, the common CYP1A sequence was determined from cDNA and genomic DNA after intraperitoneal injection with benzo-apyrene (BaP). The full-length cDNA was 2530 bp long and contained an open reading frame of 1566 bp encoding a protein of 521 amino acids and a stop codon. The sequence exhibited 5' and 3' noncodingregions of 134 and 830 bp, respectively. The deduced amino acid sequence of O. niloticus CYP1A shows similarities of 80.5, 79.3, 79.1, 77.8, 77.6, 74.3, 72.4, 77.2, 71.8, 70.7 and 50.8% with Europeanflounder CYP1A, scup CYP1A, killifish CYP1A, butterfly fish CYP1A, European sea bass CYP1A, rainbow trout CYP1A, Japanese eel CYP1A, toad fish CYP1A, European eel CYP1A, red sea bream CYP1A and common carp CYP1A, respectively. The phylogenetic tree based on the amino acid sequences clearly shows tilapia CYP1A and killifish CYP1A to be more closely related to each other than to the other CYP1A subfamilies. Sequence analysis of 3727 bp of genomic DNA showed that the clone obtained was the structural gene of CYP1A which consists of seven exons and six introns, the initiation codon was not found in the first exon but in the second one as was reported for the CYP1A genes of fish and mammals

Development and application of a real-time quantitative PCR assay for determining expression of benzo-apyrene- Inducible cytochrome P450 1A in Nile tilapia (Oreochromis niloticus)

Cytochrome P4501A’s (CYP1A) constitute a ubiquitous family of proteins associated with the detoxification of organic compounds such as PCB (polychlorinated biphenyl), PAH (polyaromatic hydrocarbons) and dioxin. These compounds are documented to induce the CYP1A gene in a variety of tissues of many fish species. Consequently, changes in CYP1A gene expression have been used as a biomarker for contaminant exposure in fish populations using a variety of techniques. Of all of thesemethods, quantitative PCR appears to be the most sensitive. It has been used to assess impact of environmental pollution in marine ecosystems using different fish models. Subsequently, for measuring benzo-a-pyrene (BaP) induction of CYP1A mRNA in different organs of tilapia (Oreochromis niloticus), ribosomal protein large P0-like protein (RPLP0-like protein) and -actin genes as internal controls were selected based on previous studies to assess their expression variability. Real-time polymerase chainreaction (real-time PCR) analysis of liver, intestine, gills and kidney revealed a distinct induced expression in liver and intestine (127.1 and 79.3 in liver, 26 and 56.1 in intestine using RPLP0 and -actin genes respectively as internal controls) with no detectable expression in the other organs studied

Multilevel System of Studying Plague Microbe Strains Proprties in the Republic of Kazakhstan

The study of freshly isolated cultures is necessary to form an objective idea of the properties of plague microbe natural populations. The analysis of the levels of investigating the properties of strains has been carried out and the characteristics of Yersinia pestis in Kazakhstan are presented. The results of studying the phenotypic and genetic properties of plague microbe natural strains are provided. Following the epizootiological survey of natural plague foci, the museums of live cultures at plague control stations annually receive strains of plague pathogen, which are transferred to the National Collection of Microorganisms of the National Scientific Center of Particularly Dangerous Infections (NSCPDI). One of the main points of Y. pestis strains analysis is the determination of their typicality/atypicality. The study of strains begins at the moment of their isolation by anti-epidemic units. The primary identification of strains is carried out in laboratories of anti-epidemic units by morphology, sensitivity to plague and pseudotuberculosis bacteriophages, fermentation of glycerol, rhamnose and sucrose. In the laboratories of plague control stations and departments, fermentation of maltose and arabinose, denitrification, amino acid requirements, virulence, sensitivity to antibiotics are additionally investigated. Analysis of strains virulence includes determination of calcium dependence, the presence and amount of F1, pesticinogenicity and sensitivity to pesticin 1 and virulence for white mice. The assessment and preservation of the collected gene pool in the NSCPDI National Collection includes various activities, one of the main ones is an in-depth study of all features using standard microbiological methods, molecular methods for complete identification and creation of a data bank containing information about the genome of strains at different intensity of the epizootic process. The NSCPDI has a digital database on the registration and movement of strains, equipment for molecular research. The collection evaluates properties, systematizes information, and ensures the viability of plague pathogen strains for longterm storage

Genotypic properties of collection plague microbes strains from the natural plague foci of Kazakhstan

Background. Epidemiological and epizootological monitoring of natural plague foci requires an integrated approach to solving problems, taking into account the phenotypic and genetic variability of Y. pestis and zoning of natural plague foci. The introduction of a new molecular genetic methodology aimed at studying the genomic polymorphism of the plague pathogen provides reliable results for the differentiation of not only groups, but also individual strains.The aim. To determine the genotypes of the plague microbe from different autonomous foci of the Republic of Kazakhstan.Materials and methods. 105 strains of Y. pestis isolated from various natural plague foci of Kazakhstan in 1951–2015 were studied. The phenotypic properties of the strains were studied using standard microbiological methods. A polymerase chain reaction (PCR) was used to detect fragments of the cafl, pst and YPO2088 genes. Multilocus variable number tandem repeat (VNTR) analysis (MLVA) was performed for 25 VNTR loci.Results. The phenotypic properties of the strains were preliminarily studied and the strains of the plague microbe were tested for specificity using the Pest-Quest test system (Kazakhstan). The PCR study confirmed the species-specific affiliation of Y. pestis strains. A variety of strains with typical phenotypic characteristics was revealed. MLVA for 25 key loci (MLVA25) revealed that the studied strains of the plague microbe are phylogenetically closest to the Mediaevalis biovar representatives. A phylogenetic tree of the studied strains has been obtained. It was found that 9 genotypes circulate on the territory of Kazakhstan, and their distribution in certain natural plague foci was determined.Conclusions. The resulting clustering indicates the relationship between the strain groups obtained on the dendrogram by the MLVA25 method and the territories of certain natural plague foci

ASSESSMENT OF THE VULNERABILITY OF THE POPULATION ON SARI-DZHAS AUTONOMOUS FOCUS OF PLAGUE, KYRGYZSTAN

This work is devoted to the assessment of the vulnerability of the Sary-Jaz autonomous focus of plague by using of GIS technologies. This focus of the plague covers a very large and difficult to access territory. The annual survey and disarming of this territory is very expensive. Therefore, it is necessary to select sectors that need more attention. That why, it is necessary to assess the vulnerability of the population in various sectors. Now in the world literature there is a lot of work on the assessment of vulnerability from these or those dangers, but there is no definition of hazard, risk and vulnerability indices for the focus of plague. The paper presents the results of the determination of these indices and calculates indices for each sector of the natural focus of the plague. As a result of the work, the final map of the Index of the epidemiological vulnerability of the Sary-Jaz autonomous focus of the plague was obtained. Detail results of the assessment and the final map for the study area are given. The authors hope that the calculations and results obtained in the course of this work will find their application both for theoretical assessments of the vulnerability of various territories from plague and for practical actions to reduce vulnerability

International Cooperation of Kazakhstan in the Prevention of Particularly Dangerous Infections

The dependence of the incidence of various nosologies on the epidemic situation in neighboring countries has been a feature of the epidemic process in recent years. In this regard, it is of particular importance to carry out joint anti-epidemic and preventive measures in border areas in order to prevent the importation of dangerous infections into the territory of neighboring states. The aim of the work was to analyze the results of international cooperation in the prevention of particularly dangerous infections. Presented are the areas of cooperation and outcomes of joint research activities. Measures for cooperation between the relevant institutions of the Ministry of Health of the Republic of Kazakhstan and the Rospotrebnadzor on the operational exchange of information in case of emergencies, joint research work on the monitoring of particularly dangerous and other natural-focal infectious diseases in the border areas, joint seminars, scientific and practical conferences on the introduction of modern methods of laboratory diagnostics into practice, internships on the exchange of experience in epizootiological survey in foci of particularly dangerous infections are described in the paper. Examples of Russian-Kazakh cooperation are provided. The results of a joint epizootiological survey of the territory of the Kazakh part of the Altai Mountains are presented. On a global scale, cases of plague and other particularly dangerous infections in any geographic region can constitute international public health emergencies and this type of threat requires international cooperation

Demarcation of the Boundaries of the Central Asian Desert Natural Focus of Plague of Kazakhstan and Monitoring the Areal of the Main Carrier, <I>Rhombomys opimus</I>

The aim of the study was to clarify the boundaries of the Central Asian natural plague focus of Kazakhstan and the modern boundaries of the areal of the great gerbil (Rhombomys opimus) in order to improve epizootiological monitoring and increase the effectiveness of preventive (anti-epidemic) measures.Materials and methods. Data from the epizootiological monitoring of the great gerbil populations in 14 autonomous foci of the Central Asian desert natural plague focus in the Republic of Kazakhstan between 2010 and 2020 were used for the analysis. An epizootiologic survey of an area of 875350 km2 was carried out. When processing the data, epidemiological, epizootiological, statistical research methods, as well as GIS technologies were used.Results and discussion. An increase in the total area of the Central Asian desert natural plague focus of the Republic of Kazakhstan by 79710 km2 (9.98 %) has been established for the period of 1990–2020. It is noted that the change in the area of plague-enzootic territory was a consequence of the ever changing areal of the main carrier of plague pathogen – the great gerbil – under the influence of climatic and anthropogenic factors. The most significant changes were found in the southeastern part of the plague-enzootic territory, including those for the Betpakdala (50 %), Balkhash (34.3 %), Taukum (13.3 %) and Mojynkum (0.32 %) autonomous foci. The area of the Aryskum-Dariyalyktakyr autonomous focus decreased by 2100 km2 (4 %). In 2000–2002, new Alakol’sky and Ili intermountain autonomous foci with a total area of 26759 km2 were discovered. It is shown that due to the regression of the Aral Sea, the areal of the great girbil expanded and the area of the North Aral and Kyzylkum natural plague foci increased by 10500 km2 (29.2 %) and 560 km2 (0.4%), respectively. The areas of the Aral-Karakum and UralEmba desert autonomous foci, on the contrary, decreased by 2000 km2 (2.6 %) and 12300 km2 (17.6 %), respectively. Passportization and landscape-epizootiologic zoning of the territory of the Central Asian desert natural plague focus of the Republic of Kazakhstan has been completed

Zoning of the Territory of the Republic of Kazakhstan by the Degree of Intensity of the Epizootic Situation on Plague in Camels

The aim of the study was to improve epizootiological monitoring and increase the effectiveness of preventive (anti-epidemic) measures for camel plague control in Kazakhstan.Materials and methods. We used the data on epizootiological and epidemiological monitoring in natural plague foci of Kazakhstan, long-term measurements and indicators for the period of 2000–2020 of the anti-plague and veterinary services of the Republic for the analysis. To process the evidence, epidemiological, epizootiological, microbiological, and statistical research methods, as well as GIS technology were applied.Results and discussion. The number of camels has increased by 2.2 times in Kazakhstan over the past 20 years.  Where there were 98.2 thousand heads in 2000, it amounted to 216.4 thousand heads in 2020. Over the past 10 years, 152 camels died of unknown causes in the focal area of the country, but laboratory tests for plague turned out negative. According to the hazard criteria, the territory of the country has been conditionally divided into three zones: five regions with a high degree of hazard with a total area of 953.15 sq. km, five regions with medium degree of hazard with a total area of 1230.72 sq. km, and with a low degree of hazard – four regions and three cities of republican significance with a total area of 541.1 sq. km. Constant epizootiological monitoring over plague in camels is a necessity for the system of preventive measures

Development of Non-Natural Flavanones as Antimicrobial Agents

With growing concerns over multidrug resistance microorganisms, particularly strains of bacteria and fungi, evolving to become resistant to the antimicrobial agents used against them, the identification of new molecular targets becomes paramount for novel treatment options. Recently, the use of new treatments containing multiple active ingredients has been shown to increase the effectiveness of existing molecules for some infections, often with these added compounds enabling the transport of a toxic molecule into the infecting species. Flavonoids are among the most abundant plant secondary metabolites and have been shown to have natural abilities as microbial deterrents and anti-infection agents in plants. Combining these ideas we first sought to investigate the potency of natural flavonoids in the presence of efflux pump inhibitors to limit Escherichia coli growth. Then we used the natural flavonoid scaffold to synthesize non-natural flavanone molecules and further evaluate their antimicrobial efficacy on Escherichia coli, Bacillus subtilis and the fungal pathogens Cryptococcus neoformans and Aspergillus fumigatus. Of those screened, we identified the synthetic molecule 4-chloro-flavanone as the most potent antimicrobial compound with a MIC value of 70 µg/mL in E. coli when combined with the inhibitor Phe-Arg-ß-naphthylamide, and MICs of 30 µg/mL in S. cerevesiae and 30 µg/mL in C. neoformans when used alone. Through this study we have demonstrated that combinatorial synthesis of non-natural flavonones can identify novel antimicrobial agents with activity against bacteria and fungi but with minimal toxicity to human cells

Observation of associated near-side and away-side long-range correlations in √sNN=5.02 TeV proton-lead collisions with the ATLAS detector

Two-particle correlations in relative azimuthal angle (Δϕ) and pseudorapidity (Δη) are measured in √sNN=5.02  TeV p+Pb collisions using the ATLAS detector at the LHC. The measurements are performed using approximately 1  μb-1 of data as a function of transverse momentum (pT) and the transverse energy (ΣETPb) summed over 3.1<η<4.9 in the direction of the Pb beam. The correlation function, constructed from charged particles, exhibits a long-range (2<|Δη|<5) “near-side” (Δϕ∼0) correlation that grows rapidly with increasing ΣETPb. A long-range “away-side” (Δϕ∼π) correlation, obtained by subtracting the expected contributions from recoiling dijets and other sources estimated using events with small ΣETPb, is found to match the near-side correlation in magnitude, shape (in Δη and Δϕ) and ΣETPb dependence. The resultant Δϕ correlation is approximately symmetric about π/2, and is consistent with a dominant cos⁡2Δϕ modulation for all ΣETPb ranges and particle pT