scholarly journals BACTERIOPHAGE PREPARATION ENGINEERING FOR BIOCONTROL OF PSEUDOMONAS SYRINGAE IN CROP SCIENCE

2020 ◽  
pp. 130-137
Author(s):  
D. А. Vasiliev ◽  
◽  
А.К. Bekkalieva ◽  
N. А. Feoktistova ◽  
Е. V. Suldina ◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Biocontrol Agent ◽  
Molecular Genetic ◽  
Living Cells ◽  
Lytic Activity ◽  
Cultivated Plants ◽  
Phage Lysate ◽  
Genetic Characteristics ◽  
Phytopathogenic Bacteria ◽  
Crop Science

Phytopathogenic bacteria Pseudomonas syringae cause diseases of many cultivated plants, causing tumor neoformation, rot, chlorosis, necrosis, etc . The advanced biological mean to control bacteriosis in crop science is bacteriophages. In this work full biological characteristic of 8 bacteriophages is shown, active according to Pseudomonas syringae. The studied phages formed similar negative colonies- clear, rounded, in diameter of 5-9 mm. Lytic actifity of phages Pseudomonas syringae by Appelman from 10-4 to 10-8; by Gratia from 1,0±0,1×106 to 2,0±0,1×109 (BFU/ml). Bacteriophages Ps.s-7 UlGAU, Ps.s-13 UlGAU and Ps.s-27 UlGAU did not change lytic activity when storing in fridge during 12 months. Lytic activity of phages Ps.s-1 UlGAU, Ps.s-8, Ps.s-15 UlGAU, Ps.s-30 UlGAU, Ps.s-77 UlGAU in the same conditions fell within 1-2 orders. Spectrum of lytic activity of phages varied from 21,4% (Ps.s-13 UlGAU) to 85,7% (Ps.s-7 UlGAU, Ps.s-27 UlGAU). The study of phage specificity on 15 species og heterologous cultures showed that phages are specific for Pseudomonas syringae. Phages are moderately stable to heating and lose activity during 30 –minute temperature effect above 62°С. The optimal way to relieve phage lysate from living cells of Pseudomonas syringae was thrichloromethane at a ratio of 10:1and temporal exposition 45 minutes. On the basis of obtained data we determined capacity of each bacteriophage for the use as biocontrol agent. For further research for the study of molecular genetic characteristics we selected advanced strains of phages Pseudomonas syringae - Ps.s-7 UlGAU и Ps.s-27 UlGAU.

scholarly journals OPTIMIZATION OF PROCESS PARAMETERS FOR CULTIVATION OF BACTERIOPHAGES PROMISING FOR BIOLOGICAL CONTROL OF PHYTOPATHOGENIC BACTERIA OF GENUS PSEUDOMONAS

2021 ◽  
Author(s):  
T.A. Pilipchuk ◽  
A.D. Gerasimovich ◽  
I.N. Ananyeva ◽  
E.I. Kolomiets
Keyword(s):  
Biological Control ◽  
Pseudomonas Syringae ◽  
Process Parameters ◽  
Fish Meal ◽  
Aeration Rate ◽  
Lytic Activity ◽  
Agitation Rate ◽  
Phytopathogenic Bacteria ◽  
Optimization Of Process Parameters ◽  
Optimized Conditions

Cultural parameters of bacteriophages screened for lytic activity toward host bacterial strain P. helmanticensis BIM В-582 D were investigated. The following conditions were shown to be optimal for cultivation of indicator culture and it’s lysis by Pseudomonas phages BIM ВV-45 D, BIM ВV-46 D, BIM ВV-47 D, BIM ВV-50 D, BIM ВV-53 D, BIM ВV-61 D in laboratory fermenters ANKUM-3M: nutrient medium – fish meal hydrolysate broth, temperature – 28°C, stirrer agitation rate – 160 rpm, aeration rate – 1 l air/ l medium per min. Biopreparation produced under optimized conditions and composed of six afore-mentioned phages display high lytic activity both in regard to the host strain and to phytopathogenic bacteria Pseudomonas syringae.

Clinical application of polymorphic variants of the genes ESR1 and CYP2D6*4 in patients with breast and endometrial cancer

2017 ◽  
pp. 132-138
Author(s):  
O.V. Paliychuk ◽  
◽  
L.Z. Polishchuk ◽  
Z.I. Rossokha ◽  
◽  
...  
Keyword(s):  
Breast Cancer ◽  
Endometrial Cancer ◽  
Hormone Therapy ◽  
Molecular Genetic ◽  
Cancer Syndrome ◽  
Genetic Characteristics ◽  
Multiple Tumors ◽  
Receptor Status ◽  
Genetic Examination ◽  
The Impact

The objective: determining gene polymorphism features ERS1, CYP2D6 in patients with breast cancer (RHZ) and endometrial cancer (EC) and the impact assessment studied genetic characteristics compared to receptor status (immunohistochemical determination of expression levels of ER, PR) tumors and the results of the treatment. Patients and methods. article presents the results of complex clinical, morphological, clinical-genealogical, and molecular-genetic examination of 28 females: 19 patients with breast cancer (BC), 9 patients with endometrial cancer (EC), including 5 patients with primary-multiple tumors (PMT) with and without tumor pathology aggregation in families. Results. The It was determined that in patients’ families malignant tumors of breast, uterine body and/or ovaries prevail that corresponds to Lynch type II syndrome (family cancer syndrome). Molecular-genetic examination of genomic DNA of peripheral blood and histological sections for the presence of SNPs of ESR and CYP2D6*4 genes comparing with the results of immunohistochemical study of tumors for receptors ER and PR status have not found associations between these characteristics; although among EC patients the occurrence of genotypes 397ТТ and 351АА was significantly higher comparing with BC patients (55.55% and 10.5% for genotype 397ТТ,and 15.8% for genotype 351АА, respectively). At the same time the patients with BC and primary-multiple tumors (PMT) of female reproductive system organs (FRSO) that carried mutations in BRCA1 in all the cases demonstrated positive ER and PR receptor status and adverse combinations of polymorphous variants of the genes ESR1 (397СС, 397ТС) and CYP2D6*4 (1846G, 1846GA), suggesting combined effect of these factors on the development of malignant neoplasias of FRSO in families with positive family cancer history. In BC patients, receiving standard hormone therapy with tamoxifen, those, who had genotype 1846GG of the gene CYP2D6*4, in 3 patients (15.8%) of 19 (100%) patients disease recurrence was diagnosed. Conclusion. The obtained results allow clinical use of the assessment of polymorphism frequency of the genes ESR1 and CYP2D6*4 for selection of individual hormone therapy regimens schemes for BC patients, to increase efficacy of dispensary observation after finishing of special therapy for such patients, and also personalization of complex and combined treatment regimens. Key words: breast cancer, endometrial cancer, family cancer syndrome, single nucleotide polymorphisms (SNPs) of the genes ESR1, CYP2D6*4.

Thickness of endometrium: predictor of the effectiveness of IVF/ICSI programs (literature review)

GYNECOLOGY ◽  
2018 ◽  
Vol 20 (1) ◽  
pp. 113-116
Author(s):  
L A Bagdasaryan ◽  
I E Korneyeva
Keyword(s):  
Assisted Reproductive Technologies ◽  
Endometrial Thickness ◽  
Molecular Genetic ◽  
Uterine Cavity ◽  
Reproductive Technologies ◽  
Genetic Characteristics ◽  
Vitro Fertilization ◽  
Need To Evaluate ◽  
The Relationship

The aim of the study is to systematically analyze the data available in the modern literature on the relationship between endometrial thickness and the frequency of pregnancy in the program of assisted reproductive technologies (ART). Materials and methods. The review includes data from foreign and domestic articles found in PubMed on this topic. Results. The article presents data on the relationship between the thickness of the endometrium and the frequency of pregnancy in ART programs. The greatest number of studies is devoted to the evaluation of the relationship between the thickness of the endometrium and the frequency of pregnancy on the day of the ovulation trigger. Data are presented on the existence of a correlation between the thickness of the endometrium measured on the day of the ovulation trigger and the frequency of clinical pregnancy, as well as data on the need to evaluate the structure of the endometrium and the state of subendometric blood flow. The importance of multilayered (three-layered) endometrium as a prognostic marker of success in in vitro fertilization/intracytoplasmic sperm injection programs in the ovum is emphasized. The conclusion. The thickness of the endometrium can not be used as an argument for canceling the cycle or abolishing embryo transfer to the uterine cavity. Further studies in this direction are needed with a study of the morphological and molecular genetic characteristics of the endometrium, which in the future will allow us to evaluate the relationship between the thickness of the endometrium and the probability of pregnancy.

The Leucine-Rich Repeat Domain Can Determine Effective Interaction Between RPS2 and Other Host Factors in Arabidopsis RPS2-Mediated Disease Resistance

Genetics ◽  
2001 ◽  
Vol 158 (1) ◽  
pp. 439-450 ◽  
Author(s):  
Diya Banerjee ◽  
Xiaochun Zhang ◽  
Andrew F Bent
Keyword(s):  
Disease Resistance ◽  
Pseudomonas Syringae ◽  
Plant Disease Resistance ◽  
Effective Interaction ◽  
Molecular Genetic ◽  
Defense Responses ◽  
Host Factors ◽  
Leucine Rich Repeat ◽  
Domain Swap ◽  
Allele Specific

Abstract Like many other plant disease resistance genes, Arabidopsis thaliana RPS2 encodes a product with nucleotide-binding site (NBS) and leucine-rich repeat (LRR) domains. This study explored the hypothesized interaction of RPS2 with other host factors that may be required for perception of Pseudomonas syringae pathogens that express avrRpt2 and/or for the subsequent induction of plant defense responses. Crosses between Arabidopsis ecotypes Col-0 (resistant) and Po-1 (susceptible) revealed segregation of more than one gene that controls resistance to P. syringae that express avrRpt2. Many F2 and F3 progeny exhibited intermediate resistance phenotypes. In addition to RPS2, at least one additional genetic interval associated with this defense response was identified and mapped using quantitative genetic methods. Further genetic and molecular genetic complementation experiments with cloned RPS2 alleles revealed that the Po-1 allele of RPS2 can function in a Col-0 genetic background, but not in a Po-1 background. The other resistance-determining genes of Po-1 can function, however, as they successfully conferred resistance in combination with the Col-0 allele of RPS2. Domain-swap experiments revealed that in RPS2, a polymorphism at six amino acids in the LRR region is responsible for this allele-specific ability to function with other host factors.

MOLECULAR GENETIC CHARACTERISTICS OF COMPONENTS OF THE COMPLEMENT SYSTEM IN SEVERE ACNE

2021 ◽  
Vol 100 (2) ◽  
pp. 40-48
Author(s):  
A.G. Rumyantsev ◽  
◽  
A.G. Rumyantsev ◽  
O.M. Demina ◽  
◽  
...  
Keyword(s):  
Nucleotide Sequence ◽  
Complement System ◽  
Molecular Genetic ◽  
Factor H ◽  
Host Cells ◽  
Pathophysiological Mechanism ◽  
Genetic Characteristics ◽  
Severe Acne ◽  
C3 Gene ◽  
The Complement System

It has been shown that the inflammatory response in acne develops at the early subclinical stages of the disease, sometimes before the formation of comedones. It is known that an important component of the innate immune system is the complement system, which includes more than 60 components, including 9 basic proteins (C1-C9), a variety of activation products (C3a, C3b, iC3b, C3d and C3dg), regulatory and inhibitory molecules [factor H, fH-like protein 1 (FHL1), CR1 (CD35), C4b-binding protein (C4BP), C1inh and vitronectin], proteases and secreted enzymes (factor B, factor D, C3bBb and C4bC2b), as well as receptors for effector molecules [C3aR, C5aR, C5L2 and C1q receptor (C1qR)]. The compliment is the central part of innate immunity, which is the first line of protection against alien and altered host cells. The objectives of this study were to determine and analyze the variants of the nucleotide sequence of the genes of the complement system C1QA, C1S, C2, C3, C5, C6, C7, C8A, C8B, C8G, C9 in patients with severe acne. Materials and methods of research: To achieve the target a prospective open non-randomized one-center study was carried out in 2017–2020. Under our supervision in the clinical setting at the Department of Skin Diseases and Cosmetology of the Pirogov Russian National Research Medical University, there were 50 patients in the main group and 20 participants in the comparison group (70 people in total) (42/60% men and 28/40% women) aged 15 to 46 years (median – 22,1 years). Molecular genetic diagnostics was performed in all 70 patients of the main and control groups by the method of high-throughput DNA sequencing – next-generation sequencing (NGS). Results: when analyzing the nucleotide sequence variants of the complement system genes identified in our study, it is shown that the severe form of acne probably has an association (4 SNPs of the C8A gene, 1 SNPs of the C8B gene, 2 SNPs of the C1S gene, 3 SNPs of the C3 gene, 2 SNPs of the C9 gene, 1 SNPs of the C7 gene, 1 SNPs of the C6 gene, 1 SNPs of the C2 gene, 2 SNPs of the C5 gene, 2 SNPs of the C8G gene), 13 SNPs of the complement system genes in introns (1 SNPs of the C8A gene, 1 SNPs of the C8B gene, 2 SNPs of the C1S gene, 1 SNPs of the C3 gene, 1 SNPs of the C7 gene, 2 SNPs of the C6 gene, 4 SNPs of the C5 gene, 1 SNPs of C8G gene), 6 SNPs of the complement system genes (2 SNPs of the C8B gene: one SNPs each in the 3'UTR and 5'UTR zones; 3 SNPs of the C3 gene in the 5'UTR zone, 1 SNPs of the C7 gene in the 3'UTR zone). Two mutations of the frame shift of the C2 gene (frameshift deletion) and the C9 gene (rs748464075, frameshift insertion) seem to have a protective effect in the development of acne. Conclusion: the obtained variants of the nucleotide sequence of the genes of the complement system C1QA, C1S, C2, C3, C5, C6, C7, C8A, C8B, C8G, C9, apparently, are associated with the formation of severe acne and cause an imbalance of the components of the complement system. It can cause a defect in chemotactic and phagocytic reactions, and as a result a disturbance of the regulation of the inflammatory reaction with chronization of the skin process occures. Thus, results of studies carried out, revealed – for the first time – polymorphic loci of genes of components of the complement system, the imbalance of which is the pathophysiological mechanism of acne.

Clinical and genetic characteristics of rare pathogenic variants of the CFTR gene in Russian patients with cystic fibrosis

2021 ◽  
Vol 31 (2) ◽  
pp. 148-158
Author(s):  
A. Yu. Voronkova ◽  
Yu. L. Melyanovskaya ◽  
N. V. Petrova ◽  
T. A. Adyan ◽  
E. K. Zhekaite ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Genetic Variants ◽  
Pancreatic Insufficiency ◽  
Protein Energy Malnutrition ◽  
Molecular Genetic ◽  
Clinical Manifestations ◽  
Cftr Gene ◽  
Genetic Characteristics ◽  
Missense Variants ◽  
Pathogenic Variants

The variety of clinical manifestations of cystic fibrosis is driven by the diversity of the CFTR gene nucleotide sequence. Descriptions of the clinical manifestations in patients with the newly identified genetic variants are of particular interest.The aim of this study was to describe clinical manifestations of the disease with the newly identified genetic variants.Methods. Data from Registry of patients with cystic fibrosis in the Russian Federation (2018) were used. The data review included three steps — the search for frequent mutations, Sanger sequencing, and the search for extensive rearrangements by MLPA. 38 pathogenic variants were identified that were not previously described in the international CFTR2 database. We selected and analyzed full case histories of 15 patients with 10 of those 38 pathogenic variants: p.Tyr84*, G1047S, 3321delG, c.583delC, CFTRdele13,14del18, CFTRdele19-22, c.2619+1G>A, c.743+2T>A, p.Glu1433Gly, and CFTRdel4-8del10-11.Results. A nonsense variant p.Tyr84* was found in 5 patients (0.08 %). Two missense variants c.3139G>A were found in 2 siblings (0.03 %). The c.4298A>G was found in 1 patient. Other variants were detected in a single patient (0.02 %) each. They included two variants of a deletion with a shift of the reading frame 3321delG and c.583delC, two splicing disorders c.2619+1G>A and c.743+2T>A, three extended rearrangements CFTRdele19-22, CFTRdele13,14del18, and CFTRdel4-8del10-11. The last two variants include 2 rearrangements on one allele, which cause the severe course in two young children. 8 of the 10 variants are accompanied by pancreatic insufficiency (PI). Among patients with p.Tyr84*, one had ABPA, one had liver transplantation, and all had Pseudomonas aeruginosa infection. Nasal polyps were diagnosed in 2 patients with p.Tyr84*, 1 with G1047S, 1 with CFTRdel4-8del10-11, and 1 patient with 3321delG, who also had osteoporosis and cystic fibrosis-related diabetes (CFRD). 2 patients with PI with 3321delG and CFTRdel4-8del10-11 genetic variants, and 1 with PI with p.Glu1433Gly genetic variant had severe protein-energy malnutrition (PEM).Conclusion. Clinical manifestations of previously undescribed CFTR genetic variants were described. 5/10 genetic variants should be attributed to class I, 3/10 – to class 7 of the function classification of pathogenic CFTR gene variants associated with transcription and translation disruptions. Class of the identified missense variants c.3139G>A and c.4298A>G has not been established and requires further functional, cultural, and molecular genetic studies.

Clinical and Genetic Characteristics of Phenylketonuria in Ryazan Region

2021 ◽  
Vol 29 (1) ◽  
pp. 5-12
Author(s):  
Grigorii I. Yakubovskii ◽  
Olga B. Serebriakova ◽  
Alina G. Yakubovskaya ◽  
Nadezhda V. Ruban ◽  
Angelina A. Lyakhovets
Keyword(s):  
Blood Level ◽  
Phenylalanine Hydroxylase ◽  
Clinical Course ◽  
Molecular Genetic ◽  
Residual Activity ◽  
Speech Development ◽  
Genetic Characteristics ◽  
Frequent Mutations ◽  
R408w Mutation ◽  
Pah Gene

Aim. This investigation seeks to determine the incidence of phenylketonuria in the Ryazan region, assess the spectrum of mutations in the PAH gene (phenylalanine hydroxylase), investigate the interrelationship between the diseases clinical course, the phenylalanine blood level, and the patients genotype. Materials and Methods. The incidence of phenylketonuria was studied based on the data of massive neonatal screening for the period from 2000 to 2019. Molecular genetic examination of mutations was conducted in 39 patients using the allele-specific multiplex ligation method. The interrelationship between the phenylalanine blood level on the fifth day of life and retest, the diseases clinical course, and the patients genotype was assessed according to the medical record data of 33 patients under dispensary observation in a medico-genetic clinic. The patients were divided into two groups. The first group (n=21) had two severe mutations (residual activity of phenylalanine hydroxylase 10%). The second group (n=12) had one severe and one mild mutation (the residual activity of the enzyme 10%). Results. The incidence of phenylketonuria in the Ryazan region was one in 5054 newborns, exceeding the Russian Federations average parameters. Eighteen mutations were discovered in the PAH gene. The most frequent was the R408W mutation (56.4% alleles). The second most frequent mutations were the IVS10-11GA (6.4%) and P281L (5.1%). The R158Q and Y418C mutations occurred with a frequency of 4.1% and Е280К mutation of 2.7%. All the rest of the mutations occurred as single cases. Investigation of the interrelationship between the phenylalanine blood level, the diseases clinical course, and the patients genotype revealed a reliably higher content of amino acid in the first group on retest (32.11.7 mg/% vs. 17.71.5 mg/% in the second group, р0.001) and predomination of more severe forms of phenylketonuria (90.5% vs. 41.7%, respectively, р0.001). Disorders in neuropsychic and speech development were present in 28.6% of patients in the first group but were absent in the second group. Conclusion. By conducting the study, the incidence of phenylketonuria was determined in the Ryazan region. The spectrum of mutations in the PAH gene was defined. The interrelationship between the diseases clinical portrait, the phenylalanine blood level, and the patients PAH genotype was revealed.

Molecular Genetic Characteristics of Medieval Populations of the Russian North

2004 ◽  
Vol 40 (1) ◽  
pp. 1-9 ◽  
Author(s):  
E. E. Kulikov ◽  
A. P. Buzhilova ◽  
A. B. Poltaraus
Keyword(s):  
Molecular Genetic ◽  
Genetic Characteristics ◽  
Russian North

scholarly journals Lytic Exoenzymes of Soil Strains of Bacillus Representatives and Manifestations of their Biological Activity

2021 ◽  
Vol 83 (4) ◽  
pp. 54-62
Author(s):  
Yu.V. Korzh ◽  
◽  
I.V. Dragovoz ◽  
L.V. Avdeeva ◽  
◽  
...  
Keyword(s):  
Biological Activity ◽  
Soil Bacteria ◽  
Antagonistic Activity ◽  
Lytic Activity ◽  
Bacillus Sp ◽  
Genus Bacillus ◽  
Phytopathogenic Bacteria ◽  
Petri Dishes ◽  
Bacillus Strains ◽  
Test Cultures

Bacteria of the genus Bacillus make up a significant (8–12%) part of the soil microbiome. Manifestation of their biological activity, in particular, the antagonistic and lytic activity against other microorganisms directly depends on their exometabolites. According to the literature, such properties of soil bacteria of the genus Bacillus thus can be related to their various lytic exoenzymes. Aim. To evaluate the role of lytic exoenzymes of the studied soil bacteria strains of the genus Bacillus in the manifestation of their biological (antagonistic, lysing) activity. Methods. The antagonistic activity of bacteria strains of the genus Bacillus against phytopathogenic micromycetes was determined by the method of double culture in Petri dishes on potato-glucose agar. For qualitative analysis of the presence of extracellular enzymes, strains of bacteria of the genus Bacillus were plated on Petri dishes with solid mineral-salt medium and a suitable substrate inducer. The ratio of the diameter of substrate hydrolysis zone to the diameter of the colony was taken as the relative enzymatic activity of the culture. Bacteriolytic activity of the studied strains was determined by the change in optical density of living cells of phytopathogenic bacteria suspension at 540 nm. Results. Six strains of bacteria of the genus Bacillus were selected by the results of preliminary screening, with at least five types of lytic activity, namely proteolytic, chitinase, amylolytic, cellulase, and xylanase of different levels (low, average, high). Analysis of the antagonistic activity of the selected strains of bacteria of the genus Bacillus to the main groups of phytopathogenic bacteria (six test cultures) singled out the strain Bacillus sp. 41 for a careful study of the nature and spectrum of its antagonism. Analysis of the level of antagonistic activity of the selected Bacillus strains against the phytopathogenic micromycetes showed that the minimum decrease of antagonism (the decrease of growth inhibition zones) during the observation period (at the 3rd and 7th days) was in Bacillus sp. 41 strain. Therefore, only this strain showed a stable and relatively wide range of antagonistic activity against phytopathogens of bacterial and fungal etiology. The nature of this antagonism is probably complex and conditioned by the participation of various biochemical mechanisms, in particular, the synthesis of a complex of lytic exoenzymes. To assess the lysing activity of Bacillus strains, three strains with the highest proteolytic and cellulolytic activity of exoenzymes were taken from the six previously chosen. Only Bacillus sp.1913 strain showed high (70%) lytic activity against gram-negative polyphagous phytopathogen Pseudomonas syringae pv. syringae UCM B-1027T. Such activity of the strain did not manifest against the rest of the phytopathogenic test cultures. The high lytic activity of Bacillus sp. 1913 strain may be associated with high activity of exogenous proteases and cellulases of the lytic complex, which is quite consistent with the literature data on the lytic activity of bacteria of the genus Bacillus. Conclusions. The spectrum and activity of lytic exoenzymes of strains of the studied soil bacteria of the genus Bacillus indicate the indirect participation of these enzymes in the manifestation of biological activity (antagonistic and lytic).

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