O. Efimova, M. Krapivin, S. Parfenyev, I. Mekina, E. Komarova, M. Ishchuk, A. Tikhonov, I. Kogan, Arina V Golubeva, E. Daev, Aleksander M. Gzgzyan, O. Bespalova, A. Pendina
Background. The epigenome of gametes is formed under the control of the developmental programme and the influence of environmental factors. How cytosine oxidation patterns are formed and altered in human spermatogenesis remains obscure so far. The aim of the study was to assess 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC) patterns in human spermatogenic cells and spermatozoa. Materials and Methods. The study was performed on testicular biopsy samples of 10 azoospermic patients and ejaculate samples of 5 sperm donors and 8 patients from infertile couples. The microscope slides were prepared for further indirect immunofluorescence to detect 5fC and 5caC and FISH to determine spermatogenic cell ploidy. Results. 5fC and 5caC were undetectable in mitotic and meiotic chromosomes of spermatogenic cells, and was present exclusively in some spermatogonia and spermatid interphase nuclei as well as in some ejaculated spermatozoa. The frequency of spermatozoa with 5fC and 5caC varied in a wide range and was higher in patients than in sperm donors (p=0,007, p=0,028). The increase in frequency of spermatozoa with 5fC and 5caC was accompanied with the decrease in frequency of morphologically normal and progressively motile spermatozoa. Conclusions. 5fC and 5caC are differentially distributed in human spermatogenic cells and spermatozoa. The immunocytochemically detected increase of 5fC and 5caC in individual spermatozoa is most likely induced by oxidative stress caused by effects of internal and external factors rather than developmental programme. The evaluation of 5fC and 5caC in spermatozoa can be potentially used as an additional criterion of ejaculate quality.
{"title":"Differential distribution of 5-formylcytosine and 5-carboxylcytosine in human spermatogenic cells and spermatozoa","authors":"O. Efimova, M. Krapivin, S. Parfenyev, I. Mekina, E. Komarova, M. Ishchuk, A. Tikhonov, I. Kogan, Arina V Golubeva, E. Daev, Aleksander M. Gzgzyan, O. Bespalova, A. Pendina","doi":"10.17816/ecogen120080","DOIUrl":"https://doi.org/10.17816/ecogen120080","url":null,"abstract":"Background. The epigenome of gametes is formed under the control of the developmental programme and the influence of environmental factors. How cytosine oxidation patterns are formed and altered in human spermatogenesis remains obscure so far. \u0000The aim of the study was to assess 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC) patterns in human spermatogenic cells and spermatozoa. \u0000Materials and Methods. The study was performed on testicular biopsy samples of 10 azoospermic patients and ejaculate samples of 5 sperm donors and 8 patients from infertile couples. The microscope slides were prepared for further indirect immunofluorescence to detect 5fC and 5caC and FISH to determine spermatogenic cell ploidy. \u0000Results. 5fC and 5caC were undetectable in mitotic and meiotic chromosomes of spermatogenic cells, and was present exclusively in some spermatogonia and spermatid interphase nuclei as well as in some ejaculated spermatozoa. The frequency of spermatozoa with 5fC and 5caC varied in a wide range and was higher in patients than in sperm donors (p=0,007, p=0,028). The increase in frequency of spermatozoa with 5fC and 5caC was accompanied with the decrease in frequency of morphologically normal and progressively motile spermatozoa. \u0000Conclusions. 5fC and 5caC are differentially distributed in human spermatogenic cells and spermatozoa. The immunocytochemically detected increase of 5fC and 5caC in individual spermatozoa is most likely induced by oxidative stress caused by effects of internal and external factors rather than developmental programme. The evaluation of 5fC and 5caC in spermatozoa can be potentially used as an additional criterion of ejaculate quality.","PeriodicalId":11431,"journal":{"name":"Ecological genetics","volume":"38 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72867163","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BACKGROUND: The causal relationship between mutagenesis and carcinogenesis is well known. Hence the wide interest in the study of the mutagen-modifying effects of natural and synthetic compounds. Particular attention is drawn to widespread compounds. One of them, metformin, is widely used as a hypoglycemic drug. AIM: evaluation of the influence of metformin on the cytogenetic effects of doxorubicin and cyclophosphamide in mouse bone marrow cells. MATERIALS AND METHODS: Male F1 CBAxC57Bl/6 hybrid mice were used. Cyclophosphamide (20 mg/kg) or doxorubicin (10 mg/kg) was administered intraperitoneally, metformin was given orally once or for 4 consecutive days. The latter administration of metformin was combined with mutagen administration. Cytogenetic preparations of bone marrow cells were prepared 18 hours after metformin administration and 24 hours after its combined administration with mutagens. Chromosomal aberrations were analyzed according to accepted protocols. RESULTS: Metformin per se showed no cytogenetic activity at doses of 500, 1000, and 2000 mg/kg. At a dose of 500 mg/kg, but not 100 or 250 mg/kg, metformin reduced the cytogenetic effects of doxorubicin. Metformin administered once and for 4 days at doses of 100, 250, and 500 mg/kg or once at doses of 10 and 20 mg/kg increased the number of metaphases with chromosome aberrations induced by cyclophosphamide by a factor of 2 to 3. At doses of 2.5 and 5 mg/kg, metformin had no modifying effect on the mutagen effect. CONCLUSION: Metformin attenuates the cytogenetic effects of doxorubicin and enhances the cytogenetic activity of cyclophosphamide in mouse bone marrow cells. This allows us to conclude that metformin has mutagen-modifying properties.
{"title":"Modifying action of metformin on the cytogenetic effects of doxorubicin and cyclophosphamide in mice","authors":"A. Zhanataev, A. Kulakova, A. Durnev","doi":"10.17816/ecogen133621","DOIUrl":"https://doi.org/10.17816/ecogen133621","url":null,"abstract":"BACKGROUND: The causal relationship between mutagenesis and carcinogenesis is well known. Hence the wide interest in the study of the mutagen-modifying effects of natural and synthetic compounds. Particular attention is drawn to widespread compounds. One of them, metformin, is widely used as a hypoglycemic drug. \u0000AIM: evaluation of the influence of metformin on the cytogenetic effects of doxorubicin and cyclophosphamide in mouse bone marrow cells. \u0000MATERIALS AND METHODS: Male F1 CBAxC57Bl/6 hybrid mice were used. Cyclophosphamide (20 mg/kg) or doxorubicin (10 mg/kg) was administered intraperitoneally, metformin was given orally once or for 4 consecutive days. The latter administration of metformin was combined with mutagen administration. Cytogenetic preparations of bone marrow cells were prepared 18 hours after metformin administration and 24 hours after its combined administration with mutagens. Chromosomal aberrations were analyzed according to accepted protocols. \u0000RESULTS: Metformin per se showed no cytogenetic activity at doses of 500, 1000, and 2000 mg/kg. At a dose of 500 mg/kg, but not 100 or 250 mg/kg, metformin reduced the cytogenetic effects of doxorubicin. Metformin administered once and for 4 days at doses of 100, 250, and 500 mg/kg or once at doses of 10 and 20 mg/kg increased the number of metaphases with chromosome aberrations induced by cyclophosphamide by a factor of 2 to 3. At doses of 2.5 and 5 mg/kg, metformin had no modifying effect on the mutagen effect. \u0000CONCLUSION: Metformin attenuates the cytogenetic effects of doxorubicin and enhances the cytogenetic activity of cyclophosphamide in mouse bone marrow cells. This allows us to conclude that metformin has mutagen-modifying properties.","PeriodicalId":11431,"journal":{"name":"Ecological genetics","volume":"11 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76229062","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
T. A. Nikitina, Mariya A. Konyashkina, F. Ingel, L. V. Akhaltseva
The expansion of the spectrum of use of food additives and, in particular, food dyes (FD), increases the risk of increasing human exposure to genotoxicants. Since in real life, not pure substances with proven genetic safety are in contact with a person, but complex mixtures of unknown composition, even minor impurities in which can become an additional source or modifier of genome instability effects. Of particular concern in this aspect are synthetic FD azo and diazo compounds which can be transformed by human intestinal microflora to some forms of genotoxicants. The purpose of the work is to evaluate the genotoxic effects of 02 mg/mL of Tartrazine FD (E102) purchased in a retail network in a micronucleus test on human blood cells cultured under cytokinetic block conditions in parallel in presence and without rat S9 hepatocyte metabolic activation system. Genotoxic effects were found in cultures without metabolic activation at 0.00002560,00064 mg/mL and 0.4 mg/mL of tartrazine, and in the presence of S9 at 0.0000256 mg/mL, 0,000128 mg/mL and 0.16 mg/mL of tartrazine. For the first time, a dose-dependent suppression of mitotic and proliferative activity of lymphocytes induced by the tested tartrazine sample was revealed, as well as a dose-dependent U-shaped curve in the frequency of apoptosis. The data obtained indicate the presence of genotoxic activity of the studied sample. We discuss the necessity to create the system for evaluation the genotoxic safety of FD real mixtures from a retail network.
{"title":"Evaluation of the genotoxic effect of tartrazine using a metabolic activation system in human lymphocyte culture under cytokinetic block conditions","authors":"T. A. Nikitina, Mariya A. Konyashkina, F. Ingel, L. V. Akhaltseva","doi":"10.17816/ecogen117502","DOIUrl":"https://doi.org/10.17816/ecogen117502","url":null,"abstract":"The expansion of the spectrum of use of food additives and, in particular, food dyes (FD), increases the risk of increasing human exposure to genotoxicants. Since in real life, not pure substances with proven genetic safety are in contact with a person, but complex mixtures of unknown composition, even minor impurities in which can become an additional source or modifier of genome instability effects. Of particular concern in this aspect are synthetic FD azo and diazo compounds which can be transformed by human intestinal microflora to some forms of genotoxicants. The purpose of the work is to evaluate the genotoxic effects of 02 mg/mL of Tartrazine FD (E102) purchased in a retail network in a micronucleus test on human blood cells cultured under cytokinetic block conditions in parallel in presence and without rat S9 hepatocyte metabolic activation system. \u0000Genotoxic effects were found in cultures without metabolic activation at 0.00002560,00064 mg/mL and 0.4 mg/mL of tartrazine, and in the presence of S9 at 0.0000256 mg/mL, 0,000128 mg/mL and 0.16 mg/mL of tartrazine. For the first time, a dose-dependent suppression of mitotic and proliferative activity of lymphocytes induced by the tested tartrazine sample was revealed, as well as a dose-dependent U-shaped curve in the frequency of apoptosis. The data obtained indicate the presence of genotoxic activity of the studied sample. \u0000We discuss the necessity to create the system for evaluation the genotoxic safety of FD real mixtures from a retail network.","PeriodicalId":11431,"journal":{"name":"Ecological genetics","volume":"71 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75357470","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The dynamics of the composition of the populations of Adalia bipunctata L. in St. Petersburg and Yalta (Crimean Peninsula) for 4732 years has been studied. The proportion of black individuals in them decreased by almost 2 times. Comparison of the composition of populations with climatic features of habitats (average annual temperature) showed that the proportion of black individuals in the population negatively correlates with the average annual temperature of the previous year. The observed change in the composition of geographically remote populations is probably the effect of global warming.
{"title":"Ecological genetics of beetles of the genus Adalia: restructuring of A. bipunctata populations as a global warming effect","authors":"I. Zakharov, A. Rubanovich","doi":"10.17816/ecogen317164","DOIUrl":"https://doi.org/10.17816/ecogen317164","url":null,"abstract":"The dynamics of the composition of the populations of Adalia bipunctata L. in St. Petersburg and Yalta (Crimean Peninsula) for 4732 years has been studied. The proportion of black individuals in them decreased by almost 2 times. Comparison of the composition of populations with climatic features of habitats (average annual temperature) showed that the proportion of black individuals in the population negatively correlates with the average annual temperature of the previous year. The observed change in the composition of geographically remote populations is probably the effect of global warming.","PeriodicalId":11431,"journal":{"name":"Ecological genetics","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83128674","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Vladivir G. Golgshtein, L. P. Nosovskaya, L. V. Adikaeva, M. Bazgiev, K. Badurgova, Aslanbek I. Buzurtanov, V. Khoreva, Vladislav N. Boyko, A. Grushin, Selminaz F. Israfilova, I. V. Fil, E. Khatefov
BACKGROUND: The study of the VIR corn collection in order to search for economically valuable sources and donors is topical. MATERIALS AND METHODS: Studies of the biochemical components of grain during its deep processing were carried out on 27 hybrids from Germany, presented in the VIR collection. Valuable results have been obtained using the method of IR spectrometry and deep processing of grain in laboratory conditions, which make it possible to identify samples of interest for the production of native starch and its by-products. RESULTS: The starch yield of more than 65% solids (% DM) grain dry matter is established for the following hybrids: KHV 7262, KHV 6431, KHV 6331, KHV 5440, Karpatis, DS 21209C, DS 21215B, DS 21212A, DS 21205B, DS 22188D. The maximum starch yield, more than 70% grain DM, is set for DS 21205B hybrids. The maximum yield of the embryo was set for the hybrid KHV 4126 up to 10% DM of the grain, the yield of starch during the processing of the grain of this hybrid was 61.2% DM. The highest yield of gluten, 18% or more, was established during the processing of hybrids DS 23190B and DS 21203B. The pulp yield of more than 15% CB was obtained by processing grain DS 22182C. Based on the results obtained, the following hybrids are proposed as the starting material for corn breeding for deep grain processing: KHV 7262, KHV 5440, DS 21209C, DS 21215B, DS 21212A, DS 21205B, DS 22188D. CONCLUSIONS: Of greatest interest as a starting material is hybrid DS 21205B, during the processing of which starch was extracted in an amount of more than 70% grain DM.
{"title":"The productivity potential of some corn hybrids of the VIR collection for starch extraction during deep grain processing","authors":"Vladivir G. Golgshtein, L. P. Nosovskaya, L. V. Adikaeva, M. Bazgiev, K. Badurgova, Aslanbek I. Buzurtanov, V. Khoreva, Vladislav N. Boyko, A. Grushin, Selminaz F. Israfilova, I. V. Fil, E. Khatefov","doi":"10.17816/ecogen111879","DOIUrl":"https://doi.org/10.17816/ecogen111879","url":null,"abstract":"BACKGROUND: The study of the VIR corn collection in order to search for economically valuable sources and donors is topical. \u0000MATERIALS AND METHODS: Studies of the biochemical components of grain during its deep processing were carried out on 27 hybrids from Germany, presented in the VIR collection. Valuable results have been obtained using the method of IR spectrometry and deep processing of grain in laboratory conditions, which make it possible to identify samples of interest for the production of native starch and its by-products. \u0000RESULTS: The starch yield of more than 65% solids (% DM) grain dry matter is established for the following hybrids: KHV 7262, KHV 6431, KHV 6331, KHV 5440, Karpatis, DS 21209C, DS 21215B, DS 21212A, DS 21205B, DS 22188D. The maximum starch yield, more than 70% grain DM, is set for DS 21205B hybrids. The maximum yield of the embryo was set for the hybrid KHV 4126 up to 10% DM of the grain, the yield of starch during the processing of the grain of this hybrid was 61.2% DM. The highest yield of gluten, 18% or more, was established during the processing of hybrids DS 23190B and DS 21203B. The pulp yield of more than 15% CB was obtained by processing grain DS 22182C. Based on the results obtained, the following hybrids are proposed as the starting material for corn breeding for deep grain processing: KHV 7262, KHV 5440, DS 21209C, DS 21215B, DS 21212A, DS 21205B, DS 22188D. \u0000CONCLUSIONS: Of greatest interest as a starting material is hybrid DS 21205B, during the processing of which starch was extracted in an amount of more than 70% grain DM.","PeriodicalId":11431,"journal":{"name":"Ecological genetics","volume":"192 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75533388","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BACKGROUND: Due to broad geographical and ecological distribution of Scots pine we witness the shaping of a significant species heterogeneity. There is a demand in researching the features of the genetic structure and differentiation of Scots pine populations in different parts of the range. MATERIALS AND METHODS: 12 populations were scrutinized with the use of ISSR markers. The genetic structure was assessed by estimating basic indicators of genetic diversity (the number of alleles per locus, the number of effective alleles, and the expected heterozygosity) and by the analysis of molecular variance (AMOVA). Genetic differentiation was assessed by Neis GST statistic, Mantel test, Principal Coordinates Analysis (PCoA), and creating a tree diagram. RESULTS: Populations that grow on the right bank of the Volga in the northern and central parts of the Volga Uplands are characterized by a higher genetic diversity (Na = 1.841.89; Ne = 1.341.39; He = 0.2170.241) and a lower subdivision (GST = 0.092). Populations that grow on the left bank proved lower rates of genetic variability (Na = 1.681.81; Ne = 1.271.35; He = 0.1740.218) while the divergence was higher (GST = 0.179). Much of the genetic variability is within the populations (more than 80%). CONCLUSIONS: The study determined differences in the genetic structure and the degree of differentiation of Scots pine populations, that grow on different banks of the Volga in the Middle and Upper Volga Regions.
{"title":"Genetic structure and differentiation of Scots pine (Pinus sylvestris L.) populations in the Middle and Upper Volga Regions","authors":"Sheikina V. Sheikina","doi":"10.17816/ecogen110866","DOIUrl":"https://doi.org/10.17816/ecogen110866","url":null,"abstract":"BACKGROUND: Due to broad geographical and ecological distribution of Scots pine we witness the shaping of a significant species heterogeneity. There is a demand in researching the features of the genetic structure and differentiation of Scots pine populations in different parts of the range. \u0000MATERIALS AND METHODS: 12 populations were scrutinized with the use of ISSR markers. The genetic structure was assessed by estimating basic indicators of genetic diversity (the number of alleles per locus, the number of effective alleles, and the expected heterozygosity) and by the analysis of molecular variance (AMOVA). Genetic differentiation was assessed by Neis GST statistic, Mantel test, Principal Coordinates Analysis (PCoA), and creating a tree diagram. \u0000RESULTS: Populations that grow on the right bank of the Volga in the northern and central parts of the Volga Uplands are characterized by a higher genetic diversity (Na = 1.841.89; Ne = 1.341.39; He = 0.2170.241) and a lower subdivision (GST = 0.092). Populations that grow on the left bank proved lower rates of genetic variability (Na = 1.681.81; Ne = 1.271.35; He = 0.1740.218) while the divergence was higher (GST = 0.179). Much of the genetic variability is within the populations (more than 80%). \u0000CONCLUSIONS: The study determined differences in the genetic structure and the degree of differentiation of Scots pine populations, that grow on different banks of the Volga in the Middle and Upper Volga Regions.","PeriodicalId":11431,"journal":{"name":"Ecological genetics","volume":"6 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74573683","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
V. Zhukov, A. Zhernakov, Maria Yu. Belozerova, I. Dodueva, M. Lebedeva, L. Lutova, I. Tikhonovich
BACKGROUND: N.I. Vavilov Institute of Plant Genetic Resources (VIR) (Saint Petersburg, Russia) maintains a large collection of pea (Pisum sativum L.). Earlier, several growth and yield parameters were recorded for plants of 99 accessions grown under inoculation with nodule bacteria and arbuscular mycorrhizal fungi. MATERIALS AND METHODS: Polymorphism of genes encoding symbiotic receptor kinase PsSym29 [participating in the autoregulation of nodulation (AON) system] and closely related receptor kinase PsNRLK1 (with yet unknown function in symbiosis) was assessed in 99 pea genotypes from the VIR collection. Nucleotide diversity, Tajimas D, and Fay and Wus H statistics were calculated using DNAsp 5.0 software. The significance of associations of allelic state of the sequenced genes with the growth and yield parameters was tested by two-way ANOVA followed by FDR correction and by regression analysis. RESULTS: Nucleotide diversity and the ratio of synonymous to non-synonymous substitutions was greater in PsNRLK1 as compared to PsSym29. The analysis of Fay and Wus H in sliding window revealed signatures of positive selection in one site of PsSym29 and in three sites of PsNRLK1 gene sequences located in 1st exons encoding LRR (leucine rich repeat) domains. No significant associations of allelic state of neither PsSym29 nor PsNRLK1 genes was found with plant growth and yield parameters. CONCLUSIONS: The sequences of both PsSym29 and PsNRLK1 genes undergo positive selection, but the conditions in which specific allelic states of the genes become adaptive are to be elucidated in future.
{"title":"Diversity of PsSym29 and PsNRLK1 genes in the VIR germplasm collection of pea (Pisum sativum L.)","authors":"V. Zhukov, A. Zhernakov, Maria Yu. Belozerova, I. Dodueva, M. Lebedeva, L. Lutova, I. Tikhonovich","doi":"10.17816/ecogen111959","DOIUrl":"https://doi.org/10.17816/ecogen111959","url":null,"abstract":"BACKGROUND: N.I. Vavilov Institute of Plant Genetic Resources (VIR) (Saint Petersburg, Russia) maintains a large collection of pea (Pisum sativum L.). Earlier, several growth and yield parameters were recorded for plants of 99 accessions grown under inoculation with nodule bacteria and arbuscular mycorrhizal fungi. \u0000MATERIALS AND METHODS: Polymorphism of genes encoding symbiotic receptor kinase PsSym29 [participating in the autoregulation of nodulation (AON) system] and closely related receptor kinase PsNRLK1 (with yet unknown function in symbiosis) was assessed in 99 pea genotypes from the VIR collection. Nucleotide diversity, Tajimas D, and Fay and Wus H statistics were calculated using DNAsp 5.0 software. The significance of associations of allelic state of the sequenced genes with the growth and yield parameters was tested by two-way ANOVA followed by FDR correction and by regression analysis. \u0000RESULTS: Nucleotide diversity and the ratio of synonymous to non-synonymous substitutions was greater in PsNRLK1 as compared to PsSym29. The analysis of Fay and Wus H in sliding window revealed signatures of positive selection in one site of PsSym29 and in three sites of PsNRLK1 gene sequences located in 1st exons encoding LRR (leucine rich repeat) domains. No significant associations of allelic state of neither PsSym29 nor PsNRLK1 genes was found with plant growth and yield parameters. \u0000CONCLUSIONS: The sequences of both PsSym29 and PsNRLK1 genes undergo positive selection, but the conditions in which specific allelic states of the genes become adaptive are to be elucidated in future.","PeriodicalId":11431,"journal":{"name":"Ecological genetics","volume":"11 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84194682","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Based on the selected criteria data from studies of the genotoxic activity of antiepileptic drugs in eukaryotic test systems in vitro and in vivo, performed by DNA comet assay, chromosomal aberrations and micronuclei assays, published in the period 19952022, were selected and summarized. Among the 20 drugs reviewed, for one drug (N03AA05 Benzobarbital), there are no data on studies of genotoxic activity; for 7 drugs information is presented only as a summary on the FDA and EMA websites without primary data and information on experimental designs. Among the remaining 12 drugs, only three drugs (phenobarbital, valproic acid and levetiracetam) have information on in vivo studies, both by DNA damage assay and by cytogenetic methods. Based on known publications, it is impossible to draw reasonable conclusions about the genotoxic potential of individual drugs. The available data are fragmentary, incomplete and contradictory. It remains to state the facts of detection of genotoxic effects in individual drugs in separate studies. In general, there is no doubt about the potential genotoxic hazard of this group the drugs in. Additional studies are needed to clarify the data on the genotoxicity of antiepileptic drugs including beyond the standard protocols. In the course of their implementation, one should take into account the possible tissue-specific manifestation of antiepileptics genotoxicity, as indicated by the facts of genotoxic effects detection in tissue cells that are not targets in classical genotoxic studies. The expediency of objectifying approaches when choosing a drug for safe therapy, taking into account information about its genotoxicity, is emphasized, and the prospects for possible studies on antigenotoxic prophylaxis in patients with epilepsy are pointed out.
{"title":"Genotoxic effects of antiepileptic drugs. Literature review","authors":"N. Eremina, A. Zhanataev, A. Durnev","doi":"10.17816/ecogen109400","DOIUrl":"https://doi.org/10.17816/ecogen109400","url":null,"abstract":"Based on the selected criteria data from studies of the genotoxic activity of antiepileptic drugs in eukaryotic test systems in vitro and in vivo, performed by DNA comet assay, chromosomal aberrations and micronuclei assays, published in the period 19952022, were selected and summarized. \u0000Among the 20 drugs reviewed, for one drug (N03AA05 Benzobarbital), there are no data on studies of genotoxic activity; for 7 drugs information is presented only as a summary on the FDA and EMA websites without primary data and information on experimental designs. Among the remaining 12 drugs, only three drugs (phenobarbital, valproic acid and levetiracetam) have information on in vivo studies, both by DNA damage assay and by cytogenetic methods. Based on known publications, it is impossible to draw reasonable conclusions about the genotoxic potential of individual drugs. The available data are fragmentary, incomplete and contradictory. It remains to state the facts of detection of genotoxic effects in individual drugs in separate studies. In general, there is no doubt about the potential genotoxic hazard of this group the drugs in. \u0000Additional studies are needed to clarify the data on the genotoxicity of antiepileptic drugs including beyond the standard protocols. In the course of their implementation, one should take into account the possible tissue-specific manifestation of antiepileptics genotoxicity, as indicated by the facts of genotoxic effects detection in tissue cells that are not targets in classical genotoxic studies. The expediency of objectifying approaches when choosing a drug for safe therapy, taking into account information about its genotoxicity, is emphasized, and the prospects for possible studies on antigenotoxic prophylaxis in patients with epilepsy are pointed out.","PeriodicalId":11431,"journal":{"name":"Ecological genetics","volume":"105 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85897500","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Miklhail A. Tsygankov, A. Rumyantsev, Anastasiya Makeeva, M. Padkina
BACKGROUND: Yeast display is an effective technology for exposure target proteins to the cell surface by fusing them with cell wall proteins. This technique, among other things, makes it possible to obtain vaccine preparations based on yeast by exposing antigen proteins on their cell surface. Finding and selecting proteins that allow effective exposure of target proteins on the surface of yeast cells is an urgent task. AIM: The aim of this work was to evaluate the efficiency of cell wall proteins ScAG1p, KpCW51p, KpCW61p for displaying the reporter protein on the Komagataella phaffii cell surface, including the study of several variants of the ScAG1 gene coding sequence. MATERIALS AND METHODS: The studied gene sequences were cloned under the control of the AOX1 gene promoter in the same reading frame as the eGFP reporter protein gene and integrated into the genome of the K. Phaffii yeast strain X-33. RESULTS: Cytoimmunochemical analysis and confocal microscopy of strains displaying the eGFP protein on their surface under conditions of induction of the AOX1 gene promoter made it possible to identify the most effective anchor protein. The best efficiency was demonstrated for the sequence of the ScAG1 gene without the native 3' non-coding region. CONCLUSIONS: The plasmids obtained in the work will make it possible to obtain a yeast strain K. phaffii that effectively exposure proteins, including antigens, on its surface, which can be used as a vaccine preparation.
{"title":"Comparasion of the effectiveness of anchor proteins ScAGα1p, KpCW51p, KpCW61p for surface display in yeast Komagataella phaffii","authors":"Miklhail A. Tsygankov, A. Rumyantsev, Anastasiya Makeeva, M. Padkina","doi":"10.17816/ecogen112509","DOIUrl":"https://doi.org/10.17816/ecogen112509","url":null,"abstract":"BACKGROUND: Yeast display is an effective technology for exposure target proteins to the cell surface by fusing them with cell wall proteins. This technique, among other things, makes it possible to obtain vaccine preparations based on yeast by exposing antigen proteins on their cell surface. Finding and selecting proteins that allow effective exposure of target proteins on the surface of yeast cells is an urgent task. \u0000AIM: The aim of this work was to evaluate the efficiency of cell wall proteins ScAG1p, KpCW51p, KpCW61p for displaying the reporter protein on the Komagataella phaffii cell surface, including the study of several variants of the ScAG1 gene coding sequence. \u0000MATERIALS AND METHODS: The studied gene sequences were cloned under the control of the AOX1 gene promoter in the same reading frame as the eGFP reporter protein gene and integrated into the genome of the K. Phaffii yeast strain X-33. \u0000RESULTS: Cytoimmunochemical analysis and confocal microscopy of strains displaying the eGFP protein on their surface under conditions of induction of the AOX1 gene promoter made it possible to identify the most effective anchor protein. The best efficiency was demonstrated for the sequence of the ScAG1 gene without the native 3' non-coding region. \u0000CONCLUSIONS: The plasmids obtained in the work will make it possible to obtain a yeast strain K. phaffii that effectively exposure proteins, including antigens, on its surface, which can be used as a vaccine preparation.","PeriodicalId":11431,"journal":{"name":"Ecological genetics","volume":"5 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78970621","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ousama Al Shanaa, A. M. Rumyantsev, E. Sambuk, M. Padkina
BACKGROUND: RNA aptamers are short, single-stranded oligonucleotides, with remarkable binding ability to target molecules characterized by high specificity and affinity. Such targets are vastly diverse and range from specific ions to entire cells. RNA aptamers are widely used in biology and medicine for basic research, as well as for practical purposes as in therapy and diagnostics. At present, chemical or in vitro methods of synthesis are mainly used to obtain RNA aptamers. However, such methods are expensive and time-consuming with low productivity. Therefore, in vivo methods are becoming more attractive to researchers working on optimizing high-scale production of RNA aptamers. AIM: The aim of this work is to develop a reporter system for optimizing the synthesis of small RNA molecules in Saccharomyces cerevisiae yeast cells. MATERIALS AND METHODS: We used the Broccoli fluorescent RNA aptamer to develop a reporter system allowing us to optimize the conditions for in vivo short RNA synthesis in yeast cells. This aptamer is about 112 bp in size and binds to the fluorogenic dye DFHBI-1T. Only upon binding, the aptamer-dye complex exhibits fluorescence properties. After excitation using light with a wavelength of 482 nm, the aptamer-dye complex emission is observed with a peak at 505 nm. RESULTS: We have designed a reporter system providing the synthesis of the fluorescent Broccoli RNA aptamer in S. cerevisiae yeast cells. Transcription of RNA molecules containing the aptamer is carried out by the regulated promoter of the GAL1 gene. The synthesized transcripts contain the HH and HDV ribozymes to ensure precise cleavage of the RNA aptamer sequences. CONCLUSIONS: This reporter system is based on the Broccoli RNA aptamer, and it can be used to optimize the in vivo synthesis of RNA aptamers in S. cerevisiae yeast cells. This work serves an urgent task in connection with the active use of such aptamers in scientific research, biotechnology and medicine.
{"title":"The synthesis of Broccoli RNA fluorescent aptamer in Saccharomyces cerevisiae yeast cells","authors":"Ousama Al Shanaa, A. M. Rumyantsev, E. Sambuk, M. Padkina","doi":"10.17816/ecogen111012","DOIUrl":"https://doi.org/10.17816/ecogen111012","url":null,"abstract":"BACKGROUND: RNA aptamers are short, single-stranded oligonucleotides, with remarkable binding ability to target molecules characterized by high specificity and affinity. Such targets are vastly diverse and range from specific ions to entire cells. RNA aptamers are widely used in biology and medicine for basic research, as well as for practical purposes as in therapy and diagnostics. At present, chemical or in vitro methods of synthesis are mainly used to obtain RNA aptamers. However, such methods are expensive and time-consuming with low productivity. Therefore, in vivo methods are becoming more attractive to researchers working on optimizing high-scale production of RNA aptamers. \u0000AIM: The aim of this work is to develop a reporter system for optimizing the synthesis of small RNA molecules in Saccharomyces cerevisiae yeast cells. \u0000MATERIALS AND METHODS: We used the Broccoli fluorescent RNA aptamer to develop a reporter system allowing us to optimize the conditions for in vivo short RNA synthesis in yeast cells. This aptamer is about 112 bp in size and binds to the fluorogenic dye DFHBI-1T. Only upon binding, the aptamer-dye complex exhibits fluorescence properties. After excitation using light with a wavelength of 482 nm, the aptamer-dye complex emission is observed with a peak at 505 nm. \u0000RESULTS: We have designed a reporter system providing the synthesis of the fluorescent Broccoli RNA aptamer in S. cerevisiae yeast cells. Transcription of RNA molecules containing the aptamer is carried out by the regulated promoter of the GAL1 gene. The synthesized transcripts contain the HH and HDV ribozymes to ensure precise cleavage of the RNA aptamer sequences. \u0000CONCLUSIONS: This reporter system is based on the Broccoli RNA aptamer, and it can be used to optimize the in vivo synthesis of RNA aptamers in S. cerevisiae yeast cells. This work serves an urgent task in connection with the active use of such aptamers in scientific research, biotechnology and medicine.","PeriodicalId":11431,"journal":{"name":"Ecological genetics","volume":"20 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91064798","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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