http://cytgen.com "TSitologiya i Genetika" (Cytology and genetics) Sat, 05 Oct 2024 09:59:35 +0300 Sat, 05 Oct 2024 09:59:35 +0300 http://blogs.law.harvard.edu/tech/rss cytgen.com RSSgeneraton 0.9b Copyright 2013 cytgen.com cytgen@cytgen.com cytgen@cytgen.com ru http://cytgen.com/en/
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/NV.htm Mon, 01 Nov 1999 00:00:00 +0200 SUMMARY. γ-aminobutyric acid (GABA) is considered a molecule that combines the properties of a stress metabolite and a signaling molecule. At the same time, the importance of its functional interaction with other signaling mediators, in particular, reactive oxygen species (ROS) and calcium ions, for the implementation of stress-protective action on plant cells remains poorly researched. We studied the effect of GABA on the resistance of wheat seedlings (Triticum aestivum L., variety Doskonala) to potentially lethal heat stress and the participation of ROS and calcium in the manifestation of the effects of GABA. Treatment of seedlings with GABA in concentrations of 0.5 and 1 mM caused a significant increase in their survival after damaging heating in a water thermostat (10 min at a temperature of 45 ºC). Under the influence of GABA, there was a transient increase in the content of hydrogen peroxide in the roots of seedlings, followed by an increase in the activity of antioxidant enzymes — superoxide dismutase, catalase, and guaiacol peroxidase. The specified effects of GABA were completely eli-minated by the preliminary application of the hydrogen peroxide scavenger dimethylthiourea (DMTS) to the root
incubation medium and were significantly suppressed in the presence of the NADPH oxidase inhibitor imidazole. At the same time, the treatment of seedlings with the chelator of extracellular calcium EGTA only partially eliminated the increase in the content of hydrogen peroxide and almost did not affect the increase in the activity of antioxidant enzymes in the roots under the influence of GABA. Treatment with neomycin, an inhibitor of calcium uptake from intracellular compartments, caused a partial reduction in the effect of GABA on indicators of the state of the pro-/antioxidant system in wheat roots, but did not eliminate these effects completely. Under the influence of GABA, damage to root cell membranes caused by heat stress was significantly reduced, which was manifested in a decrease in the release of UV-B-absorbing compounds from the cells and a decrease in the content of lipid peroxide oxidation products. At the same time, the stress-protective effect of GABA was completely eliminated by DMTS treatment and changed in the presence of calcium antagonists. A conclusion was made about the important role of ROS generated with the participation of NADPH oxidase in the implementation of the protective effect of GABA on wheat seedlings under conditions of heat stress and the partial dependence of its protective effects on calcium homeostasis.

]]> http://cytgen.com/en/2024/_3-15N2V58.htm http://cytgen.com/ru/2024/_3-15N2V58.htm Fri, 01 Mar 2024 00:00:00 +0200 SUMMARY. For the first time, the effect of 5- and 10-day soil flooding on the ultrastructure of the leaf  mesophyll cells of the psammophyte desert beetroot (Alyssum desertorum L.) was investigated. The seeds for the experiments were collected from plants of dry sandy areas of the gully slopes of the ravine forest in the steppe zone of the Dnipropetrovsk region. It is shown that a characteristic feature of the leaf photosynthetic cells of this species is the presence of single and large up to 6 μm peroxisomes, which are in close contact with chloroplasts and mitochondria, playing a key role in photorespiration. The general organization of palisade parenchyma cells on the 5^th and 10^th days of soil flooding is basically similar to that in the control. A slight decrease in the size of peroxisomes on the 5^th day of flooding and its increase on the 10^th day, and more often formation of multivesicular structures (assembly of endomembranes) in the vacuole, which is considered as an autophagy enhancement of the cytoplasm under hypoxia, were noted. Differences in the ultrastructure of chloroplasts under the influence of soil flooding consisted in a significant, almost two-fold increase in transient starch, the size and number of plastoglobules, especially on the 10^th day, swelling of granal and stromal thylakoids on the 10^th day. Changes in the ultrastructure of desert beetroot chloroplasts under the influence of soil flooding coincide with those of mesophytes studied in this respect. The obtained data on the chloroplast ultrastructure of de-sert beet psammophyte prove the functioning of the photosynthetic apparatus in conditions of short-term soil flooding, that contributes to the survival of seedlings. The subsequent yellowing of leaves and death of plants indicates, as is assumed, the lack of systemic adaptation, primarily metabolic, that is, the transition to anaerobic metabolism, in this species to long-term hypoxia.

]]> http://cytgen.com/en/2024/16-23N2V58.htm http://cytgen.com/ru/2024/16-23N2V58.htm Fri, 01 Mar 2024 00:00:00 +0200 SUMMARY. Currently, there are about six and a half thousand species of viruses known in the world, among which more than one and a half thousand are phytoviruses. Most plant viruses are capable of causing epiphytoties, which lead to decreased yields, reduced product quality, and sometimes put valuable commertial varieties or even entire plant species at risk of extinction. The glo-bal spread of viruses leads to the need to strengthen phytosanitary and quarantine restrictions, which re-quires additional financial costs. Understanding of the viral biology and the principles of their propagation is a key factor in the formation of strategies and methods of combating these pathogens. Among the newest ap-proaches are the genetic engineering technologies. Their use made it possible to create a number of plant varieties with increased resistance to viruses. However, the problem of creating virus-resistant plants still remains one of the most urgent, since with time viruses acquire the ability to bypass defense mechanisms and there is a need to obtain new resistant varieties. There are several main approaches for obtaining of transgenic plants with increased resistance to viruses. They are based on: RNA interference, resistance associated with viral capsid proteins, RNA-satellites, antisense RNAs, replicases, RNA-dependent RNA polymerase, the ac-tion of ribonucleases, ribosome-inactivating proteins, hammerhead ribozymes, miRNAs, plant antibodies, ets. One of the approaches to creating of virus-resistant plants is the use of ribonucleases genes. The genes encoding ribonucleases have different natural origin and belong to a wide range of hosts: bacteria, fungi, plants, animals. In particular, extracellular ribonucleases are able to cut non-specifically molecules of viral RNA in apoplast, that allows to create plants with increased resistance to various phytoviruses. This review is focused on the study of various genetic engineering approaches and the prospects of their use for the creation of virus-resistant plants. Emphasis is placed on the study of heterologous ribonuclease genes influence.

]]> http://cytgen.com/en/2024/24-43N2V58.htm http://cytgen.com/ru/2024/24-43N2V58.htm Fri, 01 Mar 2024 00:00:00 +0200 SUMMARY. The review examines the main stages of cytokinin biosynthesis and metabolism with an emphasis on the important role of cytokinin oxidase/dehydrogenase (CKO/CKX) in cytokinin degradation. In this context, arguments are made for the crucial importance of this enzyme in maintaining a balanced level of cytokinins in plants. The role of CKX genes encoding cytokinin oxidase/dehydrogenase in determining plant resistance to abiotic stress factors and their yield was analyzed. The molecular genetic ways of regulating the activity of CKX genes are characterized. The results of research on the regulation of CKO/CKX activity in increa-sing the resistance to abiotic stress and crop yield are summarized and the biotechnological ways of realizing such opportunities are described. Prospects for finding substances that inhibit CKO/CKX activity with the aim of creating preparations for agriculture are outlined separately. Prospective chemical inhibitors of CKO/CKX and their effects on cultivated plants are considered.

]]> http://cytgen.com/en/2024/44-57N2V58.htm http://cytgen.com/ru/2024/44-57N2V58.htm Fri, 01 Mar 2024 00:00:00 +0200 Clematis tomentella 2001 (Ranunculaceae) is a typical drought-tolerant sand-fixing plant in desert ecosystem in northwest China. To elucidate the phylogenetic status of C. tomentella and its related species, we determined the complete chloroplast (cp) genome and analyzed their interspecific relationships. The result showed [Rev1] that the cp genome of C. tomentella was 159,816 bp in length, including two inverted repeats of 31,045 bp, a large single copy region of 79,535 bp, and a small single copy region of 18,191 bp. 136 genes were annotated across the whole cp genome, including 92 protein-coding genes, 8 rRNA genes, and 36 tRNA genes, as well as the GC content accounted for 38 %. Crucially, we found that the regions of psbE-petL, trnG_UCC-atpA, ndhF-rpl32, and rps8-infA were highly divergent, which could be marked as DNA barcodes for the identification of C. tomentella in Ranunculaceae. A maximum likelihood phylogenetic tree revealed that C. tomentella was closely related to C. fruticosa. Our results provide the references and implications for the phylogenetic study of Clematis in Ranunculaceae in the future.

]]> http://cytgen.com/en/2024/58-60N2V58.htm http://cytgen.com/ru/2024/58-60N2V58.htm Fri, 01 Mar 2024 00:00:00 +0200 Apis cerana abansis is a unique ecological type of the Asian bees, Apis cerana, distributed mainly over China’s western Sichuan plateau. We used Illumina sequencing to obtain the complete mitochondrial genome of A. c. abansis and determined the phylogenetic relationships between A. c. abansis and other Apis cerana ecotypes. The mitogenome of A. c. abansis contains 15,694 bps and includes 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes and 1 A+T-rich control region. All protein-coding genes are initiated by ATT or ATG codons and terminated by the typical stop codons, TAA or TAG, but the start codon of the ATP8 gene (one of the 13 protein-coding genes) is ATC. The ML phylogenetic tree based on the 13 protein-coding genes showed that A. c. abansis formed a sister group with the Yun-Gui Plateau Chinese bee and the Central China Chinese bee. This study provides a scientific basis for the protection and breeding of A. c. abansis.

]]> http://cytgen.com/en/2024/61-62N2V58.htm http://cytgen.com/ru/2024/61-62N2V58.htm Fri, 01 Mar 2024 00:00:00 +0200 Worldwide, Pigeonpea (Cajanus cajan L.) is a protein source. SBP-box transcription factors are crucial for plant development and regulation of stress resistance. The SBP genes in Pigeonpea were examined utilizing genomic information. Using databases, PlantTFDB and NCBI, SBP-box family genes of Pigeonpea were identified and then characterized in silico using bioinformatics tools. In this study, 5 major chromosomes out of 11 and an unplaced scaffold of the Pigeonpea were found to have 24 SBP genes. Significant differences in CcaSBPs protein length, molecular weight, GRAVY value (grand average of hydropathicity), and theoretical isoelectric point were observed. It was shown by Gene Structure Display Server (GSDS) that all CcaSBP genes contain one or more introns. CcaSBP proteins and SBP proteins from other species (A. thaliana and O. sativa) were analyzed phylogenetically and grouped into seven major groups (I, II, III, IV, V, VI, VII). Through this, an effort has been made to present unique information on CcaSBP genes to study Pigeonpea growth and stress mechanisms.

]]> http://cytgen.com/en/2024/63-65N2V58.htm http://cytgen.com/ru/2024/63-65N2V58.htm Fri, 01 Mar 2024 00:00:00 +0200 We obtained the full-length cDNA clone of KAP26.1 in Liaoning cashmere goat, and then we further investigated biological functions of KAP26.1. First, we discovered KAP26.1 was specifically expressed in internal root sheath of skin hair follicles by semi-quantitative reverse transcription and polymerase chain reaction (semi-quantitative RT-PCR), hybridization in situ; immunohistochemistry revealed KAP26.1 was located in the internal and external root sheaths. Next, quantitative real-time polymerase chain reaction (qRT-PCR) results showed relative KAP26.1 expression quantity was significantly different between primary and secondary follicles during anagen, catagen, and remarkably increased during telogen. Moreover, after inhibiting Noggin expression, we found relative KAP26.1 expression quantity significantly declined; after KAP26.1 overexpression, we found relative Noggin expression quantity highly significantly declined. Finally, we found MT played a positive role in KAP26.1 and KAP26.1 expression; FGF5 and IGF-1 palyed a negative role in KAP26.1 and blocked the degradation of KAP26.1. The results revealed KAP26.1 played an important role in regulating fine hair development.

]]> http://cytgen.com/en/2024/66-67N2V58.htm http://cytgen.com/ru/2024/66-67N2V58.htm Fri, 01 Mar 2024 00:00:00 +0200 In this study, dose-dependent effects of lunularic acid (LA) on some physiological, cytogenetic, biochemical and anatomical parameters were investigated in Allium cepa L. bulbs. For this purpose, physiological parameters to be analyzed experimentally: germination percentage, root length, root number and fresh weight; cytogenetic parameters: micronucleus (MN) frequency, chromosomal aberrations (CAs) and mitotic index (MI); biochemical parameters were determined as catalase (CAT), superoxide dismutase (SOD) activities, malondialdehyde (MDA) level and free proline (Pr) content. In addition, cross-sections were taken from the roots and structural changes in meristem cells were examined. Onion bulbs were divided into four groups as one control and three treatments. The bulbs of the control group were kept in cuvettes containing tap water and the bulbs of the treatment group were kept in cuvettes containing 1 mM, 5 mM and 10 mM LA for 7 days. LA administrations caused a decrease in all investigated physiological parameter values, an increase in the frequency of MN and CAs, and reduce in MI compared to control group. In addition, LA application caused dose-related increases in CAT and SOD activities and MDA and Pr levels compared to control group. LA application promoted CAs such as sticky chromosome, spindle fiber damage, vagrant chromosome, reverse polarization in root meristem cells. After all LA applications, root anatomical structure changes such as epidermis cell deformations, flattened cell nucleus and unclear transmission tissue were observed and it was determined that these changes reached a maximum at 10 mM LA dose. As a result, it has been understood that high doses of LA promote multi-directional toxicity and the Allium test is a very reliable test in determining this toxicity.

]]> http://cytgen.com/en/2024/68-71N2V58.htm http://cytgen.com/ru/2024/68-71N2V58.htm Fri, 01 Mar 2024 00:00:00 +0200 SUMMARY. Ivermectin influence on phytopathogenic strains of Fusarium graminearum (F-55644, F-55748) and Fusarium oxysporum f. sp. lycopersici (F-52897, F-55547) was analysed. As the result, it was determined that ivermectin has antifungal effect on the growth of colonies of these strains at high concentrations (2–3 mg/ml). Moreover, the F. oxysporum strains in general were more susceptible to ivermectin than F. graminearum strains. As it is known that ivermectin can cause the microtubules stabilization through binding to β-tubulin, the 3-dimensional model of the interaction of this compound with β-tubulin of F. graminearum was developed to identify of ivermectin induced changes in β-tubulin molecular conformation including the stabilization and spiralization of M-loop. This structural element is important for the establishment of lateral contacts between tubulin subunits of adjacent microtubular protofilaments. As the M-loop stabilization reflects a very important feature of microtubules sta-bilizing agents binding to the taxane site of β-tubulin, it can be supposed, that ivermectin possesses the same effects on Fusarium microtubules. Consequently, the obtained results allow to consider fungal tubulin as a potential target for realization of antifungal activity of ivermectin and its derivatives.

]]> http://cytgen.com/en/2024/_3-13N1V58.htm http://cytgen.com/ru/2024/_3-13N1V58.htm Mon, 01 Jan 2024 00:00:00 +0200 SUMMARY. Garden asparagus contains unique compo-unds that can stop the development of cancerous tumors and even destroy cancer cells. However, there is still insufficient information about the features of the biosynthesis of these compounds and the conditions for their production. In this paper, the influence of an inorganic nitrogen source on the growth of a suspended culture of garden asparagus, the accumulation of phenolic compounds in the culture and culture liquid, as well as the synthesis of saponins is studied. A callus and suspension culture of garden asparagus Asparagus officinalis L. was obtained under in vitro conditions. For further studies, the concentrations of growth regulators 1 mg/l of kinetin and 1.5 mg/l of 2.4-D were selected. The fastest growth of the suspension culture was established on a medium with a concentration of potassium nitrate 1,900 mg/l and ammonium sulfate 250 mg/l. Individual explants showed spontaneous differentiation of callus tissue, which affected the increase in the mass of callus and the ratio of the concentration of exo-phenols to the total mass of the explant. The highest concentrations of endo- and exo-phenols were obtained on a medium without ammonium nitrogen sources. A positive effect of a medium with an ammonium sulfate concentration of 500 mg/l on the synthesis of the main steroid saponins of Asparagus officinalis L. was determined. As a result of the study, a modified MS medium was developed that promotes the accumulation of steroid saponins in a suspension culture of garden asparagus.

]]> http://cytgen.com/en/2024/14-24N1V58.htm http://cytgen.com/ru/2024/14-24N1V58.htm Mon, 01 Jan 2024 00:00:00 +0200 SUMMARY. To study the effect of brassinosteroids and salinity stress on the fatty acid metabolism in the seeds of oil plants, we analyzed the turnover of fatty acids in the seeds of Crambe tatarica under the mentioned conditions. The results of gas-liquid chromatography and mass-spectrometry demonstrated the decrease in the level of palmitic and linoleic fatty acids along with the increase in the level of oleic, eicosenoic, and docosenoic fatty acids in C. tatarica seeds in response to 28-homobrassinolide under salinity stress on some stages of seed germination. The found regularities allowed for the assumption that 28-homobrassinolide promotes the stabilization of fatty acid composition of C. tatarica seeds, disrupted under salinity stress. It may indicate the possibility of modulating the activity or expression of desaturase genes and the enzymes of fatty acid oxidation under the effect of brassinosteroids.

]]> http://cytgen.com/en/2024/25-33N1V58.htm http://cytgen.com/ru/2024/25-33N1V58.htm Mon, 01 Jan 2024 00:00:00 +0200 SUMMARY. The genetic structure of the Holstein cattle population raised in Ukraine was studied by beta-casein and tumor necrosis factor alpha gene polymorphism. Polymorphism of the beta-casein gene (CSN2) was analyzed by A¹ and A² allelic variants using allele-specific PCR (AS-PCR) and the tumor necrosis factor alpha (TNF-α) gene using PCR with restriction fragment length polymorphism analysis (PCR-RFLP) that included the SacI-polymorphism in the promoter region of the gene (marker mutation –824A > G) and the RsaI-polymorphism in the fourth exon. It was demonstrated that both loci are polymorphic in the research cattle population. For each of the polymorphic loci, main genetic parameters of the Holstein cattle population were determined. For the CSN2 locus, a significant prevalence of allele A² frequency over A¹ was established (0.78 versus 0.22). In the case of the SacI- and RsaI-polymorphisms of TNF-α, maximum values (1.980 and 1.988) were found for the locus’s polymorphism level (number of effective alleles). For both TNF-α gene mutations, actual parity was established by distribution of allele frequencies (0.55 and 0.45 for SacI-polymorphism; 0.54 and 0.46 for RsaI-polymorphism). Regarding the RsaI-polymorphism in the fourth exon of TNF-α, there was a deviation from the state of genetic equilibrium according to Hardy-Weinberg, with a significant excess the number of heterozygous individuals (25 %) in the research population. Analysis of the milk production of individuals with different CSN2 genotypes revealed that individuals with the A²A² genotype had higher standard milk yields compared to individuals with the A¹A¹ genotype (p = 0.042). There were no significant differences in milk yield for two lactations between individuals with different genotypes by SacI- and RsaI-polymorphisms of the TNF-α gene.

]]> http://cytgen.com/en/2024/34-45N1V58.htm http://cytgen.com/ru/2024/34-45N1V58.htm Mon, 01 Jan 2024 00:00:00 +0200 SUMMARY. Cytogenetic studies were performed in bone marrow cells of 24 patients in the relapse of acute lymphoblastic leukemia (ALL), admitted to the laboratory during 2012–2022. A significant percentage of mosaic karyotypes (75.0 %) was noted, among which the combination of an abnormal clone and a normal karyotype (33.3 %) dominated. Trisomies of chromosomes led to the formation of hyperdiploid clones, among which trisomies of chromosomes 6, 21, 15 and 5 were most often found. Most often, additional marker chromosomes were recorded (22.2 %). According to the mechanisms of formation of abnormal clones in the relapses of ALL, structural abnormalities of chromosomes were recorded, among which balanced (translocations, inversions) and unbalanced (deletions, isochromosomes, additional material of unknown origin, duplications, unbalanced translocations) were determined. Losses of genetic material (deletions) (24.1 %) and translocation (33.3 %) were more often detected. The total number of cases with translocation t(9;22)(q34;q11.2) was 20.4 %. The evolution of clonal chromosome abnormalities was due to the appearance of additional quantitative and unbalanced structural abnormalities. Comparison of chromosome abnormalities at diagnosis and relapse in B-ALL did not reveal common mechanisms for the formation of chemotherapy  resistant clones. Trisomy of chromosome 5, deletion del(6)(q23), and translocation t(9;22)(q34;q11.2) were involved in the formation of clones that were insensitive tochemotherapy. The group of unfavorable prognosis included 95.8 % of karyotypes.

]]> http://cytgen.com/en/2024/46-53N1V58.htm http://cytgen.com/ru/2024/46-53N1V58.htm Mon, 01 Jan 2024 00:00:00 +0200 Fennel is considered as a very important spice crop with powerful therapeutic potential. An improvement in this valuable crop, selected physical mutagen (Gamma irradiation) on the seeds with five selective doses viz., 50 Gy, 100 Gy, 150 Gy, 200 Gy and 250 Gy for break genetic consistency in narrow genetic base in Fennel crop, remunerative phenomenon of syncytes was seen in some cases, where complete chromatin was transmitted to the recipient PMC, that generates dimorphic pollen grain. Such pollen grain with varying genetic content plays a significant role in the emergence of intraspecific polyploidization of species. A curious biological process which is often observed in microsporogenesis of higher plants like, development of syncytes, cytomixis between plant cells and due to this the creation of big pollen has evolutionary relevance. The cytomictic behaviour of Foeniculum vulgare Mill. has been reported in this experiment.

]]> http://cytgen.com/en/2024/54-55N1V58.htm http://cytgen.com/ru/2024/54-55N1V58.htm Mon, 01 Jan 2024 00:00:00 +0200 Worldwide, endometrial cancer is one of the most frequently diagnosed malignancies in women and a notable cause of death. The aim of this study was to perform image cytometric DNA ploidy analysis on a prospective material of endometrial adenocarcinomas in order to determine potential correlation between ploidy status and their histological features. The analysis was carried out in fresh tissue samples resected by implementing complete hysterectomy in a series of patients (n=126). We found that ploidy status using image cytometry correlate with histologic type, grade and stage in endometrial cancer and aneuploid tumor samples are associated with aggressive phenotype statistics. Furthermore, DNA ploidy should be used as a reliable and applicable prognostic marker in the routine clinical practice.

]]> http://cytgen.com/en/2024/56-58N1V58.htm http://cytgen.com/ru/2024/56-58N1V58.htm Mon, 01 Jan 2024 00:00:00 +0200 We have hypothesized that the TAP2 gene is associated with lipid metabolism. Here, 10 Min-pig tissues were collected to detect the expression of TAP2 in different tissues. We obtained dorsal subcutaneous structural vascular fraction (SVF) cells from the Min-pig’s back adipose tissue and induced SVF cells into mature adipocytes. By overexpression and interference, the effect of TAP2 on fat deposition in Min-pig SVF cells was studied. Recombinant human insulin, dexamethasone, indomethacin, 3-isobutyl-1-methylxanthine, triiodothyronine, and rosiglitazone could successfully induce SVF cells into mature adipocytes, and the induction efficiency was above 50%. The tissue expression profile showed that TAP2 was expressed in different tissues, and the highest expression was found in back fat, spleen, and back muscle. Overexpression of the TAP2 gene in SVF cells could significantly increase the expression of adipose differentiation-related genes. The expression of TAP2 in SVF cells was reduced to 0.6 times after transfection of the TAP2 gene interference fragment. The adipogenic transcription factor gene C/EBPα, fatty acid synthase gene FSA, and adipocyte directional differentiation factor ADD1 were downregulated, while the expression of lipolysis-related gene LPL was inhibited. In conclusion, TAP2 expression can promote the deposition of subcutaneous fat on Min-pig’s back adipose tissue.

]]> http://cytgen.com/en/2024/59-61N1V58.htm http://cytgen.com/ru/2024/59-61N1V58.htm Mon, 01 Jan 2024 00:00:00 +0200 The pea is an important cool-season pulse crop cultivated for animal and human consumption. However, the presence of ‘flatulence-causing factors’ hinders its consumption and acceptance worldwide. The raffinose family oligosaccharides (RFOs) have been identified as the principal ‘flatulence causing factors’. Hence, reducing RFO level is the major goal to promote pea consumption and acceptance worldwide. However, very little is known about the genes involved in RFO metabolism at the genome-wide scale in pea. In the present study, genes for five key enzymes (galactinol synthase, raffinose synthase, stachyose synthase, alpha-galactosidase_Acid/Alkaline and beta-fructofuranosidase) involved in RFO metabolism pathway were identified at the genome-wide scale in pea. A total of two galactinol synthase, two raffinose synthase, one stachyose synthase, six alpha-galactosidase_ Alkaline and three alpha-galactosidase_Acid and ten beta-fructofuranosidase genes were identified in the pea genome. Phylogenetic relationships analysis, exon/intron structure as well as conserved domain within each enzyme family and their chromosomal location were also determined to establish their relationship with the known proteins. In silico anlaysis showed that pea RFO genes contain 26 microsatellite loci. Taken together, this study provides useful candidate genes for improving the nutritional quality of pea through genetic engineering approaches as well as microsatellite loci for the development of SSR markers for the introgression of low RFO trait through marker assisted selection.

]]> http://cytgen.com/en/2024/62-63N1V58.htm http://cytgen.com/ru/2024/62-63N1V58.htm Mon, 01 Jan 2024 00:00:00 +0200 SUMMARY. Stem rust caused by the fungus Puccinia graminis Pers. is a dangerous disease of wheat which occurs in all regions of its cultivation. New highly virulent races causing severe yield losses have appeared in recent decades. Sr33 introgressed from Aegilops tauschii is one of the genes conferring resistance against most races of stem rust including Ug99. To develop winter common wheat lines with the gene Sr33 and evaluate a possible effect of the gene on yield traits, we made a cross between the spring line DH31 carrying the Sr33 gene and the winter cultivar Myrkhad followed by marker-assisted selection of winter genotypes with Sr33 beginning from F₃. To identify the Sr33 gene, we used PCR with the gene-specific marker Sr33A. Alleles at the storage protein loci of the parental forms were identified using acid polyacrylamide gel electrophoresis and SDS-electrophoresis, as well as the molecular marker MAR for Glu-B1al. As a result of marker-assisted selection, winter F₅ lines with the Sr33 gene were developed from the cross DH31 × Myrkhad. The yield traits of F₅ spikes of families derived from single F₃ spikes with and without Sr33 were analyzed considering that the line DH31 has a specific allele at Gli-D1 from Ae. tauschii, dark glimes, and high molecular weight glutenin subunit alleles associated with high dough strength, in particular Glu-B1al. Comparison of means of yield traits of spikes from families with Sr33 and without it did not reveal significant differences between these two groups. Thus, winter F₅ lines with the Sr33 gene from the cross DH31 × Myrkhad may be used in the breeding practice to develop cultivars with high bread-making qualities of flour and stem rust resistance.

]]> http://cytgen.com/en/2023/__3-10N6V57.htm http://cytgen.com/ru/2023/__3-10N6V57.htm Wed, 01 Nov 2023 00:00:00 +0200 SUMMARY. Tandemly arranged repetitive regions (repeats) that encode 5S rRNA (5S rDNA) are a vital component of eukaryotic genomes. Typically, 5S rDNA repeats are highly similar within a genome due to the concerted nature of the evolution of this type of repeats. Each 5S rDNA repeat consists of an evolutionarily conserved coding sequence (CDS) and a variable intergenic spacer (IGS). 5S rDNA is a popular model for studying the molecular evolution of repetitive sequences, and the high rate of IGS mutations determines its wide use in phylogenetic analysis of closely related taxa. Nevertheless, 5S rDNA still remains unexplored for many groups of higher plants, in particular, the family Plumbaginaceae. Some taxa of this family are endemic to southern Ukraine and are included in the Red Book. However, their taxonomic status is controversial, and its clarification requires the use of molecular phylogenetic methods. In this work, we investigated the molecular organization of 5S rDNA for representatives of four genera of the tribe Limonieae, which is the largest one in the Plumbaginaceae family. It was shown that the CDS of 5S rDNA of representatives of the genera Limonium, Armeria, and Ceratolimon possess single mutations that do not interfere with the formation of the secondary structure of 5S rRNA. In contrast, in the genomes of Goniolimon species, in addition to functionally normal 5S rDNA repeats, numerous pseudogenes that evolve in a non-concert manner and contain numerous mutations in the CDS that disrupt the secondary structure of 5S rRNA were found. A significant phylogenetic distance between representatives of the subgenera Pteroclados and Limonium of the genus Limonium indicates that Pteroclados can be considered a separate genus. The high rate of molecular evolution makes 5S rDNA IGS a convenient tool for the reconstruction of phylogenetic relationships within the studied genera of the tribe Limonieae and barcoding of Ukrainian endemics of the genus Limonium.

]]> http://cytgen.com/en/2023/_11-26N6V57.htm http://cytgen.com/ru/2023/_11-26N6V57.htm Wed, 01 Nov 2023 00:00:00 +0200 SUMMARY. The Crucifers family (Brassicaceae) includes a large number of economically important crops, particularly Brassica rapa, which is a widely-used model plant for molecular genetic studies of oilseeds. B. rapa is a highly polymorphic species that includes a large number of genetically distinct subspecies. Considering this fact, intraspecific hybridization of B. rapa subspecies is considered a promising breeding approach aimed on in-creasing genetic diversity of the crop. Previously, we have shown that one of such hybrids, oil tyfon (B. rapa subsp. oleifera f. biennis × (subsp. rapifera × pekinensis)), could be a valuable oil feedstock due to its increased productivity. However, obtaining hybrids and their subsequent breeding would require the involvement of diverse molecular marker systems. So far, the method of estimating the length polymorphism of the first (TBP) and second (cTBP) introns of β-tubulin has demonstrated its high accuracy and reliability in the identification (DNA-barcoding) of flowering plants taxonomic units at different levels. In the present study, we evaluated the productivity of such hybrid oilseed crop as tyfon, as well as carried out DNA-barcoding of various hybrid lines of tyfon and its parental B. rapa subspecies using on β-tubulin intron length polymorphism assessment approach. Based on the data of the molecular genetic analysis, which included the assessment of length polymorphism of 1^st and 2^nd introns of β-tubulin genes, we were able to confirm the origin of oil tyfon hybrid from Dutch leaf tyfon (B. rapa subsp. rapifera × pekinensis) and winter turnip rape (B. rapa subsp. oleifera) with a high confidence. Along with that, it was possible to differentiate var. glabra and var. laxa accession of napa cabbage (B. rapa subsp. pekinensis) for the first time using the combined TBP and cTBP analyses. A variation in the number of amplified regions of β-tubulin introns was noted in different genotypes, however these differences did not appear to be a specific feature a particular subspecies/hybrid. This suggests that B. rapa hybrids most likely do not differ in ploidy, compared to their parental genotypes. In addition, it was shown that the mentioned oil tyfon hybrid lines of Ukrainian breeding show a significant level of morphological variation, despite their common breeding pedigree.

]]> http://cytgen.com/en/2023/_27-40N6V57.htm http://cytgen.com/ru/2023/_27-40N6V57.htm Wed, 01 Nov 2023 00:00:00 +0200 SUMMARY. Comparative analysis of the rates of molecular evolution, transition bias and its evolutionary compensation is carried out on the example of mitochondrial (D-loop, Cytb, COI, 12S RNA) and nuclear (IRBP, Fv) DNA markers in the Murinae subfamily. According to the levels of variability, markers can be divided into three classes: 1) hypervariable (D-loop), 2) rapidly evolving (Cytb, COI), 3) conservative (12S RNA, IRBP, Fv). The nature of nucleotide substitutions appears in accordance to the levels of variability. In the situation with D-loop, there is a maximum initial bias, which is already partially compensated at the early stages of speciation, and completely compensated at the stages of species divergence. The pronounced bias within the Cytb, COI genes is only partially compensated, moreover at the genus levels. The 12S RNA, IRBP, and Fv genes with a low level of transition bias do not show evolutionary compensation as such, and the decrease of the ts/tv index in the evolutionary lineage has a technical character and is a consequence of a relative decrease of the difference in the frequencies of transitions and transversions against the background of an absolute increase in the frequencies of substitutions. The positive relationship between the intensity of nucleotide substitutions, the level of transition bias and the rates of its evolutionary compensation proves that these phenomena have the same primary basis.

]]> http://cytgen.com/en/2023/_41-47N6V57.htm http://cytgen.com/ru/2023/_41-47N6V57.htm Wed, 01 Nov 2023 00:00:00 +0200 SUMMARY. One of the way to enhance the efficiency of alkylating chemotherapy is reducing the level of the repair enzyme MGMT (O⁶-methylguanine-DNA methyltransferase) in cancer cells. The standard MGMT inhibitor, O⁶-benzylguanine (BG), has exhibited cytotoxicity towards hematopoietic cells in the third stage of clinical trials, making the search for new alternative inhibitors relevant. In this study, we have conducted research to determine the cytotoxicity and efficacy of new potential MGMT inhibitors, which were modeled using molecular flexible docking. At the first stage of the study, MTT and clonogenic assays were performed to assess cytotoxicity, in which HEp-2 cells were cultured with the tested compounds at a concentration of 10 µM. At the second stage, the efficacy of the compounds was evaluated. One of the methods used was a clonogenic assay, in which the cell treatment consisted of combinations of the tested compounds (10 µM) and the alkylating agent N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) at different con-
centrations. Another method was Western blot analysis, for which proteins were extracted from HEp-2 cells treated with potential inhibitors in combination with MNNG. The obtained results were analyzed in Microsoft Excel 2016, Origin8.1 and ImageLab. As a result, 4 of the 5 examined compounds demonstrated low cytotoxicity at a concentration of 10 µM in HEp-2 cells compared to the standard inhibitor BG. According to the clonogenic assay, compound 41B (5-Benzo[1,3]dioxol-5-ylmethylene-thiazolidin-2,4-dione) was the most effective, and compounds 41 (5-(5-Chloro-2-hydroxy-benzylidene)-4-thioxo-thiazolidin-2-one) and 89 (2-[5-(4-Bromo-phenyl)-pyrimidin-4-yl]-5-ethoxy-phenol) also showed high efficiency. The results of Western blot analysis showed a significant decrease in MGMT protein after treatment with compounds 41, 41B and 89, thereby confirming the inhibitory properties of these compounds.

]]> http://cytgen.com/en/2023/_48-59N6V57.htm http://cytgen.com/ru/2023/_48-59N6V57.htm Wed, 01 Nov 2023 00:00:00 +0200 SUMMARY. Using morphological traits and molecular-genetic research methods, we identified 24 species of naked amoeba from natural biotopes. The sequences of gene 18S rRNA were obtained for the following naked amoeba: Amoeba proteus isolate AP07 (ON907618), Saccamoeba limax isolate SLU_22 (OP894078), Saccamoeba limax isolate SL_Uk19 (OQ520144), Saccamoeba sp. strain IDL777 (MZ079370), Thecamoeba striata isolate THS19 (OQ134482), Thecamoeba striata isolate THS20 (OQ134483), Thecamoeba similis isolate from the Prut river (OL604177), Thecamoeba similis isolate from Baggersee Innsbruck (Baggersee Rossau) (OL604178), Thecamoeba quadrilineata isolate THQD2 (ON398269), Thecamoeba quadrilineata isolate THQA1 (ON398268), Thecamoeba sp. strain THS203 (MZ079371), Stenamoeba stenopodia isolate UKSS7 (OP375108), Stenamoeba stenopodia isolate POLSS7 (OP419588), Korotnevella stella isolate KSD2 (ON398267), Korotnevella stella isolate KSA1 (ON398266), Vexillifera bacillipedes isolate from the Dnieper river (OK649262), Vannella lata isolate from the Kamenka river (OL305063), Vannella lata isolate from the Varta river (OL305064), Vannella sp. strain VLS303 (MZ079372), Vannella simplex isolate from the Black Sea (OM403052), Vannella simplex isolate from the Mediterranean Sea (OM403053), Ripella sp. strain RPL100 (MZ079369), Mayorella vespertilioides isolate MV_7 (OP739500), Mayorella sp. isolate MY_7 (OP729930), Acanthamoeba sp. strain ATM123 (MZ079366), Acanthamoeba sp. isolate from the Elbe river (OK649261), Acanthamoeba polyphaga isolate AcPoly01 (ON908497), Acanthamoeba polyphaga isolate AcPoly15 (ON908496), Acanthamoeba griffini isolate from the Black sea (OM522832), Acanthamoeba griffini isolate from the Mediterranean Sea (OM522833), Cochliopodium actinophorum strain COP101 (MZ079367), Cochliopodium minus isolate from the Stokhid river (OK649264), Cochliopodium sp. strain COP102 (MZ079368), Vahlkampfia avara isolate VA7 (OP179657), Willaertia magna isolate from the Teteriv river (OK649263). All the species of naked amoeba in the phylogenetic tree, built using gene 18S rRNA, are within Amoebozoa, united into groups Tubulinea and Discosea. There are separate groups of freshwater, sea and land biotopes; these groups are sister species with low results of bootstrap analysis, which demonstrates low probability of remoteness of some amoeba species, found in different natural biotopes.

]]> http://cytgen.com/en/2023/_60-72N6V57.htm http://cytgen.com/ru/2023/_60-72N6V57.htm Wed, 01 Nov 2023 00:00:00 +0200 SUMMARY. The coronavirus disease (COVID-19), caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), was detected in December 2019. At the beginning of the pandemic, it was believed that children were less susceptible to COVID-19 than adults, but further research has shown that children are also vulnerable to SARS-CoV-2 infection. In recent years, there have been a lot of studies about the role of genetic factors in the course of COVID-19. This fact is suggestive of possible factors of a hereditary predisposition of individuals to SARS-CoV-2 infection. Recently, some obtained evidence has demonstrated that certain genetic polymorphisms, notably the genotypes, differing by the rs12979860 polymorphic variant of the IFNL gene, may predict the severity of respiratory viral infections among children, including COVID-19. The aim of our study was to investigate the specificities of genotype distribution by the rs12979860 polymorphism of the IFNL gene in a cohort of children with COVID-19, and to evaluate the association of this polymorphism with the risk of SARS-CoV-2 virus infection, the development of pneumonia during coronavirus disease, and the course of the disease among children with recurrent respiratory infections (RRI). To this end, the study group of 70 children with laboratory-confirmed COVID-19 was genotyped for the polymorphism of the rs12979860 locus of the IFNL gene. The study found that the C allele was more common in children with RRI compared to those with episodic viral infections (p < 0.05, OR 3.2; CI 1.52–6.71), so this variant can be considered a risk allele for more frequent viral infections. Also, the C allele was predominant in the subgroup of children with pneumonia (p < 0.05, OR 2.36 CI 1.19–4.68), indicating that the C allele can be considered a risk allele for more severe COVID-19 due to pneumonia. The results suggest that the C allele may be a predictive marker of the risk of pneumonia in children with COVID-19. In addition, the carriage of the C allele is associated with cases of RRI among children.

]]> http://cytgen.com/en/2023/_73-81N6V57.htm http://cytgen.com/ru/2023/_73-81N6V57.htm Wed, 01 Nov 2023 00:00:00 +0200 SUMMARY. RNA interference (RNAi) is a new potential tool for plant breeding by introducing small non-coding RNA sequences that can silence gene expression in a sequence-specific manner. The ability to reduce the expression of a specific gene provides the possibility of acquiring a new characteristic by eliminating or accumulating certain plant traits, which leads to biochemical or phenotypic changes that the original plants do not have. This literature review describes the progress achieved over the past decades in the application of RNAi for the creation of cereal crops with improved economically valuable traits. The main stages of the gene silencing mechanism, mediated by short interfering RNAs (siRNAs), the features of their biogenesis, the mode of action, and distribution are briefly presented. Numerous examples of the development of various biotechnological approaches to improving cereals using gene transformation and exogenous double-stranded RNA molecules are summarized. The possibility of using RNAi technology to change the agronomic characteristics of plants, enhance the nutritional value and quality of the grain, and reduce the number of toxic compounds and allergens is highlighted. Considerable attention is paid to the practical results of various applications of RNAi to increase the resistance of grain crops to biotic stress factors, in particular, viruses, bacteria, fungi, insect pests, and nematodes. Examples of the use of siRNA-mediated RNAi to improve cereal resistance to abiotic stresses, including drought and salinity, are given.

]]> http://cytgen.com/en/2023/_82-109N6V57.htm http://cytgen.com/ru/2023/_82-109N6V57.htm Wed, 01 Nov 2023 00:00:00 +0200 HER2 expression is associated with 30 % of breast cancer patients with a poor prognosis. Though Trastuzumab is approved for HER2 targeted therapy, its use is limited because of its systemic toxicity and resistance in most patients. This study evaluated the synergistic effects of Phenethyl isothiocyanate (PEITC) and Curcumin (CUR) in HER2 overexpressing SK­BR­3, BT­474, and AU­565 breast cancer cells. The cytotoxic effect of PEITC: CUR against breast cancer cells was evaluated using an MTT assay, and the Loewe additivity model was used to evaluate the synergistic effect. Apoptosis induction and cell cycle arrest over the treatment of PEITC: CUR in breast cancer cells were examined using the flow cytometric annexin­V/Propidium iodide method. Downregulation of HER2­mediated signaling was deduced from protein expression analysis using western­blot. Our results showed that treatment of PEITC: CUR at varying levels of combinations in all three breast cancer cells extensively reduced the survival of the cells with the lowest inhibitory concentrations (IC₅₀). Cytotoxic data revealed that the 3:1 ratio of PEITC: CUR was the best among several (1:1, 3:1, and 1:3) combinations, with the maximum cytotoxicity. PEITC: CUR (3:1) displayed the lowest combination index (CI) against SK­BR­3, and AU­565 cells indicated its potential synergistic effect. At twice the concentration of its IC₅₀, the 3:1 combination elicited 3.5 to 4.5 fold apoptosis in HER2 overexpressing cells, approximately double the effect of the individual drugs alo­ne. In addition, the selected combination induced the G2/M cell cycle arrest in HER2 expressing cells over the treatment. Western blot protein expression analysis revealed that the PEITC: CUR combination suppressed the HER2/PI3K/Akt signaling, eventually connected to various apoptotic biological events. Our results showed the specificity of PEITC: CUR combination in inducing apoptosis and G2/M cell cycle arrest in HER2­expressing tumor cells in­vitro and enhancing the anti­cancer effect. For a subset of breast cancer patients who overexpress HER2, this combination of PEITC and CUR could be a potential treatment option.

]]> http://cytgen.com/en/2023/110-112N6V57.htm http://cytgen.com/ru/2023/110-112N6V57.htm Wed, 01 Nov 2023 00:00:00 +0200 SUMMARY. Genus Muscari Mill. (Asparagaceae Juss.) includes about 80 species distributed in Eurasia, mainly in the Mediterranean region. Recent molecular phylogenetic studies have shown that the taxa belonging to this group form a monophyletic clade and are closely related. However, the phylogeny and status of some taxa of Muscari sensu lato remain controversial. So far, most phylogenetic stu-dies of the genus Muscari have used almost exclusively Mediterranean plant material, while representatives of the genus from the Carpathian region still remain unexplored. In this work, we used the sequencing of three regions of chloroplast DNA, psbA-trnH, trnT-L and trnL-F, to clarify the phylogenetic relationships in the genus Muscari, to assess the genetic polymorphism and taxonomic status of Ukrainian populations of M. botryoides, as well as specimens of other Muscari species from the Carpathian region. According to the results of the phylogenetic analysis, the genus Muscari is a monophyletic group that includes three subgenera: Muscari, Muscarimia and Pseudomuscari. Leopoldia species have been placed in the subgenus Muscari. Specimens of M. botryoides from Ukraine and Austria together with M. transsilvanicum from Romania and M. serpentinicum/M. sandrasicum from Turkey form the clade «Botryoides», one of the three main clades identified in the subgenus Muscari. A significant genetic distance between Ukrainian specimens of M. botryoides, specimens of this species from other habitats, and other species of the genus Muscari allows us to consider the Ukrainian specimens of M. botryoides as a new, previously undescribed species. A comparison of the sequences of the investigated regions of the chloroplast genome revealed genetic differences between two groups of Ukrainian populations of M. botryoides, which can be interpreted as the existence of two intraspecific forms.

]]> http://cytgen.com/en/2023/__3-16N5V57.htm http://cytgen.com/ru/2023/__3-16N5V57.htm Fri, 01 Sep 2023 00:00:00 +0300 SUMMARY. The intergenic spacer (IGS) of 5S ribosomal RNA genes (5S rDNA), which are present in the genome of all living organisms, is characterized by high variability, which makes it a convenient and widely used tool for studying issues of genome evolution, population genetics, systematics, etc. The objective of this study was to study the IGS of 5S rDNA of Antarctic pearlwort Colobanthus quitensis. Using molecular genetic methods, IGS region was amplified, cloned, and sequenced, followed by the analysis of structural organization. The IGS of 5S rDNA of C. quitensis was shown to contain basic regulatory elements typical of other vascular plants. At least two classes of 5S rDNA repeats, which differ significantly in length and nucleotide sequence of the spacer, were found in the individual genome. In addition, based on the differences in length and nucleotide sequence, 2 subclasses of the repeats with a long IGS and 3 subclasses of repeats with a short IGS were distinguished. Comparison of the sequences of the IGS of 5S rDNA of C. quitensis and Silene latifolia Poir., a species of another section of Caryophyllaceae, showed significant differences in the structure of the spacer of 5S rRNA genes, excluding its parts that contain regulatory elements. In general, the results indicate a significant level of intragenomic polymorphism of MGS 5S rDNA in C. quitensis.

]]> http://cytgen.com/en/2023/_17-24N5V57.htm http://cytgen.com/ru/2023/_17-24N5V57.htm Fri, 01 Sep 2023 00:00:00 +0300 SUMMARY. The morphological study was aimed at determining the role of fibrillar organization of the collagen-containing connective tissue of prostate cancer at the stage of neoplastic proliferation, including me-tastatic spreading into bone tissue. The histological material of 55 patients with prostate cancer, Glea-son 6–9 malignancy score, without neoplastic proliferation and with metastases into bones, was used. The architecture specificities of collagen-containing connective tissue of neoplasms were determined using Van Gieson’s method. It was found that the remodelling of collagen-containing connective tissue around the epithelial tumor structures with aggressive prostate cancer (Gleason score of 8–9) resulted in the increase in the percentage of extended and flattened fibrils as compared to curved fibrils; the adjacent and surrounding stroma was notable for the enlarged total area of collagen-containing fibrils, manifestations of desmoplasia, compactization of the location, widening, flattening, and extending. The data obtained demonstrate that the remodelling of collagen-containing connective tissue component of prostate cancer conditions unrestricted migration and invasion of tumor cells, including the ones, expressing the proteins, involved in bone tissue remodelling.

]]> http://cytgen.com/en/2023/_25-32N5V57.htm http://cytgen.com/ru/2023/_25-32N5V57.htm Fri, 01 Sep 2023 00:00:00 +0300 SUMMARY. The phenomenon of the positive influence of introns on the expression of a corresponding gene, which is called intron-mediated enhancement (IME), is characteristic of a wide variety of organisms, including nematodes, insects, mammals, fungi, and plants, and occurs due to an as-yet-undefined fundamental mechanism. IME introns have been used for a long time, in particular, in plant biotechnology. Understanding the mechanisms of this phenomenon allows predicting and easily generating stimulatory introns with the given properties and creating highly advantageous phenotypes. It will also greenlight the use of IME in gene therapy and to improve the production of pharmaceutical proteins. In this review, we analyzed previously proposed models of IME functioning mechanisms and identified factors that can directly or indirectly determine IME under different conditions and at different levels of gene expression, such as experimental methods of IME research, regulatory RNAs, sequence properties, intron position and orientation, factors at the levels of DNA, transcription, splicing, mRNA, translation, genes in which IME is detected, tissue specificity, repression and how some factors relate to each other by importance. Since there is no single mechanism of IME, and the effect may differ in different species, when modeling this process, only the cases of IME affecting the same level of expression should be compared with each other, taking into account the experimental conditions. Identifying the biological factors that can determine IME and the relationship between them will help in the future to create a corresponding dataset suitable for machine learning and to try to solve the mystery of the IME phenomenon with the help of machine learning.

]]> http://cytgen.com/en/2023/_33-57N5V57.htm http://cytgen.com/ru/2023/_33-57N5V57.htm Fri, 01 Sep 2023 00:00:00 +0300 SUMMARY. The members of the Brassicaceae family are important oilseed, vegetable and forage crops. The Brassicaceae species are also used for phytoremediation of heavy metal polluted soils, or as siderates and ornamental plants. The widening of biodiversity of already existing Brassicaceae cultivars by genetic engineering manipulations can increase tolerance to biotic and abiotic stresses, increase plant productivity and production quality. Protoplast fusion method makes possible artificial synthesis of various somatic hybrids between different plant species. The review presents advances in protoplast fusion of Brassica crops and their wild relatives; main directions of somatic hybridisation in Brassicaceae; the conditions for obtaining and selection of the somatic hybrids; the peculiarities of genetic material inheritance, introgression of economically important traits by somatic hybridization to cultivated Brassicaceae, resynthesis of allotetraploid species; production of cytoplasmic male sterility (CMS) alloplasmic lines, creation of somatic hybrids for phytoremediation of contaminated with heavy metals soils, and production of transplastomic plants by transfer of genetically modified plastids through protoplast fusion.

]]> http://cytgen.com/en/2023/_58-79N5V57.htm http://cytgen.com/ru/2023/_58-79N5V57.htm Fri, 01 Sep 2023 00:00:00 +0300 Long noncoding RNAs (lncRNAs) have recently been identi­fied as crucial biomarkers of papillary thyroid cancer (PTC). In this study, we aimed to investigate the biological function and potential clinical role of lncRNA ENST00000494165 in PTC. From January 2019 to December 2019, 226 PTC patients who underwent preoperative thyroid US­fine needle aspiration biopsy(US­FNAB) and confirmed by postoperative pathology were enrolled at our hospital. Thyroid tissues were collected from FNAB samples and stored in the refrigerator at –80 ºC. qRT­PCR (quantitative reverse transcriptasepolymerase chain reaction) analysis was performed to detect the relative expression level of ENST00000494165. CCK­8 (Cell Counting Kit­8) and colony formation assay were performed to detect the cell proliferation ability. Cell migration and invasion abilities were evaluated by transwell and scratch assay. We also evaluated the relation between the expression level of ENST00000494165 and the clinicopathological features of PTC. Functional assays demonstrated that the cell proliferation, migration and invasion abilities were all promoted in PTC cell lines when ENST00000494165 expression was overexpressed. The expression of ENST00000494165 in the lymph node metastasis group was significantly higher than that in nonlymph node metastasis group. High level expression of ENST00000494165 was significantly associated with lymph node metastasis of PTC(P < 0.001). The overexpression of ENST00000494165 promoted the progression, migration and invasion abilities of PTC and was significantly correlated with lymph node metastasis in PTC. ENST00000494165 could act as a possible promoter gene and a potential biomarker for PTC.

]]> http://cytgen.com/en/2023/_80-81N5V57.htm http://cytgen.com/ru/2023/_80-81N5V57.htm Fri, 01 Sep 2023 00:00:00 +0300 MicroRNA(miRNA), a small non­coding class of RNA that regulates the gene expression, is conserved among several plant species. In the present study, an in­silico approach was adopted to identify miRNA from the known expressed sequences tags (ESTs) of Citrus reticulata L. A total of 17 miRNAs from 23 different ESTs along with their secondary structures and targets were predicted. The identified 63 targets include several transcription factors, proteins that regulate plant growth, development, flowering, and seed development together with stress response. The cis­regulatory element present at the promoter region of the MIR genes of C. reticulata showed relevance towards light responsiveness, auxin, gibberellins, abscisic acid (ABA), anthocyanin responsiveness, salicylic acid responsiveness, anaerobic induction, circadian control, nitrate dependent regulation of the cell cycle and DNA replication, defense, and stress responsiveness. The present study identifies the miRNAs along with their regulatory elements in C. reticulate. The study will also support the research on miRNAs identification from the genomic data of different plants and prediction of the possibilities of expression of identified MIR genes based on the presence of upstream promoter and other regulatory elements.

]]> http://cytgen.com/en/2023/_82-85N5V57.htm http://cytgen.com/ru/2023/_82-85N5V57.htm Fri, 01 Sep 2023 00:00:00 +0300 To obtain information of Periplaneta americana, we analyzed the distribution characteristics of microsatellite sequences in the P.americana transcriptome (229 MB) by using MSDBv2.4. The total number of perfect microsatellite sequences was 38,082 and covered about 0.3 % of P. americana transcriptome. The cumulative length of microsatellites was 618,138 bp, and the density of microsatellites was 2978.54 bp/Mb. In the different repeat types of the microsatellites, the number of the mononucleotide repeats was 20,002 (accounting for 52.52 %), which obviously was the most abundant type. While the trinucleotide, tetranucleotide, dinucleotide,pentanucleotide and hexanucleotide repeats accounted for 24.51, 12.97, 8.1, 1.61 and 0.26 %, respectively. The kind of different repeat copy categories in each repeat type was also quite dif­ferent, such as the A in mononucleotide repeat type, the AG in dinucleotide, the AAT in trinucleotide, AAAT in tetranucleotide, the AAGAA in pentanucleotide, and the CAGTAG in hexanucleotide were the most of each cate­gory. The A, T, AC, AG, AT, GT, AAG, AAT, ATC, ATG, ATT, CTT, AAAG and AAAT were the dominant repeat copy categories, the total number of all these types was 29,933, accounting for 78.6 % in the total number of microsatellite sequences. These results based on a foundation for developing high polymorphic microsatellites to research the functional genomics, population genetic structure and genetic diversity of P. americana.

]]> http://cytgen.com/en/2023/_86-88N5V57.htm http://cytgen.com/ru/2023/_86-88N5V57.htm Fri, 01 Sep 2023 00:00:00 +0300 The year 2022 paid a bicentennial tribute to the phenomenal work of the father of Genetics, Gregor Johann Mendel for deciphering the genetic logic behind the phenotypes. His principles were distilled as the law of segregation and law of independent assortment. His work was rediscovered 34 years later by H. De Vries, C. Correns, and E. Tschermak and popularized by W. Bateson. While C. Darwin accounted for similarities among organisms through the differences in the form of evolution, G. Mendel accounted for similarities through heredity; the ideological gaps were bridged mathematically by R. Fisher. Later with the test of time, the interaction among researchers paved Mendelian principles into different branches of genetics viz., cytogenetics, molecular genetics, population genetics, quantitative genetics, etc. At present we have landed in the era of genomics and the emerging field of phenomics which have potential to bridge the huge gap between demand and supply in different agro­industrial and allied goods. In order to connect the budding researchers in the field of genetics with Mendelism and its significance, catalyzed our concentrated effort to link Mendelism across the centuries, highlighting its importance and extrapolating the concept of heredity and variation from garden peas to different life forms. In conclusion, as our knowledge on genetics deepens, more insights on underlying mechanisms and subsequent applications will be witnessed.

]]> http://cytgen.com/en/2023/_89-94N5V57.htm http://cytgen.com/ru/2023/_89-94N5V57.htm Fri, 01 Sep 2023 00:00:00 +0300 SUMMARY. Chronic myelogenous leukemia (CML) is a myelo-proliferative neoplasm arising from the appearance of abnormal hematopoietic stem cells that carry the Bcr-Abl oncoprotein, which results from a reciprocal translocation between chromosomes 9 and 22. The main elements of the disease pathogenesis are due to both increased tyrosine kinase activity of the Abl protein and the role of the Bcr part of the hybrid protein. The presence of the PH domain in Bcr determines its interaction with PI(3)P on the phagosomal membrane. We have shown that this interaction is accompanied by colocalization with the FNBPI protein in the phagosomes of J774 macrophage cells. A model of the impact of the Bcr-Abl oncoprotein on the ROS excess formation in CML due to the uncontrolled expression of phagosomal NADPH oxidase is presented.

]]> http://cytgen.com/en/2023/__3-10N4V57.htm http://cytgen.com/ru/2023/__3-10N4V57.htm Sat, 01 Jul 2023 00:00:00 +0300 SUMMARY. Puroindolines (puroindoline a and puroindoline b) determine the texture of the wheat grain, which affects milling properties of the grain and water absorption properties of the flour. The level of common wheat hardness is controlled by the allelic composition at the Ha locus on the short arm of chromosome 5D, which contains closely linked Pina-D1 and Pinb-D1 genes. Aegilops species can be a source of novel variants of puroindoline genes for enriching the wheat gene pool. Among them is the tetraploid species Ae. biuncialis Vis. (UUMM). In our study, the polymorphism of puroindoline gene length was analyzed in Ae. biuncialis using PCR amplification with gene-specific primers. We analyzed a collection of Ae. biuncialis accessions originating from the Crimean Peninsula. Polymorphism with respect to the number of amplicons produced with gene-specific primers to the puroindoline b gene was revealed: there were one (about 520 bp) or two amplification products (about 520 and 500 bp). The frequency of accessions with two amplicons in the collection was 12.5 %. Samples with two amplicons were found in the Eastern and Southern parts of the area of the species on the Crimean Peninsula. Probably, in the Ae. biuncialis accessions with two amplicons, the fragment of about 520 bp corresponds to the Pinb-U1 gene; the 500-bp fragment, to the Pinb-M1 gene, and this allele may be similar to the rare Pinb-M1-III allele of Ae. comosa.

]]> http://cytgen.com/en/2023/_11-18N4V57.htm http://cytgen.com/ru/2023/_11-18N4V57.htm Sat, 01 Jul 2023 00:00:00 +0300 SUMMARY. Ribosomal protein kinase S6 (S6K1) is one of the key components of the PI3K/mTOR/S6K1-dependent signaling cascade, which plays an important role in the regulation of translation, cell growth, survival, proliferation, and cell migration. Disturbances in the func-tioning of the PI3K/mTOR/S6K1 signaling cascade, in particular changes in the expression and activity of S6K1, accompany the development of a number of human pathologies, including oncological, neurodegenerative and cardiovascular diseases, metabolic disorders, as well as pathologies associated with aging. Recently, in addition to the already known p70 and p85, the existence of a new isoform of the S6K1 kinase – p60 – was confirmed. In addition, it was established that a change in the balance in the expression of various S6K1 isoforms towards an increase in p60-S6K1 leads to a multiple increase in the mobility of model malignant cells, which is accompanied by significant changes in the expression of molecules involved in cell adhesion and intercellular interaction and may indicate that the cells acquire tumor characteristics important for invasion and metastasis. To find out whether the expression of the p60 isoform of S6K1 correlates with invasion and metastasis at the tumor level, the expression level of S6K1 isoforms was analyzed in three groups of human papillary thyroid carcinoma tumors, namely: without signs of invasion, with signs of invasion, and with lymph node metastases. Conditionally normal tissues adjacent to the tumors were also analyzed at the same time. The analysis was carried out by real-time PCR. A trend towards an increase in the expression level of all S6K1 isoforms, as well as the p60 isoform of S6K1 in tumors with manifestations of invasion and metastasis, was established. However, changes in the ratio of the total expression level of all S6K1 isoforms to p60-S6K 1 in the direction of an increase in the share of the latter in tumors with manifestations of metastasis turned out to be statistically reliable. The obtained results indicate an important role of the p60S6K1 isoform in the maintenance or initiation of invasion and metastasis of thyroid tumors.

]]> http://cytgen.com/en/2023/_19-26N4V57.htm http://cytgen.com/ru/2023/_19-26N4V57.htm Sat, 01 Jul 2023 00:00:00 +0300 SUMMARY. Brassinosteroids are plant hormones that play an es-sential role in plant growth, development, and stress responses, for example, in the induction of tolerance to oxidative stress. Oxidative stress in plants that is the dramatic reactive oxygen species (ROS) production is known to be induced by methyl viologen herbicide. Since calcium plays a vital role in brassinosteroid signaling in plants and tightly interacts with the ROS network, the role of calcium was studied in bras-sinosteroid-induced modulation of ROS balance with the application of transgenic tobacco 35S::AtCAX1 plants. It was found that 35S::AtCAX1 tobacco plants harboring artificial calcium shortage in the cytosol were more sensitive to methyl viologen-induced oxidative damage. The level of lipid peroxidation and oxidized glutathione was higher, but the activation of superoxide dismutase and glutathione reductase was lower in the mutated plants. In 35S::AtCAX1 plants, the effect of brassinosteroids on the oxidative state parameters was modified. Brassinosteroid-induced partial restoration of chlorophyll level, reduction of lipid peroxidation and oxidized glutathione level, superoxide dismutase, and glutathione reductase activation were less evident in the transgenic plants. The results of gene co-expression analysis, protein-protein interactions, and post-trans-lational modifications in silico demonstrated that by applying calcium-dependent signaling events, brassinosteroids regulated the mechanisms of response to methyl viologen in tobacco plants, contributing to the balance of reactive oxygen species production.

]]> http://cytgen.com/en/2023/_27-35N4V57.htm http://cytgen.com/ru/2023/_27-35N4V57.htm Sat, 01 Jul 2023 00:00:00 +0300 Aeluropus littoralis is a valuable halophyte grass belonging to the same family of wheat and is used as forage. Although A. littoralis has the potential to become an important genetic resource for improving salt and drought tolerance in economically important crops, no SSR markers have been developed for it. The main goal was to rapidly develop a set of genic SSR markers for A. littoralis. Repeat analysis of non-redundant EST sequences of Aeluropus and transferability assessment of 110 SSR-rich loci from rice and wheat were used to identify EST-SSRs. Then selected EST-SSR loci and some physiological traits includings Na⁺, K⁺ and Ash content were utilized for marker characterization and assessment of genetic diversity among A. littoralis accessions collected from all around the country. The results showed that 6.7 % of EST records of A. littoralis comprising SSR motifs which was used for desiging 18 primer pairs (ALES). In addition 48 SSR loci (GDES) from 110 of the gramineae were shown to be transferable to A. littoralis based on the PCR profiles. Finally, genotypic clustering based on EST-SSR markers divided the accessions into seven groups. The accessions were also categorized into six groups according to the physiological traits. Our finding indicated that there are remarkable variations about 33 % in coding regions of Iranian Aeluropus accessions. The results of both genotypic and physiologic clustering were partially consistent and most groups corresponded to geographic regions.

]]> http://cytgen.com/en/2023/_36-38N4V57.htm http://cytgen.com/ru/2023/_36-38N4V57.htm Sat, 01 Jul 2023 00:00:00 +0300 In this study, the complete mitochondrial genome (mt genome) of Triosteum pinnatifidum was investigated for the first time. The mt genome consisted of 803,609 bp, comprised of 62 genes including 19 tRNA genes, 3 rRNA genes and 40 protein­coding genes. We detected protein­coding genes and codon usage, RNA editing sites and repeat sequences. The protein­coding genes substitution rates and DNA segments migration were generated, the comparison of six species genomic features was performed. The phylogenetic analysis of 32 species was also taken. A total of 463 RNA editing sites were found in 40 protein­coding genes. Being slightly positive, the GC­skew and AT­skew were 0.0060 and 0.0004, respectively. Most of PCGs had Ka/Ks ratio less than 1, indicating the existence of purifying or negative selection in these genes. This is the first report of the mt genome in the Caprifoliaceae family and could provide a useful foundation for evolutionary analysis, molecular biology and taxonomy in genus Triosteum and other higher plants.

]]> http://cytgen.com/en/2023/_39-41N4V57.htm http://cytgen.com/ru/2023/_39-41N4V57.htm Sat, 01 Jul 2023 00:00:00 +0300 Nonaka myopathy (MIM 605820) is caused by homozygous pathogenic variants in the GNE gene. It is a recessively inherited early adult­onset myopathy that usually preserves the quadriceps and presents with bilateral foot drop, usually caused by anterior tibialis weakness. In patients with Nonaka myopathy, serum creatine kinases are slightly elevated, muscle weakness progresses slowly, and ambulation loss develops after 15–20 years. The current study aims to raise awareness of Nonaka myopathy that occurs as a rare phenotype due to pathogenic variants in GNE gene. Detailed family histories and clinical data were recorded. Whole exome sequencing was performed and co­segregation analysis of the family were done by Sanger sequencing. Also the homology model of the mutant protein was created with the ProMod3 algorithm. We identified a bialelic pathogenic variant (c.830G>A) in GNE gene, which explain the patients’ clinical status. We present the main findings of two siblings with Nonaka myopathy together with detailed clinical and genetic profiles of the patients together with a three­dimensional mutant GNE protein model. We think that the clinical characteristics and the effect of the (c.830G>A) variant will facilitate our understanding of GNE gene in Nonaka myopathy pathogenesis.

]]> http://cytgen.com/en/2023/_42-44N4V57.htm http://cytgen.com/ru/2023/_42-44N4V57.htm Sat, 01 Jul 2023 00:00:00 +0300 Ceylon oak (Schleichera oleosa (Lour.) Oken), belonging to Sapindaceae family, is a widely used plant for its medicinal properties. Currently, few studies have been conducted on Ceylon oak’s genetic background and its relationship with other closely related plants in Thailand. Therefore, this study focused on the analysis of nucleotide sequences to understand the genetic diversity and relationship of Ceylon oak with longan and lychee. The nucleotide sequencing of six loci, namely ITS2, matK, rbcL, trnH­psbA, trnL­i and trnL­ trnF were analyzed in 5 Ceylon oak samples collected from various locations in Thailand and then were additionally aligned with nucleotide sequences of 36 longan samples and 2 lychee samples (7 species) (GenBank accession numbers KY174077­KY174314). The sequencing results were then used to construct a phylogenetic tree using the maximum likelihood criteria. Multiple sequence alignment revealed the highest InDel polymorphism of trnH­psbA fragment which can be developed as a DNA molecular marker in the identification of Ceylon oak. Interestingly, the K2P pairwise distance analysis revealed a high degree of genetic variation between Ceylon oak, longan and lychee samples and the combination of either matK and trnH­psbA or matK and ITS2 provided the most potential candidate DNA barcoding region for discrimination of Ceylon oak from longan and lychee. The phylogenetic tree showed that Ceylon oak is completely different from longan and lychee. This is the first report of phylogenetic information among Ceylon oak, longan and lychee in Thailand which might be used to assist Ceylon oak conservation and breeding program in the future.

]]> http://cytgen.com/en/2023/_45-46N4V57.htm http://cytgen.com/ru/2023/_45-46N4V57.htm Sat, 01 Jul 2023 00:00:00 +0300 Global warming threatens human life on many aspects, including heat stress on crop production. Breeding heat­tolerant varieties is a fundamental way to meet the chal­lenge. To study the genetic basis of seedling tolerance to heat stress in rice (Oryza sativa L.), we performed QTL mapping based on a large F₂ population consisting of 4450 individuals derived from a cross between a japonica rice variety Huaidao 5 (HD5) and an indica rice variety 1892S using the method of bulked segregant analysis coupled with whole­genome sequencing (BSA­seq). HD5 was more tolerant to high temperature than 1892S at the seedling stage. By analyzing a pair of opposite DNA pools made from 124 extremely­sensitive seedlings and 178 extremely­tolerant seedlings from the F₂ population using the block regression mapping (BRM) method, we mapped a QTL on chromosome 12, of which the additive effect was estimated to explain 3.75 % of the phenotypic variance. We named the QTL qSLHT12.1, which must be a novel QTL, because no QTLs for rice seedling tolerance to heat stress have been mapped on chromosome 12 before. The information obtained in this study will facilitate marker­assisted breeding of heat­resistant lines and positional cloning of the gene conferring heat tolerance in rice.

]]> http://cytgen.com/en/2023/_47-50N4V57.htm http://cytgen.com/ru/2023/_47-50N4V57.htm Sat, 01 Jul 2023 00:00:00 +0300 This review describes eight genes and 21 loci for resistance to pests localized in rye chromosomes of wheat­rye and triticale genetic stocks. Detailed information is given for the rye chromosome donor, the type of tchromatin inserted, the molecular marker, if present, and resulting wheat and/or triticale lines for deployment of the resistance in breeding. The main insect resistance factors are determined in chromosome 1R, followed by 6R, in the form of wheat­rye chromosome translocations or substitutions. Most of the genes provide resistance to Russian wheat aphid and Hessian fly. The recorded genetic stocks can efficiently serve as important bridges for wheat and triticale improvement. The data thus provided will help researchers to competently use resistances of rye chromatin through classical and marker­assisted breeding.

]]> http://cytgen.com/en/2023/51-55N4V57.htm http://cytgen.com/ru/2023/51-55N4V57.htm Sat, 01 Jul 2023 00:00:00 +0300 SUMMARY. Insects are one of the most successful groups of multicellular organisms with more than 1 million species. Among them, Belgica antarctica Jacobs (Diptera: Chironomidae) is an endemic species of Antarctica that exist in extremely cold conditions. A significant number of microorganisms colonize most groups of insects, which may lead to a symbiotic interaction, in many cases resulting in an increase of the adaptability of the host organism, for example to cold conditions. Using PCR methods and metagenomic analysis, we demonstrated that the endosymbiotic bacteria Spiroplasma and Wolbachia are probably absent in Belgica antarctica. Nevertheless, we identified 14 bacterial species which are potentially associated with Belgica antarctica and/or with the substrate in which this Diptera species lives by screening whole-genome sequences available in the open databases. Further analysis is needed to elucidate to what extent this association is persistent with Belgica antarctica, and whether identified microorganisms confer any adaptive advantage to this species.

]]> http://cytgen.com/en/2023/__3-9N3V57.htm http://cytgen.com/ru/2023/__3-9N3V57.htm Mon, 01 May 2023 00:00:00 +0300 SUMMARY. Genetic divergence and evolutionary bias of transitions and transversions ratio were analyzed based on examples of 61 sequences of the mtDNA control region of 27 species of mice (Murinae) of the Palearctic region. Representatives of Cricetinae and Arvicolinae were taken as control samples. The constructed phenogram corresponds to the accepted phylogenetic scheme and
is clustered according to population, subspecies, allospecies, species, genus and family levels of divergence. In the phyletic order, there is a clearly defined evolutionary shift in the ratio of transitions and transversions (evolutionary transition:transversion rate bias). Its distinction is an extremely abrupt transition from dynamic to stable phase. The dynamic phase is a rapid decrease in the ts/tv ratio and refers to the population and semi-species levels of divergence. The stable phase refers to species levels of divergence and higher levels of divergence and is connected to the attainment of a state of genetic saturation, moreover in a situation of a predominance of transversions. The extreme expressivnes of the evo-lutionary bias in this case can be explained by the removal of the pressure on the selection caused by restrictions in amino acid substitutions. This means that the causes of transition/transversion bias are purely DNA level biochemical mechanisms. Simultaneously, the stability of the ts/tv ratio at species and higher levels of organization amidst the further accumulation of total number of nucleotide substitutions indicates a fundamentally different nature of genetic processes at the intraspecies and interspecies levels of divergence.

]]> http://cytgen.com/en/2023/10-18N3V57.htm http://cytgen.com/ru/2023/10-18N3V57.htm Mon, 01 May 2023 00:00:00 +0300 SUMMARY. The coronavirus disease of the year 2019 (COVID-19) poses a serious threat to human health. Nowadays the relevant topic appears to be the prognosis of human chronic illnesses progress and development after the coronavirus infection beating. Osteoarthritis is a chronic degenerative joint disease. It was shown that this malady may be induced by various factors, such as oxidative stress impact, hypercholesterolemia, increased aggrecan-degradative activity of specific proteases during certain endocytosis pathway disruption, and so on. Hence the aim of this work was to analyze the expression of OLR1, ACAN and LRP1 genes in synovial membrane cells of osteoarthritis patients having beaten SARS-CoV-2 infection. The research included 60 humans (males) aged 50–55 years. The volunteers were separated into following groups: the first group (n = 20) – conditionally healthy individuals, the second group (n = 20) – patients with II–III degree knee joint osteoarthritis, the third group consisted of 20 patients with II–III degree knee joint osteoarthritis having successfully beaten COVID-19. The human blood plasma cholesterol concentration was measured by the enzymatic method using a diagnostic set of reagents. The intensity of superoxide anion ra-dical generation in synovial liquid was estimated by the HTT-formazan accumulation. The level of OLR1, ACAN and LRP1 gene expression in knee joint synovial membrane cells was assessed by RT-qPCR. The OLR1 gene expression increase was demonstrated mostly in osteoarthritis patient synovial membrane cells having beaten COVID-19, if compared to the group with knee joint osteoarthritis, against the background of more intensive increase of both blood plasma cholesterol concentration and free radical process activation (the superoxide anion radical amount increase) in synovial liquid of osteoarthritis patients after SARS-CoV2-infection. This may be due to the increase in system-wide inflammation while а body responds to the virus. At the same time the more prominent decrease of the ACAN gene expression was shown in synovial membrane cells of patients with osteoarthritis having beaten COVID-19, in comparison with the group of patients with knee joint osteoarthritis. Such findings indicate more pronounced activation of destructive processes within cells after carried out infection, and can be mediated by the decrease of the LRP1 gene expression, which, in its own turn, is capable of propagating the illness further progress. The understanding of a more severe osteoarthritis progress and possible complication development in patients with the post-COVID-19 syndrome, taking both LOX1/ox-LDL and LRP1-endocytosis pathway into consideration requires additional research attempts.

]]> http://cytgen.com/en/2023/19-27N3V57.htm http://cytgen.com/ru/2023/19-27N3V57.htm Mon, 01 May 2023 00:00:00 +0300 SUMMARY. Since the nanoscale combined with luminescent pro-perties and promising applications in various fields of optoelectronics and biomedicine leads to a growing interest in studying the features of cadmium telluride quantum dots, we have developed a method for the «green» synthesis of CdTe quantum dots using the mycelium culture of Pleurotus ostreatus as a biological matrix. The study of their physical and chemical characteristics revealed that the synthesized CdTe quantum dots have a predominantly spherical morphology and a size of 3–8 nm, a crystal structure, and a maximum luminescence in the range of 340–370 nm. When studying their effects on various types of mammalian cells, it was found that CdTe quantum dots have a dose-dependent effects on mouse endothelial cells, erythrocytes, human and rat T and B lymphocytes, colon cancer cells (Colo 205) and human breast cancer cells (MCF-7). In particular, we observed inhibition of endothelial cell proliferation and an increase in dead cells, indicating the cytotoxic effect of nanocrystalline CdTe and its antiproliferative effect on endothelial cells. CdTe quantum dots at a con-centration of 5 µM exhibited hemolytic activity when exposed to erythrocytes, affected adhesive contacts and survival of cancer cells. At the same time, human breast cancer cells (MCF-7) were more sensitive to their action. The data obtained are extremely important for understanding the mechanisms of toxicity of CdTe quantum dots for their further use in biological and biomedical research.

]]> http://cytgen.com/en/2023/28-39N3V57.htm http://cytgen.com/ru/2023/28-39N3V57.htm Mon, 01 May 2023 00:00:00 +0300 SUMMARY. Biobutanol is one of the types of biofuel obtained by microbial synthesis (acetone-butanol-ethanol fermentation). Using adaptive cell changes, it is possible to increase the level of accumulation of the target product due to the effect of stress factors. For this aim, the effect of the following stress factors: allopurinol, furfural, zinc, oxygen and glycerol as a non-specific substrate on the functioning of Clostridium sp cells was investigated. Stress factors were added to the medium in adapting doses. It was found that the concentration of butanol was the greatest (1.2 g/l) when 0.5 g/l of furfural was added to the medium compared to the control (0.6 g/l). It was shown that the use of adaptive dose (0.001 g/l) of ZnSO₄ resulted in the greatest concentration of butanol (1.0 g/l), and the highest concentration of butanol (1.3 g/l) was obtained when using glucose and glycerol in the ratio of 1 : 3 as a carbon source. It was demonstrated that the concentration of butanol in the culture medium increa-sed (0.8 g/l) at a concentration of allopurinol in the medium of 0.025 g/l. The possibility of using adaptive stress to increase the accumulation of the target product due to changes in metabolic pathways in the cells of microorganisms, indirectly due to the action of stress factors, is substantiated.

]]> http://cytgen.com/en/2023/40-47N3V57.htm http://cytgen.com/ru/2023/40-47N3V57.htm Mon, 01 May 2023 00:00:00 +0300 Potato (Solanum tuberosum L.), fourth largest food crop in the world and is the major vegetable produced in India. New varieties with improved agronomic traits are released every year and characterizing these using both morphological and molecular techniques is not only vital for registration and univocal identification but also to quantify the diversity present within. To estimate the diversity present and to establish distantness among the 55 Indian potato cultivars, 50 morphological descriptors of distinctiveness, uniformity and stability (DUS) including 22 quantitative parameters were used along with 24 highly informative microsatellite or simple sequence repeat (SSR) markers. The cultivars were grouped into 5 and 3 different clusters based on morphological parameters and molecular analysis respectively. Varieties, Kufri Kashigaro and Kufri Anand recorded the highest diversity value of 1.95 based on morphology, whereas two indigenous cultivars Phulwa and Jeevan Jyoti which are used as parents in breeding programs showed the highest diversity using molecular markers. As expected, SSR markers showed detailed and in-depth diversity with 294 polymorphic alleles and PIC (polymorphism informative content) value ranging from 0.57 to 0.91.  The heterozygosity expected varied from 0.57 to 0.92 with an average Rp (resolving power) value of 4.57. Considerable diversity was observed in the Indian potato cultivars and these distinct cultivars may be used as parents for breeding of potato for wider environments and changing climatic scenario. Moreover, the genotypic data would serve as reference for distinguishing different cultivars of potato.

]]> http://cytgen.com/en/2023/48-50N3V57.htm http://cytgen.com/ru/2023/48-50N3V57.htm Mon, 01 May 2023 00:00:00 +0300 The purpose of this study is to determine the genetic association and compare the distribution of ABCA4 gene variants in patients with various inherited retinal dystrophies, including autosomal recessive cone-rod dystrophy (AR-CRD) autosomal recessive retinitis pigmentosa (AR-RP) in the South Indian cohorts. This study included nineteen probands, ophthalmic examination of all the probands were carefully evaluated by the Paediatric Ophthalmologist. Eleven of the nineteen probands had the clinical phenotype of AR-CRD, eight showed AR-RP-like clinical phenotype. Genomic DNA was extracted from their peripheral blood, the exon and intronic boundaries of the ABCA4 gene were screened using the Sanger sequencing to identify the genetic association of these two retinal dystrophies. Sanger sequencing results revealed that only 18 % (2/11) of AR-CRD probands had disease-causing ABCA4 mutations, while the remaining 9 AR-CRD, 8 AR-RP were negative for the pathogenic variant associated with ABCA4. Furthermore, this study identified 18 common SNPs of the ABCA4 (2 missense, 3 synonymous, 13 intronic variants) and found them associated with AR-CRD and AR-RP probands. This is the first study to show that two missense variants in the ABCA4 (c.302T>C, c.1319A>G) are associated with AR-CRD probands and two rare NNCS variants (c.3191-10G>T, c.3814-5G>A) associated with AR-RP patients from the South Indian population.

]]> http://cytgen.com/en/2023/51-53N3V57.htm http://cytgen.com/ru/2023/51-53N3V57.htm Mon, 01 May 2023 00:00:00 +0300 The complete mitochondrial genome (mitogenome) of the Rhus gall aphid Schlechtendalia peitan in China was obtained using the shotgun genome-skimming method on an Illumina platform. The complete mitogenome of S. peitan is 15,494 bp in length with a high A+T content of 84.1 %. This mitogenome consists of 13 protein-coding genes, 22 tRNA genes, two rRNA genes, and a control region. All the protein-coding genes initiate with a typical ATN codon and terminate with TAA codon except for COX1, ND4 and ND5 with a single T. The 22 tRNAs range from 59 to 72 bp in length, and each one is predicted as a clover-leaf secondary structure except for tRNA-Ser (AGN), which loses a dihydrouridine (DHU) arm. The ML phylogenetic tree of Fordini aphids constructed using 13 protein-coding genes and two rRNAs showed that S. peitan was sister to the group clustered by the two species Schlechtendalia chinensis and Nurudea ibofushi.

]]> http://cytgen.com/en/2023/54N3V57.htm http://cytgen.com/ru/2023/54N3V57.htm Mon, 01 May 2023 00:00:00 +0300 With this study, we aim to summarize and assess the activity and performance of the Cytogenetic sector of the Laboratory of Medical Genetics – Varna, regarding the conventional cytogenetic analysis of bone marrow samples from patients with (onco)hematological diagnoses. Another purpose is to evaluate the tendencies noticed over a period of eleven years to draw conclusions and share our experience. We have performed the analysis with the G-banding technique on 2,653 samples from patients of age 0–93 years by the current European recommendations and the International System for Human Cytogenomic Nomenclature. The greater part of these samples (90.9 %) was with an indication of а hematological malignancy, most commonly Acute myeloid leukemia, Myelodysplastic syndrome, Acute lymphoid leukemia, Chronic myeloid leukemia, and Multiple myeloma. Analysis was successful in 2,215 (83.5 %) – from those normal karyotypes were found in 1492 (67.4 %) and pathology in 723 (32.6 %). Regarding the latter, the most common were complex karyotypes (30.6 %), Philadelphian chromosome (21.3 %), trisomy 8 (5.9 %), and deletion in the long arm of chromosome 5 (4.3 %). Cytogenetic analysis is a method with great impact on the evaluation of many hematological malignancies and for this reason, it remains an essential part of routine assessment of these diseases. The disadvantages of it mainly in the field of oncohematological diseases, recognized by the scientific society and confirmed in our own experience, suggest a need for an additional genetic method to overcome these limitations.

]]> http://cytgen.com/en/2023/55-56N3V57.htm http://cytgen.com/ru/2023/55-56N3V57.htm Mon, 01 May 2023 00:00:00 +0300 Osteosarcoma is a malignant bone tumor that is common in children and adolescents. The tumor microenvironment is highly effective in the development and progression of osteosarcoma. Transforming growth factor-β (TGF-β) is one of the most abundant cytokines in the tumor microenvironment, and can regulate tumor initiation, progression, and metastasis promoting extracellular matrix (ECM) remodeling and epithelial-mesenchymal transition (EMT). ADAMTS (ADAM Metallopeptidase With Thrombospondin Motifs) proteases have critical functions in normal and tumor microenvironments by processing individual proteins in the ECM. ADAMTSs contribute to tissue remodeling, inflammation, cell migration and, angiogenesis. Among the family members, ADAMTS-2 is a well-known example for ECM remodeling which cleaves the N-terminal propeptide of procollagen and promotes correct collagen fibrillogenesis. Cytokines can regulate normal and tumor microenvironments by affecting ECM proteins. In this study, the effect of TGF-β1, on the transcriptional regulation of the ADAMTS-2, which is an essential enzyme for ECM remodeling was investigated in Saos-2 cells. TGF-β1 upregulated ADAMTS-2 expression both at mRNA and protein levels. Transient transfection assays revealed that TGF-β1 was also induced ADAMTS-2 promoter activity. According to the pathway inhibition studies, both canonical and non-canonical signaling pathways and post-translational mechanisms were responsible for the induction. These studies will contribute to future research on ADAMTS-2 mediated ECM remodeling in osteosarcoma.

]]> http://cytgen.com/en/2023/57-59N3V57.htm http://cytgen.com/ru/2023/57-59N3V57.htm Mon, 01 May 2023 00:00:00 +0300 SUMMARY. Currently, melatonin (N-acetyl-5-methoxytryptamine) is considered as a multifunctional bioregulator not only in mammals, but also in plants. The aim of the work was to study the effect of melatonin on the resistance of wheat (Triticum aestivum L., var. Doskonala) and rye (Secale cereale L., var. Pam’yat Khudoyerka) seedlings to high temperatures and the functioning of key cellular defense systems – antioxidant and osmoprotective. Rye seedlings differed from wheat seedlings in higher heat resistance, which was manifested in less inhibition of growth after 6-hour heating at the temperature of 44 °С and in less manifestation of the effects of oxidative stress. Treatment of wheat grains with melatonin at concentrations ranging from 20–100 µM significantly reduced shoot and root growth inhibition caused by high temperature. Rye seedlings were less affected by melatonin, reducing only the inhibition of shoot growth. Treatment with melatonin prevented the development of oxidative stress caused by the effect of high temperature, which was manifested in decrease of the rate of superoxide radical generation, the content of hydrogen peroxide and malondialdehyde in the shoots of wheat and rye seedlings. Treatment of grains of both types of cereals with melatonin caused an increase in catalase activity under the heat stress. Treatment with melatonin also contributed to the stabilization of peroxidase activity in wheat under the stress conditions and caused its increase in rye. In addition, treatment of grains with melatonin caused an increase in the content of soluble carbohydrates under the stress conditions, but did not significantly affect the content of proline in the shoots of seedlings of both species. In general, a less noticeable effect of melatonin treatment on the functioning of protective systems of rye was noted. The key effects of melatonin under the stress conditions are the reduction of oxidative damage to cells, increased activity of antioxidant enzymes, and increased accumulation of soluble carbohydrates.

]]> http://cytgen.com/en/2023/__3-14N2V57.htm http://cytgen.com/ru/2023/__3-14N2V57.htm Wed, 01 Mar 2023 00:00:00 +0200 SUMMARY. Nowadays the possible influence of the coronavirus infection onto cartilage degeneration and synovial mem-brane inflammation during joint chronic pathology – osteoarthritis – remains largely unelucidated. The aim of the presented work is to analyze the ТGFB1, FOXO1 and COMP gene expression and free radical generation intensity in blood of patients suffering from osteoarthri-tis after beating the SARS-CoV2 infection. The work was carried out using molecular genetics and bio-chemistry methods. The decrease of the ТGFB1 and FOXO1 expression level was shown to be more evident in the osteoarthritis patients having beaten COVID-19 if compared to the group with knee joint osteoarthritis, during simultaneous and more prominent diminishing of both superoxide dismutase and catalase activity (possibly indicating cell redox state disruption and TGF-β1-FoxO1 signaling attenuation) in patients with osteo-arthritis having beaten SARS-CoV2 disease. At the sa-me time the more prominent decrease of COMP gene expression level was demonstrated in patients with osteoarthritis having beaten COVID-19 comparing to the patient group with knee joint osteoarthritis in the event of more intense increase of the COMP concentration in patients with osteoarthritis after the SARS-CoV2 in-fection. This data indicates more significant activation of cell destructive processes after beating the infection as well as further pathology progression.

]]> http://cytgen.com/en/2023/15-21N2V57.htm http://cytgen.com/ru/2023/15-21N2V57.htm Wed, 01 Mar 2023 00:00:00 +0200 Шпильковий зонд порівняно з лінійним в умовах проведення ПЛР-РЧ характеризується вищою ефективністю гасіння флуоресценції, що веде до нижчого фонового рівня флуоресценції та, отже, більшого співвідношення сигнал/шум при проведенні ПЛР у реальному часі. Проведено експериментальне порівняння ефективності гасіння флуоресценції двох олігонуклеотидних зондів в різних конформаціях – шпилькового у форматі молекулярного маяка та лінійного у форматі TaqMan. Існує різниця у взаємодії гасника з флуорофором для зондів різної конформації. Для лінійного зонда гасіння відбувається через механізм індуктивно-резонансного переносу енергії (ФРПЕ, або FRET), а для шпилькового зонда – за допомогою контактного гасіння через більш близьке розташування флуорофора та гасника, але можливим є й резонансний перенос енергії за механізмом Ферстера. Показано, що спектр поглинання для лінійного зонда практично збігається зі спектром поглинання олігонуклеотида, який представляє зонд без гасника, що вказує на динамічний (Ферстеровський) механізм переносу енергії. Навпаки, спектри поглинання для шпилькового зонда та олігонуклеотида, який представляє зонд без гасника, значно відрізняються, що свідчить про контактний механізм переносу енергії між флуорофором та гасником флуоресценції. Спектри флуоресценції зондів та їхніх комплексів з олігонуклеотидом, що є комплементарним лінійному зонду (та петлі шпилькового зонда), та ампліконом (довжиною 200 п.н., який містить ДНК-мішень для зондів) дозволили порівняти ці два зонди через порівняння радіусів міграції енергії, ефективності гасіння флуоресценції донора. Розрахований з експериментальних даних радіус міграції енергії R для шпилькового зонда становив 32,4 Å, а для лінійного зонда – 47,3 Å.

]]> http://cytgen.com/en/2023/22-32N2V57.htm http://cytgen.com/ru/2023/22-32N2V57.htm Wed, 01 Mar 2023 00:00:00 +0200 SUMMARY. Orchids are one the most widespread groups of flowering plants with a wide geographical range and species diversity. A number of tropical and subtropical species are used as decorative, medicinal and edible. The increased demand for plant material, while a large number of species are under threat of extinction in nature, makes growing of orchids in culture relevant. Traditionally, new interesting forms have been obtained through hybridization and selection, which require considerable time. Not all requirements for elite varieties can be solved by traditional breeding methods. The application of the achievements of modern molecular biology significantly expands the possibilities of breeders. The development of genetic engineering methods allows introducing both new heterologous genes to orchids and editing their own genes, which can significantly speed up and increase the success of the traditional selection process. Members of the Orchidaceae family can be used not only for introduction of valuable heterologous genes, but also as saurce of unique genes for the improvement of cultivated species of other families. The review examines the current state and prospects of genetic engineering of orchids, their use as recipients and donors of genes for genetic transformation.

]]> http://cytgen.com/en/2023/33-53N2V57.htm http://cytgen.com/ru/2023/33-53N2V57.htm Wed, 01 Mar 2023 00:00:00 +0200 Strobilanthes virendrakumarana Venu & P. Daniel karyotype was found to be a mixoploid one with 2n = 20+0­1B and reveals the karyotype formula as 2nsm(­)+18nm+0­1B. The plant is endemic to Southern Western Ghats of Kerala. The basic set of chromosomes was found to be x = 10. The plant shows mixoploidy with four different chromosome counts from hypoploid (10+0­1B), diploid (20 = 0­1B) and hyperploid chromosome complements (30+0­2B, 40+0­2B). The karyotype formula of hyperploid cell was 2nsm(­)+8nm+0­1B. In hypoploid cells with 30 and 40 chromosomes the resulted karyotype formula were 30nm+0­2B and 6M+34nm+0­2B respectively. The total chromosome length of diploid complement was 45.34 µm with an average chromosome length of 2.2 µm. The detailed karyomorphometrical analysis of the plant reveals that is a primitive one with symmetric karyotype. The presence of B­chromosomes in the genus Strobilanthes is reporting for the first time. Ploidy level changes may be due to the prolonged flowering periodicity of the plant which enables them to survive by propagating through vegetative mode.

]]> http://cytgen.com/en/2023/54-55N2V57.htm http://cytgen.com/ru/2023/54-55N2V57.htm Wed, 01 Mar 2023 00:00:00 +0200 The karyotype characteristics of 24 taxa of Iranian Astragalus genus from different geographic origins were studied and their relation with a systematic classification of studied taxa was discussed. Chromosome numbers based on x = 8 were found in all of the taxa. Diploid (2n = 2x = 16) and tetraploid (2n = 4x = 36) chromosome number, were recorded in 12 taxa representing nine species, and in 12 taxa representing six species, respectively. The degree of karyotype asymmetric is set by values of A₁, AR, CI, TF%, SA%, and LA%. So, the most value of A₁, AR , LA% and the most value of CI, TF%, SA% were shown in A. stevianus (accession 20105) and A. cyclophyllon (accession 18375), respectively. The size of chromosomes in the studied taxa, ranging between 1.930µ in A. cyclophyllon (accession 18375) and 4.095µ in A. bombycinus (accession 20114). By using Ward cluster analysis, the relation of studied taxa based on karyotype characteristic was shown. In this plot, all of the taxa were located in four groups on karyotype characteristics similarity. Then this grouping was compared with their traditional systematic classification.

]]> http://cytgen.com/en/2023/56-57N2V57.htm http://cytgen.com/ru/2023/56-57N2V57.htm Wed, 01 Mar 2023 00:00:00 +0200 Stellaria dichotoma L. var. lanceolata Bunge is a typical rare medicinal plant commonly used in therapeutic formulations. To reveal the structural arrangents and variation of complete chloroplast genomes between S. dichotoma var. lanceolata and its related species is of great significance for the study of its evolutionary status. In this study, evolutionary relationships between S. dichotoma var. lanceolata and its related species of Caryophyllaceae were documented based on the complete chloroplast genome sequence of S. dichotoma var. lanceolata. The result showed that the whole circular genome of S. dichotoma var. lanceolata was 150,461 bp in length, annotated 129 genes, possessing RSCU of 21 types of amino acids and 64 codons encoding. By comparing and analyzing the SSR and variation region of the chloroplast gene of S. dichotoma var. lanceolata and its related genus pseudostellaria, we found that the divergent regions of trnk­rps16, atpH­atpI, rpoC1­rpoB, rbcL­accD, trnS­trnG, psaA­ycf3, trnV­trnM, ycf4­cemA, petL­petG, trnL­ccsA, ndhF, ndhA, and ycf1 fragments were highly obvious, which could be used as DNA barcodes for the taxonomic evidence of S. dichotoma var. Lanceolata and Pseudostellaria in Caryophyllaceae. A maximum likelihood (ML) phylogenetic tree elucidated that S. dichotoma var. lanceolata was closely related to pseudostellaria, and cluster into a branch with Cerastium. Our results lay a robust foundation for future phylogenetic and evolutionary status of S. dichotoma var. lanceolata and among relatives within Caryophyllaceae.

]]> http://cytgen.com/en/2023/58-61N2V57.htm http://cytgen.com/ru/2023/58-61N2V57.htm Wed, 01 Mar 2023 00:00:00 +0200 Phospholipase Cγ2 has a promotive effect on hepatic carcinoma development. Meanwhile, lncRNAs play a critical role in the pathology. Therefore, to determine whether phospholipase Cγ2 could enhance HCC cell proliferation by regulating lncRNA transcription, hepatic carcinoma cells RH35 were infected with Ad­phospholipase Cγ2 constructed previously, followed by lncRNA sequencing by high­throughput technology. Differently expressed lncRNAs (DElncRNAs) and their target genes were identified according to strict criteria. GO and KEGG, Reactome pathway analyses were performed to analyze biological processes and the related pathways of DElncRNAs. lncRNA/mRNA coexpression pairs were screened according to expression profiling combined with bioinformatics analysis. The results showed that 231 DElncRNAs were identified in Ad­phospholipase Cγ2­overexpressing cells compared to control, containing 60 up­ and 171 down­regulated ones. Target genes prediction analysis showed that 61 cis­ and 30 trans­acting DElncRNAs were matched to 55 and 26 targets, respectively. Coexpression analysis found 33 lncRNA/mRNA coexpression pairs including 24 pairs in cis. GO analysis showed that these cis­mode lncRNA/mRNA pairs were involved in cytoskeleton organization, cell adhesion, and multiple signaling pathways related to apoptosis, proliferation, and metastasis. Collectively, phospholipase Cγ2 caused significant alterations in the expression of many lncRNAs in liver cancer cells, providing valuable insight into the precise mechanism of phospholipase Cγ2­promoting liver cancer cell growth.

]]> http://cytgen.com/en/2023/62-64N2V57.htm http://cytgen.com/ru/2023/62-64N2V57.htm Wed, 01 Mar 2023 00:00:00 +0200 Lung cancer is one of the most common types of cancer that causes death. In this study the effects of cisplatin, taurine and combination of these two compounds on A549 cell line were examined. A549 Cells were treated with different concentrations of taurine, cisplatin and the combination of two compounds. MTT assay, flow cytometry analysis of apoptosis and flow cytometry analysis of cell cycle were carried out. The expression of genes was examined by real­time PCR. Cisplatin and taurine reduced the viability of the A549 cell line but this effect was greater in taurine and cisplatin combination. Cells that were in G0/G1 stage increased in all treated group and this inhibition was notable in the combination group. The expression of some genes such as P53, Bax, caspase 3, caspase 9 and P14 increased. Our studies also showed that cisplatin and taurine combination was moderately synergic with Cl values ranging from X to Y for Fa 0.5. Combination of cisplatin and taurine may be effective in the cancer therapy and it can be a suitable choice for reduction of drug resistance problems and other side effects of cisplatin high dosage, but more researches are needed to be performed in this field.

]]> http://cytgen.com/en/2023/65-66N2V57.htm http://cytgen.com/ru/2023/65-66N2V57.htm Wed, 01 Mar 2023 00:00:00 +0200 SUMMARY. The article describes new genetic variants of Gli-/Glu-loci encoding the biosynthesis of gluten proteins, according to their influence on the basic indicators of baking quality of wheat flour used in breeding programs of high baking quality varieties. The methods of obtaining wheat breeding material by remote crosses were used, new alleles of Gli-/Glu-loci were identified by A-PAGE, mini-SDS-PAGE, PCR test, SDS-30K sedimentation, dough elasticity was determined by the Chopin alveograph test. The effect of new introgressions of Gli-D1ts, Gli-D1cyl and Gli-B1null alleles, Gpc-B1 gene from wild emmer T. dicoccoides, extra-expressions of Glu-A1x2* and Glu-D1x5 and deletions of Glu-D1x5-null on basic breeding traits of wheat quality, the value of W ‘strength’ of flour, and the elasticity index of the dough Ie% was studied. The positive effect of rye locus substitution Sec-1 on wheat cluster Gli-B1/Glu-B3 in the short arm of the modified chromosome-engineered central rye-wheat translocation 1RSm.1BL and 1RSm.1BLal on the basic breeding characteristics of bakery quality was shown. Extra-expression and deletion of the HMW-GS Glu-D1x5 subunit indicate the role of the latter as a critical determinant of the baking quality of wheat flour. Modified rye-wheat translocation 1RSm.1BL (1RSm.1BLal) with the eliminated locus Sec-1 should be used in wheat breeding for quality and resistance to leaf diseases. New genetic factors of positive effect on the characteristics of flour quality are recommended for use in breeding programs to create wheat varieties with high baking quality.

]]> http://cytgen.com/en/2023/__3-16N1V57.htm http://cytgen.com/ru/2023/__3-16N1V57.htm Sun, 01 Jan 2023 00:00:00 +0200 SUMMARY. T-cell receptor excision rings (TRECs) and kappa-deleting excision rings (KRECs) are circular DNA molecules that are formed during the maturation of human T- and B-cells and may serve as markers of immune response cell development. To determine the TREC and KREC quantities in different biological material and their informative value for the screening of patients with inborn errors of immunity. The analysis of TREC and KREC molecules was performed by real-time polymerase chain reaction followed by the analysis of melting curves in neonatal dry spots and DNA bank of patients with inborn errors of immunity. The proposed method showed that determining the TREC copies shows 100 % sensitivity for the detection of Nijmegen syndrome and 84 % for ataxia-telangiectasia. The determination of KREC showed considerably lower sensitivity, 75 and 37 %, respectively. The calculations performed with 1,500 tests indicate the specificity of 97.6 % for the first blood spot and 97.3 % after re-collection. It would be expedient to use the method on a cohort of over 10,000 samples to clarify the specificity and to set the TREC and KREC threshold levels in our population. Determining the number of TRECs and KRECs copies is a quick and affordable method for implementation in screening programs on condition of the replication of the obtained results using a larger cohort. The determination of the TREC quantity by the proposed method showed the sensitivity of 93.6 and the 97.6 % specificity for the detection of inborn errors of immunity accompanied by T-lymphopenia.

]]> http://cytgen.com/en/2023/_17-25N1V57.htm http://cytgen.com/ru/2023/_17-25N1V57.htm Sun, 01 Jan 2023 00:00:00 +0200 SUMMARY. The effectiveness of immediate implantation of the fibrin matrix associated with mesenchymal stromal cells of Wharton’s jelly was investigated in a spinal cord injury (SCI) model. The study was conducted using white adult outbred male rats (~260 g, 4–5 months old). The trauma model is a left-sided section of a half of the spinal cord at the level of T₁₃–L₁ segments. The rehabilitation involved the immediate transplantation of the human fibrin matrix associated with mesenchymal stromal cells of human Wharton’s jelly (mesenchymal stromal cells, MSC, n = 9) into the injury area. The reference groups had the isolated SCI (trauma, Tr, n = 7) and the implantation of the human fibrin matrix (fibrin, Fb, n = 6) in the area of injury. The motor activity and spasticity of the paretic limb were evaluated on the BBB scale and the Ashworth scale in our own modifications, respectively. The morphological picture in the area of injury was studied in the remote period using the impregnation of longitudinal sections of the spinal cord with silver nitrate. Mesenchymal stromal cells of human Wharton’s jelly in the presence of fibrin matrix showed the signs of active vivality, growth, and migratory potential in the culture. The intense increase in the motor activity of the paretic limb in the Fb group was limited to the first 2 weeks of follow-up, in the MSC group – 3 weeks of follow-up. Throughout the experiment, the level of function in the MSC group was inferior to the level of the Fb group, but only in the first week of observation it was significant. Five months later, the index of motor function was 10.4 ± 1.0 points of BBB (MSC) and 11.6 ± 2.0 points of BBB (Fb), and in the group Tr – 5.9 ± 1.6 points of BBB. However, a significant difference in the values of the indicator was found for the MSC and Tr groups – 6 weeks, 3 and 5 months after the implantation. A significant advantage of the level of spasticity in the Tr group over the MSC group was found 6 and 7 weeks and 5 months after the injury, and an advantage over the Fb group – 7 weeks after injury. No significant differences in the level of spasticity between the MSC and Fb groups were found throughout the experiment. Immediate fibrin matrix implantation to the spinal cord injury area has a positive effect on the restoration of motor function of the paretic limb of the animal, especially in the presence of mesenchymal stromal cells of human Wharton’s jelly.

]]> http://cytgen.com/en/2023/_26-43N1V57.htm http://cytgen.com/ru/2023/_26-43N1V57.htm Sun, 01 Jan 2023 00:00:00 +0200 SUMMARY. Carotenoids as precursors for the vitamin A synthesis are important micronutrients for food and feed. β-Carotene is a carotenoid which converts into vitamin A in animal organisms most effectively. The increase in its content in mature maize grain is possible via marker-associated selection by the identification of genotypes with favorable allelic state of the key genes of carotenoid biosynthesis and utilization. The gene of β-carotene hydroxylase 1 is important for the accumulation of β-carotene in mature maize grain. One of its alleles blocks the transition of β-carotene into β-cryptoxanthin and thus ensures the accumulation of β-carotene in mature grain. This allele is detected by the marker crtRB1-3′TE using the polymerase chain reaction as an amplicon of 543 bp unlike two others, 296 bp and 296 + 875 bp, which are not associated with an increase in β-carotene content in the grain of complete maturity. It was established that 26.7 % among 15 well-known inbreds of foreign breeding and 21.6 % of inbreds within 153 perspective inbreds of the Dnipro breeding program carried the allele of β-carotene hydroxylase 1 gene (543 bp) favorable for the accumulation of β-carotene. Allele 543 bp of β-carotene hydroxylase 1 gene by the marker crtRB1-3′TE was found among most of the analyzed maize subspecies, germplasms, and maturity groups. The tendency to its increased frequency for inbreds with flint grain type, Lancaster and Lacon germ-plasms as well as early and middle early inbreds was noticed. Modern perspective inbreds – the carriers of allele 543 bp on gene of β-carotene hydroxylase 1 by marker crtRB1-3′TE, are recommended for the application in special programs of marker-assisted se-lection to increase the content of β-carotene in maize subspecies, groups of maturity, and different types of germplasm.

]]> http://cytgen.com/en/2023/_44-54N1V57.htm http://cytgen.com/ru/2023/_44-54N1V57.htm Sun, 01 Jan 2023 00:00:00 +0200 SUMMARY. The genetic diversity of the marsh frog Pelophylax ridibundus populations and the hemiclonal structure of the hybrid form Pelophylax esculentus-ridibundus within the drainages of Prypiat, Dniester and Southern Buh rivers were analyzed. The absence of a single evolutionary scenario for this hybrid form within the borders of the region has been revealed. The conservation of the basic level of parental species evolutionary potential and the interpopulation differention of the hybrid form within the drainages of Dniester and Southern Buh rivers was demonstrated. At the same time, in the populations of P. esculentus-ridibundus from the Prypiat basin, a loss of evolutionary potential was revealed (in the southern part by 31 %, in the northern part by 69 %). It was revealed that the reason for this was the tendency to the extinction of rare haplotypes and the expansion of the mass ones. It was also demonstrated that there was a significant increase (9–10 times) in the interpopulation differentiation of the hybrid form from the Prypiat river drainage compared with sympatric populations of the parental species P. ridibundus. It was shown that the evolutionary potential loss of the hybrid form P. esculentus-ridibundus accelerated in the absence of parental species, which confirms the hypothesis about regular hybridization as an effective mechanism to compensate for the loss of evolutionary potential.

]]> http://cytgen.com/en/2023/_55-67N1V57.htm http://cytgen.com/ru/2023/_55-67N1V57.htm Sun, 01 Jan 2023 00:00:00 +0200 SUMMARY. Extreme temperatures are among the most dangerous environmental factors, the impact of which on plants has been increasing significantly in last few decades. The review analyses the latest information about the perception mechanisms of hyperthermia signal by plant cells. The emphasis is placed on the role of membrane fluidization, changes in calcium channels, and increased generation of reactive oxygen species in the perception of heat stress signal. The significance of gasotransmitters (NO and H₂S) and their interaction with other me-diators in the transduction of hyperthermia signal into the genetic apparatus is discussed separately. The role of key transcription factors (HSF, MBF, NAC, and WRKY) in formation of plant adaptive responses to high temperatures is analysed. The present-day concepts on the mechanisms of activation and functioning of main stress-protective systems that provide plant resistance to hyperthermia (synthesis of heat shock proteins, antioxidant and osmoprotective systems) are summarized. Examples of genetic improvement of plants heat resistance by transformation of genes involved in the control of these systems are presented.

]]> http://cytgen.com/en/2023/_68-92N1V57.htm http://cytgen.com/ru/2023/_68-92N1V57.htm Sun, 01 Jan 2023 00:00:00 +0200 Breast Cancer BC is the extreme most frequent cancer among women. Death from BC is around 15 % of all cancer deaths among women. BC risk factors are classified into non-modifiable risk factors and modifiable risk factors. Excessive studies on folate pathway have been done as a possible mechanism for rising cancer. The present work aimed to study genetic polymorphisms in cystathionine beta-synthase CBS (CBS 844ins68) and the cytosolic serine hydroxy-methyltransferase-1 SHMT1 (C1420T) genes and their expression in patients with breast cancer . Methodology the study included 100 diagnosed breast cancer patient and 100 healthy as a control. Conclusion of this study, results obtained indicated that; there is a significant association between CBS 844ins68 polymorphism and increased risk of breast carcinoma as There was a significant difference in the frequency of the mutant insertion allele between breast cancer patients and control subjects, Also for SHMT1 C1420T SNP, there was a significant difference in the frequency of the mutant T allele between breast cancer patients and control subjects that reflected that SHMT1 C1420T may has a protective effect against breast cancer. On The level of gene expression of CBS and SHMT1 genes, there was insignificant difference between malignant and adjacent normal tissue samples as a control.

]]> http://cytgen.com/en/2023/_93-96N1V57.htm http://cytgen.com/ru/2023/_93-96N1V57.htm Sun, 01 Jan 2023 00:00:00 +0200 In the present investigation, optimization of the best concentration combinations of BAP (Benzyl Amino Purine), TDZ (Thidiazuron) and Silver Nitrate (AgNO₃) for micropropagation of carnation (Dianthus caryophyllus L) cv. Irene was performed. Genetic fidelity in the microplants of different generation was determined with the help of ISSR markers. Results of the present investigation showed that the MS gelled media supplemented with BAP (0.5 mgl^–1) and AgNO₃ (1.5 mgl^–1) showed maximum shoot proliferation, good shoot length and reduced hyperhydricity significantly. Our study also indicates that hyperhydricity decreased by substantially by increasing the concentration of AgNO₃ compared to control. High level of AgNO₃ in MS medium did not helped to reduce hyperhydricity. Another chemical combination of TDZ (1.0 mgl^–1) and AgNO₃ (0.5 mgl^–1) in MS gelled media gave the maximum shoot proliferation, shoot length with minimum hyperhydricity which showed normal growth and development compared to control and other treatments. This concentration combination proved effective to reduce hyperhydricity and to enhance shoot proliferation. Because TDZ with AgNO₃ had pronounced effect to reduce hyperhydricity, therefore, this concentration combination was used to advance the progeny up to subculture (SC10) using donor mother microplants (SC1) selected from the best concentration response to know the genetic fidelity among the different generations. Microplants which were non hyperhydrified were used to assess the genetic fidelity using ISSR markers. Fourteen ISSR primers were used that gave good amplification and  produced 616 bands with an average of 44 amplicon per primer from in vitro raised microplants including mother plant where the banding patterns exhibited uniform and homogeneous banding patterns. This study may help us to know applicability of ISSR markers to establish genetic fidelity among in vitro – raised microplants of carnation.

]]> http://cytgen.com/en/2023/_97-99N1V57.htm http://cytgen.com/ru/2023/_97-99N1V57.htm Sun, 01 Jan 2023 00:00:00 +0200 In this study, morphotoxic and cytogenotoxic potency of trade formulation of Propineb were analyzed using Allium cepa assay. The root tips were treated with different concentrations of fungicide Propineb for 24, 48, 72 and 96 h. The effects of propineb on morphological parameters and cytological effects were determined. It was found that Propineb significantly increased the percentage of total abnormality at all concentrations of the Propineb and these increases were dependent on the increasing concentrations for 24, 48 and 72 h. It was determined mitotic chromosomal abnormalities such as irregular metaphase, stickiness, c­mitosis, micronucleus, vagrant chromosomes and bridges. These results lead us to the conclusion that Propineb may have genotoxic and cytotoxic properties due to induction in the frequency of total abnormality and a reduction in mitotic index in root tip cells of A. cepa L. It should be necessary to be careful when using pesticides in agricultural areas and should take precautions.

]]> http://cytgen.com/en/2023/100-102N1V57.htm http://cytgen.com/ru/2023/100-102N1V57.htm Sun, 01 Jan 2023 00:00:00 +0200 Numerous experimental data shows crucial involvement of miRs in skeletal development in embryos, osteogenic differentiation, and maturation. However, molecular me­chanisms of miRs action, in other words their target signaling pathways and transcriptional factors that specific drives osteogenic differentiation is far from being understood.  With meta­analysis we identified miRs significantly involved in hMSCs osteogenic differentiation. Statistical analysis revealed significant trend of upregulation of let­7a, mir­21, mir­26a, mir­29b, mir­101, mir­143, mir­218 during hMSCs differentiation into osteoblast. And the opposite trend was shown for mir­17, mir­31, mir­138 and mir­222: their content was significantly lower during osteogenic differentiation. Using bioinformatics approaches we identified predictable genes­target for each miRs, analyzed signaling networks and biological process enriched by these genes. Bioinformatic assay shows that miRNAs specifically involved in hMSCs transition into osteogenic differentiation via microenvironment formation (i.e. let­7a, mir­17, mir­21, mir­29b and mir­101), TGF­β/BMP–SMAD dependent pathway (i.e. let­7a, mir­17, mir­21, mir­26a mir­101) and MAPK signaling pathway (i.e. let­7a, mir­21, mir­26a, mir­29b, mir­143 and mir17). Yap­dependent expression of osteogenic transcriptional factors modulated by let­7a, mir­31mir­101, mir­138 and mir­222. We predicted that mir­17, mir­26a, mir­29b, mir­101, mir­138 and mir­222 are specifically involved in canonical Wnt signaling dependent osteogenesis as well as in osteoblast maturation together with let­7a, mir­29b and mir­218 which modulate AMPK signaling. Additionally, identified mir­101 is likely involved into osteoblast homeostasis via Hedgehog signaling. Presented here data expands knowledge in the field of hMSCs fate and osteogenesis orchestration by miRs, points to pro­osteogenic and anti­osteogenic miRs and their potential molecular pathways.

]]> http://cytgen.com/en/2023/103-105N1V57.htm http://cytgen.com/ru/2023/103-105N1V57.htm Sun, 01 Jan 2023 00:00:00 +0200 SUMMARY. Peach is one of the most important fruit crops, cultivation of which held the third largest areas among all fruit crops, grown in temperate climate zone. Cultivation of this crop under less favourable clime conditions would require the creation of new resistant genotypes via the intra- or interspecific hybridization, including crossing with almond. Efficient breeding of hybrids and their long-term selection will require a rapid and accurate method of genetic barcoding that would be able to distinguish closely-related genotypes or to detect interspecific hybrids. One such approaches is TBP-analysis, which is based on the evaluation of intron length polymorphism of β-tubulin genes. However, the correct interpretation of the results of such an analysis should be based on data on the diversity of β-tubulin genes panel in the genomes of the analyzed species. Thus, here we report on the successful whole-genome identification and on comprehensive analysis of the phylogeny and synteny of the β-tubulin genes of P. persica and P. dulcis, as well as we demonstrate the the possibility to use such data of the genomic search for interpretation of data of TBP genotyping of intra- and interspecific hybrids of peach and almond species. In general, 11 β-tubulin genes were indentified within the P. persica genome, and 10 genes, in the P. dulcis genome, accounting pseudogenes. Additionally, phylogenetic and synteny analyses of the identified genes made it possible to identify the orthologues in the genomes of A. thaliana and A. lyrata, as well as to designate identified β-tubulins to specific isotypes. Genotyping via the TBP-method allowed obtaining distinct molecular profiles for 11 investigated accessions, among which 8 were intra- or interspecific hybrids. Based on obtained results of genotyping, a cluster analysis was carried out, the results of which correlated well with the breeding history of the analysed genotypes, what additionally confirmed the effectiveness and accuracy of the genotyping approach used.

]]> http://cytgen.com/en/2022/__3-18N6V56.htm http://cytgen.com/ru/2022/__3-18N6V56.htm Tue, 01 Nov 2022 00:00:00 +0200 SUMMARY. The genus Aconitum L. includes a large number of toxic and pharmaceutical important plants. One of the centers of diversity of this genus is located on the territory of the Eastern Carpathians. In this region there are many representatives of the genus with unclear taxonomic status, in particular, members of the complex A. anthora s. l. The taxonomic position of this complex within the genus also remains controversial, as the regions of the chloroplast and nuclear genomes previously used for phylogenetic analysis appeared to be insufficiently variable. Therefore, the search for an optimal molecular marker with a high level of polymorphism within the genus Aconitum remains a relevant task. The 5S rDNA IGS (intergenic spacer) is an evolutionarily variable region of the nuclear genome, which was previously successfully used for phylogeny reconstruction in many groups of angiosperms. In this work, using methods of molecular genetics and bioinformatics, we obtained 5S rDNA IGS sequences for representatives of the A. anthora complex and phylogenetically distant species of the genus Aconitum. Analysis of IGS sequences showed that this region is relatively long in species of the genus, 574–619 bp. The IGS variability is due to numerous nucleotide substitutions, while short oligonucleotide indels occur only at the 5′ end of the spacer. Four conserved regions were found in the IGS of Aconitum, two of which correspond to the external promoter and terminator elements of RNA polymerase III, while the function of the other two regions remains unknown. First of them shows homology to the 5S rRNA coding region, while the second one demonstrates high similarity to the sequences from the genomes of representatives of taxonomically distant families of monocots and dicots, suggesting horizontal gene transfer. The phylogenetic analysis applying the 5S rDNA IGS supports the inter-pretation of A. anthora s. l. as a separate subgenus within the genus Aconitum.

]]> http://cytgen.com/en/2022/19-30N6V56.htm http://cytgen.com/ru/2022/19-30N6V56.htm Tue, 01 Nov 2022 00:00:00 +0200 SUMMARY. The high-yielding Cabernet Sauvignon grape variety is susceptible to viral diseases, which may affect its productivity and taste quality. Our aim was to identify a Cabernet Sauvignon variety of clonal origin from the south of Ukraine; to detect the contamination of plants of this variety with harmful viruses included in the planting material certification system; to identify the causative agents of viral diseases using molecular methods and to establish the nucleotide sequence of the 2C^CP coat protein gene of the detected viruses. Plants with the possible symptoms of Grapevine Fanleaf Virus (GFLV) and Grapevine Leaf Roll-Associated Virus (GLRaV) viruses were found out during the phytosanitary survey of Cabernet Sauvignon grapevine plantations. But only GFLV presence was confirmed using a real-time reverse transcription-polymerase chain reaction (RT-PCR) and sequencing of the 2C^CP coat protein gene (the GenBank accession number MN072356.1). It was established that the nucleotide sequence of the Cabernet Sauvignon plant isolate is very close to samples from regions geographically distant from Ukraine, primarily the USA, Iran and France. Identity of the Cabernet Sauvignon variety of clone origin was also proved by the microsatellite analysis.

]]> http://cytgen.com/en/2022/31-41N6V56.htm http://cytgen.com/ru/2022/31-41N6V56.htm Tue, 01 Nov 2022 00:00:00 +0200 SUMMARY. The associations between SNP in leptin gene (LEP g.2845A>T, LEP g.3996T>C, LEP g.3469T>C) and lep-tin receptor gene (LEPR c.232A>T, LEPR c.2856C>T, LEPR c.915C>T) and the meat quality traits of Ukrainian Large White pigs (ULW) were studied. The computational analysis (SIFT, PolyPhen-2 and I-Mutant bioinformational services) was conducted to investigate the effect of mutations, causing SNP, for both selected SNP, and those missense SNP, for which they can be potential linkage disequilibrium (LD) markers, on the structure and functions of leptin and leptin receptor. LEP SNP c.3469T>C (exon 3, rs45431504) is associated with the protein content in meat and the content of moisture in the back fat. LEP SNP g.2845A>T (rs344615147), which is associated with moisture-retaining ability of meat and the content of moisture in the back fat, is located in the second intron and, similar to SNP c.3469T>C, may be viewed as an LD marker of missense-polymorphism of SNP LEP rs701423985. LEPR SNP c.232A>T (exon 4, rs45435517) is a missense mutation, causing the amino acid replacement S52I. The evaluations, made using SIFT and PolyPhen-2, demonstrate a considerable impact of rs45435517 on the functional characteristics of leptin receptor. Our study determined the association between SNP c.232A>T and fat thickness. At the same time, SNP c.2856C>T (exon 20, rs694660564, a synonymic replacement), which in our work demonstrated the association with the loss of moisture in meat for ULW pigs, may be a potential LD marker and be in disequilibrium with the linkage to missense SNP LEPR rs1113972516, rs792804682 and rs1109261799. SNP, for which in our work the associations were found with meat quality traits, may be viewed as potential markers for the breeding of ULW pigs and possibly other breeds, directed at obtaining genotypes with improved characteristics of meat quality.

]]> http://cytgen.com/en/2022/42-56N6V56.htm http://cytgen.com/ru/2022/42-56N6V56.htm Tue, 01 Nov 2022 00:00:00 +0200 SUMMARY. Coronavirus disease (COVID-19) is an acute infectious disease of the respiratory tract, the causative agent of which is the new coronavirus SARS-CoV-2.The global vaccination program against SARS-CoV-2 is still on-going, and the incidence of COVID-19 worldwide is significantly decreasing. However, among the millions of survivors of COVID-19, large groups will require assistance due to increased clinical consequences fol-lowing COVID-19. Currently, there is a need to find molecular biomarkers to monitor the occurrence and progression of the post-covid syndrome. For this pur-pose, we were measuring the relative average length of telomeres in groups of women of reproductive age: in a group of patients (n = 64) who contracted COVID-19 and in a control group (n = 42) of women of the same age. The analysis was performed by monochrome quantitative PCR in real time on DNA samples isolated from peripheral blood leukocytes. According to the study’s results, it was established that the relative average length of telomeres in peripheral blood leukocytes in the group of patients with COVID-19 was lower than in the control group (p < 0.05). Our results suggest that the observed shortening of the relative average length of telomeres in a group of patients infected with COVID-19 may be evidence in favor of the fact that SARS-CoV-2 infection, may directly cause erosion of telomeres  in blood cells, in particular, in leukocytes. Accordingly, the determination of the relative average length of telomeres can be an informative prognostic marker for assessing the risk of the severity of the course of the disease in COVID-19 and the development of the post-covid syndrome.

]]> http://cytgen.com/en/2022/57-62N6V56.htm http://cytgen.com/ru/2022/57-62N6V56.htm Tue, 01 Nov 2022 00:00:00 +0200 SUMMARY. Cytokeratins are a large group of intermediate filament proteins that form the cytoskeleton of epithelial cells and their appendages (hair, nails). Biochemically, cytokeratins are divided into two main types: acidic and basic. Each cytokeratin pair necessarily contains both acidic and basic cytokeratins in equimolar amounts. This significantly distinguishes cytokeratins from other intermediate filament proteins and is essential for proper organization of the cytoskeleton. Cytokeratins also provide signaling in the cell and participate in cell-cell adhesion, and apoptosis. Today, the general principles of cytokeratin expression at different stages of epithelial cell development are known. The expression of cytokeratins is organ-specific, depending on the type of epithelial cells, the degree of differentiation, and tissue development. Therefore, the cytokeratins profile can be used to diagnose various pathological processes. Special attention in the review is paid to cytokeratins 8, 18, and 19 as possible biomarkers of carcinogenesis.

]]> http://cytgen.com/en/2022/63-75N6V56.htm http://cytgen.com/ru/2022/63-75N6V56.htm Tue, 01 Nov 2022 00:00:00 +0200 Barbus pergamonensis Karaman, 1971 and Luciobarbus lydianus (Boulenger, 1896) were studied karyologically. Karyotypes and chromosomal banding techniques with C-banding and silver staining were determined. Diploid chromosome numbers (2n) were invariably 100; karyotypes were composed of 26 metasentric (m), 20 submetacentric (sm) and 54 subtelo-acrocentric (st-a) chromosomes in B. pergamonensis and 24 m, 22 sm and 54 st-a chromosomes in L. lydianus. No heteromorphic sex chromosomes were determined. C-bands were observed on the pericentromeric regions of some of the chromosomes in the studied species. Multiple nucleolus organizer regions were detected in both species. This study shall contribute to barbels cytotaxonomy.

]]> http://cytgen.com/en/2022/76-77N6V56.htm http://cytgen.com/ru/2022/76-77N6V56.htm Tue, 01 Nov 2022 00:00:00 +0200 Barley is the fourth most important grain crop in the world which is well­adapted to marginal and stress­prone environments, and hence a more reliable plant than wheat or rice in regions located in colder or higher altitude. Genetic diversity is a key component in breeding programs. We have analyzed the genetic diversity of 106 barley accessions using RAPD and ISSR molecular marker systems. The accessions were composed of wild and domesticated barley representing genotypes from fifteen different countries. A total of 40 polymorphic primers (20 RAPD and 20 ISSR) were used. The RAPD primers generated 331 amplification products and the ISSR primers produced 295 DNA fragments, out of which 225 and 247 were polymorphic, respectively. By RAPD markers the polymorphic banding patterns with the number of amplified fragments varied from 10 (AF­14 and OPL­09) to 26 (OPA­04).  The percentage of polymorphism ranged from 53.84 % (BY­15) to 100 % (OFG­14, OPF­03 and W­07) with an average of 77.47 %. Contrary to this, the number of bands amplified per primer by ISSR markers varied between 10 (UBC­872, ISSR­4, 6 and 7) and 21 (UBC­845) and the percentage of polymorphism ranged from 57.14 % (UBC­814) to 100 % (ISSR­5 and 7) with 83.97 % polymorphism. Dendrogram was constructed based on RAPD and ISSR polymorphism dividing the barley accessions into eleven however by combined RAPD+ISSR all samples were grouped into ten main distinct clusters. High correlation between the RAPD and ISSR marker systems was shown using a Mantel test (r = 0.852). The data obtained confirmed and indicated that Azerbaijan cultivated barley accessions distinctly diverged from wild types.

]]> http://cytgen.com/en/2022/78-80N6V56.htm http://cytgen.com/ru/2022/78-80N6V56.htm Tue, 01 Nov 2022 00:00:00 +0200 Nasopharyngeal carcinoma (NPC) is the most common cancer of in the areas of head and neck cancer, which gravitates toward Asia. NPC is tightly associated with the gene expression of Epstein Barr Virus (EBV). Among them, Latent membrane protein 1 (LMP-1) gene, and its 30-bp deleted variant have profound effects on many pathways of NPC pathogenesis leading to the tumorigenesis. The study of the 30-bp deleted variant was not performed in Vietnam, the high endemic NPC country. Therefore, the aim of the current study is to analyze the genetic characteristics of whether present a 30-bp deletion in NPC biopsy tumors is associated with EBV. Twenty biopsy NPC samples were collected from local patients, analyzed by PCR-sequencing and compared to the prototype B95-8 30-bp deletion variant. As the results, the 30-bp deletion polymorphism was recorded in 55.00% samples. The 30-bp deletion polymorphism yielded from nucleotide 168,285 to nucleotide 168,256, different from the prototype B95-8 and other previously reported sequences. Therefore, we termed as the new 30-bp deletion* observed in the region of CDS LMP-1 gene. Three patterns of amino acid substitution/deletion observed in 30-bp deletion/no 30-bp deletion in LMP-1 C-cytoplasmic terminus were recorded. In conclusion, the new 30-bp deletion* variant, which was different from previously reported 30-bp deletion, as well as the pattern of amino acid substitution/deletion was investigated in the Vietnamese nasopharyngeal carcinoma biopsy samples.

]]> http://cytgen.com/en/2022/81-82N6V56.htm http://cytgen.com/ru/2022/81-82N6V56.htm Tue, 01 Nov 2022 00:00:00 +0200 The role of the GFRα1 receptor in the regulation of cell differentiation and spermatogenesis is poorly investigated. The influence of GFRα1 on the apoptosis and proliferation of spermatogenic cells are not fully elucidated. In this study, GC­1 spg cells were cultured and then assigned to a control cell (control group) transfected with empty adenovirus AdH1­siRNA and an inhibited cells (inhibited group) with GFRα1 interfering adenovirus (AdH1­siRNA/GFRα1). The GFRα1 inhibition was determined by employing western blot and qRT­PCR analysis. Furthermore, we used CCK8 for determining the proliferation of the GFRα1 interfering adenovirus (AdH1­siRNA/GFRα1). Transfected cells, employing flow cytometry and Caspase 3, Bax and Bcl­2 expression analysis to determine their apoptosis. This study demonstrated that after transfection, the inhibited cells presented lower proliferation and invasion and higher apoptosis. According to these findings, the inhibition of GFRα1 might reduce spermatogenesis and remarkably affect spermatogenic cells proliferation.

]]> http://cytgen.com/en/2022/83-84N6V56.htm http://cytgen.com/ru/2022/83-84N6V56.htm Tue, 01 Nov 2022 00:00:00 +0200 http://cytgen.com/en/2022/85-86N6V56.htm http://cytgen.com/ru/2022/85-86N6V56.htm Tue, 01 Nov 2022 00:00:00 +0200 SUMMARY. The organization of promoters of DREB2B TF orthologous genes involved in the response to abiotic stress was studied in extremophile plant D. antarctica E. Desv and 12 other grass species with different cold and drought resistance. The average evolutionary distances were 0.621 between the promoter sequences and 0.442 between coding sequences, including introns; the values of nucleotide diversity (π) for these regions were 0.410 and 0.274, respectively. Clustering of sequences was generally consistent with the accepted taxonomy of the Poaceae family. In total, 54 cis-elements involved in the response to abiotic and biotic stresses, light, hormones, such as abscisic acid, auxin, methyl jasmonate, ethylene, gibberellin and salicylic acid, and tissue-specific cis elements were identified. A large proportion of these cis elements were associated with abiotic stress response that is consistent with known functions of DREB2B TF. Except for a few isolated differences, the studied grass species of different subfamilies and D. antarctica had a similar set of cis elements in the DREB2B promoter, a finding that indicates the similarity in the control of this gene expression and its potential functions in these species.

]]> http://cytgen.com/en/2022/__3-15N5V56.htm http://cytgen.com/ru/2022/__3-15N5V56.htm Thu, 01 Sep 2022 00:00:00 +0300 SUMMARY. Seasonality affects the morphofunctional characteristics of reproductive cells, which makes the natural reproduction of dairy goats possible only in certain seasons. Cryopreservation of sperm, as part of assisted reproductive technologies, provides their flexibility, which increases the chances of increasing the number of livestock. However, it can cause changes in the morphofunctional characteristics and genetic material of sperm. Therefore, the aim of this study was to determine seasonal changes in viability, motility and DNA fragmentation level of native and cryopreserved goat sperm. The experiment was conducted using ejaculates of sexually mature male goats of Saanen breed, obtained in breeding and non-breeding seasons. To detect the effect of seasonal differences of seminal plasma on the characteristics of cryopreserved sperm, cells were cryopreserved in ejaculate and following isolation. The results of the study showed that the motility of native ejaculate sperm in the breeding season was higher than in the non-breeding season (p ≤ 0.05). Cryopreservation led to a decrease in the number of motile sperm of ejaculate and the selected fraction of sperm in the non-breeding season, and the selected fraction of cells in the breeding season (p ≤ 0.05). When comparing the viability and integrity of sperm DNA, there was a significant decrease in all groups in the non-breeding season compared to the breeding season (p ≤ 0.05). It was found that the level of DNA fragmentation of goat ejaculate sperm in the breeding season after cryopreservation did not change compared to the native sample, while in cryopreserved sperm of the selected fraction of the same season increased (p ≤ 0.05). Cryopreserved spermatozoa of the non-breeding season in the ejaculate and isolated fraction had an increased level of DNA fragmentation compared to baseline before cryopreservation. Thus, it can be concluded that the composition of semen liquid varies depending on the mating season, which affects its cryoprotective properties against spermatozoa during the ejaculate freezing. Therefore, it is recommended to collect whole ejaculate by freezing in autumn and early winter to improve the effectiveness of artificial insemination using cryopreserved sperm of male goats of Saanen breed.

]]> http://cytgen.com/en/2022/16-24N5V56.htm http://cytgen.com/ru/2022/16-24N5V56.htm Thu, 01 Sep 2022 00:00:00 +0300 SUMMARY. The elucidation of molecular genetic mechanisms in the development of rare inherited diseases is crucial in understanding the processes of pathogenesis, improving the diagnosis and treatment of patients. The aim of our study was to determine the nature of genetic defects in a patient who showed signs of sexual dysfunction in combination with neuromuscular pathology. The study included: clinical and genealogical examination, ultrasonographic examination, electroneuromyography, study of biochemical parameters in serum: hormones and enzymes, molecular cytogenetic study using the FISH method, whole exome sequencing of the patient's DNA analysis, bioinformatiс analysis. The patient showed signs of primary (hypergonadotropic) hypogonadism and polyneuropathic changes in the lower extremities (axonal-demyelinating). It was found that these clinical characteristics of the patient's phenotype do not agree with any description of the clinical phenotype registered in the OMIM International Database. According to the results of molecular cytogenetic study in the patient's karyotype, an unbalanced translocation of the Yp11.3 region containing the SRY gene to the short arm region of the X chromosome Xp22.33 with the formation of a derivative X chromosome was detected. On another side, the results of whole exome sequencing of the patient's genomic DNA revealed the presence of a homozygous mutation chr5: 131165096 C> G (c.110G> C, p.Arg37Pro) in the HINT1 gene. Thus, it can be concluded that the phenotype with signs of disorders of sex differentiation in combination with axonal neuropathy with myotonia is due to two independent pathogenetic factors - unbalanced translocation of the SRY gene sequence on the X chromosome, resulting in paternal spermatogenesis and inheritance  of the HINT1 gene homozygous missence mutation (c.110G>C, p.Arg37Pro). These results support the effectiveness of a combination of molecular cytogenetic research methods and the results of high-performance WES to elucidate the molecular mechanisms of pathogenesis in patients with rare clinical phenotypes.

]]> http://cytgen.com/en/2022/25-31N5V56.htm http://cytgen.com/ru/2022/25-31N5V56.htm Thu, 01 Sep 2022 00:00:00 +0300 SUMMARY. Comparison of cytogenetic and molecular-cytogenetic rearrangements of bone marrow cells in 251 patients at diagnosis of myelodysplastic syndromes (MDS) and in 7 - in the transformation into secondary acute myeloid leukemia was carry out. Significant heterogeneity of karyotypes in the structure of clones in all isolated forms of MDS and in transformed MDS into secondary acute myeloid leukemias was established and indicating a different genetic composition of bone marrow cells. It is shown that the evolution of clonal chromosome abnormalities is a universal mechanism for the formation of abnormal clones. An increase in the frequency of pseudodiploid and hypodiploid clones depending on the complexity of the form of MDS were marked: pseudodiploid from 4.5 % in MDS with single-line dysplasia (RA) to 27.3 % in MDS with excess blasts (RAEB 1–2), hypodiploid – from 4.5 in RA to 18.2 at RAEB 1–2)were shown, which is associated with the loss of genetic material. In the group of RA losses of genetic material in the form of deletions (57.9 %) more often recorded. Chromosome 11 (31.6 %) was more often involved in structural rearrangements.

]]> http://cytgen.com/en/2022/32-40N5V56.htm http://cytgen.com/ru/2022/32-40N5V56.htm Thu, 01 Sep 2022 00:00:00 +0300 SUMMARY. The relationship between stimulation of low-molecular-weight antioxidant accumulation in pharmaceutical plant raw material (inflorescences) and the markers of radiation-induced genome instability at the stage of plant flowering under X-ray exposure were investigated. The study of rearrangements of DNA primary structure under different dose exposure was carried out by PCR using eight ISSR and ten RAPD primers. Dose – dependent changes in amplicon spectra during ISSR – RAPD – PCR were analyzed using the Jacquard similarity index. It was found that the largest rearrangements of the primary DNA structure of both genotypes, which was indicated as a decrease in similarity with the control spectra of amplicon, was observed under exposure with doses 5–10 Gray. There was a tendency to approach this indicator to the control one under 15 Gray dose, which mint increased efficiency of reparative processes. The relationship between the polymorphism of the primary structure of DNA by ISSR-RAPD-sequences through different genotypes and the nature of its rearrangement under radiation exposure was shown. Comparison of the results with no monotonic dose curves of the specific flavonoids’ and phenols’ content allowed us to conclude that the stimulation of antioxidant protection was shown under doses corresponding to low efficiency of repair processes and, accordingly reduced it under genetic material repair. The interpretation of the identified phenomenon is based on the known connection between the effects of genomic instability and the increase in the level of reactive oxygen species and the general principles of antioxidant protection. The significance of the obtained results through the development of the scientific basis for the implementation of small radiation exposure doses in biotechnology, particularly in pharmacology is discussed.

]]> http://cytgen.com/en/2022/41-51N5V56.htm http://cytgen.com/ru/2022/41-51N5V56.htm Thu, 01 Sep 2022 00:00:00 +0300 SUMMARY. Wheat-rye translocations involving arm 1RS are widespread introgressions among commercial bread wheat cultivars: 1BL.1RS from the rye Petkus (as in the cultivar Kavkaz) and 1AL.1RS from the rye Insave (as in Amigo). Genotypes with recombinant arm 1RS involved in the translocation can carry new combinations of resistance genes in the chromosome region flanked by the loci Sec-1 (Gli-R1) and Sec-N, which is located distal to Sec-1 at a distance of 10–20 cM. The aim of this investigation was to study the frequency of genotypes with recombinant 1RS arms involved in wheat-rye translocations in segregating F₅ populations from crossing bread wheat carriers of translocations 1BL.1RS and 1AL.1RS. Samples of F₅ seeds from the crosses Myronivska 67 (M67) × Kolumbiia, Kolumbiia × M67, B-16 × Smuhlianka, and Smuhlyanka × B-16 served as the material for the investigation. The cultivar M67 and the line B-16 carry 1BL.1RS as in the cultivar Kavkaz; Smuhlianka and Kolumbiia carry 1AL.1RS as in the cultivar Amigo. Electrophoresis of alcoholsoluble grain proteins and identification of alleles at the Gli-A1/Gli-R1, Gli-B1/Gli-R1 loci were used to identify translocations involving 1RS and their position (on 1AL or 1BL). Sec-N alleles were identified by SDS electrophoresis. The presence of recombination products between the 1RS arms was determined based on the change of the chromosomal position of the respective allele at Gli-R1 (Sec-1) and Sec-N. In the segregating populations, there were five of six theoretically possible variants of genotypes with recombinant arm 1RS with respect to the secalin loci. The total frequency of genotypes with the simultaneous presence of two translocations ranged from 3.3 to 18.0 %, and the total frequency of genotypes with one identified recombinant arm 1RS was 5.7–30.5 %. The highest frequency of genotypes with recombinant arm 1RS was found in the population M67 × Kolumbiia.

]]> http://cytgen.com/en/2022/52-60N5V56.htm http://cytgen.com/ru/2022/52-60N5V56.htm Thu, 01 Sep 2022 00:00:00 +0300 Methylation of DNA promoter sequences at the CpG islands has become a molecular tool for gene regulation. NAC6D gene is induced by different biotic and abiotic stimuli. The proximal promoter sequence of NAC6D was investigated for the impact of CpG methylation on its expression in response to high temperature in wheat. Gene expression was estimated by real time PCR and methylation of NAC6D promoter sequence was investigated by bisulfite sequencing. Results showed that NAC6D was highly induced by high temperature, whereas DNA methylatransferase 3 (Met3) was highly reduced by high temperature. Close investigation of NAC6D promoter methylation revealed that high temperature caused hypomehtylation of the proximal promoter sequence. Twelve CpGsites showed low difference in methylation compared to the control (normal temperature, 25 ºC), while 3 CpGs(–59, –169, –204) were extremely hypomethylated in response to high temperature compared to their methylation status under the normal condition. The induction of NAC6D was negatively correlated with Met3 suppression and methylation level at the CpG sites in the promoter region. Results prove that methylation greatly contribute to the regulation of NAC6D in response to high temperature. This will improve our current understanding of how plants respond to abiotic stresses at the molecular level and the integration of DNA methylation and epigenetics in the next generation plant breeding.

]]> http://cytgen.com/en/2022/61-64N5V56.htm http://cytgen.com/ru/2022/61-64N5V56.htm Thu, 01 Sep 2022 00:00:00 +0300 Calcium supersaturation in urine during hyperoxaluria can lead to renal calcium oxalate (CaOx) stone deposition; a condition termed as nephrolithiasis. Recent genetic studies indicate the possibility of epigenetic alterations during nephrolithiasis. We aimed to study the influence of renal CaOx stone formation on the global levels of 5-methylcytosine (5 mC) and 5-hydroxymethycytosine (5 hmC) in hyperoxaluric rat kidney. In this study, twelve male Wistar rats were divided equally into two groups (control and experimental). Animals in the experimental group received 2.5 % (w/v) sodium oxalate in 5 ml of sterile water, every day, orally for four weeks to induce hyperoxaluria. At the end of four weeks, urine parameters, serum biochemistry and renal histopathological changes were evaluated. Global 5mC and 5hmC levels were analysed using enzyme-linked immune sorbent assay (ELISA), and renal mRNA expression of DNMTs and TET genes were also determined. Urine parameters and serum biochemistry showed that rats fed with 2.5 % (w/v) sodium oxalate orally for four weeks developed hyperoxaluria (p < 0.05). Histopathological evaluation of hyperoxaluric rat kidneys showed the deposition of CaOx crystals and marked tubular injury. ELISA showed significantly high levels of 5 hmC (p < 0.0001) in the kidneys of rats with renal CaOx stones, whereas 5 mC was only slightly significant (p < 0.05) as compared to the control rats. mRNA expression of TET 2, a regulator of DNA demethylation process, was found to be significantly upregulated (p < 0.01) in the rats with CaOx kidney stones. This preliminary study showed some early evidence of epigenetic modifications being influenced by hyperoxaluria-induced renal CaOx stones, which may likely contribute to the transcriptional regulation during CaOx nephrolithiasis.

]]> http://cytgen.com/en/2022/65-66N5V56.htm http://cytgen.com/ru/2022/65-66N5V56.htm Thu, 01 Sep 2022 00:00:00 +0300 Present investigation deals with the reporting of 1B and 2B chromosomes in Coriandrum sativum L. that had previously not reported. Study deals with the influence of B chromosomes on chiasma frequency, distribution pattern and pollen fertility in carrier and non-carrier MMCs of carrier plants and MMCs of non-carrier diploid and autotetraploid plants of coriander. Results envisaged that the presence of B chromosomes enhances the chiasma frequency in carrier MMCs of carrier diploid and autotetraploid plants in comparison to non-carrier plants. However, there is a moderate reduction in pollen fertility due to presence of B chromosomes. It was noticed that the higher number of B- chromosomes have greater influence on rate of chiasma distribution. In autotetraploid plants, the rate of bivalents and multivalent associations is greatly influenced due to the presence of B chromosomes in comparison to diploid ones. The probable mechanism of B chromosome machinery of action has been discussed in accordance with the light of earlier works. B chromosomes, we noticed to behave as univalents when unpaired or behave as bivalent when present as paired but do not aligned at the equator or either of the poles however they remain as lagging chromosomes at periphery of the cells. Further, the study of Bs might be used to explore the unrevealed facts regarding their organisation, function, evolution of genomes and how they retain themselves as selfish elements by using machinery of host genomes.

]]> http://cytgen.com/en/2022/67-68N5V56.htm http://cytgen.com/ru/2022/67-68N5V56.htm Thu, 01 Sep 2022 00:00:00 +0300 Epigenetics is the study of modifications in DNA expression without changing the sequences in deoxyribonucleic acid. Epigenetic mechanisms are specific “control” modifications responsible for the activity or inactivity of selected genes. Researchers are revealing a strong impact of epigenetic mechanisms on various general diseases in human. It gives clinicians great hope to understand pathomechanisms and start causal treatment. The possibility for dental clinicians is also wide and consists of diagnosis and treatment of diseases occurring in the oral cavity. This review presents the role of epigenetic mechanisms and the growing interest in their possible associations with dental pathologies such as periodontal diseases, craniofacial malformations, and tooth agenesis.

]]> http://cytgen.com/en/2022/69-71N5V56.htm http://cytgen.com/ru/2022/69-71N5V56.htm Thu, 01 Sep 2022 00:00:00 +0300 SUMMARY. At higher order levels chromatin is organized into loops, and this looping plays an important role in transcription regulation. In our previous works we investigated the kinetics of DNA loop migration during single cell gel electrophoresis (the comet assay) of nucleoids obtained from lysed cells. It was shown that there are three parts of DNA in nucleoids: DNA in rather small loops which migrate rapidly; DNA in the loops up to ~150 kb, the migration of which is retarded; and larger loops that cannot migrate. Here we applied, for the first time, the pulse-field electrophoresis in the comet assay. Our results show that the first rapid step during the usual comet assay can be attributed to loops on the nucleoid surface while the second slow component represents loops inside the nucleoid.

]]> http://cytgen.com/en/2022/__3-9N4V56.htm http://cytgen.com/ru/2022/__3-9N4V56.htm Fri, 01 Jul 2022 00:00:00 +0300 SUMMARY. The results of the study of exon 2 polymorphism of the BoLA-DRB3 gene in four domestic cattle breeds: Ukrainian black-and-white dairy (UBWM, n = 293), Ukrainian red-and-white dairy (URWM, n = 131), Gray Ukrainian (n = 93) and Ukrainian white-headed (UWH, n = 49) are presented. Allele typing was performed by the PLR-PDRF method. Genetic profiles of the breeds were formed according to the following indices: allele (P_a) and genotype (G_a) frequencies, observed (H_o) and expected (H_e) heterozygosity, Wright fixation index (F_IS), total (N_a) and effective number (A_e) of alleles, indices Shannon (I) and Pielu (J). The maximum number of alleles was in the UBWM breed (N^a = 37). For the URWM, UWH, and Gray breeds, 31, 29, and 28 va-riants were identified, respectively. Two breeds were ty-ped with «no established nomenclature» alleles: Gray – *jba, *jab, *jbb, *nad and *nda, UWH – *nab, *mdb, *iab, *gbb, *fbd, *naa, whose proportion was 8.9 and 7.1 %, respectively. The number of «informative» alleles was: UBWM 55.8 % (7 alleles), URWM 60.4 % (6), Gray 69.9 % (4), and UWH 65.3 % (7). The expected heterozygosity and Wright index had the following values: UBWM – H_e = 0.942; F_IS = 0.022; URWM – H_e = 0.93; F_IS = 0.032; Gray – H_e = 0.774; F_IS = 0.041; WHU – H_e = 0.927; F_IS = –0.035. The following biodiversity indices were obtained for the studied rocks: UBWM – A_e = 17.2; I = 3.13; J = 0.6; WHU – A_e = 13.7; I = 2.93; J = 0.61 and URWM – A_e = 11.5; I = 2.87; J = 0.558, and Gray – A_e = 4.42; I = 2.21; J = 0.455. Analysis of dendrograms based on genetic distances showed that domestic breeds and Holstein are combined in the same cluster, indicating their genetic similarity behind the BoLA-DRB3 gene. Exon 2 polymorphism data of BoLA-DRB3 gene showed that the greatest genetic diversity among Ukrainian cattle is characteristic of UBWM breed. The results obtained contribute to better understanding of genetic diversity and distribution of BoLA-DRB3 gene alleles in cattle in Ukraine and increase our knowledge about global variability of BoLA-DRB3 gene, which forms immune response to protect against pathogens.

]]> http://cytgen.com/en/2022/10-23N4V56.htm http://cytgen.com/ru/2022/10-23N4V56.htm Fri, 01 Jul 2022 00:00:00 +0300 SUMMARY. Heterogenity of the tumor population, the presence of tumor stem cells in it, is one of the reasons for the resistance of tumors to antitumor therapy, recurrence and metastasis, as well as the complexity of treatment of the cancer. The aim of the study was to enrich of the breast adenocarcinoma cells, MCF-7, multicellular tumor spheroids (MTS) with cancer stem cells, enriched MTS (eMTS). Other aim was to investigate the resulting subpopulation of CSC in MTS by biochemical, immunological and cytological methods. According to the results of the study, it was found that in the conditions of lack of nutrient medium, with the addition of certain growth factors, the percentage of CSC in the cell population of MTS increased significantly. The obtained results demonstrated increasing of the CSC subpopulation. It was indicated according to biochemical, cytological and immunological methods simultaneously. Thus, the percentage of CD133+ cells increased from 12.47 to 82.08 %, Nestin+ from 31.3 to 82.58 %. According to immunohistochemical staining data the expression of
other markers of CSC: CD44, CD133, bmi1, also increased. The activity of aldehyde dehydrogenase in MCF-7 cells in monolayer culture was 0.07 mol/mg protein per minute and increased to 1.58 mol/mg of protein per minute in eMTS. The activity of glucose-6-phosphate dehydrogenase (G6FDG) in MCF-7 under conditions of monolayer growth was 934.6 ± 148.3 × 10^–6 mol/mg of protein per minute. At the same time in the enriched by CSC MTS the activity of G6FDG increased more than in 1.5 times. The activity of the lactate dehydrogenase (LDH) in MCF-7 cells in monolayer culture was 65.12 ± 1.28 µmol/mg of protein per minute, and in eMTS, decreased in 5.5 times. Thus, based on the analysis of the obtained data, we can assume that under conditions of enrichment of the tumor population by CSC the receptor and energy profile of MCF-7 cells changed. So, MTS are approaching to characteristics of metastatic micronode, and tumor cells are approaching to cancer stem cells.

]]> http://cytgen.com/en/2022/24-38N4V56.htm http://cytgen.com/ru/2022/24-38N4V56.htm Fri, 01 Jul 2022 00:00:00 +0300