Molecular Genetic Studies Research Articles

Pathogenetic role of a number of factors in the development and progression of preeclampsia with varying severity in pregnant women.

Context Preeclampsia is a common pregnancy complication, posing significant risks to both the mother and fetus. Predicting and determining the risks of this disease is crucial. Aims This research aims to understand the pathogenetic role of several factors in the development and progression of preeclampsia, particularly in relation to its severity in pregnant patients. Methods The study included 60 pregnant women diagnosed with either mild or severe preeclampsia and 40 healthy pregnant women for comparison. Blood plasma was analysed using biochemical methods, and blood microcirculation parameters were determined to identify homeostatic abnormalities in early preeclampsia. Key results A molecular genetic study revealed the frequency of the endothelial nitric oxide gene eNOSC774T . Homeostatic abnormalities were statistically correlated with polymorphic genotypes of the eNOSC774T gene. Conclusions The research found a correlation between the T774T eNOS genotype mutation and the severity of preeclampsia, alongside significant homeostasis abnormalities in patients. Implications The T774T mutant genotype of the eNOS gene and higher levels of lipid peroxidation products are strongly linked to the severity and progression of preeclampsia. This highlights a significant connection between genetic predisposition and biochemical abnormalities in the disease's development.

Humoral immune response to shiga toxin 2 (Stx2) in children with escherichiosis with hemolytic-uremic syndrome

Shiga toxin-producing Escherichia coli (STEC) causes acute intestinal infections and also causes acute renal failure, especially in children. Shiga toxins (Stx) occupy a central place in the pathogenesis of hemolytic uremic syndrome (HUS) in Escherichiosis. The presented work analyzes the effectiveness of laboratory diagnostics of enterohemorrhagic escherichiosis in patients at the stage of manifestation of HUS and/or acute renal failure using microbiological, immunological research methods and PCR analysis. The study used clinical material from 30 patients in the pediatric intensive care unit of the St. Vladimir Children’s City Clinical Hospital in Moscow with symptoms of HUS aged from 8 months to 5 years. Blood sera from 20 healthy donors were used as control. As a result of PCR analysis, stx2 DNA was detected in 23.3% of cases. Bacteriological research made it possible to sow a pure culture of Escherichia coli O157:H7 in only 3.3% of cases. Since the development of HUS begins in patients with acute intestinal infection caused by Shiga toxin-producing microorganisms starting from the 5th day of the disease, when antibiotic therapy is already carried out, the bacteria can be completely destroyed, which makes it difficult to identify them by bacteriological methods, as well as to detect genes encoding Shiga toxin in PCR analysis. Typically, patients with HUS are admitted to the intensive care unit 5–7 days after the onset of the disease, when class G immunoglobulins specific to the pathogen are already beginning to circulate in the blood. In this regard, the use of immunological tests can be effective to confirm the diagnosis of STEC infection. In our studies, enzyme immunoassay allowed us to detect antibodies to Stx2A in 63.3% and to Stx2B in 43.3% of patients. Using immunoblotting, antibodies to Stx2A were detected in all sera obtained from patients and in 66.7% of cases to Stx2B. Immunoblot analysis was characterized by higher sensitivity for detecting antibodies to Stx2, however, due to the presence of an immunological layer among healthy people, it is preferable to use ELISA analysis. In healthy donors with antibodies to Stx2, the antibody titer was significantly lower than in patients. Laboratory confirmation of the diagnosis of STEC infection is difficult when conducting microbiological and molecular genetic studies, which is confirmed in this work. The effectiveness of laboratory diagnostics can be expanded by performing an ELISA aimed at detecting antibodies to Stx2A.

The Ability of the Indigenous Bacteria Chromobacterium haemolyticum strain W15 to Degrade Chromium (Cr) and Lead (Pb) in Liquid Coal Waste

The use of coal as a raw material for power plants has a good economic impact, but it also has a detrimental environmental impact, particularly due to the presence of Cr and Pb, heavy metals with bioaccumulation and biomagnification qualities. Efforts to control Pb and Cr in liquid coal waste can be achieved by bioremediation. The goal of this study is to screen indigenous bacteria, identify, and test biodegradation on the best bacteria capable of degrading Cr and Pb. Bacterial screening is done experimentally in the lab. Bacterial identification is done using morphological, biochemical, and molecular genetic methods. Using atomic absorption spectroscopy to validate Cr and Pb biodegradation research. Biodegradation experiments revealed that the efficacy of indigenous bacteria reduced Pb by 216% (0.238 ppm to 0.11 ppm) and Cr by 195% (0.34 ppm to 0.174 ppm). The findings of biochemical, morphological, and molecular genetic studies revealed that the top bacterial strains were up to 96% related. using Chromobacterium haemolyticum strain W15. Chromobacterium haemolyticum strain X, an indigenous bacteria capable of degrading Cr and Pb, was successfully isolated from liquid waste.

Whole-genome sequencing of 13 Arctic plants and draft genomes of Oxyria digyna and Cochlearia groenlandica

To understand the genomic characteristics of Arctic plants, we generated 28–44 Gb of short-read sequencing data from 13 Arctic plants collected from the High Arctic Svalbard. We successfully estimated the genome sizes of eight species by using the k-mer-based method (180–894 Mb). Among these plants, the mountain sorrel (Oxyria digyna) and Greenland scurvy grass (Cochlearia groenlandica) had relatively small genome sizes and chromosome numbers. We obtained 45 × and 121 × high-fidelity long-read sequencing data. We assembled their reads into high-quality draft genomes (genome size: 561 and 250 Mb; contig N50 length: 36.9 and 14.8 Mb, respectively), and correspondingly annotated 43,105 and 29,675 genes using ~46 and ~85 million RNA sequencing reads. We identified 765,012 and 88,959 single-nucleotide variants, and 18,082 and 7,698 structural variants (variant size ≥ 50 bp). This study provided high-quality genome assemblies of O. digyna and C. groenlandica, which are valuable resources for the population and molecular genetic studies of these plants.

Enzymatic testing for mucopolysaccharidosis type I in Kuwaiti newborns: a preliminary study toward newborn screening.

Mucopolysaccharidosis type I (MPS I) is an autosomal recessive lysosomal storage disorder characterized by deficient or absent α-L-iduronidase (IDUA) enzyme activity due to pathogenic variants in the IDUA gene. Early treatment with hematopoietic stem cell transplantation and/or enzyme replacement therapy is associated with improved outcomes in this progressive multisystem disease. The diagnosis is usually delayed due to late presentation and non-specific symptoms, which result in high morbidity and mortality. The incidence of MPS I is unknown in Kuwait. This pilot study was undertaken to screen MPS I in all Kuwaiti neonates born at Farwaniya Hospital (FH), a major center in Kuwait, over 12 months. This study examined the incidence of MPS I for inclusion in the national newborn screening (NBS) to enable its early detection and adequate treatment. All Kuwaiti neonates born at FH between December 2021 and December 2022 were screened for MPS I. The screening consisted of determining IDUA enzyme activity in dried blood spot-derived samples using tandem mass spectrometry. A follow-up genetic analysis of the IDUA gene has been planned to screen the cases with diminished IDUA enzyme activity as second-tier testing. A total of 618 newborns, including 331 (54%) boys and 287 (46%) girls, were screened. Of them, 20 had deficient IDUA enzyme activity but showed negative genetic testing. However, we have diagnosed one additional female infant with MPS I who belonged to FH, but the parents chose to deliver in a private hospital. The molecular genetic study revealed the presence of a previously reported pathogenic nonsense variant in the IDUA c.1882C>T, which is associated with severe phenotype. That being included, MPS I is estimated to be approximately 0.2% of all screened cases in Kuwait. Our study is the first to evaluate the incidence of MPS I in Kuwait. Given the single center, small number of screened infants, and the short study duration thus far, it is premature to calculate the incidence. It is anticipated that as the study continues, we would be able to estimate the incidence in our population correctly. Screening newborns in all maternity hospitals in Kuwait is necessary to calculate the actual incidence of this severe disorder. Still, our preliminary data support the inclusion of MPS I in national NBS program to allow early initiation of treatment and thus improve disease outcome.

Clinical and radiological variants of the nail–patella syndrome: Why is it important to make a correct diagnosis?

BACKGROUND: Nail–patella syndrome is an autosomal dominant disease caused by LMX1B mutations, characterized by a combination of dermatological and musculoskeletal abnormalities. The classic tetrad of the nail–patella syndrome includes hypoplasia of the nail plates, absence or hypoplasia of the patella, elbow joint contractures, often accompanied by posterior subluxation of the radial head, presence of “iliac horns” on radiographs of the pelvic bones, and renal changes. However, its diagnosis and treatment are not described in sufficient detail in domestic and foreign literature. AIM: This study aimed to analyze the variability of clinical and radiological manifestations in children with nail–patella syndrome. MATERIALS AND METHODS: Nine patients with nail–patella syndrome were examined and treated (aged 4 months to 14 years). All children presented to the H. Turner National Medical Research Center for Children’s Orthopedics and Trauma Surgery with a diagnosis of congenital multiple arthrogryposis. The examination results excluded this diagnosis. The diagnosis of nail–patella syndrome has been verified. RESULTS: Various variants of the nail–patella syndrome have been identified, with lesions to only the elbow joints (n = 3), damage to the upper and lower extremities (n = 5), and damage to only the lower extremities (n = 1). All patients had hypoplasia of fingernail plates. However, the pathognomonic sign – “iliac horns” on radiographs of the pelvic bones – was noted only in 33% of cases. Regarding functional disorders of the kidneys, proteinuria was observed in one patient, which appeared at the age of 15 years. In two cases, the parents suffered from this disease. A genetic examination was performed in three patients: a pathogenic variant in LMX1B was detected in a heterozygous state. All patients underwent conservative treatment for knee and elbow joint contractures and foot deformities. When casting, the presence of dislocation of the radial head or patella was considered. Surgical treatment was performed on five patients. The results of surgery on the lower extremities (knee contractures, foot deformities, and hip dislocation) were good in 87% of the cases. Recurrence of flexion contractures in the elbow joints was observed both after conservative and after surgical treatment in all cases; however, their severity varied. CONCLUSIONS: Clinical observations were considered to attract the attention of doctors of different specialties to a rare pathology. Moreover, a molecular genetic study is recommended for the timely diagnosis of the nail–patella syndrome, determining the treatment techniques and disease prognosis.

Alström Syndrom: A Clinical Case

Background. Alström syndrome is a rare autosomal recessive disease characterized by multiple organ dysfunction. The prevalence in the population is less than one person per 1 million. Genomic mutation analysis study identified 109 new mutations increasing the number of known mutations of the ALMS1 protein to 239 which highlights the allelic heterogeneity of this disease and its phenotypic diversity.The aim of the work. Increasing awareness among medical professionals regarding Alström syndrome.Materials and methods. The analysis of the medical record of an outpatient patient, data from paraclinical research methods of the child of patient P., born in 2013, who was examined at the Children’s Regional Clinical Hospital (CRCH; Krasnodar).Results. The disease debuted with an acute respiratory infection at the age of 4 months, the primary clinical manifestations were acute respiratory failure, and therefore, the child was hospitalized at CRCH. The results of additional research methods revealed dilated cardiomyopathy with reduced global contractility of the left ventricle. For further disease diagnosis and treatment correction, the child was referred to the National Medical Research Center for Children’s Health (NMRCCH), where he was observed for several years by a multidisciplinary team of specialists. Based on results of molecular genetic studies (pathogenic homozygous mutation in the ALMS1 gene), the child was diagnosed with Alström syndrome at the age of 8 years.Discussion. This report illustrates difficulty diagnosis and lack of specific treatment.Conclusion. This clinical case demonstrates a wide range of phenotypic features of the rare hereditary Alström syndrome.

Diagnosis of metastatic melanoma by fine-needle aspiration biopsy and primary melanoma by using the skin and mucousa imprints

BACKGROUND: Determination of the cytogenetic origin and morphological type of metastatic or primary tumor determines the prescription of therapy and affects the efficiency of patient’s treatment. Currently, a number of publications evaluate the potential of cytological diagnostics of melanoma. AIM: To evaluate the significance of diagnosing metastatic melanoma by fine-needle aspiration biopsy and primary melanoma by using the skin and mucosa imprints. METHODS: In a retrospective study, a comparative analysis of cytological melanoma samples, in comparison with clinical and anamnestic information and the results of histological, immunohistochemical, and molecular genetic studies, was carried out. Information about 109 patients from the cancer registry of the Altai Regional Oncology Dispensary (Barnaul, Russia) for 2022 was used in the study. Traditional and liquid-based methods for preparing samples were used. The samples were stained using Pappenheim and Papanicolaou methods. In some observations, cytological material was used for molecular genetic studies. Using the cancer registry data of the dispensary, the results of histological and of molecular genetic studies, and a final conclusion was given about each patient. RESULTS: Fine-needle aspiration biopsy was carried out in 80 patients. Tumor smears were obtained from 29 patients. The cytological diagnose “melanoma” was consistent with the data of histological and immunohistochemical studies (p 0.001) for all 109 patients. Melanoma was diagnosed for the first time in 66 (60.5%) patients. In other cases, the process of progression was noted within the period from one year to 20 years. Epidermal melanoma was noted in 101 (92.7%) cases, including 9 patients with acral melanoma, and 2 cases with localization on the vulva. Melanomas of mucosa were found in 5 cases (4.5%): in the rectum and anal canal, vagina, and in 2 cases on the hard palate. Non-epidermal (uveal) melanomas metastases were diagnosed in liver by fine-needle aspiration biopsy in 3 patients (2.8%). Based on the cellular composition, epithelioid cell melanoma was determined in 81 (74.3%) patients, mixed cell in 14 (12.8%) cases, spindle cell in 9 (8.2%), pigmentless in 2 (1.8%) cases and nevoid melanoma in 1 (0.9%) case. The mutation status was determined in 96 patients (88.1%). Of these, in 8 patients it was determined using cytological material. In epidermal melanomas, mutations in codon 600 of exon 15 of the BRAF gene were found in 43 (44.8%) patients, including 2 cases of acral melanoma. Mutations V600K, V600E/Ec were found in one patient each. In mucousal melanomas: Q61R mutation was found in exon 3 of the NRAS gene (vaginal melanoma), G12C mutation was identified in exon 2 of the NRAS gene (anal canal melanoma). In uveal melanomas, the assessed mutations were absent (it is necessary to determine mutation is GNAQ11 and BAP1 genes). CONCLUSION: Cytological diagnosis of melanoma by fine-needle aspiration biopsies and imprints from the tumor mass is a highly informative method that allows diagnosing melanoma and verifying the tumor subtype. The obtained results indicate tumor heterogeneity and differences in mutational status depending on the location of melanomas. The molecular classification of melanoma is important when choosing individualized therapy.

Genetic Confirmation of the Striped Eel Catfish Plotosus lineatus (Thunberg, 1787) from Iskenderun Bay (Eastern Mediterranean, Türkiye)

The records of the striper eel catfish Plotosus lineatus have been assigned based on morphological characters in the Mediterranean Sea. However, molecular and genetic analyses are needed to confirm the morphological taxonomic determination and avoid misidentification. In this study, the mitochondrial cytochrome c oxidase subunit I (COI) gene region was used for genetic confirmation of Plotosus lineatus in the Mediterranean Sea. The obtained P. lineatus sequences were found to have a 100% identity match with the Red Sea and Mediterranean records. The present molecular genetic study confirms the presence of P. lineatus in Turkish marine waters. Also, this study is the first genetic confirmation of P. lineatus on the coast of Türkiye and in the Mediterranean Sea.

Antibiotic Resistance in Acetic Acid Bacteria Originating from Vinegar.

Acetic acid bacteria (AAB) are major contributors to the production of fermented vinegar, offering various cultural, culinary, and health benefits. Although the residual unpasteurized AAB after vinegar production are not pathogens, these are necessary and require safety evaluations, including antibiotic resistance, before use as a starter. In this research, we investigated the antibiotic resistance profiles of 26 AAB strains, including various species of Komagataeibacter and Acetobacter, against 10 different antibiotics using the E-test method. All strains exhibited resistance to aztreonam and clindamycin. Komagataeibacter species demonstrated a 50% resistance rate to ciprofloxacin, analogous to Acetobacter species, but showed twice the resistance rates to chloramphenicol and erythromycin. Genomic analysis of K. saccharivorans CV1 identified intrinsic resistance mechanisms, such as multidrug efflux pumps, thereby enhancing our understanding of antibiotic resistance in acetic acid-producing bacteria. These findings enhance understanding of antibiotic resistance in AAB for food safety and new antimicrobial strategies, suggesting the need for standardized testing methods and molecular genetic study.

Characterization and novel gene rearrangements in the mitochondrial genome of Chelonus munakatae (Hymenoptera: Braconidae): Insights into evolutionary dynamics and genomic variability

Chelonus munakatae Matsumura (Hymenoptera: Braconidae) is an important parasitoid of several lepidopteran species. The mitochondrial genome of this species is a crucial source for molecular identification and genetic studies. Here, we used next-generation sequencing to obtain the C. munakatae mitochondrial genome, which was found to be 14,868 bp in length and contained 13 protein-coding genes, 22 tRNAs and two rRNAs. The nucleotide composition exhibits a significant bias towards A + T content, with a value of 86.8 %. A large number of PCG and tRNA gene rearrangement events were detected (I-V-M-C-W-Q and A-N-S1-E-R-F), which represent a novel rearrangement type in Hymenoptera. Furthermore, synteny analysis based on the whole mitochondrial genomes showed that a significant rearrangement of homologous blocks had occurred within this Braconidae species. This study not only delineates the mitochondrial characteristics of C. munakatae but also significantly contributes to the repository of mitochondrial genetic information for the Braconidae family. Moreover, it highlights the occurrence of gene rearrangement within Hymenoptera species, offering valuable insights into the evolutionary dynamics and genomic variability of these insects.

Clinical characterizations and molecular genetic study of two co-segregating variants in PDZD7 and PDE6C genes leading simultaneously to non-syndromic hearing loss and achromatopsia

BackgroundAutosomal recessive non-syndromic hearing loss (NSHL) and cone dystrophies (CODs) are highly genetically and phenotypically heterogeneous disorders. In this study, we applied the whole exome sequencing (WES) to find the cause of HL and COD in an Iranian consanguineous family with three affected individuals.MethodsThree members from an Iranian consanguineous family who were suffering from NSHL and visual impairment were ascertained in this study. Comprehensive clinical evaluations and genetic analysis followed by bioinformatic and co-segregation studies were performed to diagnose the cause of these phenotypes. Data were collected from 2020 to 2022.ResultsAll cases showed congenital bilateral NSHL, decreased visual acuity, poor color discrimination, photophobia and macular atrophy. Moreover, cornea, iris and anterior vitreous were within normal limit in both eyes, decreased foveal sensitivity, central scotoma and generalized depression of visual field were seen in three cases. WES results showed two variants, a novel null variant (p.Trp548Ter) in the PDE6C gene causing COD type 4 (Achromatopsia) and a previously reported variant (p.Ile84Thr) in the PDZD7 gene causing NSHL. Both variants were found in the cis configuration on chromosome 10 with a genetic distance of about 8.3 cM, leading to their co-inheritance. However, two diseases could appear independently in subsequent generations due to crossover during meiosis.ConclusionsHere, we could successfully determine the etiology of a seemingly complex phenotype in two adjacent genes. We identified a novel variant in the PDE6C gene, related to achromatopsia. Interestingly, this variant could cooperatively cause visual disorders: cone dystrophy and cone-rod dystrophy.

Expanding the Spectrum of NUTM1 -Rearranged Sarcoma : A Clinicopathologic and Molecular Genetic Study of 8 Cases.

Apart from the lethal midline carcinoma (NUT carcinoma), NUTM1 translocation has also been reported in mesenchymal tumors, but is exceedingly rare. Here, we describe a series of 8 NUTM1 -rearranged sarcomas to further characterize the clinicopathologic features of this emerging entity. This cohort included 2 males and 6 females with age ranging from 24 to 64 years (mean: 51y; median: 56y). Tumors occurred in the colon (2), abdomen (2), jejunum (1), esophagus (1), lung (1) and infraorbital region (1). At diagnosis, 6 patients presented with metastatic disease. Tumor size ranged from 1 to 10.5cm (mean: 6cm; median: 5.5 cm). Histologically, 4 tumors were composed of primitive small round cells to epithelioid cells intermixed with variable spindle cells, while 3 tumors consisted exclusively of small round cells to epithelioid cells and 1 tumor consisted predominantly of high-grade spindle cells. The neoplastic cells were arranged in solid sheets, nests, or intersecting fascicles. Mitotic activity ranged from 1 to 15/10 HPF (median: 5/10 HPF). Other features included rhabdoid phenotype (4/8), pronounced nuclear convolutions (2/8), prominent stromal hyalinization (2/8), focally myxoid stroma (1/8), foci of osteoclasts (1/8), and necrosis (1/8). By immunohistochemistry, all tumors showed diffuse and strong nuclear staining of NUT protein, with variable expression of pancytokeratin (AE1/AE3) (2/8), CK18 (1/8), CD99 (3/8), NKX2.2 (2/8), cyclin D1 (2/8), desmin (2/8), BCOR (2/8), S100 (1/8), TLE1 (1/8), and synaptophysin (1/8). Seven of 8 tumors demonstrated NUTM1 rearrangement by fluorescence in situ hybridization analysis. RNA-sequencing analysis identified MXD4::NUTM1 (3/7), MXI1::NUTM1 (3/7), and MGA::NUTM1 (1/7) fusions, respectively. DNA-based methylation profiling performed in 2 cases revealed distinct methylation cluster differing from those of NUT carcinoma and undifferentiated small round cell and spindle cell sarcomas. At follow-up (range: 4 to 24mo), 1 patient experienced recurrence at 8.5 months, 4 patients were alive with metastatic disease (5, 10, 11, and 24mo after diagnosis), 3 patients remained well with no signs of recurrence or metastasis (4, 6, and 12mo after diagnosis). Our study further demonstrated that NUTM1 -rearranged sarcoma had a broad range of clinicopathologic spectrum. NUT immunohistochemistry should be included in the diagnostic approach of monotonous undifferentiated small round, epithelioid to high-grade spindle cell malignancies that difficult to classify by conventional means. DNA-based methylation profiling might provide a promising tool in the epigenetic classification of undifferentiated sarcomas.

Associations of the TNFSF11 gene polymorphisms with knee osteoarthritis in post-menopausal women

Objective: to determine the role of the TNFSF11 gene polymorphisms rs9594738 and rs9594759 in the development of knee osteoarthritis in postmenopausal women. Material and methods. The case-control study included 483 postmenopausal women. Of these 157 were diagnosed with primary knee osteoarthritis. The remaining 326 women without signs of joint disease were included in the control group. Molecular genetic studies included determination of TNFSF11 rs9594738 and TNFSF11 rs9594759 gene polymorphisms using the real-time polymerase chain reaction. Results. Analysis of genetic markers distribution in two groups of women showed a decrease in the frequency of rs9594738 polymorphism TT genotype among the patients with osteoarthritis (odds ratio 0.59; 95% confidence interval 0.36-0.97; p=0.049). The distribution of this polymorphism alleles, as well as alleles and genotypes of TNFSF11 rs9594759 gene polymorphism in the group of women with osteoarthritis did not differ significantly from the results of the control group (p>0.05). Conclusion. An association of TNFSF11 rs9594738 gene polymorphism with knee osteoarthritis in postmenopausal women has been established. Further research of TNFSF11 gene polymorphic variants role in the knee osteoarthritis etiopathogenesis are needed to develop individual approaches to the prevention and treatment of the above disease.

Search for molecular markers of joint hypermobility

Background: Joint hypermobility (JH) is a condition that can be either benign or accompanied by pain and early development of osteoarthritis. For early verification of the risk of complications, it is necessary to study the molecular pathogenesis of various JH variants. The aim of the study: The aim of this study was to search for candidate genes involved in connective tissue metabolism and causing joint hypermobility in a form isolated or combined with connective tissue dysplasia (CTD). Materials and methods: The study involved young (21,86±0,22 y.o.) people (n=181). A clinical study was conducted for the presence of signs of JH (Beighton scale) and CTD (tables Kadurina T.I.). Further, a molecular genetic study was carried out and the search for associations of polymorphic variants of genes (vitamin D receptor (VDR), lumican (LUM), tenascin (TNXB), growth/differentiation factor 5 (GDF5), magnesium ion channel TRPM6), bone morphogenetic protein type 5 (BMP5)) and JH, CTD in isolated and combined forms. Statistical data processing was carried out by comparing qualitative features using the χ2 test, Fisher's exact test, and the χ2 test with Yates' correction for contingency tables 2x2. The strength of associations was assessed in terms of the odds ratio (Odds Ratio, OR, at p<0.05), the correction for multiple comparisons was carried out using the FDR method (Benjamin- Hochberg). Quantitative data were evaluated in pairs using the Student's t-test, the normality of distribution according to the Kolmogorov-Smirnov test. Results: Associations of the G allele and the GG genotype of the rs3734444 locus of the BMP5 gene with the presence of JH (OR=3.70 and OR=5.10), the A allele of the rs1470527 locus of the BMP5 gene and the presence of JH+CTD- (OR=8.00), the T allele and the TT genotype of the rs11144134 locus of the TRPM6 gene with JH in isolated (OR=3.00 and OR =10.19) and in the forms combined with CTD (OR=3.17 and OR=11.28), the TT genotype was also associated with the isolated form of CTD (OR=3.74). The GT genotype of the rs73611720 locus of the GDF5 gene was associated with isolated CTD (OR=4.15). No statistically significant associations were found for polymorphic variants rs11540149 (VDR), rs2268578 and rs3759222 (LUM), rs3130342 (TNXB). Conclusion: The G allele of locus rs3734444 of the BMP5 gene is a potential risk marker for JH in general, the A allele of locus rs1470527 is an isolated JH, the GT genotype of locus rs73611720 of the GDF5 gene is a marker for isolated CTD, the T allele of locus rs11144134 of the TRPM6 gene is a marker for JH combined with CTD.

Identity of the pathogenesis, genetic and epigenetic mechanisms of osteoarthritis and rheumatoid arthritis development

Osteoarthritis is characterized by heterogeneity of clinical manifestations and, in some cases, a severe progressive course. In this regard, it is important to identify new molecular targets for the treatment of the disease. To determine the role of autoimmune processes, general genetic and epigenetic changes in osteoarthritis and rheumatoid arthritis, as well as to identify osteoarthritis-specific ribonucleic acids (microRNAs), potential targets for targeted therapy, information was searched using scientific platforms PubMed, Scopus, ResearchGate, RSCI over the past 10 years. Although the pathogenesis of rheumatoid arthritis and osteoarthritis differs, evidence has been obtained that identical pathological immune reactions are involved in the mechanism of osteoarthritis and disruption of the expression of 26 identical genes with identical changes in the levels of 13 of them. Changes in the expression of the same microRNAs (miR-140, miR-149, miR-25, miR-146a, miR-16, miR-23b) were detected in osteoarthritis and rheumatoid arthritis. Molecular genetic studies make it possible to find new markers of pathological immune reactions in osteoarthritis, which can be used to treat the disease and prevent its rapid progression, as well as to design targeted therapy using gene expression products as targets. MicroRNAs associated with osteoarthritis and rheumatoid arthritis and involved in the pathogenesis of both diseases may become promising targets for targeted therapy of osteoarthritis and rheumatoid arthritis.

The first Genetic data on <i>Lernaea Cyprinacea</i> Linnaeus, 1758 from the European North of Russia (The Pinega River)

Morphological and molecular genetic data about Lernaea cyprinacea Linnaeus, 1758, collected from pike (Esox lucius Linnaeus, 1758) in the European North of Russia (Pinega river) in 2022, were presented. Molecular genetic studies were carried out based on the analysis of COI mtDNA, 18S rDNA and 28S rDNA gene sequences. A significant genetic distance for the COI gene from individuals of the same species collected in other territories (China, Australia, South Africa, Canada, Austria, Argentina, Pakistan, Iran, India, Japan, Romania, Egypt and Western Siberia) was revealed, which indicates the existence of a cryptic species L. cyprinacea in the European North of Russia. Analysis of 28S rDNA sequences showed the presence of two substitutions fixed in the population, which may indicate a fairly long-standing divergence of sister species. The 18S rDNA gene fragment is less suitable for identifying L. cyprinacea due to low variability compared to the other two gene regions.

Generation of rescued Japanese encephalitis virus genotype 1 from infectious full-size clone using reverse genetics

Japanese encephalitis virus (JEV) is a pathogen responsible for high mortality and morbidity rates among children with encephalitis. Since JEV genotype 1 (GI) is the most prevalent strain in South Korea these days, corresponding research and vaccine development is urgently required. Molecular genetic studies on JEV vaccines can be boosted by obtaining genetically stable full-length infectious JEV complementary DNA (cDNA) clones. Furthermore, the significance of the reverse genetics system in facilitating molecular biological analyses of JEV properties has been demonstrated. This study constructed a recombinant JEV-GI strain using a reverse genetics system based on a Korean wild-type GI isolate (K05GS). RNA extracted from JEV-GI was used to synthesize cDNA, a recombinant full-length JEV clone, pTRE-JEVGI, was generated from the DNA fragment, and the virus was rescued. We performed in vitro and in vivo experiments to analyze the rescued JEV-GI virus. The rescued JEV-GI exhibited similar characteristics to wild-type JEV. These results suggest that our reverse genetics system can generate full-length infectious clones that can be used to analyze molecular biological factors that influence viral properties and immunogenicity. Additionally, it may be useful as a heterologous gene expression vector and help develop new strains for JEV vaccines.

Drought Has a Greater Negative Effect on the Growth of the C3Chenopodium quinoa Crop Halophyte than Elevated CO2 and/or High Temperature.

Plant growth and productivity are predicted to be affected by rising CO2 concentrations, drought and temperature stress. The C3 crop model in a changing climate is Chenopodium quinoa Willd-a protein-rich pseudohalphyte (Amaranthaceae). Morphophysiological, biochemical and molecular genetic studies were performed on quinoa grown at ambient (400 ppm, aCO2) and elevated (800 ppm, eCO2) CO2 concentrations, drought (D) and/or high temperature (eT) treatments. Among the single factors, drought caused the greatest stress response, inducing disturbances in the light and dark photosynthesis reactions (PSII, apparent photosynthesis) and increasing oxidative stress (MDA). Futhermore, compensation mechanisms played an important protective role against eT or eCO2. The disruption of the PSII function was accompanied by the activation of the expression of PGR5, a gene of PSI cyclic electron transport (CET). Wherein under these conditions, the constant Rubisco content was maintained due to an increase in its biosynthesis, which was confirmed by the activation of rbcL gene expression. In addition, the combined stress treatments D+eT and eCO2+D+eT caused the greatest negative effect, as measured by increased oxidative stress, decreased water use efficiency, and the functioning of protective mechanisms, such as photorespiration and the activity of antioxidant enzymes. Furthermore, decreased PSII efficiency and increased non-photochemical quenching (NPQ) were not accompanied by the activation of protective mechanisms involving PSI CET. In summary, results show that the greatest stress experienced by C. quinoa plants was caused by drought and the combined stresses D+eT and eCO2+D+eT. Thus, drought consistently played a decisive role, leading to increased oxidative stress and a decrease in defense mechanism effectiveness.

Genetic characteristics of producing bulls of the Kholmogorsky breed, depending on their linear affiliation and the level of Holstein

Relevance. Genetic analysis using STR markers makes it possible to effectively use various techniques to control the breeding process in a population.Methods. The material for DNA research was the deep-frozen sperm of 103 bulls-producers of the Kholmogorsky breed, with different blood levels of Holstein cattle belonging to five main genealogical lines. Molecular genetic studies were conducted in the laboratory of DNA technologies at VNIIPlem.Results. The animals of the VBA (I = 1,557) and Nl (I = 1,556) lines had the maximum genetic diversity, and the Chl lines had the lowest (the indicator is lower by 0.229 (р ≤ 0.05) relative to VBA). A pronounced discrepancy between the observed and expected heterozygosity towards heterozygote deficiency was found among animals of the VBA line (F = 0.173), which is higher by 0.220–0262 relative to RS and Chl (р ≤ 0.01–0.001). The bulls of the Chl and RS lines have an equilibrium shift towards an excess of heterozygosity (F = -0.111 and F = -0.054 respectively). The most related lines are Nl and VBA (DN = 0.097), as well as Nl and Lm (DN = 0.101), and the least VBA and Chl (DN = 0.247), as well as VBA and RS (DN = 0.218). When differentiating the samples by the level of Holstein, it was found that the greatest genetic diversity was characterized by bulls with a blood density of 25–49% (1,633), and the minimum was animals with a degree of Holstein above 74% (which is lower by 0.299–0.334 relative to other samples, р ≤ 0.05–0.01). With an increase in the degree of Holstein, an increase in the level of genetic inbred was observed (in crossbreeds with a bloodline above 74%, the indicator was 0.142–0.229 more (р ≤ 0.05–0.001) compared with other samples).