Genetic Research Methods Research Articles

Syndromic obesity in children (using the example of clinical cases)

Obesity is a heterogeneous group of hereditary and acquired diseases associated with excessive accumulation of adipose tissue in the body. One example of syndromic obesity in children is Bardet-Biedl syndrome. This is a rare autosomal recessive disease from the group of ciliopathies, characterized by retinal dystrophy, obesity, polydactyly, mental retardation, hypogonadism, and renal dysfunction. The article presents two clinical cases of Bardet-Biedl syndrome. Diagnostic criteria for the disease are given, and the need for molecular genetic research methods in the early stages of the diagnostic search is shown. Promising directions in the treatment of the syndrome are considered.

Characteristics of risk factors and molecular genetic parameters in children with chronic kidney disease

Introduction. Information about the health status of children with nephrological pathology includes many manifestations of functional system disorders. However, early diagnosis of chronic kidney disease (CKD) is rarely carried out due to the lack of reliable predictors of this process.The aim: To analyze the association of anamnestic signs, laboratory and genetic research results with chronic kidney disease in children to identify potential predictors of the disease.Material and Methods. The data were obtained from a single-center retrospective catamnestic cohort study (2011–2022). The main group included 128 children with chronic kidney disease aged 1 to 17 years. The comparison group consisted of 30 children without diagnosed kidney pathology aged from 1 to 17 years. There were no statistically significant differences in sex and age between the two groups. The informative value of more than 150 features, including anamnestic ones, the state of the mother’s health during pregnancy, antenatal and perinatal factors, features of the child’s development and manifestation of the disease, the results of clinical, laboratory, and genetic research methods were evaluated. An integrated approach to the study included methods for hypotheses testing on statistical significance of indicators differences, regression analysis, and ROC analysis.Results. Hereditary burden of nephrological pathology in the family, unfavorable course of pregnancy (anemia, toxicosis of the first half), obstetric and somatic pathology in the mother, the serial number of childbirth are associated with the subsequent development of CKD. Erythrocytes in the urine and daily protein loss were detected only in the group of children with CKD. In this group, a decrease in the number of red blood cells, increased erythrocyte sedimentation rate (ESR), allergies at an early age, otitis media, persistent viral and bacterial infections, and a number of signs of connective tissue dysplasia were more common. The main group statistically significantly differed from the comparison group by a number of polymorphic genetic markers. It can be assumed that the polymorphisms AGT1 Thr174Thr, AGT2 Thr235Thr, NO3 C786T are associated with a predisposition to CKD, and the products of polymorphisms AGT1 Thr174Met, AGT2 Met235Met, R1 AGT2 C1166C, NO3 T786T may have a protective effect according to development of CKD.Conclusion. The risk factors for CKD identified in this study can be used to develop software to support medical decision making for early detection of children at high risk of disease progression.

Хвороба Байлера: консервативні і хірургічні стратегії лікування

Byler's disease (BD) is rare and difficult for diagnosing hereditary cholestatic liver disease. Proper diagnosis, early correction and treatment (including liver transplantation (LT)) improves the quality of life (QoL), prevents irreversible liver changes, multiple organ failure and death. Aim - to study the features of differential diagnostics, feasibility, terms, strategies, methods of conservative treatment, surgical correction (palliative surgery (PS), LT) in five children with BD. Materials and methods. Five clinical episodes of chronic disease with data on hematological indicators during treatment, physical development (PD) (harmony, body mass index - BMI), neuropsychiatric development (NPD), QoL before treatment, after PS, after LT. Results. Five clinical episodes of chronic disease were analyzed, one of which was fatal. All indicators (hematological, FD, NPD, QoL) improved after PS, but completely normalized after LT. Conclusions. Differential diagnosis of severe hereditary rare disease progressive familial intrahepatic cholestasis (BD) is possible using genetic research methods as a marker of this disease among all cholestatic diseases in young children. The first clinical signs of the disease appear in the neonatal period and progress in the first months of life. Early modern diagnosis and diversion of bile prevent liver damage and death. Palliative surgical intervention is performed in the case of rapid progression of cirrhosis and the threat of liver failure as a temporary measure to better prepare the patient for transplantation and obtain the results of genetic analysis, carry out the necessary vaccinations and correct concomitant congenital and acquired diseases, improve PD, NPD, QoL, biochemical indicators, ultrasound data. LT is an effective method of radical treatment of BD, normalization of FD, NPD, QoL, biochemical and sonographic indicators. The research was carried out in accordance with the principles of the Declaration of Helsinki. Informed consent of the child and child's parents was obtained for the research. The author declares no conflict of interest.

Possibilities of using nested PCR-RFLP for taxonomic identification of L3 larvae of the family Trichostrongylidae, Leiper, 1912

The purpose of the research is to apply molecular genetic research methods to identify the taxonomic affiliation of gastrointestinal parasitic sheep nematodes of the family Trichostrongylidae using nested PCR followed by the restriction fragment length polymorphism (RFLP) analysis.Materials and methods. Parasitic nematodes, L3 Strongylata larvae obtained from incubated fecal samples of sheep. The genomic DNA was isolated using a commercial kit for DNA extraction from micro-quantities of tissues (Synthol, Moscow) as per the manufacturer’s guidelines. For DNA amplification, a T-100 Bio-Rad thermal cycler and a commercial Eurogen Master Mix reagent kit were used. The PCR regime was performed according to the WAAVP guidelines, 2006. The restriction endonuclease Rsa I of amplified Trichostrongylidae fragments was performed according to guidelines of the enzyme manufacturer (Sibenzyme, Novosibirsk).Results and discussion. To determine the taxonomic affiliation of Strongylata larvae isolated after incubation of feces from sheep, molecular genetic studies were performed using nested PCR followed by the restriction fragment length polymorphism (RFLP) analysis. This method makes it possible to identify, with the least effort, the genotypes of three species of Strongylata Haemonchus contortus, Trichostrongylus colubriformis, and Teladorsagia circumcincta at the larval stage.

THE FREQUENCY OF DETECTED PATHOGEN GENOMES IN CHILDREN WITH ACUTE RESPIRATORY INFECTIONS

Due to the high prevalence of acute respiratory infections in children, it was of interest to determine the composition of bacterial and viral-bacterial associations for respiratory tract inhabiting microorganisms. To identify respiratory tract pathogens, especially viruses and unculturable bacteria, it is advisable to use molecular genetic research methods particularly PCR. The work was aimed at comparatively analyzing rate of genome detection for respiratory tract pathogens in children with acute respiratory infections and apparently healthy controls. Material and methods. Children (97 people) were examined, of which 35 people – with acute respiratory infections of the upper respiratory tract (nasopharyngitis, pharyngitis, laryngitis) (ARI URT), 32 people - with acute respiratory infections of the lower respiratory tract (acute bronchitis, acute obstructive bronchitis) (ARI NDP) and 30 people - apparently healthy control at the time of examination. RNA and DNA of viral and bacterial pathogens were assessed in the oropharyngeal smears using PCR. Test systems produced by the Federal Budgetary Institution Central Research Institute of Epidemiology of Rospotrebnadzor and JSC Vector-Best were used. Results. An analysis of detection rate for DNA and RNA of bacterial and viral pathogens showed the presence of a wide range of them (20 different species/strains out of 27 identified) in oropharyngeal biomaterial from all examined children. The peak diversity and detection rate of pathogen genomes were found in apparently healthy subjects and children with ARI UDP (DNA of members from Herpesviridae family, S.pneumoniae, H.influaenzae, MSSA, MRCoNS). Associations of 4-6 pathogens prevailed in the microflora, and the “core” of both bacterial and viral-bacterial associations was the combination of S. pneumoniae and H. influaenzae (in 93.3% and 60.0% of cases, respectively). In bacterial associations, staphylococci (MRSA and MRCoNS) and P. aeruginosa were also detected in healthy children, MSSA and other very rarely bacterial types were detected in children with ARI UDP. In children with NPD infections, the majority of whom (75.0±7.7%) were classified as long-term ill and had a complicated disease course, a noticeable depletion (decrease in taxonomic diversity) of the nasopharyngeal microbiota was found. Conclusion. The presented data indicate the presence in the biomaterial of the examined children of molecular genetic markers of a wide range of pathogens presented as viral-bacterial and bacterial associations, with varying composition being related to patient clinical status. Detection of pathogen genomes is important for choosing proper etiotropic therapy at early disease stages as well as prescribing probiotic drugs that can restore a balance in respiratory tract microflora.

Морфофункциональные изменения СОЖ откормочных свиней при хеликобактериозе

Abstract. The purpose is to study the features of morphofunctional changes in the gastric mucosa of fattening pigs in gastric ulcer disease and to establish a possible correlation between pathological changes and colonization of the gastric mucosa of H. suis. Methods. The study is based on pathoanatomic, microscopic, molecular genetic and histological research methods. Results. In 86 % of cases, significant pathomorphological changes in the gastric mucosa of pigs were detected during a pathoanatomical autopsy. Spiral-shaped microorganisms in smears were found in 28 %, H. suis DNA on the gastric mucosa – in 76 %. In the examined samples in which H. DNA was found. suis in the gastric mucosa, 76.3 % revealed various pathologies in the esophagus, 92% revealed various pathologies on the gastric mucosa. In 24 %, when H. suis DNA was not detected, 33.3 % showed no visible damage and 66.7 % revealed various pathologies. The results of the correlation between morphological changes in the gastric mucosa and its infection with helicobacter reached the limits of statistical reliability. Histological methods revealed that active chronic gastritis was detected in 48.0 %, inactive chronic gastritis was detected in 34.0 % and the norm (almost the norm) was found in 8.0 %. Chronic gastritis during exacerbation was reliably diagnosed in 55.2 % of cases when H. suis DNA was detected on the gastric mucosa. Chronic gastritis during remission was diagnosed in 34.2 % of cases when H. suis DNA was detected on the gastric mucosa. The results presented in the work reflect the undoubted connection between the presence of H. suis and the development of gastritis in pigs.

Hospital environment microbiome

The aim of the review is to give a brief description of the biodiversity and structure of the hospital environment microbiome based on molecular genetic research methods. Until a certain time, studies of the hospital environment microbiota for the purposes of epidemiological surveillance and control of healthcare-associated infections (HAIs) were based on routine microbiological identification of clinically relevant bacterial taxa. Discovery of DNA, the development of sequencing technologies, PCR and cloning techniques enabled the investigation of microbial communities using cultivation-independent, DNA and RNA-based approaches. At the current level of knowledge, the hospital environment can be considered as a superorganism with its own microbiome. Multiomic technologies, including meta-transcriptomic, meta-proteomic and metabolomic approaches, provide detailed information about microbial activity in the environment. Now it has been established that there is a stable core of the hospital microbiome where the vast majority of microorganisms are necessary for the functioning of the hospital ecosystem and are not classified as human pathogens. The hospital microbiome has a homogeneous structure composed by a massive dominance of a few taxa and microbial network with low connectivity forming a clustered topology. A keystone species is a taxon whose importance for maintaining community structure is relatively higher than others and its identification is of paramount importance. Due to the lack of knowledge of the hospital environment microbiome by molecular genetic technologies, there is no single shared point of view on the microbial diversity in different healthcare facilities. But there is no doubt that molecular genetic technologies will shed light on the evolution of hospital strains and determine which indicators are the most informative for monitoring and prognosis of HAIs.

Clinical course of respiratory tuberculosis relapses in the Kazakh population depending on HLA-DRB1 gene alleles.

The study aimed to ascertain the prevalence and frequency of relapses of respiratory tuberculosis (TB) and examine the characteristics of its clinical progression in members of the Kazakh population based on the alleles of the HLA-DRB1 gene. Methods of clinical and genetic research, statistical processing and analysis of the obtained data were used to achieve this goal. The research led to an analysis of the statistical processing of clinical and genetic investigations that found out how often TB in the respiratory tract is found and how different HLA-DRB1 gene alleles affect the disease's progression. To find out how exposure to certain HLA-DRB1 gene alleles affects the chance of relapse, the number of times they were found was compared between people who had relapsed and people who had just been diagnosed with TB. The impact of these alleles on the progression of the disease was assessed based on their frequency of detection of different clinical forms of TB (infiltrative, fibro-cavernous, generalised, disseminated), unilateral and bilateral lung damage, lung tissue deterioration, and the presence of bacterial secretions. The highest detection rate for all comparisons had gene alleles HLA-DRB1*01 (9.5%), *08 (4.2%), *15 (3.9%), *09 (1.6%), *12 (1.3%), *13 (0.9%), *11 (0.2%). The study found that Kazakhs with the HLA-DRB1*01, *08, and 15 gene alleles are more likely to develop recurrent respiratory TB. The study's practical value lies in its potential to utilise its findings for the prompt identification and eradication of genetic variables contributing to the recurrence of TB.

РОЛЬ МОЛЕКУЛЯРНО-ГЕНЕТИЧЕСКИХ МЕТОДОВ ИССЛЕДОВАНИЙ В ЭТИОПАТОГЕНЕЗЕ ОСТЕОМИЕЛИТА ЧЕЛЮСТЕЙ

The subject of the study is the importance of molecular genetic research methods in the study of the etiopathogenesis of osteomyelitis of the jaws. The purpose of the work is to provide up-to-date information to researchers, dental surgeons, and maxillofacial surgeons on the possibilities of molecular genetic research in identifying bacterial pathogens in osteomyelitis of the jaws, as well as to reflect genetic markers of pathogenicity factors for a number of the main causative agents of the disease. Methodology. International scientific databases PubMed, ScienceDirect, Scopus, Cochrane Collaboration, Elsevier, as well as electronic catalogs eLIBRARY.RU and CyberLeninka.ru were used. Results. A review of publications demonstrated that S. aureus and S. Epidermidis dominate the etiological spectrum of causative agents of bone tissue infections. The participation of these microorganisms is determined by a whole range of pathogenicity factors. Toxins and Panton-Valentine leukocidin (PVL) genes play a major role in the pathogenesis of osteomyelitis and disease progression. It has been shown that the pathogenic bacteria Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans are capable of inducing differentiated production of cytokines. The most attention has been attracted to E. faecium, which exhibits multidrug resistance to a wide range of antibiotics. The proportion of infections mediated by S. epidermidis and S. Saprophyticus is, on average, about 25% of cases. The proportion of representatives of gram-negative flora Escherichia, Klebsiella, Enterobacter, Citrobacter, Proteus, Providencia, Serratia reaches 23% of cases. Pathogenic nosocomial strains of P. aeruginosa are also involved in the formation of chronic inflammation in osteomyelitis. According to the results of published studies, more than a third of cases of chronic osteomyelitis are mediated by microbial associations, which are dominated by S. aureus, S. epidermidis and, less commonly, E. faecalis. Conclusions. The use of PCR analysis to identify the causative agents of osteomyelitis and gene amplification using specific primers has a huge advantage over routine microbiological tests, being an informative method for studying the pathogenicity factors of the main pathogens. The high importance of molecular genetic methods in the study of the etiopathogenesis of osteomyelitis of the jaws requires their widespread use in the clinic of surgical dentistry and maxillofacial surgery to successfully solve complex problems in the rehabilitation of patients with this disease.

Эпидемиологические аспекты формирования новых рисков заболеваемости и лечения туберкулеза среди населения.

In accordance with the instruction of the Minister of Health of the Russian Federation No. 19 dated 02/07/2023, specialists currently face several important tasks: from 2015 to 2025, to reduce the incidence of tuberculosis, as well as the number of deaths from tuberculosis by 75%. To do this, it is necessary to identify all the risk factors that may create difficulties in achieving these goals. Purpose of the study is to characterize the epidemiological aspects of the formation of new risks of morbidity and treatment of tuberculosis among the population. Materials and methods. An analysis was carried out among newly identified patients with tuberculosis by molecular genetic (n=1682) and bacterial (n=1630) research methods in the period from 2017 to 2020 according to drug resistance indicators. Results. It was found that in the period from 2015 to 2017, the indicators of drug resistance in tuberculosis patients were almost at the same level. The leading antimicrobial drugs in demand among the population were ceftriaxone, azithromycin, levofloxacin, moxifloxacin. The maximum increase in sales in 2020 relative to 2019 data to 150% was determined among quinolone derivatives of moxifloxacin and levofloxacin, which began to be used in the treatment of coronavirus infection. At the same time, these drugs are traditionally used to treat tuberculosis, but in 2020, an alarming trend was revealed, which was that drug resistance was detected in patients with tuberculosis to these drugs. Among newly diagnosed tuberculosis patients, from 2018 to 2020, an increase in the proportion of patients with primary drug resistance to levofloxacin and moxifloxacin was established by 189-480%; whereas for other antibiotics, the increase averaged up to 60.8%. Findings. The findings suggest an alarming scenario in which drug resistance develops towards highly virulent and highly resistant to antibacterial drugs genotypes.

РАСПРЕДЕЛЕНИЕ ГЕНОТИПОВ ГЕНА SLC2A9, УРОВЕНЬ МОЧЕВОЙ КИСЛОТЫ И МЕТАБОЛИТЫ ПУРИНОВОГО ОБМЕНА У ПАЦИЕНТОВ С АРТЕРИАЛЬНОЙ ГИПЕРТЕНЗИЕЙ И ФИБРИЛЛЯЦИЕЙ ПРЕДСЕРДИЙ

The aim is to investigate the frequency of genotypes and alleles of the SLC2A9 gene rs734553 polymorphism, the level of serum uric acid (sUA) and metabolites of purine metabolism in patients with arterial hypertension (AH) and atrial fibrillation (AF), as well as in healthy individuals. Material and methods. The study included 154 patients: 50 were healthy individuals (group 0), of which 22 (44%) were men and 28 (56%) were women aged 50 [45;53] years and 104 were patients with AH and AF (main group), of which 94 (90.4%) were men and 10 (9.6%) were women aged 55 [45; 61] years. The main group was divided into subgroups: subgroup I – patients with AF without a history of AH and other rhythm disorders (n=13); subgroup II – patients with AH in combination with AF (n=68); subgroup III – patients with AH without a history of AF or other rhythm disturbances (n=23). Hyperuricemia was detected in 34 (22.1%) patients, normal uric acid levels were revealed in 120 (77.9%) patients. All patients were studied using clinical, laboratory, instrumental and molecular genetic research methods. The level of sUA was determined by the enzymatic colorimetric method. Serum xanthine oxidase was measured using a method based on a solid phase sandwich-type enzyme-linked immunosorbent assay. Metabolites of purine metabolism in blood plasma were measured using the method of high-performance liquid chromatography. The SLC2A9 gene rs734553 polymorphism was determined using the polymerase chain reaction with real-time detection of results. Results. The patients with AH and AF, as compared to healthy individuals, had more severe disturbances of purine metabolism, characterized by higher concentration of sUA (330 [283; 412] µmol/l and 197 [161; 229] µmol/l (p<0.001), respectively). Also, in contrast to the group of healthy individuals, the group of patients with AH and AF demonstrated an increase in the level of adenosine (p=0.001), and a decrease in the levels of hypoxanthine and xanthine (p<0.001). There were no statistically significant differences in xanthine oxidase activity level (p>0.05), however, in 54% of patients in the main group it was higher than normal values. The dominant allele A and the dominant genotype A/A of the SLC2A9 gene rs734553 polymorphism (75%, p=0.005; 64%, p=0.001, respectively) occurred significantly more often in healthy individuals, while the recessive allele C and the heterozygous genotype A/C were found significantly more often in the group of patients with AH and AF (41.3%, p=0.005; 48.1%, p=0.003, respectively). The C/C genotype (41.7%, p=0.001) was significantly more common in patients with AH in combination with AF and hyperuricemia, compared to patients with AH combined with AF without hyperuricemia as well as healthy individuals (6.8%; 14%, p=0.001, respectively). Patients with AH in combination with AF and the C/C genotype were characterized by a significantly higher level of sUA (p=0.003) compared to patients with the A/A genotype. Conclusions. A statistically significant predominance of the recessive allele C of the SLC2A9 gene rs734553 polymorphism was established in patients with AH and AF compared with healthy individuals (p=0.005). In patients with AH in combination with AF and hyperuricemia, the C/C genotype was significantly more common (41.7%, p=0.001). Patients with AH in combination with AF and the C/C genotype were characterized by a significantly higher level of sUA (p=0.003) compared to patients with the A/A genotype.

Творческая история комедии Наки Исанбета «Рыжий чичян и Черноволосая красавица»

The purpose of the study. The purpose of our research is to study the genesis of Naki Isanbet’s comedic text “Jiran chichan belan Karachach sylu” (“Red-haired chichyan and Black-haired beauty”, 1942). Materials and methods of research: The material for the study was preserved in the archives of Naki Isanbet’s “Red-haired chichyan and Black-haired beauty” comedy versions. Its study allows us to reconstruct the nature of the author’s idea. The study was conducted using genetic, cultural-historical, and comparative research methods, the choice of which is determined by the nature of the analyzed text. The novelty of the research consists in a comprehensive study of the history of the creation and existence of this comedy in Tatar culture and the identification of its role in popularizing information about the Golden Horde among the Tatars. Research results. Naki Isanbet’s comedy “Jiren chichan belan Karachach sylu” (“Red-haired chichyan and Black-haired beauty”, 1942) is a vivid evidence of the valorization of national cultural values during the Great Patriotic War. During the short period the playwright managed to perpetuate national images of heroes on the stage: Idegey, Tulyak, Jiren chichyan and Karachech sylu. Their roots go back to the glorious history of medieval Tatar states. During the Great Patriotic War, the voices of Jiren chichyan and Karachech sylu (14th century) were heard from the Tatar stage, and people got acquainted with the instructions of Jamukha chichyan (12th century). The performance and the work were banned by censorship. The work was published in 1963 and returned to the stage in 1999 (directed by Farid Bikchantaev).

ДИАГНОСТИКА МЕЛАНОМЫ: СОВРЕМЕННОЕ СОСТОЯНИЕ И ПЕРСПЕКТИВЫ (обзор литературы)

The article reflects the main factors influencing the survival of patients.
 The basic principles of diagnosing melanoma are presented. Methods for diagnosing skin melanoma, including invasive and non-invasive methods, are presented in detail and with justification.
 Among non-invasive methods for diagnosing skin melanoma, the role of physical examination and dermatoscopy in cases of suspected melanoma is emphasized. Among instrumental examination methods, the use of such research methods as ultrasound, computed tomography, magnetic resonance imaging, positron emission tomography is considered.
 Among laboratory methods, methods of molecular genetic research and determination of biomarkers of skin melanoma are presented.
 Among the invasive methods, the use of such biopsy methods as excisional and incisional biopsy is considered, with clarification of the indications and contraindications for their use.
 At the end of the work, the basic requirements for the early diagnosis of skin melanoma are listed. In conclusion, the latest trends in the diagnosis of skin melanoma, new non-invasive methods and an algorithm for diagnosing skin melanoma are identified.

KNOWLEDGE AS A SOURCE OF ENTERPRISE INTELLECTUALISATION

The purpose. The aim of scientific research is to deepen the methodological foundation regarding the essence of knowledge as a source of the enterprise intellectualization and the innovative enterprise intellectual potential effective use ensuring priority. Methodology. The methodological basis of the study consists of: knowledge-based economy theories; entrepreneurship, institutionalism; microeconomic theory of knowledge management; knowledge transformation theories, obtaining new knowledge; of self-learning organization concept. General scientific and formal-logical methods of comparison, analysis, synthesis, abstraction, as well as historical, genetic, linguistic, hermeneutic and heuristic research methods are used. Results. The article is devoted to the justification of the theoretical and methodological principles regarding the essence, place and role of knowledge as the enterprise intellectualization source and the priority of ensuring the innovative enterprises intellectual potential effective use in order to develop their competitive advantages in the knowledge-based economy paradigm. The main stages that change the role of knowledge in society are characterized. Emphasis is placed on the fact that the enterprise can face active competition only if the intellectual component of its resource portfolio is improved. In this context, the information revolution caused new requirements for economic knowledge, and not so much for their volume, but for quality, that reflects the imperatives of progressive, highly moral management. Accordingly, the “center of gravity” of the modern reproduction process objectively shifts from the sphere of market economic transactions to the knowledge reproduction economic space of a high moral and creative level. Modern approaches to understanding information and knowledge are considered, and their comprehensive characteristics are given. The relationship between the categories “data”, “information” and “knowledge” is studied. A new essence understanding of categories “knowledge” and “enterprise intellectualization” is proposed. The knowledge types in the context of epistemological approach are singled out. Scientific novelty. The methodological knowledge essence basis is developed in the work. A classification of knowledge is proposed for better explicit and hidden knowledge management effectiveness differentiation and increasing in the context of innovative development is proposed. Practical significance of the obtained results. The author's developments and recommendations can be used for the purpose of managing the enterprise intellectual potential in order to increase the economic added value due to the generation of organizational knowledge.

Selective list of VIR’s methodological recommendations (guidelines) for 2000–2022

The N.I. Vavilov All-Russian Institute of Plant Genetic Resources (VIR) releases almost all types of publications traditional for research institutions. Since the 1920s, VIR has been regularly publishing methodological recommendations, or guidelines.This kind of publications has won its own specialized audience and retained it for over a hundred years. All recommendations (guidelines) are an assemblage of documents dedicated to the study, conservation, and replenishment of the unique global plant genetic resources collection held by VIR; plant breeding; seed production; cultivation; resistance of individual crops to diseases and pests; herbarization; long-term seed storage; biochemical, mycological, phytopathological, genetic, and biotechnological research methods; and many other issues. Such publications are widely used by the staff of VIR and the network of its branches in their practical work, and are also in demand with various scientists, experts, lecturers and teachers, postgraduate and undergraduate students all over Russia.Keeping in mind the ever increasing interest in such information, the Editorial Board of the journal Proceedings on Applied Botany, Genetics and Breeding decided to publish a selective list of VIR’s methodological recommendations (guidelines) for 2000–2022. For the sake of convenience, the material is arranged in chronological order. This list partially includes recommendations and guidelines published by other research institutions where VIR’s staff members were among the authors. It is our hope that the list will be found useful by our readers and authors of scientific publications. The list was compiled by the staff of the Library and Publishing Department of VIR.

Cytological origination of the first found allotriploid tree peony, Paeonia × lemoinei ‘Oukan’ (AAB), reveled by molecular karyotype comparation

Polyploid breeding is one of the most important approaches to improve the traits of cultivated plants, but there has been no feasible method to get polyploid tree peony. In this study, we revealed a sexual triploidization approach from a diploid intersubsectional hybrid tree peony, Paeonia × lemoinei ‘High Noon’, to an allotriploid one, P. × lemoinei ‘Oukan’, by accurate karyotyping based on genomic in situ hybridization (GISH) and fluorescence in situ hybridization (FISH). The ancestor genomes from P. suffruticosa and P. delavayi, which were defined as A-genome and B-genome respectively, were clearly distinguished by GISH in ‘High Noon’ and ‘Oukan’. Nonhomologous chromosomes were successfully recognized by means of 5S and 45S rDNA FISH. Karyotype comparation based on GISH and FISH indicated that the B-genome and one of the two A-genomes in ‘Oukan’ (2n=3x=15, AAB) were highly consistent to the corresponding genomes in ‘High Noon’ (2n=2x=10, AB), implying the triploid origination via 2n gametes produced by first division restitution (FDR). In addition, it was demonstrated for the first time that the dual in series sites of 5S rDNA located on the short arm of chromosome 3 in both A and B-genomes. Besides the familiar strong 45S rDNA signals observed at terminal region of short arms, single weak 45S rDNA signal was newly detected at terminal region in the short arm of certain chromosome 1 in A-genome, and at interstitial-distal region in the long arm of chromosome 5 in both genomes. These findings provide a proof and theoretical support for tree peony polyploid breeding, and an accurate karyotyping method for genetic research of plants in Paeonia.

Genetic Monitoring of Grey Wolves in Latvia Shows Adverse Reproductive and Social Consequences of Hunting.

Nowadays, genetic research methods play an important role in animal population studies. Since 2009, genetic material from Latvian wolf specimens obtained through hunting has been systematically gathered. This study, spanning until 2021, scrutinizes the consequences of regulated wolf hunting on population genetic metrics, kinship dynamics, and social organization. We employed 16 autosomal microsatellites to investigate relationships between full siblings and parent-offspring pairs. Our analysis encompassed expected and observed heterozygosity, inbreeding coefficients, allelic diversity, genetic distance and differentiation, mean pairwise relatedness, and the number of migrants per generation. The Latvian wolf population demonstrated robust genetic diversity with minimal inbreeding, maintaining stable allelic diversity and high heterozygosity over time and it is not fragmented. Our findings reveal the persistence of conventional wolf pack structures and enduring kinship groups. However, the study also underscores the adverse effects of intensified hunting pressure, leading to breeder loss, pack disruption, territorial displacement, and the premature dispersal of juvenile wolves.

Features of the profile of drug resistance to first-line and second-line anti-tuberculosis drugs in patients with human immunodeficiency virus infection

BACKGROUND: Drug resistance to anti-tuberculosis drugs directly affects the disease course. However, a high percentage of multidrug resistance in patients with HIV infection not only worsens the disease course but also forces the development of new strategies for managing such patients.
 AIMS: To characterize drug resistance to anti-tuberculosis drugs in patients with HIV infection.
 MATERIALS AND METHODS: The study recruited patients with newly diagnosed tuberculosis by molecular genetic research methods and bacterial research methods in the Orenburg Regional Clinical Tuberculosis Dispensary.
 RESULTS: For 20182022, the proportion of people with HIV infection who are resistant to isoniazid by 14.6%, rifampicin by 28.85%, moxifloxacin by 717.39%, and amikacin by 104.25% has increased. The reduction in the choice of drugs and possibility of effective anti-tuberculosis therapy in people with HIV infection are also complicated by the detection of a multidrug-resistant culture in 20.34% of the examined patients and multidrug resistance in 39.6%. In over 5 years, the percentage of gene mutations of mycobacteria responsible for resistance to anti-tuberculosis drugs has increased.
 CONCLUSIONS: The results of this study indicate the need to search for new solutions and prevent resistance to existing anti-tuberculosis drugs and for the development of new drugs effective for use as first- and second-line drugs.

Phylogenetic processes in dog domestication and breeding

Aim. The significance of the problem of origin and selection of animals for practice is that by studying the main stages of formation of modern domestic animals in the past, scientists and breeders develop even more effective methods of improving existing breeds by targeted changes in their hereditary properties in purebred breeding, using hybridization and modern biotechnological technologies. Methods. Using archaeological, morphological, biochemical and genetic research methods, scientists managed to establish the karyotype and genome of the wild European wolf and domestic dog. Results. Domestication is the process of turning wild animals into domesticated ones. Over many generations, people have gradually accumulated experience in keeping animals, learned the laws of animal heredity, which helped to accelerate the domestication process and the formation of the original breed populations. The dog is the first domesticated animal, and the history of its domestication dates back to the Mesolithic period (12-15 thousand years BC). The wild European wolf is considered to be the wild ancestor of the dog, since wolves and dogs have 78 chromosomes in their karyotype, and when they mate, they produce fertile offspring. In the process of dog domestication, both morphological and physiological changes occurred compared to the wild European wolf. Since the domestication of dogs involved adaptation to new conditions in captivity, important genetic mechanisms such as mutations, recombination, a hidden reserve of heredity, and the direct effect of selection and selection were important. Conclusions. The representatives of two families of carnivores were domesticated – Canidae and Felidae. Dogs, as already mentioned, were the first domesticated animals tamed and domesticated by humans. The taming and domestication of the wild ancestor of dogs, the wolf, took place in the Mesolithic period, as is known from bone remains discovered by scientists. In different centres of domestication, different forms of wolves have been found that could be domesticated and gave rise to many varieties of dogs since ancient times. Further selection by humans has led to an even greater diversity of domestic dog breeds that serve a variety of purposes (guarding, hunting, sledding, service, and decorative breeds).

Clinical case of diagnosis of generalized tuberculosis in a child with a positive result of an intradermal test

At present day, generalized forms are rarely found in the structure of tuberculosis in children. A significant risk factor for the development of generalized lesions is prolonged contact with a tuberculosis patient. The diagnosis of a specific etiology is often complicated by the absence of pathognomonic symptoms. Immunodiagnostics, microbiological and molecular genetic research play an important role in the diagnosis of tuberculosis infection in children. The purpose of this study was to analyze the case of the development and diagnosis of generalized tuberculosis in a child of primary school age (7 years and 11 months) with lesions of the lungs, larynx, middle ear. The examination methods included immunological intradermal tests (Mantoux test, RTA test), in vitro test (QuantiFERON-TB), multispiral computed tomography, bronchoscopy and laboratory methods of examination. The manifestation of the disease occurred gradually with the occurrence of hypochromic anemia with a further increase in symptoms of intoxication. A comprehensive examination established a disseminated process in the lungs, larynx and middle ear lesions. Family tuberculosis contact was established when the disease was detected in a child. During the examination, the evaluation result of the Mantoux test showed an increase in sensitivity to tuberculin. The hyperergic result of the RTA test indicated the activity of MBT in the body. At the same time, the irregular conduct of immunodiagnostics using the Mantoux test, the monotony of the test did not allow timely measures to be taken in relation to the in-depth examination of the child for tuberculosis. The specific etiology of the disease was reliably confirmed by cultural and molecular genetic research methods. The child was diagnosed with generalized tuberculosis, disseminated pulmonary tuberculosis in the infiltration phase, MBT (+) with drug resistance to isoniazid, streptomycin, paraaminosalicylic acid, tuberculosis of the larynx and vocal apparatus, PCR (+) with drug resistance to isoniazid, tuberculous right-sided otitis media, MBT (+) with drug resistance to isoniazid, streptomycin, paraaminosalicylic acid. The presented clinical case indicates the need for timely detection of tuberculosis contacts, regular immunodiagnostics, and compliance with phthisiological alertness of doctors of all specialties in relation to children with long-lasting symptoms of various diseases. The complete physical examination using an intradermal RTA test (Diaskintest), microbiological and molecular genetic research methods allows to establish the tuberculous etiology of the lesion.