Molecular Genetic Techniques Research Articles

The finding of black currants from sect. Eucoreosma (Grossulariaceae) in the central part of southern Primorsky Territory, Russia

Background. The ever increasing anthropogenic impact on natural populations of rare and, especially, endangered plant species has recently become even more forceful. Therefore, the finding of a small black currant population representing sect. Eucoreosma (Spach) Jancz. within a natural environment in the south of Primorsky Territory appears highly important. Previously, none of the black currant species was recorded as occurring in this area. Materials and methods. Plants from the new locality were compared with accessions of Ribes nigrum L., R. ussuriense Jancz., R. pauciflorum Turcz. ex Pojark., and R. fontaneum Boczkarn. using morphological and molecular genetics techniques. Results and conclusion. A molecular genetics analysis of two chloroplast DNA fragments excluded any association of the discovered plants with R. nigrum or R. fontaneum. Meanwhile, no differences in chloroplast DNA fragments were found between these plants and R. ussuriense and R. pauciflorum accessions. A number of morphological features attested to a possibility to attribute the finding to the rare species R. ussuriense: its isolated localities had previously been reported in southern Primorsky Territory. The differences in chloroplast DNA fragments between the Siberian/Far Eastern species R. pauciflorum and the European/Siberian species R. nigrum make doubtful their possible fusion, based only on morphological characters, and require further studies.

Identification of SNP and SilicoDArT Markers and Characterization of Their Linked Candidate Genes Associated with Maize Smut Resistance

The implementation of biological advancements in agricultural production is the response to the needs of the agricultural sector in the 21st century, enabling increased production and improved food quality. Biological progress in the maize breeding and seed industries is unique in terms of their social and ecological innovation aspects. It affects agricultural productivity and the adaptation of cultivated maize varieties to market demands and changing climate conditions without compromising the environment. Modern maize resistance breeding relies on a wide range of molecular genetic research techniques. These technologies enable the identification of genomic regions associated with maize smut resistance, which is crucial for characterizing and manipulating these regions. Therefore, the aim of this study was to identify molecular markers (SilicoDArT and SNP) linked to candidate genes responsible for maize smut resistance, utilizing next-generation sequencing, as well as association and physical mapping. By using next-generation sequencing (NGS) and statistical tools, the analyzed maize genotypes were divided into heterotic groups, which enabled the prediction of the hybrid formula in heterosis crosses. In addition, Illumina sequencing identified 60,436 SilicoDArT markers and 32,178 SNP markers (92,614 in total). For association mapping, 32,900 markers (26,234 SilicoDArT and 6666 SNP) meeting the criteria (MAF > 0.25 and the number of missing observations < 10%) were used. Among the selected markers, 61 were highly statistically significant (LOD > 2.3). Among the selected 61 highly statistically significant markers (LOD > 2.3), 10 were significantly associated with plant resistance to maize smut in two locations (Smolice and Kobierzyce). Of the 10 selected markers, 3 SilicoDArT (24016548, 2504588, 4578578) and 3 SNP (4779579, 2467511, 4584208) markers were located within genes. According to literature reports, of these six genes, three (ATAD3, EDM2, and CYP97A3) are characterized proteins that may play a role in the immune response that develops in response to corn smut infection. In the case of genotypes belonging to the same origin groups, markers linked to these genes can be used to select varieties resistant to corn smut. These markers will also be tested on genotypes belonging to other maize origin groups to demonstrate their universality.

Molecular Characterization of Anaplasma spp. in Cattle from Kazakhstan.

Bovine anaplasmosis is an infectious vector-borne disease caused by bacteria of the genus Anaplasma, which have a wide global distribution and represent a high economic burden for agriculture. The use of molecular genetic techniques has increased our knowledge of the species diversity of Anaplasma spp. and naturally susceptible animals. Monitoring studies allow us to assess the level of infection in herds, as well as the involvement of natural vectors in the processes of maintaining and spreading infection. Despite the high prevalence of Theileria and Babesia in cattle in Kazakhstan, there is no information on the distribution and species diversity of Anaplasma spp in this country. As part of this work, 7027 DNA samples isolated from the whole blood of cattle from 175 settlements in all 17 Kazakhstan regions were PCR-tested for the presence of Anaplasma spp. Anaplasma carriers were found in 1.3% (90 out of 7027) of the tested animals in 9 of the 17 regions of Kazakhstan. The highest percentage of infected animals was recorded in Turkistan (South Kazakhstan) and North Kazakhstan with 4.46% and 2.48% positive samples, respectively. The partial sequencing of 16S rRNA and the groEL gene allowed us to identify five species of Anaplasma: A. centrale, A. marginale, Candidatus Anaplasma Mongolica, A. ovis, and Unknown Anaplasma with infection rates of 0.63%, 0.44%, 0.13%, 0.01%, and 0.01%, respectively.

Epidemiological and clinical characteristics of cases and detection of pathogens’ markers of tick-borne infections in the Tomsk region

BACKGROUND: Among the spectrum of the tick-borne infections, Lyme borreliosis and tick-borne encephalitis are most often recorded in the Tomsk region, however it is obviously that possible range of the pathogens is not limited to these ones. AIM: To make epidemiological and clinical characteristics of cases and detect frequencies of pathogen’s markers of tick-borne infections among adult patients applying serological and molecular genetic techniques in the Tomsk region during 2020–2023. METHODS: A single-center prospective sample study was conducted applying a comprehensive approach in peripheral blood specimens obtained from 197 patients with suspected tick-borne infections hospitalized during 2020–2023. To verify the diagnoses of tick-borne encephalitis or Lyme borreliosis, specific immunoglobulins of classes M and G to the antigens of virus of tick-borne encephalitis and Borrelia burgdorferi s.l. were determined in paired blood serum samples applying enzyme-linked immunosorbent assay. Additionally, the polymerase chain reaction was used to detect genetic markers of virus of tick-borne encephalitis, B. burgdorferi s.l., B. miyamotoi, Rickettsia spp., Coxiella burnetii, and Francisella tularensis. Epidemiological surveillance data of incidence rates of tick-borne infections obtained from official reports were analyzed as well. MS Office Excel 2010 software was used to calculate frequency morbidity measures. RESULTS: We verified the final diagnosis of tick-borne encephalitis in 52 (26.4%) patients, Lyme borreliosis in 116 (58.9%) patients, co-infection of tick-borne encephalitis and Lyme borreliosis 15 (7.6%) patients, hard tick relapsing fever caused by B. miyamotoi in 8 (4.1%) patients, coinfection of Q fever caused by C. burnetii with tick-borne encephalitis or Lyme borreliosis in 4 (2.0%) patients, and coinfection of tularemia with tick-borne encephalitis or Lyme borreliosis in 2 (1.0%) cases. We identified markers of tick-borne encephalitis and Lyme borreliosis pathogens by polymerase chain reaction in 3.6% and 4.1% of the patients. Most number of individuals with tick bites, and the highest incidence rates of Lyme borreliosis and tick-borne encephalitis were recorded in the suburbs of Tomsk. The age groups of 40–59 years, and elderly people 60 years and older were predominated among the patients. CONCLUSIONS: It was found out that, in addition to Lyme borreliosis and tick-borne encephalitis, there were such tick-borne infections as tick relapsing fever, Q fever, and tularemia in the Tomsk region in the 2020–2023 period.

7775 Clinical Features of NKX2-1 Gene-Related Diseases

Abstract Disclosure: S. Fukui: None. E. Tanimoto: None. N. Ujita: None. H. Miyagi: None. K. Yoshii: None. N. Yasuhiro: None. R. Horikawa: None. Background: Congenital hypothyroidism (CH) is the most common congenital endocrine disorder. The utilization of molecular genetic techniques has facilitated the detection of CH resulting from various single-gene mutations; NKX2-1 gene-related disease stands out as one etiology of symptomatic CH. We report the clinical course of three cases with CH caused by NKX2-1 pathogenic variants. Case 1: An 8-year-old boy. He was born at 40 weeks and six days of gestation, weighing 2845g, and was presented to our department by high TSH (76.1 µIU/mL) at newborn screening (NBS), and LT4 treatment was initiated. He showed motor developmental delay, and choreiform movements were exhibited at two years and 11 months without respiratory symptoms. Case 2: An 18-year-old female. She was born at 40 weeks and five days gestation, weighing 2680g, and was admitted to her previous hospital following poor weight gain at approximately six months. During hospitalization, she was initiated LT4 treatment with a TSH elevation(14.7μIU/mL). After receiving a diagnosis of idiopathic interstitial pneumonia, she was subsequently referred to our hospital. At the age of 11, an NKX2-1 mutation was identified. Case 3: An 18-year-old female. She was born at 41 weeks and three days of gestation, weighing 2364g.TSH elevation (27.9 µIU/ml) was detected at NBS, and LT4 treatment was started. Thyroid hormone levels were maintained within normal range. However, wheezing and tracheomalacia were noted around five months. Hypotonia and delayed motor development were evident, and an NKX2-1 mutation was identified at the age of 12 years. Discussion: Although NKX2-1 gene-related disorders can be readily suspected when typical symptoms of central nervous system, pulmonary, and thyroid dysfunction manifest concurrently, diagnosis is sometimes established later because of the diversity of the occurrence of these symptoms. Despite reports of choreiform movements being inevitable, they did not appear in Case 2.3 until the age of 18 years. Conversely, muscle hypotonia and delayed motor development were observed in all our cases. When atypical neurological symptoms combined with CH are observed, NKX2-1 gene-related disease should be considered since the disease is known to develop malignant tumors such as lung cancer and thyroid cancer, which can significantly affect the prognosis of patients. Conclusion: When muscle hypotonia and delayed motor development are observed in addition to hypothyroidism and respiratory symptoms, NKX2-1 gene-related disease should be suspected, and follow-up for complications, including malignancy, is essential after diagnosis. Presentation: 6/2/2024

Monogenic Kidney Diseases in Adults With Chronic Kidney Disease (CKD).

According to current evidence, every 10th to 11th adult with chronic kidney disease (CKD) has a monogenic disease of the kidney. This review is based on reported studies in which molecular genetic diagnostic techniques were used to investigate monogenic kidney diseases in adults with CKD. The studies were identified by a selective literature search using predefined criteria. In 12 selected studies, diagnostic variants of 179 different genes were identified in 1467 out of 6607 study participants with CKD (22.2%). More than 60% of these variants affected 8 genes (PKD1, PKD2, COL4A3, COL4A4, COL4A5, UMOD, MUC1, HNF1B). Three diseases are associated with these genes: autosomal dominant polycystic kidney disease (ADPKD), Alport syndrome, and autosomal dominant tubulo-interstitial kidney disease (ADTKD). Physicians treating patients with CKD should be alert to the presence of any red flags, such as onset at a young age, a positive family history, or hematuria of unknown cause. When a genetic etiology is suspected, a specialized work-up is indicated, often including a molecular genetic investigation. A positive genetic finding usually leads to a modification of the patient's specific diagnosis and/or treatment. Awareness of the high prevalence of monogenic kidney diseases in adults with CKD and alertness to their suggestive clinical features are crucial for the timely initiation of targeted diagnostic testing. The molecular genetic identification of these diseases is a prerequisite for appropriate patient management.

Consanguinity and Occurrence of Monogenic Diseases in a Single Tertiary Centre in Riyadh, Saudi Arabia: A 2 Years Cross-Sectional Study.

Consanguinity, or the practice of marrying close relatives, is a common cultural tradition in Saudi Arabia, with rates among the highest in the world. This practice has significant implications for the prevalence and distribution of major single genetic defects and chromosomal abnormalities within the Saudi population. Herein, using the BESTCare electronic medical record system (designed to streamline hospital operations, enhance patient care, and improve the overall efficiency of healthcare services; bestcare.ezcaretech.com) in a single tertiary centre, King Abdullah Specialized Children Hospital (KASCH) in Riyadh, Saudi Arabia, we performed a cross-sectional study for all patients referred to the hospital from the 1st January 2020 until 1st January 2022. The present study, which included 1100 individuals, found a high prevalence of consanguinity (64%) and a significant proportion of third-degree relatives (69%). The mean age of participants was 12.24 years, and the diagnostic rate using advanced molecular genetics techniques was 45%, with whole exome sequencing (WES) being the most common method (43%). The study also noted a significant delay in diagnosis for more than a year in 16% of cases, with a common neurodevelopmental phenotype (18%). In conclusion, we revealed the prevalence of consanguineous marriages in the KASCH hospital in Riyadh, Saudi Arabia. We also highlighted the most frequently referred phenotype. These findings are consistent with previous research on the prevalence and impact of consanguinity on rare genetic disorders.

Unusual familial cystic kidney disease: combining fine radiologic and genetic evaluation to solve the case

BackgroundAutosomal dominant polycystic kidney disease (ADPKD) is the most prevalent hereditary kidney disease, characterized by enlarged kidneys with numerous cysts, high blood pressure, and a variety of extrarenal complications. This disease is a significant cause of renal failure and requires accurate differentiation from other cystic kidney diseases, especially when family history does not clearly indicate ADPKD. This is crucial due to differences in prognosis, treatment, and familial implications. Advanced molecular genetics and imaging techniques are employed to diagnose and assess the prognosis of patients and their families.Case presentationThe case study revolves around three patients from the same family—two sisters and one daughter—referred to a nephrology department for ADPKD management. The initial proband, a 42-year-old woman, experienced abdominal discomfort leading to an ultrasound that suggested ADPKD. However, MRI findings indicated standard-sized kidneys with bilateral parapelvic cysts, and no genetic markers for ADPKD were found. Her sister, presenting with controlled hypertension and similar ultrasound findings, also had her initial ADPKD diagnosis refuted by MRI and genetic testing, which revealed no significant mutations. The daughter, however, exhibited a different scenario with enlarged kidneys and multiple cysts characteristic of early-stage ADPKD. Genetic testing confirmed a deleterious PKD1 mutation, suggesting a de novo mutation, as her father showed no signs of the disease.ConclusionThis study highlights the complexity and necessity of thorough diagnostic processes in suspected ADPKD cases to prevent misdiagnosis. The initial symptoms and imaging might misleadingly suggest ADPKD, as seen in the cases of the two older patients. Still, further detailed imaging and genetic analyses revealed no ADPKD, preventing inappropriate treatment and stress. In contrast, the younger patient's distinctive clinical and genetic profile confirmed ADPKD, illustrating the variability within even closely related individuals. Such detailed assessments are crucial in guiding correct treatment decisions and providing accurate familial counseling, emphasizing the importance of considering a broader spectrum of renal cystic disorders before confirming a diagnosis of ADPKD.

GENETIC DIVERSITY AND DIGITAL DOCUMENTATION OF KAZAKHSTAN'S FLORA

Kazakhstan, with its expansive and diverse climate zones, boasts a rich and varied flora, featuring around 6,000 vascular species, including approximately 10% that are endemic. Effective management and study of this diverse flora requires continuous monitoring and inventory using advanced botanical, molecular genetic and bioinformatics techniques. This research focuses on exploring the genetic diversity within Kazakhstan's flora and establishing a comprehensive information database. Under Program 0237/PTF (2015-2017), efforts began to assess the genetic diversity of endemic, rare, endangered, and economically significant plant species of Kazakhstan. As part of grant АР05131621, an open-access web resource was developed, offering botanical and genetic data on Kazakhstan's flora. This database includes detailed botanical information on around 500 species. Ongoing research into the genetic diversity of different genera such as Juniperus, Tulipa, Allium, Iris, Salsola, and etc. promises to provide valuable insights into their population structure, phylogenetic relationships, and conservation strategies. Notably, this research has also initiated the first comparative analysis of plastid genomes in Kazakhstan. This research has been funded by the Science Committee of the Ministry of Science and Higher Education of the Republic of Kazakhstan (AP23490860, AP23483825, AP14870612, АР14869593, AP09259027, 0237/PTF-14, and АР05131621).

OSTEOLOGICAL COLLECTIONS OF THE CHERNYAKHOVSKAYA ARCHAEOLOGICAL CULTURE IN THE FUNDS OF THE ANUCHIN RESEARCH INSTITUTE AND MUSEUM OF ANTHROPOLOGY OF LOMONOSOV MOSCOW STATE UNIVERSITY

Introduction. The cycle of publications on the historical and archaeological characteristics of osteological collections from the funds of the Anuchin Research Institute and Museum of Anthropology of Lomonosov Moscow State University continues in the article. The purpose of this work is to create an information base for paleoanthropologists studying collections from the burial grounds of the Chernyakhovskaya archaeological culture dated III – beginning. V centuries AD. Materials and methods. This section provides a general description of the collections from the burial grounds of the Chernyakhovskaya culture, stored in the museum's funds. The materials considered were unearthed during excavations by Soviet archaeologists of the 1950s-1960s (E.A. Simonovich, G.B. Fedorov) on the territory of modern Ukraine and Moldova. The work uses all available publications of the authors of the excavations, as well as subsequent summary and analytical studies. Results. The main part of the work presents information on the burial grounds of the Chernyakhovkaya culture of Zhuravka, Chernyakhov, Malaeshty. The geographical characteristics of the sites, the history of their research, and basic information about the archaeological context for the presented anthropological materials from these necropolises are given. The most common, standard features of the funeral rite are shown and burials with unusual features that fall out of the general framework are indicated. A number of burials have been identified, which are marked in the accounting materials as belonging to the Chernyakhovskaya culture, but, in fact, are not related to it. Conclusion. The analysis of information about the three burial grounds allows us to consider them, to some extent, reference, standard for the Chernyakhovskaya culture at the main stage of its existence. A large collection from Zhuravka –143 storage units, as well as fairly large ones, 21 storage units and 20 storage units, respectively, from the burial grounds of Chernyakhov and Malaeshty allows to apply a wide range of anthropological and molecular genetic techniques.

MicroRNA and renal fibrosis in autosomal dominant polycystic kidney disease: a longitudinal study.

Autosomal dominant polycystic kidney disease (ADPKD) is a hereditary kidney disorder that may progress to kidney failure, accounting for 5-10% of all patients with end-stage kidney disease (ESKD). Clinical data, as well as molecular genetics and advanced imaging techniques have provided surrogate prognostic biomarkers to predict rapid decline in kidney function, nonetheless enhanced tools for assessing prognosis for ADPKD are stillneeded. The aim of this study was to analyze specific microRNAs involved in the pathogenesis of ADPKD and in the development of renal fibrosis, evaluating their potential role as predictors of renal function loss. We evaluated kidney function by estimated glomerular filtration rate (eGFR) in 32 ADPKD patients in different stages of kidney disease at T0 and after a 24-month follow up (T1). Patients were divided into two groups: Rapid disease progression ([RP], n 15) and Non-rapid disease progression ([NRP], n 17), according to the Mayo Clinic classification criteria. At T0, ADPKD patients underwent plasma sampling for quantitative analysis of h-miR-17-5p, h-miR-21-5p and h-miR-199a-5p microRNA expression, using the quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) method and a 3T magnetic resonance imaging (MRI), using an advanced MRI imaging protocol, for the quantification of total kidney volume (TKV), total perfusion volume (TPV) and total fibrotic volume (TFV). The expression of h-miR17-5p was higher (p < 0.05) in ADPKD patients with rapid disease progression. h-miR-17-5p, h-miR-21-5p and h-mir-199-5p showed a positive and significant correlation with the eGFR slope (mL/min/1.73 m2/year) (p < 0.05) but not with the eGFR at both T0 and T1. Both total fibrotic volume (cm3) and height-adjusted total fibrotic volume (cm3/m) were positively and significantly correlated to h-miR 21-5p and h-miR 199-5p (p < 0.05), but not to total kidney volume (cm3) and height-adjusted total kidney volume (cm3/m). The microRNAs we studied were associated with fibrosis and renal damage, suggesting their possible role as biomarkers able to identify ADPKD patients at high risk of disease progression regardless of the degree of kidney function, and therefore suitable for medical therapy, and may help uncovering new molecular mechanisms underlying cystogenesis.

O-039 Con - the faster the better

Abstract Late o’clock: Are slow growing embryos clinically usable? Con - the faster the better Reproductive medicine and, within it, embryology, have experienced milestones in the understanding of pre-implantation embryonic development over the last quarter of a century. The development of blastocyst culture media for embryo culture even after embryonic genome activation and the possibility of continuous monitoring of embryonic morphokinetics throughout the pre-implantation period have been major contributors. The development of molecular genetic techniques has also played an important role. The introduction of new cryopreservation techniques has also improved the success rate of the clinical use of surplus embryos, including slow-growing embryos not otherwise selected for fresh cycle transfer. Two questions are most frequently encountered in clinical embryology today: which embryo in a cohort has the best chance of resulting in a live-born child, and how slow and morphologically suboptimal can an embryo be to still be considered clinically useful? In particular, the latter question does not have an easy answer, as embryos demonstrate varying pace of development. A further problem arises if there are no optimal embryos available in the cohort and this answer has to be given on day 2, 3 or 5, as this is when the embryo transfer is scheduled to take place. Some studies suggest that longer timing of any stage of embryonic development from pronuclear fading onwards means that the embryo is late in development due to certain abnormalities and is likely to be energetically depleted by the activation of its repair mechanisms. Such embryos not only have a lower implantation potential compared to fast embryos, but also diverge in time from the optimal receptivity of the endometrium. Additional testing and/or storage of slow-growing embryos brings the question of their clinical usability to the time of frozen embryo replacement cycles and quiets the couple's expectation of a “successful” completion of the stimulated cycle. Whether embryos are to be selected for fresh ET or FER, the “the faster - the better” principle is likely to be applied. In daily clinical practice, decisions on the use of slow-growing and/or morphologically sub-optimal embryos and the timing of embryo transfer are made on an individual basis, taking into account various medical as well as non-medical (e.g. economic, logistical) factors. These may also include portraying a positive image of the centre's performance. However, patients expect infertility specialists to make realistic assessments not only of the viability of slow growing embryos, but also of the outcome of the transfer of them. The assessment must be based not only on medical and ethical but also on rational grounds.

O-282 Detection and elimination of malignant cells from cryopreserved testis and ovarian tissue?

Abstract For young patients receiving treatments that might impair their reproductive functions, gonadal tissue cryopreservation emerges as a technique to preserve their future fertility. The reimplantation of cryopreserved prepubertal gonadal tissue is currently the most promising strategy for restoring fertility in these individuals. There have been reports of successful births after autotransplanting ovarian tissue harvested at peri-puberty. In males, although autotransplantation of testicular tissue has not yet led to the production of viable sperm, successful reproduction has been documented in primate studies following such procedures. The preservation of gonadal tissue in the setting of cancer introduces concerns regarding the potential for malignant cell contamination, leading to worries about the future reimplantation of contaminated tissue. Recent analysis shows significant contamination rates with cancer cells in cryopreserved gonadal tissues from children with leukemia and lymphoma — 37% in testicular tissues and 12% in ovarian tissues. The risk of contamination extends beyond hematological cancers with documented cases of cancer cell contamination in ovarian tissue from patients with apparently localized sarcoma, and detection of neuroblastoma cells in cryopreserved testicular tissue. Such significant contamination rates prompt discussions about harvesting tissue post-chemotherapy, although gonadotoxic treatments may adversely affect the quality of the cryopreserved germ cells. Emerging diagnostic techniques, especially molecular methods, are being investigated for their ability to more accurately detect residual maliganat disease. While traditional histology and immunohistochemistry have been the norm, their sensitivity is relatively low. More precise molecular-genetic techniques, such as Polymerase Chain Reaction (PCR) and Reverse Transcriptase-quantitative PCR, demonstrate greater sensitivity. Moreover, advanced methods like next-generation sequencing offer even higher precision in detecting minimal residual disease. Xenotransplantation of gonadal tissue into immunodeficient animals may also provide some insights as a preclinical method for relapse prediction. Future research is essential to refine these diagnostic techniques to achieve the necessary sensitivity for clinical application. This presentation aims to explore the current knowledge on gonadal tissue contamination rates for both hematological cancers and solid tumors in prepubertal patients and to review strategies for decontaminating gonadal tissues. A critical examination of existing knowledge gaps will be conducted with the objective of identifying secure methods for fertility restoration in cancer survivors.

The likelihood of detecting abnormal karyotypes in fetuses with a single major anomaly or "soft" marker on ultrasonographic scanning.

Fetuses with abnormal karyotypes often exhibit distinctive ultrasonographic markers, including major anomalies and "soft" markers, indicating potential chromosomal issues. A crucial consideration arises when a single fetal anomaly is detected, raising the question of whether karyotyping is warranted, given the associated procedural risks. Our objective was to establish correlations between single fetal anomalies identified through ultrasound and chromosomal abnormalities. A cross-sectional study analyzed the karyotype of 1493 fetuses and detected a single ultrasonographic anomaly over a 16-year period. Karyotyping was performed using the standard karyotype technique. Moreover, data regarding the type of anomaly detected ultrasonographically, karyotype results, and outcomes following interventions were collected. Among other methods, the use of positive likelihood ratios (LR+) was used to evaluate the diagnostic accuracy of ultrasound compared to karyotyping. In total, an aberrant karyotype was identified in 99 fetuses (6.6%). This was most commonly observed in cases involving a "soft" marker, occurring in 27 out of 218 fetuses (12.4%). The most frequently detected aberrant karyotype resulted from aneuploidies (80.6% of cases), notably trisomy 21 (50.5%). "Soft" markers predicted chromosomal issues (LR+ = 1.9; OR = 2.4), and isolated polyhydramnios (LR+ = 1.54; OR = 1.6) showed significance in predicting fetal chromosomal aberrations. When assessing the necessity for karyotyping in fetuses with single major anomalies or "soft" markers, it is crucial to consider individual risks for chromosomopathies, including the LR+ of the detected marker. In cases where fetuses exhibit isolated anomalies with a normal karyotype, additional diagnostic measures, such as molecular cytogenetic and molecular genetics techniques, may become necessary.

Approach to the Patient With Suspected Silver-Russell Syndrome.

Silver-Russell syndrome (SRS) is a clinical diagnosis requiring the fulfillment of ≥ 4/6 Netchine-Harbison Clinical Scoring System (NH-CSS) criteria. A score of ≥ 4/6 NH-CSS (or ≥ 3/6 with strong clinical suspicion) warrants (epi)genetic confirmation, identifiable in ∼60% patients. The approach to the investigation and diagnosis of SRS is detailed in the only international consensus guidance, published in 2016. In the intervening years, the clinical, biochemical, and (epi)genetic characteristics of SRS have rapidly expanded, largely attributable to advancing molecular genetic techniques and a greater awareness of related disorders. The most common etiologies of SRS remain loss of methylation of chromosome 11p15 (11p15LOM) and maternal uniparental disomy of chromosome 7 (upd(7)mat). Rarer causes of SRS include monogenic pathogenic variants in imprinted (CDKN1C and IGF2) and non-imprinted (PLAG1 and HMGA2) genes. Although the age-specific NH-CSS can identify more common molecular causes of SRS, its use in identifying monogenic causes is unclear. Preliminary data suggest that NH-CSS is poor at identifying many of these cases. Additionally, there has been increased recognition of conditions with phenotypes overlapping with SRS that may fulfill NH-CSS criteria but have distinct genetic etiologies and disease trajectories. This group of conditions is frequently overlooked and under-investigated, leading to no or delayed diagnosis. Like SRS, these conditions are multisystemic disorders requiring multidisciplinary care and tailored management strategies. Early identification is crucial to improve outcomes and reduce the major burden of the diagnostic odyssey for patients and families. This article aims to enable clinicians to identify key features of rarer causes of SRS and conditions with overlapping phenotypes, show a logical approach to the molecular investigation, and highlight the differences in clinical management strategies.

Treatment of Achilles tendon rupture in a patient with alkaptonuric ochronosis

The treatment result of a patient with spontaneous rupture of the Achilles tendon and features of diagnosing alkaptonuric ochronosis, a rare disease, are presented, providing doctors beneficial information for the prevention and/or delay of possible complications. On March 8, 2021, at about 20:50, while at his workplace, getting up from his chair from the table, patient K, 52 years old, felt a sharp pain in the lower third of his right shin and was urgently taken to the hospital. He was examined by a traumatologist and was initially diagnosed with rupture of the right Achilles tendon. His right lower limb was immobilized with a plaster splint. On March 11, 2021, he entered the military traumatology and orthopedics clinic of the Kirov Military Medical Academy for surgical treatment. Upon admission, the patient complained of pain in the right lower leg and impaired support function of the right lower limb. Based on complaints, medical history, laboratory results, and instrumental studies, subcutaneous rupture of the right Achilles tendon was diagnosed. On March 12, 2021, anchor suture of the right Achilles tendon was performed. Anesthesia — combined, access: two separate incisions on the posterior surface of the lower third of the right leg and in the area of the calcaneal tubercle, each 5 cm long, intraoperatively revealed a complete subcutaneous rupture of the right Achilles tendon. Notably, the tendon was black in color when cut. The postoperative period proceeded without complications, and postoperative wounds of linear shape in the right Achilles tendon healed by primary intention. No signs of disturbances were observed in peripheral circulation and innervation of the right lower limb. To confirm the diagnosis, a simple laboratory test was performed: a change in the color of urine during the day, after its additional alkalization. The color change is due to the polymerization of homogentisic acid, which is excreted in large quantities in the urine. To verify the diagnosis, a molecular genetic technique was performed: sequencing of the clinically significant HGD gene and encoding the enzyme homogentisate-1,2-dioxygenase, which are mutations that lead to alkaptonuria. The analysis results confirmed the presence of an amino acid substitution at the pGLy161Arg position, indicating pathogenicity. The use of simple methods of additional laboratory diagnostics and molecular genetic analysis enabled the diagnosis of alkaptonuric ochronosis. Thus, characteristic diagnostic signs were damage to the Achilles tendon in combination with the obtained anamnestic, clinical, and laboratory data of alkaptonuria.

Genetic and Genomic Pathways to Improved Wheat (Triticum aestivum L.) Yields: A Review

Wheat (Triticum aestivum L.) is a fundamental crop essential for both human and animal consumption. Addressing the challenge of enhancing wheat yield involves sophisticated applications of molecular genetics and genomic techniques. This review synthesizes current research identifying and characterizing pivotal genes that impact traits such as grain size, number, and weight, critical factors influencing overall yield. Key genes including TaSPL17, ABP7, TaGNI, TaCKX6, TaGS5, TaDA1, WAPO1, TaRht1, TaTGW-7A, TaGW2, TaGS5-3A, TaSus2-2A, TaSus2-2B, TaSus1-7A, and TaSus1-7B are examined for their roles in these traits. The review also explores genes responsive to environmental changes, which are increasingly significant under current climate variability. Multi-trait regulatory genes and quantitative trait loci (QTLs) that contribute to these traits are discussed, highlighting their dual influences on grain size and yield. Furthermore, the paper underscores the utility of emerging technologies such as CRISPR/Cas9, Case13, and multi-omics approaches. These innovations are instrumental for future discoveries and are poised to revolutionize wheat breeding by enabling precise genetic enhancements. Facing unprecedented challenges from climate change, the identification and utilization of these candidates is crucial. This review aims to be a comprehensive resource for researchers, providing an integrative understanding of complex traits in wheat and proposing new avenues for research and crop improvement strategies.

Diagnosis and management of non-CAH 46,XX disorders/differences in sex development.

Prenatal-onset androgen excess leads to abnormal sexual development in 46,XX individuals. This androgen excess can be caused endogenously by the adrenals or gonads or by exposure to exogenous androgens. The most common cause of 46,XX disorders/differences in sex development (DSD) is congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency, comprising >90% of 46,XX DSD cases. Deficiencies of 11β-hydroxylase, 3β-hydroxysteroid dehydrogenase, and P450-oxidoreductase (POR) are rare types of CAH, resulting in 46,XX DSD. In all CAH forms, patients have normal ovarian development. The molecular genetic causes of 46,XX DSD, besides CAH, are uncommon. These etiologies include primary glucocorticoid resistance (PGCR) and aromatase deficiency with normal ovarian development. Additionally, 46,XX gonads can differentiate into testes, causing 46,XX testicular (T) DSD or a coexistence of ovarian and testicular tissue, defined as 46,XX ovotesticular (OT)-DSD. PGCR is caused by inactivating variants in NR3C1, resulting in glucocorticoid insensitivity and the signs of mineralocorticoid and androgen excess. Pathogenic variants in the CYP19A1 gene lead to aromatase deficiency, causing androgen excess. Many genes are involved in the mechanisms of gonadal development, and genes associated with 46,XX T/OT-DSD include translocations of the SRY; copy number variants in NR2F2, NR0B1, SOX3, SOX9, SOX10, and FGF9, and sequence variants in NR5A1, NR2F2, RSPO1, SOX9, WNT2B, WNT4, and WT1. Progress in cytogenetic and molecular genetic techniques has significantly improved our understanding of the etiology of non-CAH 46,XX DSD. Nonetheless, uncertainties about gonadal function and gender outcomes may make the management of these conditions challenging. This review explores the intricate landscape of diagnosing and managing these conditions, shedding light on the unique aspects that distinguish them from other types of DSD.

Prenatal diagnosis of a trisomy 7 mosaic case: CMA, CNV-seq, karyotyping, interphase FISH, and MS-MLPA, which technique to choose?

ObjectiveThis study aims to perform a prenatal genetic diagnosis of a high-risk fetus with trisomy 7 identified by noninvasive prenatal testing (NIPT) and to evaluate the efficacy of different genetic testing techniques for prenatal diagnosis of trisomy mosaicism.MethodsFor prenatal diagnosis of a pregnant woman with a high risk of trisomy 7 suggested by NIPT, karyotyping and chromosomal microarray analysis (CMA) were performed on an amniotic fluid sample. Low-depth whole-genome copy number variation sequencing (CNV-seq) and fluorescence in situ hybridization (FISH) were used to clarify the results further. In addition, methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) was performed to analyze the possibility of uniparental disomy(UPD).ResultsAmniotic fluid karyotype analysis revealed a 46, XX result. Approximately 20% mosaic trisomy 7 was detected according to the CMA result. About 16% and 4% of mosaicism was detected by CNV-seq and FISH, respectively. MS-MLPA showed no methylation abnormalities. The fetal ultrasound did not show any detectable abnormalities except for mild intrauterine growth retardation seen at 39 weeks of gestation. After receiving genetic counseling, the expectant mother decided to continue the pregnancy, and follow-up within three months of delivery was normal.ConclusionIn high-risk NIPT diagnosis, a combination of cytogenetic and molecular genetic techniques proves fruitful in detecting low-level mosaicism. Furthermore, the exclusion of UPD on chromosome 7 remains crucial when NIPT indicates a positive prenatal diagnosis of trisomy 7.

Pathological changes of biochemical, hematological and coagulation analyses in patients with COVID-19 disease

The identification of patients with poor prognosis and early detection of COVID-19 disease complications are made possible by pathological analyses of routine hematological, coagulation, and biochemical tests. Interpreting analyses needs to be done within the framework of each patient’s unique clinical picture. It’s also critical to keep an eye on changes at the individual parameter level. From May 20th, 2021, to March 30th, 2024, a comprehensive search of literature was carried out using international databases, such as PubMed, Embase, Web of Science, Scopus, and the Cochrane Library, in compliance with the PRISMA guidelines. The research question was formulated using the PICO strategy. The following terms were used: biochemical parameters in COVID-19, hematological parameters in COVID-19, blood coagulation parameters in COVID-19, indicators of inflammation, and indicators of tissue damage in SARS-CoV-2. Routine hematological, coagulation, and biochemical tests are primarily used to monitor the progression of the disease and the effectiveness of treatment rather than being utilized for the established diagnosis of COVID-19 due to their low specificity. Molecular genetics and immunological techniques should be used to determine the COVID-19 disease diagnosis.