Cytogenetic Methods Research Articles

The Anti-Genotoxic Activity of Wastewaters Produced after Water-Steam Distillation of Bulgarian Rosa damascena Mill. and Rosa alba L. Essential Oils.

The steam distillation of valuable rose essential oil from R. damascena Mill. and R. alba L. generates large volumes of wastewaters. Although such wastewaters are bio-pollutants, they contain valuable bioactive compounds. In this study we investigated the cytotoxic/genotoxic and anti-cytotoxic/anti-genotoxic potential of these products. We used cytogenetic methods for induction of chromosome aberrations and micronuclei in two different experimental test-systems: ahigher plant and human lymphocyte cultures. Different experimental schemes of treatment with the waste products showed that the genotoxic activity of wastewater from the distillation of oils from R. alba and R. damascena was low in both test–systems. Human lymphocytes showed a higher sensitivity to the products than plant cells. Both types of waste products manifested anti-genotoxic effect against N-methyl-N′-nitro-N-nitrosoguanidine, a direct mutagen. The wastewaters obtained from steam distillation of rose essential oil have cytoprotective/genoprotective effect and could decrease DNA damage. Data are promising for further use of these products in pharmacy and other areas of human life.

Tracing the Evolution of the Angiosperm Genome from the Cytogenetic Point of View

Cytogenetics constitutes a branch of genetics that is focused on the cellular components, especially chromosomes, in relation to heredity and genome structure, function and evolution. The use of modern cytogenetic approaches and the latest microscopes with image acquisition and processing systems enables the simultaneous two- or three-dimensional, multicolour visualisation of both single-copy and highly-repetitive sequences in the plant genome. The data that is gathered using the cytogenetic methods in the phylogenetic background enable tracing the evolution of the plant genome that involve changes in: (i) genome sizes; (ii) chromosome numbers and morphology; (iii) the content of repetitive sequences and (iv) ploidy level. Modern cytogenetic approaches such as FISH using chromosome- and genome-specific probes have been widely used in studies of the evolution of diploids and the consequences of polyploidy. Nowadays, modern cytogenetics complements analyses in other fields of cell biology and constitutes the linkage between genetics, molecular biology and genomics.

EP406: Germline 16p13.1 microdeletion identified during routine hematologic testing

Inversion 16 or translocation 16;16, resulting in CBFB::MYH11 fusion, accounts for approximately 5–8% of acute myeloid leukemia (AML) and is recognized by the World Health Organization (WHO) as a distinct entity within the category of AML with recurrent genetic abnormalities. It is critical to correctly characterize AML patients with CBFB::MYH11 fusion for appropriate diagnosis and the association with a favorable clinical prognosis (a high rate of complete remission and favorable overall survival). However, germline microdeletions spanning the MYH11 gene region are associated with a rare, but recurrent, 16p13.1microdeletion syndrome. We present this case series to highlight the risk of neoplastic versus germline mutational results involving the MYH11 locus and to initiate a discussion on standardization for reporting incidental germline findings in the setting of hematologic testing. We performed a retrospective review of the Mayo Clinic Cytogenetics Database and identified four cases that were tested using the Mayo homebrew CBFB::MYH11 dual-color, dual-fusion FISH (D-FISH) probe strategy and had a probable MYH11 germline microdeletion (Table 1). In 3 of 4 cases, loss of the MYH11 probe was observed in the absence of CBFB::MYH11 gene fusion. Since the MYH11 deletion was identified in 97-100% of nuclei in each of these cases, this result strongly suggested a germline 16p deletion. In addition, a MYH11 gene deletion is not associated with diagnostic or prognostic significance in the context of myeloid neoplasia. Germline 16p13.1 microdeletion syndrome is characterized by a broad spectrum of clinical phenotypes, with the most severely affected having intellectual disability, epilepsy and additional phenotypic features. However, the 16p13.1 microdeletion has also been identified in individuals with very mild clinical phenotypes and in apparently normal individuals, suggesting either incomplete penetrance and/or variable expressivity of the phenotype. In 3 of 4 patient cases identified in this study through an initial hematologic evaluation, additional studies were recommended to document the constitutional origin of the MYH11 gene deletion. To our knowledge, none of these cases pursued follow-up germline testing. However, in one patient (patient 2) FISH performed at diagnosis showed CBFB::MYH11 fusion in addition to the MYH11 deletion. A follow-up bone marrow specimen demonstrated no evidence of CBFB::MYH11 fusion, but the MYH11 deletion persisted, representing additional evidence for a constitutional deletion of the MYH11 gene region in this patient. These cases illustrate the possibility of discovering incidental constitutional abnormalities when performing testing on hematologic neoplasms. Currently there are no defined guidelines for the detection and reporting of possible germline alterations detected by conventional cytogenetic methods in hematologic disease. Consequently, patients may not be counselled appropriately, and testing consent forms in cancer settings often do not address the possibility of the discovery of germline abnormalities. These cases highlight a need of standardization for reporting incidental germline findings from hematologic testing by clinical laboratories as well as consenting and genetic counselling from the clinical perspective.Tabled 1Age (years)SexReason for referralSpecimennuc ish(MYH11x1,CBFBx2) %Additional clonal abnormalitiesPatient 122FemaleAcute myeloid leukemiaBone marrow100.0NoPatient 259FemaleInversion 16 at diagnosisBone marrow99.6UnknownPatient 339FemaleAcute myeloid leukemia relapseBlood97.4NoPatient 459FemalePancytopeniaBone marrow100.0No Open table in a new tab

EP391: Optical genome mapping for constitutional postnatal SV, CNV, and repeat array sizing: A multi-site clinical study

Chromosomal aberrations that include structural variations (SVs), copy number variations (CNVs), and tandem repeat expansions/contractions are major causes of human genetic disorders. Current cytogenetic methods that are routinely employed for the detection of SVs and CNVs are low resolution (karyotype), targeted and require prior knowledge (FISH), or cannot detect balanced SVs and orientation of duplicated segments of genome (chromosomal microarrays). Repeat array analysis methods are time-consuming and have low accuracy (southern blotting) or low dynamic range (PCR, NGS).

The effect of Anzer honey on X-ray induced genotoxicity in human lymphocytes: An in vitro study.

Anzer honey is well known in Turkey and used for its medicinal properties, especially for pharyngitis, tonsillitis, ulcers and cancer. In this study, we investigated whether Anzer honey, which is shown to have antioxidant, anti-tumoral, and anti-inflammatory properties, has a protective effect against X-ray induced genotoxic damage by cytogenetic methods. Peripheral blood lymphocytes isolated from 20 healthy volunteers were divided into two groups and cultivated by conventional methods. Study group lymphocytes were treated with 10% diluted honey while those in the control group were not. Both groups were exposed to a high dose (2 Gy) X-ray at the 48th hour of culture. Conventional cytogenetic staining and Giemsa banding methods were applied to evaluate chromosomal breakage and ring formation. Micronucleus frequencies were determined by the cytokinesis-block micronucleus (CBMN) assay. Paired sample t test was used to compare groups. Anzer honey, which was analyzed melissopalynologically, was used. Micronucleus frequency was significantly decreased in the study group (CI=348.75 ± 31, median 326, min. 98, max. 704) compared to the control group (CI=489.10 ± 27, median 500, min. 216, max. 645) (p=.001). Chromosomal breakage was also significantly decreased in the study group (CI=118.70 ± 16, median 109, min. 12, max. 316) compared to the control group (CI=233.60± 25, median 225, min. 65, max. 492) (p < .0001). This is the first study indicating that genotoxic damage in the peripheral blood lymphocytes of healthy volunteers induced by X-radiation may be prevented or alleviated by adding Anzer honey in vitro. These results encourage further research about the protective effects of honey. RESEARCH HIGHLIGHTS: Anzer honey has a genoprotective effect against radiation-induced genotoxicity, probably by preventing oxidation damage.

Characteristics of the Edilbay sheep breed of the Republic of Kazakhstan

The article analyses material from sheep domestication to the present state. Data on the world’s leading countries, where sheep breeding is most intensively developed, are given. The largest sheep populations are concentrated in China, Australia, India and Sudan. There are 995 registered local sheep breeds, of which 100 are international cross-border sheep breeds. Many species are continually becoming extinct. Therefore, the problem of preserving the unique gene pool of aboriginal breeds is acute in many countries. More than 20 breeds of sheep are bred in Kazakhstan. Over the last ten years, the number of sheep in the Republic has been between 18.0-20.0 million. The most widespread sheep breeds are Edilbay, Kazakh Arkharmerinos and Kazakh Kurdish coarse-wool sheep. Sheep of the Edilbay breed are bred in 10 regions of Kazakhstan. Edilbay breed is adapted to breeding in areas of dry steppes, semi-desert and desert regions. The article gives a brief zootechnical characteristic of Edilbay sheep and describes the epizootic situation in the Republic of Kazakhstan. Particular attention is paid to changing the breeding strategy. In further breeding and pedigree work, the aim is to create a sheep population with reduced fat content. A plan has been outlined for comprehensive studies of the sheep gene pool and phenofund, including evaluating protein, carbohydrate, fat and mineral metabolism, chemical, physiological, cytogenetic, molecular-genetic, and other research methods. Water, soil, and feed will be monitored for heavy metals and macronutrients during the experimental work.

Genotype-phenotype correlations for the type 1 lissencephaly of eight Tunisian children

Background: Lissencephaly represents a rare subgroup of genetically distinct neurological disorders of neuronal migration characterized by a paucity or absence of cerebral gyration. The most common form of lissencephaly has been isolated and referred to as classic or type 1 lissencephaly. It is frequently related to abnormalities within LIS1 or DCX genes, with abnormalities ranging from single base pair substitutions to contiguous gene deletions. Methods: In this study, we report, for the first time, a clinical and genetic characterization of eight unrelated Tunisian children presenting type 1 lissencephaly. We screened LIS1 and DCX abnormalities thanks to a combination of molecular cytogenetic methods and next generation sequencing. Results: One deletion of DCX and three deletions of LIS1 were observed. One of these deletions was inherited from a maternal reciprocal translocation and estimated to approximately 2,9 Mb length. In addition, a 26 Kb LIS1 deletion was detected and refined between exon 3 up to exon 11 by target capture and sequencing. The last LIS1 rearrangement was a mutation (c.779T&gt;A, p.V260E). Finally, two novel DCX mutations were found out (c.910G&gt;C, p.G304R/c.436T&gt;C, p.F146L). Conclusions: Our data confirm the individuality and originality of type 1 lissencephaly on both the phenotypic and the genetic levels. Furthermore, our data confirm again that LIS1 and DCX are the most genes associated with type 1 lissencephaly and spotlight the usefulness of developing approaches and methods for detecting a large number of known causative gene mutations.

Composite Classic Hodgkin Lymphoma and Follicular Lymphoma

Composite classic Hodgkin lymphoma and follicular lymphoma (CHLFL) is a rare and poorly characterized entity. Herein, we report the clinicopathologic features of 22 cases of CHLFL from 3 institutions and we assess 27 additional cases reported in the literature. In our cohort (n=22), patients with CHLFL had a median age of 61 years and an equal male to female incidence. Most cases (95%) arose de novo with the remaining patients having a history of non-Hodgkin lymphoma. CHLFL always involved lymph nodes (100%) and most cases (95%) revealed 2 distinct areas separately diagnostic for CHL and FL. The CHL component represented a variable proportion of the overall neoplasm (5% to 90%) and was either mixed cellularity (82%) or nodular sclerosis (18%) type. The Hodgkin/Reed-Sternberg cells expressed CD30 (100%), PAX5 (100%), CD15 (62%), BCL6 (47%), BCL2 (29%), and EBER (25%), in a polymorphous inflammatory background typical of CHL. The FL component was low-grade in 55%, grade 3A in 36%, and grade 3B in 9% of cases. All 3 cases investigated by cytogenetic methods for a clonal relationship between the CHL and FL components were clonally related. These clinicopathologic features of our cohort are similar to those of cases reported in the literature. The 5-year overall survival in combined patients with CHLFL (n=49) was 48%, comparable to CHL but worse than FL in the elderly. In summary, CHLFL is a rare entity that most often occurs in older adults, involves lymph nodes, and most commonly presents de novo. In the small number of cases assessed, the CHL and FL components are usually clonally related suggesting that the CHL and FL components may share a common progenitor B-cell, likely a mutated germinal center B-cell.

Using the CRISPR/Cas9 System for Dissection of Functional Sites of the Notch Gene in Drosophila melanogaster.

The Notch gene is a key factor in the signaling cascade that allows communication between neighboring cells in many organisms, from worms and insects to humans. The relative simplicity of the Notch pathway in Drosophila, combined with a powerful set of molecular and cytogenetic methods, makes this model attractive for studying the fundamental principles of Notch regulation and functioning. Here, using the CRISPR/Cas9 system in combination with homologous recombination, for the first time at the level of the whole organism, we obtained a directed deletion of the 5'-regulatory region and the first exon of the Notch gene, which were replaced by the attP integration site of the ΦC31 phage. Based on this approach, we obtained and characterized new Notch mutations. Thus, a new powerful tool is provided for studying the genetic regulation of the Notch gene and the organization of chromatin at this locus.

ЦИТОГЕНЕТИЧЕСКИЕ МАРКЕРЫ ДЛЯ ОТБОРА МАТЕРИНСКИХ ДЕРЕВЬЕВ СОСНЫ ОБЫКНОВЕННОЙ И ЕЛИ БЕЛОЙ, ПРОДУЦИРУЮЩИХ СЕМЕННОЕ ПОТОМСТВО С ОПРЕДЕЛЕННЫМ УРОВНЕМ ГЕНЕТИЧЕСКОЙ СТАБИЛЬНОСТИ

Currently, the depletion of the gene pool of forest woody plants is observed due to the fact that in the course of selective felling, specimens valuable for economic characteristics were destroyed and seeds from the remaining specimens were used, which did not have the best qualities. It was noted that a high level of genetic heterogeneity allows the population to adapt to changing environmental conditions, ensures the stability of the population and is a resource for breeding. The selection of valuable genotypes in the creation of forest seed plantations, which is carried out according to the phenotype, is based on the heterogeneity of populations. Meanwhile, studies show that phenotypically normal trees do not always produce good offspring. The cytogenetic method allows assessing the offspring. With its help, it is possible to distinguish among the seed offspring the mother trees of the group with different levels of stability of the genetic material. The aim of the work was to identify markers among cytogenetic parameters for the selection of trees producing seed offspring with different stability of the genetic material. The objects of the study were seedlings of seeds of phenotypically normal Scots pine trees growing in the Khopersky State Nature Reserve (51°10’56.9″N 41°44’17.2″E), and white spruce growing in the Botanical Garden. prof. B.M. Kozo-Polyansky Voronezh State University (51°42’41.57 «N 39°12’17.57″E). The trees had no visible pest damage. The method of preparation and analysis of preparations of seed sprouts is described in the work of Butorina A.K. (2000). In the course of the study, 20 cytogenetic parameters were determined: the mitotic index (counted with and without taking into account cells at the prophase of mitosis), the level of mitotic pathologies (counted with and without taking into account cells at the prophase of mitotic), the proportions of cells at the stages of prophase, metaphase, anaphase , the proportions of cells with 1–10 nucleoli in the nucleus of interphase cells, the proportion of cells with micronuclei, the proportion of cells with residual nucleoli in interphase and mitosis. Statistical processing of the study results was carried out using the STADIA 7.0 and MedCalc 17.5.3 software. The diagnostic value of the indicator is characterized by the area under the ROC-curve: 0.9–1.0 – excellent; 0.8–0.9 – very good; 0.7–0.8 – good, 0.6–0.7 – average, 0.6 and less – unsatisfactory. Among the cytogenetic indicators, there are those that are suitable for separating the mutable group from the weakly mutable and intermediate, as well as the weakly mutable from the intermediate. In pine, these include the proportion of cells at the prophase stage and the average number of nucleoli in the nucleus, in spruce – the mitotic index without taking into account cells at the prophase stage, the level of mitotic pathologies with and without taking into account cells at the prophase stage and the proportion of cells at the metaphase stage. There are indicators that make it possible to distinguish the mutable group from the weakly mutable and from the intermediate, but are not suitable for separating the weakly mutable group from the intermediate. These include the mitotic index, calculated taking into account cells at the prophase stage in pine, and the proportion of cells at the metaphase and ana-telophase stages in spruce. According to some indicators, it is possible to distinguish the intermediate group from the mutable and the weakly mutable, but it is impossible to distinguish the mutable group from the weakly mutable. In pine, such indicators are the proportion of cells in the metaphase, and in spruce, the proportion of cells with residual nucleoli in the interphase and meta-telophase. Indicators have been identified that make it possible to distinguish only mutable group from weakly mutable (in pine, the proportion of cells at the ana-telophase stage), and weakly mutable group from intermediate (in pine, the mitotic index taking into account cells at the prophase stage, the level of mitotic pathologies with and without taking into account cells at prophase stage, in spruce – mitotic index, taking into account cells at the prophase stage, the proportion of cells with micronuclei).

From the ashes of "Ewing-like" sarcoma: A contemporary update of the classification, immunohistochemistry, and molecular genetics of round cell sarcomas.

Round cell sarcomas include a diverse group of bone and soft tissue tumors, which comprise well-defined entities as well as several nascent categories presented in the 2020 World Health Organization classification. The morphologic overlap yet disparate nosology, prognostic implications, and management strategies places a high value on ancillary testing, including a strategic immunohistochemical approach and directed confirmation by cytogenetic and molecular genetic methods. We review the diagnostic categories that have emerged from the former wastebasket "undifferentiated round cell sarcoma" ("Ewing-like" sarcomas), with an emphasis on algorithmic exclusion of nonsarcomatous entities, diagnostic stratification of well-defined entities (Ewing sarcoma, rhabdomyosarcomas, poorly differentiated synovial sarcoma), and a discussion of the new categories with novel genetic alterations (CIC-rearranged sarcomas, sarcomas with BCOR genetic alterations, and round cell sarcomas with EWSR1-non-ETS fusions).

Effect of UV Radiation and Other Abiotic Stress Factors on DNA of Different Wild Plant Species Grown in Three Successive Seasons in Alpine and Subalpine Regions

Plants in natural ecosystems are exposed to a combination of UV radiation, ionizing radiation (IR) and other abiotic factors. These factors change with the altitude. We investigated DNA alterations of some wild plants of different plant families in natural ecosystems at three altitudes in Rila Mountain, Bulgaria (1500, 1782, and 2925 m above sea level (a.s.l.) exposed to UV radiation, IR and other abiotic stresses, to assess the tolerance of plant species to the changing environmental conditions in three successive growth seasons. For this purpose, physicochemical, cytogenetic, and molecular methods were applied. DNA damage was assessed by micronucleus test and molecular method comet assay adapted and applied by us to wild plant species from Onagraceae, Rosaceae, Boraginaceae, Saxifragaceae, Orobanchaceae, Asteraceae and Poaceae families, growing at three different altitudes. Variability in the DNA sensitivity and the level of tolerance was observed among the plant species in response to combined abiotic factors assessed by induced DNA damage and gross beta activity. The studied representatives of Poaceae were less susceptible than the other studied species at all three altitudes and showed close level of DNA injuries to that of unaffected control plant grown in laboratory conditions. The lower levels of DNA damage of these wild plant species corresponded to their lower ability to accumulate radionuclides. There was a particularly pronounced low level of DNA injuries in the plant species at the highest altitude. The level of DNA damage showed correlation with the values of some abiotic environmental factors. The results would contribute to the elucidation of the extent of adaptation of plant species to the continuously changing environment and would be useful in selecting sensitive herbaceous monitor species for environmental impact assessment at mountain and alpine sites.

52. Optical genome mapping and SNP microarray: integrated workflow for optimizing analysis of products of conception

Conventional cytogenetic analysis of products of conception (POC) is of limited utility because of failed cultures, microbial contamination, and maternal cell contamination (MCC). Optical genome mapping is an emerging alternative to conventional cytogenetic methods but is limited to the need for ultra-high molecular weight DNA. Single nucleotide polymorphism (SNP) microarray has demonstrated higher performance compared to routine karyotype, but a high incidence of MCC precludes a valid clinical diagnosis.

7. Adoption of optical genome mapping in clinical cancer cytogenetic laboratory: A stepwise approach

Detection of chromosomal abnormalities is important for cancer patients. Compared to common cancer cytogenetics methods including karyotype, fluorescent in-situ hybridization (FISH), and chromosomal microarray (CMA), optical genome mapping (OGM) is an innovative method to detect CA. The objective of this pilot study was to determine a stepwise approach to adopt OGM in the clinical setting. Blinded study was used to compare OGM, karyotype, FISH, and CMA on five well-characterized cancer cases (four leukemia/lymphoma and one solid tumor).

Metaphase-Based Cytogenetic Approach Identifies Radiation-Induced Chromosome and Chromatid Aberrations in Zebrafish Embryos.

Metaphase-based cytogenetic methods based on scoring of chromosome aberrations for the estimation of the radiation dose received provide a powerful approach for evaluating the associated risk upon radiation exposure and form the bulk of our current knowledge of radiation-induced chromosome damages. They mainly rely on inducing quiescent peripheral lymphocytes into proliferation and blocking them at metaphases to quantify the damages at the chromosome level. However, human organs and tissues demonstrate various sensitivity towards radiation and within them, self-proliferating progenitor/stem cells are believed to be the most sensitive populations. The radiation-induced chromosome aberrations in these cells remain largely unknown, especially in the context of an intact living organism. Zebrafish is an ideal animal model for research into this aspect due to their small size and the large quantities of progenitor cells present during the embryonic stages. In this study, we employ a novel metaphase-based cytogenetic approach on zebrafish embryos and demonstrate that chromosome-type and chromatid-type aberrations could be identified in progenitor cells at different cell-cycle stages at the point of radiation exposure. Our work positions zebrafish at the forefront as a useful animal model for studying radiation-induced chromosome structural changes in vivo.

Chromosome and Genome Diversity in the Genus Trifolium (Fabaceae).

Trifolium L. is an economically important genus that is characterized by variable karyotypes relating to its ploidy level and basic chromosome numbers. The advent of genomic resources combined with molecular cytogenetics provides an opportunity to develop our understanding of plant genomes in general. Here, we summarize the current state of knowledge on Trifolium genomes and chromosomes and review methodologies using molecular markers that have contributed to Trifolium research. We discuss possible future applications of cytogenetic methods in research on the Trifolium genome and chromosomes.

B-cell acute lymphoblastic leukaemia: recent discoveries in molecular pathology, their prognostic significance, and a review of the current classification.

Acute lymphoblastic leukaemia (ALL) remains a leading cause of non-traumatic death in children, and the majority of adults diagnosed will succumb to the disease. Recent advances in molecular biology and bioinformatics have enabled more detailed genomic analysis and a better understanding of the molecular biology of ALL. A number of recurrent genomic drivers have recently been described, which not only aid in diagnosis and prognostication, but also may offer opportunities for specific therapeutic targeting. The present review summarises B-ALL genomic pathology at diagnosis, including lesions detectable using traditional cytogenetic methods as well as those detected only through advanced molecular techniques.

Conventional Cytogenetic and Molecular Analysis in Acute Myeloid Leukemia (AML) and Their Association with Overall Survival

Abstract Background: Conventional cytogenetic is one of the most important diagnostic tools for predicting the overall survival of the patients. Molecular genetics in acute myeloid leukemia (AML) has provided insights into the molecular mechanism of leukemogenesis. In this study we aimed to investigate the impact of cytogenetic and molecular methods on the survival of patients with de novo established AML in order to achieve a useful marker or test in the process of predicting the disease course. Material and methods: Eighty newly diagnosed AML patients who were treatment naive entered the study. Cytogenetic and molecular studies such as, the conventional karyotyping, sequencing and reverse transcriptase real time quantitative PCR (RT-qPCR) were included. Overall survival was calculated by Kaplan-Maier technique and the data were analyzed by SPSS.V.19. Results: Among 80 patients, 36 (45%) were female and 44 (55%) were male patients. Patients’ median age was 29 years, ranging from 1 to 76 years. The mean overall survival was 19 months (95% CI: 1523 months). The 1-year AML survival rate was 61%. There were significant differences in overall survival between the NPM1-mutated groups compared to the patients without any mutations (19% versus 61%) (p &lt; 0.032). Conclusion: This study makes a significant contribution in assessing the prognostic value of cytogenetic and molecular markers. This study showed the heterogeneity of de novo AML that involved various factors and prevalence of distinct cytogenetic subgroups. Our data in comparison with other population-based studies, confirmed a differential distribution of cytogenetic and molecular classification indicating geographic heterogeneity.

Array-Based Comparative Genomic Hybridization Analysis in Children with Developmental Delay/Intellectual Disability.

Developmental delay (DD) is a condition wherein developmental milestones and learning skills do not occur at the expected age range for patients under 5 years of age. Intellectual disability (ID) is characterized by limited or insufficient development of mental abilities, including intellectual functioning impairments, such as learning and cause–effect relationships. Isolated and syndromic DD/ID cases show extreme genetic heterogeneity. Array-based comparative genomic hybridization aCGH) can detect copy number variations (CNVs) on the whole genome at higher resolution than conventional cytogenetic methods. The diagnostic yield of aCGH was 15.0–20.0% in DD/ID cases. The aim of this study was to discuss the clinical findings and aCGH analysis results of isolated and syndromic DD/ID cases in the context of genotype-phenotype correlation. The study included 139 cases (77 females, 62 males). Data analysis revealed 38 different CNVs in 35 cases. In this study, 19 cases with pathogenic CNVs (13.6%) and five cases with likely pathogenic CNVs (3.5%) were found in a total of 139 cases diagnosed with DD/ID. When all pathogenic and likely pathogenic cases were evaluated, the diagnosis rate was 17.1%. The use of aCGH analysis as a first-tier test in DD/ID cases contributes significantly to the diagnosis rates and enables the detection of rare microdeletion/microduplication syndromes. The clear determination of genetic etiology contributes to the literature in terms of genotype-phenotype correlation.

PSX-B-13 Effect of epidermal growth factor and prolactin on the quality of bovine oocytes aging in vitro

Abstract The evaluation of factors responsible for the protection of the oocytes attained the metaphase-II stage from aging is importance for successful in vitro embryo reproduction. The aim of the present research was to study dose-dependent effects of epidermal growth factor (EGF) and prolactin (PRL) on the quality of bovine oocytes after their aging in vitro. Bovine cumulus-enclosed oocytes (CEOs) were matured in vitro for 20 h in TCM 199 containing 0.2 mM sodium pyruvate, 10% fetal calf serum (FCS), 10 μg/ml FSH and LH. At the end of in vitro maturation, oocytes were transferred to TCM 199 supplemented with 10% FCS (aging medium) and cultured for additional 24 h in the absence (Control) and in presence of EGF (10 and 50 ng/ml) and PRL (20 and 50 ng/ml). After prolonged culture oocytes were used for apoptosis detection (TUNEL staining, n=251) and the state of chromosomes evaluation (Tarkowski’s cytogenetic method, n=359). The data from 3–4 replicates were analyzed by ANOVA. At the end of prolonged culture (24 h) the rate of apoptotic oocytes in the Control group was 47.4±8.5%. EGF at concentration of 10 ng/ml and PRL at both doses decreased this rate to 15.0–22.1% (p &amp;lt; 0.05). Furthermore, PRL (not EGF) reduced the frequency of abnormal chromosome modifications (decondensation, adherence, clumping) at concentrations of 20–50 ng/ml from 58.7±2.1% (Control) to 41.2±1.9 and 45.6±2.7% respectively (p &amp;lt; 0.01). Thus, EGF and PRL is able to maintain the apoptosis resistance of bovine oocytes during their prolonged in vitro culture as well as PRL have the decelerating effect on abnormal modifications of M-II chromosomes. The research was supported by RFBR (17-29-08035) and the Ministry of Science and Higher Education of Russia.