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Macewicz L.L.,NASU Institute of Molecular Biology and Genetics | Kushniruk V.O.,NASU Institute of Molecular Biology and Genetics | Iatsyshyna A.P.,NASU Institute of Molecular Biology and Genetics | Kotsarenko K.V.,NASU Institute of Molecular Biology and Genetics | And 5 more authors.
Biopolymers and Cell | Year: 2013

Long-term cultivation of cell lines inevitably leads to genetic and epigenetic changes. Aim. A comparative analysis of karyotype and level of the expression of reparative enzyme O6-methylguanine-DNA-methyltransferase (MGMT) at different stages of establishment and stabilization of human cell line 4BL and cell line of mouse germ cells G1. Methods. The set of methods was used to research the dynamics of karyotypes changes: the differential staining of chromosomes, FISH-method and comparative genomic hybridization. The level of MGMT expression was analyzed by PCR reaction and Western blot analysis. Results. General trends of establishment of mouse and human cell lines were revealed: at the first stage, which is characterized by increased structural instability of the genome, an increase in the MGMT expression was revealed while at the second stage of stabilization - a decrease in the expression. Therefore, almost complete disappearance ofMGMT protein in unmodified form (24 kDa) is observed. Conclusions. Statistically significant correlation between MGMT repair enzyme and mutations induction processes during mammalian cell adaptation and cell line establishment to in vitro was described. © Institute of Molecular Biology and Genetics, NAS of Ukraine, 2013. Source


Pylyp L.Y.,Clinic of Reproductive Medicine Nadiya | Pylyp L.Y.,National University of Kyiv-Mohyla Academy | Zukin V.D.,Clinic of Reproductive Medicine Nadiya | Bilko N.M.,National University of Kyiv-Mohyla Academy
Journal of Assisted Reproduction and Genetics | Year: 2013

Purpose: To study meiotic segregation patterns of Robertsonian translocations in sperm of male carriers and to assess the frequencies of unbalanced sperm formation. Methods: FISH with combination of probes to detect all the variants of meiotic segregation was performed on decondensed sperm nuclei of 5 carriers of der(13;14), 3 carriers of der(14;21) and one carrier of a rare der(13;21) translocation. Results: The frequency of sperm with alternate segregation and normal/balanced chromosomal complement ranged from 68 % to 94.4 % (mean 79.2 ± 8.4). Adjacent segregation was detected in 17.9 ± 7.3 % of sperm (from 5.6 % to 29 %). No significant differences in frequencies of gametes with nullisomies and disomies of chromosomes involved in translocations were observed. The mean frequency of 3:0 segregation products was 2.5 ± 1.4 %. Conclusions: All analyzed patients showed homogenous segregation pattern with clear predominance of alternate segregation resulting in normal/balanced sperm production. Still, from 5.8-32 % (mean 20.4 ± 8.3 %) of sperm was unbalanced, which is the evidence of the increased risk of unbalanced offspring in carriers of Robertsonian translocations. Our results highlight the importance of genetic counseling of Robertsonian translocation carriers prior to ICSI or IVF. © 2013 Springer Science+Business Media New York. Source


Pylyp L.Y.,National University of Kyiv-Mohyla Academy | Spinenko L.A.,Clinic of Reproductive Medicine Nadiya | Zukin V.D.,Clinic of Reproductive Medicine Nadiya | Bilko N.M.,National University of Kyiv-Mohyla Academy
Cytology and Genetics | Year: 2014

Meiotic segregation of chromosomes 13 and 14 was assessed by fluorescence in situ hybridization on sperm of five heterozygous carriers of the most frequent Robertsonian translocation der(13;14). Alternate segregation mode was predominant (mean 78.2 ± 5.7%). The prevalence of balanced sperm varied from 69.4 to 86.5%. Adjacent segregation mode was detected in 18.64 ± 4.90% of sperm; 3:0 mode was detected in 2.48 ± 1.20% of sperm. These results are informative for reproductive counseling of Robertsonian translocation der(13;14) carriers providing information for assessment of probability of receiving normal/balanced embryos in assisted reproduction cycles. © 2014 Allerton Press, Inc. Source


Sudoma I.,Clinic of Reproductive Medicine Nadiya | Sudoma I.,National Medical Academy For Postgraduate Education Named After P Shupyk | Goncharova Y.,Clinic of Reproductive Medicine Nadiya | Zukin V.,Clinic of Reproductive Medicine Nadiya | Zukin V.,National Medical Academy For Postgraduate Education Named After P Shupyk
Reproductive BioMedicine Online | Year: 2011

The aim of this study was to investigate pinopode formation patterns in patients with a history of multiple IVF failures and to evaluate if their detection with subsequent modification of protocols using frozen-thawed embryos could help to increase the pregnancy and live-birth rates in these patients. The study included 55 women with at least three implantation failures. On-time pinopodes were present in only 12.7% of cases, the rest showed acceleration, delay, arrest or asynchronization of pinopode formation. Patients with anomalies of pinopode development were divided into two subgroups, one of which had the standard protocol and the other a modified protocol, in which the duration of progesterone administration before embryo transfer was changed according to pinopode formation pattern. In nine patients with the modified protocol, with asynchronized and arrested pinopodes, simultaneous transfer of embryos of different ages was fulfilled. The implantation, pregnancy and take-home-baby rates for the modified and standard protocols were 28.0% versus 6.0%, 52.4% versus 12.0% and 47.6% versus 12.0%, respectively. Detection of inappropriately timed pinopode formation with subsequent synchronization of embryonic development and endometrial maturation allowed improvements in the effectiveness of programmes using frozen-thawed embryos in women with multiple implantation failure. © 2011 ALPHA Scientists in Reproductive Medicine and the European Society of Human Reproduction and Embryology. Published by Elsevier Ltd. All rights reserved. Source


Pylyp L.Y.,Clinic of Reproductive Medicine Nadiya | Spinenko L.O.,Clinic of Reproductive Medicine Nadiya | Verhoglyad N.V.,Clinic of Reproductive Medicine Nadiya | Zukin V.D.,Clinic of Reproductive Medicine Nadiya
Journal of Assisted Reproduction and Genetics | Year: 2013

Purpose: To assess the frequency and types of chromosomal abnormalities in 204 Ukrainian patients with non-obstructive azoospermia and oligozoospermia and 87 men with normozoospermia. Methods: Cytogenetic studies were performed on peripheral blood lymphocyte samples of 164 men with oligozoospermia, 40 men with non-obstructive azoospermia and 87 men with normozoospermia attending infertility clinic. Results: Chromosomal abnormalities were detected in 17 % of patients with sperm disorders: in 35 % of men with azoospermia and in 12.7 % of men with oligozoospermia. The frequency of chromosomal abnormalities in patients with sperm disorders was significantly higher, than in patients with normozoospermia (P = 0.0001). An increase in the incidence of chromosomal abnormalities with the decrease of sperm count was observed. Chromosomal abnormalities were detected in 1.1 % of patients with normozoospermia, 6.5 % of patients with mild oligozoospermia (sperm count 5-15 × 106/ml), 18.4 % of patients with severe oligozoospermia (sperm count <5 × 106/ml) and 35 % of patients with azoospermia. A significant increase in the frequency of chromosomal abnormalities in patients with severe oligozoospermia was observed when compared to mild oligozoospermia (P = 0.01). A statistically significant association (P = 0.02) of chromosomal abnormalities and sex chromosome abnormalities (P = 0.0001) with azoospermia when compared to oligozoospermia was observed. Conclusions: Our results highlight the importance of cytogenetic studies in patients with oligozoospermia (both mild and severe) and non-obstructive azoospermia. The presence of chromosomal abnormalities influences significantly the fertility treatment protocols, as well as provides a definite diagnosis to couples suffering from infertility. © 2013 Springer Science+Business Media New York. Source

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