Endometrial Receptivity Analysis - a tool to increase an implantation rate in assisted reproduction.

INTRODUCTION: A successful embryo implantation is crucial for a positive outcome of in vitro fertilization. But there is only a short period during which the endometrium is receptive for embryo, this so called implantation window can be detected by a molecular diagnostic method endometrial receptivity analysis (ERA). OBJECTIVE: To find out the percentage of patients with a non-receptive endometrium in the time of ERA and to learn what part of them got pregnant after the identification of their personalized implantation window. DESIGN: A retrospective study. SETTING: REPROMEDA Biology Park, Centre of Reproductive Medicine and Preimplantation Genetic Diagnosis, Brno. METHODS: A cohort of 85 patients undergoing ERA from August 2015 to October 2018 was studied. 74 patients experienced a previous implantation failure, the average number of preceding unsuccessful frozen embryo transfers was 2,5 in this group, 11 women went through ERA due to the preventive reason before the first FET. In all women one euploid embryo was transferred. 48 patients were prepared either for ERA or FET in a natural menstrual cycle, 37 women in HRT cycle. We were interested in a percentage of non-receptive patients in the time of ERA and wanted to discover what part of non-receptive women got pregnant after the identification of their personal implantation window. The average number of frozen embryo transfers needed to achieve the pregnancy was also calculated. RESULTS: 31 of 85 patients (36.5%) were found to have a non-receptive endometrium. In the natural cycle 13 of 48 (27.1%) were non-receptive: five were pre-receptive, three early receptive, two late receptive and three post-receptive. In the HRT cycle 18 of 37 patients (48.6%) were non-receptive: 12 were pre-receptive, four early receptive, one late receptive, one post-receptive. Personalized FET was done in 26 of total 31 initially non-receptive patients, 18 of them got pregnant (69.2%). In the natural cycle 6 of 11 (54.5%) achieved the pregnancy, in the HRT cycle 12 of 15 women (80.0%) got pregnant. To achieve the clinical pregnancy 1.5 frozen embryo transfer in average was needed. CONCLUSION: A displaced implantation window was found in more than 1/3 of patients undergoing an assisted reproductive treatment. After the personalized FET the clinical pregnancy was noticed in 69.2% of them. This result supports an individual approach to patients in IVF programme besides other at the timing of embryo transfer after the identification of pWOI.

[Assisted Reproduction and Preimplantation Genetic Diagnosis in Patients Susceptible to Breast Cancer]. / Asistovaná reprodukce a preimplantacní genetická diagnostika u pacientek ohrozených karcinomem prsu.

BACKGROUND: Assisted reproduction, as well as pregnancy itself, in patients with breast cancer or other hereditary type of cancer, is a widely discussed topic. In the past, patients treated for breast cancer were rarely involved in the discussion about reproductive possibilities or infertility treatment. However, current knowledge suggests, that breast cancer is neither a contraindication to pregnancy, nor to assisted reproduction techniques. On the contrary, assisted reproduction and preimplantation genetic diagnosis methods might prevent the transmission of genetic risks to the fetus. AIM: In this review we summarize data concerning pregnancy risks in patients with increased risk of breast cancer. In addition, we introduce current possibilities and approaches to fertility preservation prior to assisted reproduction treatment as well as novel methods improving the safety of fertility treatment. In the second part of this review, we focus on karyomapping--an advanced molecular genetic tool for elimination of germinal mutations in patients with predisposition to cancer. Moreover, the rapid development of preimplantation genetic diagnosis methods contributes to detection of both chromosomal aneuploidy and causal mutations in a relatively short time-span.

Immunodetection and N-terminal sequencing of DNA replication proteins of bacteriophage BFK20 - lytic phage of Brevibacterium flavum.

Phages are excellent models for studying the mechanism of DNA replication in prokaryotes. Identification of phage proteins involved in phage DNA replication is the first prerequisite for elucidation of the phage replication module. We focused on replication proteins gp41 (a putative helicase from SF2 superfamily), gp43 (a RepA-like protein), and gp44 (a putative DNA polymerase A) of phage BFK20 grown in Brevibacterium flavum. To identify them in the phage-host system, we prepared antibodies to these proteins which were cloned and expressed in Escherichia coli as his-tagged recombinant proteins. After purification to homogeneity the recombinant proteins served for raising specific polyclonal antibodies in mice. Using these antibodies in Western blot analysis the phage proteins gp41, gp43 and gp44 were detected during the phage growth cycle. The proteins gp41 and gp43, prepared from cell lysate by ammonium sulphate precipitation, were N-terminally sequenced and found to contain the sequences N-SVKPRELR-C and N-MLGSTML-C, respectively. This means that gp41 starts with serine but not with common methionine. We consider these findings an initial but important step towards more thorough characterization of replication proteins of phage BFK20.