The role of mesenchymal-epithelial transition in endometrial function and receptivity.

OBJECTIVE: During reproductive age of a woman, endometrium undergoes frequent stuctural and functional changes. Abilities of regeneration, remodelation and differentiation are precondition of endometrial receptivity and implantation and development of an embryo. These processes are conditioned by mutual transformation between mesenchymal and epithelial fenotype of endometrial cells: epithelial-mesenchymal transition (EMT) and mesenchymal-epithelial transition (MET). The aim of this study is to present contemporary knowledge of transformation between epithelial and mesenchymal endometrial cells and its influence on human fertility. DESIGN: Review article. SETTING: Department of Obstetrics and Gynecology, Faculty of Medicine, Masaryk university and University Hospital Brno; Department of Obstetrics and Gynecology, University Hospital Faculty of Medicine, Palacky University, Olomouc. METHODS: PubMed was searched for articles in English indexed until February 2019 with terms of „endometrial receptivity“, „embryo implantation“, „endometrial regeneration“, „mesenchymal-epithelial transition/transformation“. RESULTS: It has been proved, that mesenchymal stromal cells participate on regeneration of not only the endometrial stroma, but also of the epithelium. During endometrial decidualisation under influence of ovarian steroids, the MET is under way. Stromal fibroblasts gain the morfological and functional properties of epithelial cells. During implantaion of an embryo, the trofoblast interacts with decidualised endometrium. Epithelial cells transform into mesenchymal (EMT), which mediate the growth of trofoblast. CONCLUSION: Mutual transformation between stromal and epithelial cells in essential for normal function of endometrium and implantation and development of an embryo.

Uterine microbiome and endometrial receptivity.

OBJECTIVE: Structure and correct function of endometrium is necessary for embryo implantation, pregnancy development and childbirth. It is a result of many factors - anatomical and histological structure, hormonal effects and signalling pathways at the molecular genetic level. A little known phenomenon is the presence of microorganisms on the endometrium. Traditionally, the uterine cavity was considered sterile, but new findings have been changing this view fundamentally. The aim of this work is to present new findings on endometrial microbiome and its importance for embryo implantation and development. DESIGN: Review article. SETTING: Department of Obstetrics and Gynecology, Faculty of Medicine, Masaryk University and University Hospital Brno; Department of Obstetrics and Gynecology, University Hospital Faculty of Medicine, Palacky University Olomouc. METHODS: PubMed was searched for articles in English indexed until 30th June 2018 with terms of „uterine microbiome analysis“ and „endometrial receptivity“. RESULTS: The vital information on bacterial colonization of endometrium brought new diagnostic methods for their detection based on ribosomal RNA analysis in 16S subunit, which are capable of detection and exact identification of bacteria that cannot be detected by classical cultivation methods. The endometrial microbiome is assumed to modulate the function of endometrial cells and local immunity system, it prevents growth of pathogenic microorganisms by its presence and production of protective substances. CONCLUSION: Endometrial microbiome seems to be important factor of endometrial receptivity.

Bisphenols in the pathology of reproduction.

OBJECTIVE: Bisphenols are one of the most widespread endocrine disruptors that the population of west world countries is exposed to. Objective of this study is to summarize information about influence of bisphenols on reproduction health. DESIGN: Review article, Setting: Department of Obstetrics and Gynecology, Faculty of Medicine, Masaryk University and University Hospital Brno. METHODS: PubMed was searched for articles in English indexed bisphenol and reproduction up to October 2018. RESULTS: Increased levels of bisphenol A and S have been proven in body fluids and tissues. Bisphenol molecules have effect similar to estrogens therefore they influence hormonal regulation and activity of estrogen receptors. Their negative influence on oocyte maturation, spermatogenesis and development of reproductive system has been shown. Bisphenol S, which has replaced bisphenol A, has comparable negative effects on reproduction. CONCLUSION: Bisphenols are widespread endocrine disruptors that could cause severe fertility disorders of men and women.

[Medroxyprogesteron acetate use to block LH surge in oocyte donor stimulation]. / Vyuzití medroxyprogesteron acetátu k blokáde LH pri stimulaci dárkyní vajícek.

OBJECTIVE: To compare the results of ovarian stimulation with LH surge blockade by medroxyprogesterone acetate or GnRH antagonist in oocytes donors. To present current options of exogenous and endogenous progestins instead of GnRH analogues to block LH surge during ovarian stimulation. DESIGN: Retrospective study of oocyte donor cycles and literature review. SETTING: Department of Obstetrics and Gynecology, Faculty of Medicine, Masaryk university and University Hospital Brno. METHODS: Thirteen oocyte donors (26.8 ± 2.5 years old) were stimulated with recFSH and MPA to block the LH surge during January - October 2017. The results were compared to the previous cycle stimulated with recFSH and GnRH antagonist performed during January -December 2016. Wilcoxon´s pair test was used to test the statistics. A literature search of SCOPUS was carried out. RESULTS: In cycles with MPA blockade the average number of oocytes was 14.5 ± 5.1, in cycles with GnRH anta-gonist blockade 12.0 ± 4.5 oocytes (statistical significance p = 0.025). FSH total dose (1611 ± 327 vs. 1565 ± 322 IU), days of stimulation (9.1 ± 0.8 vs. 8.5 ± 0.9) and maximum estradiol levels (5.9 ± 2.1 vs. 6.0 ± 3.0 nmol/l) were not statistically different. Progestins are effective in blocking the LH surge during ovarian stimulation and do not affect the number and quality of collected oocytes or obtained embryos. Their adverse effect on the endometrial receptivity obstructs the embryo implantation in the same cycle. Such protocol requires total freezing and delayed transfer. Progestins can be used in a variety of stimulation protocols - progestin primed follicular phase stimulation, luteal phase stimulation with endogenous progesteron, double stimulation in follicular and luteal phase of the same cycle "duostim" in low responders. CONCLUSION: Eggs donor ovarian stimulation with MPA resulted in more oocytes than stimulation protocol with GnRH antagonist, the total dose of FSH and the length of stimulation were similar. According to current experiences progestins effectively block the LH surge and do not affect the number and quality of collected eggs and obtained embryos. Their use opens new possibilities of ovarian stimulation protocols and their flexibility. Its main constraint is that it requires total freezing and delayed transfer.

[Fertility preserving methods in women with breast cancer before gonadotoxic therapy]. / Zachování fertility u zen s karcinomem prsu pred gonadotoxickou lécbou.

OBJECTIVE: To present current options and own experiences with ovarian stimulation in young women with breast cancer before gonadotoxic therapy. DESIGN: Review article with own experiencies. METHODS: Literary search, analysis of own experiences with complex therapy in women with breast cancer planning pregnancy, definitions of stimulating protocols, using methods of assisted reproduction. SETTING: Department of Obstetrics and Gynecology, Faculty of Medicine, Masaryk University and University Hospital Brno. RESULTS: Embryo and oocyte cryopreservation are important methods of fertility preservation requiring controlled ovarian stimulation before the start of chemotherapy. Current studies demonstrante effectivenes of this special stimulating protocol and did not find negative side effects such a progression of the breast cancer. Since January 2016 to June 2016 we performed controlled ovarian stimulation and oocyte retrieval in three patients with breast cancer before chemotherapy. We used short stimulation protocol with recombinant FSH, GnRH agonists and letrozole and retrieved 13, 12 and 9 oocytes. We cryopreserved 8 and 6 embryos, one woman prefered freezing of 12 oocytes. During the stimulation estradiol level did not exceed 1,6 nmol/l. CONCLUSION: Neither the normal nor the cancerous cells in the breast react to the gonadotropins FSH, LH, nor to hCG. Conversely, there is a cellular proliferation and an increase in cancer cell lines with oestrogen receptors with exposure to oestrogen, and it is dose - dependent. The special stimulation protocol with FSH, GnRH agonists and letrozole is effective in gain of efficient amount of gamets with minimal increase of estradiole level. Current study reviews did not find higher risk of progression of breast cancer in association with ovarian stimulation. It is important to respect the recomendation of oncology committee and to effectively suppress the estradiol level.

Galantamine has impact on immunity in mice exposed to keyhole limpet hemocyanin.

OBJECTIVES: In this work, we hypothesized whether galantamine could interact with the cholinergic anti-inflammatory pathway and modulate immunity this way. BACKGROUND: Galantamine is a drug used for the therapy of Alzheimer disease. The drug inhibits enzyme acetylcholinesterase in the central nervous system, which causes better availability of neurotransmitter acetylcholine. METHODS: In the experiment, we immunized BALB/c laboratory mice by keyhole limpet hemocyanin (KLH) in combination with galantamine in a dose 0.02-0.5 mg/kg. The animals were sacrificed from 1 to 7 days after the substances applications and plasma was collected in order to examine immunochemical markers by enzyme-linked immunosorbent assay. RESULTS: We found significant drop in production of immunoglobulins and interleukin (IL) 4 level while IL2, IL4 and tumour necrosis factor α remained unaltered for the whole experiment. We infer that galantamine causes better availability of acetylcholine also in blood system, where the neurotransmitter interacts with nicotinic acetylcholine receptors on macrophages and initiates cholinergic anti-inflammatory pathway. CONCLUSIONS: In a conclusion, galantamine can cause lower efficacy of vaccination or immunity response to an infectious disease and the phenomenon should be taken into consideration in the current therapy (Tab. 1, Fig. 2, Ref. 24).

The intracellular concentration of homocysteine and related thiols is negatively correlated to sperm quality after highly effective method of sperm lysis.

The aims of this study were to determine concentrations of total homocysteine, cysteine, cysteinylglycine and glutathione in spermatozoa, seminal fluid and blood plasma and to analyse their relationships with sperm parameters. For this reason, a new highly effective method of spermatozoa lysis was developed, using methanol, freezing and subsequent thawing in ultrasonic bath. An HPLC-FD assay was conducted on thiols concentrations in lysed spermatozoa, seminal fluid and blood plasma. Concentrations of thiols in spermatozoa were significantly lower in men with normozoospermia than in samples with pathological semen parameters. Statistical analysis found significant correlations between thiol concentrations in spermatozoa and semen parameters, while the same analysis with thiol concentrations in seminal fluid was substantially less powerful. Only cysteinylglycine concentrations in seminal fluid significantly correlated with pathological semen parameters. No significant differences or correlations were found with blood plasma concentrations.

[Current issues of reproductive medicine in the Czech Republic]. / Soucasná témata reprodukcní medicíny v Ceské republice.

INTRODUCTION: During the 25th symposium of assisted reproduction in Brno was lunch time organised as the lunch table discussion on the selected topics of assisted reproduction. More than 150 specialists reviewed themes related to gynecology and embryology.Discussed topics: Lunch table discussion covered the following topics: (1) Cross-border health care in assisted reproduction; (2) Indication for PGS (preimplantation genetic screening) in the context of actual information; (3) Does ovarian stimulation belong to the ambulance of registering gynecologists? (4) Therapy with clomifen - only for IVF specialists? (5) How and with whom should psychological support be directed during IVF? (6) Stimulation in women with low ovarian reserve; (7) Is basic semen analyses sufficient? (8) Time-lapse systems as relevant markers of embryonic development; (9) How to be oriented with choices of media and consumables in the IVF lab, and (10) "Freeze All" - is this new trend in cryopreservation suitable for all? CONCLUSIONS: Panel conclusions were presented during the afternoon session, which had great attendance, featured lively commentary, and produced some definitive consensus. Certain issues remained inconclusive, and these matters will be the subject of further discussion in the future. Specific summation of all deductions is presented in this paper.

Fertility status of Hodgkin lymphoma patients treated with chemotherapy and adjuvant gonadotropin-releasing hormone analogues.

PURPOSE: Aim of this prospective observational study was to analyze fertility status of Hodgkin lymphoma (HL) patients treated with different types of chemotherapy while receiving GnRH analogues to preserve ovarian function. METHODS: Fertility status was assessed among 108 females in reproductive age treated by curative chemotherapy for freshly diagnosed HL between 2005 and 2010 in university-based tertiary fertility and oncology center. All patients received GnRH analogues during chemotherapy to preserve their ovarian function. Their reproductive functions were assessed by follicle-stimulating hormone (FSH) measurement and pregnancy achievement. Ovarian function was determined separately in three groups with increasing gonadotoxicity of chemotherapy. RESULTS: One year following the treatment, normal ovarian function was found in 89 (82.4%) of patients. Two years after chemotherapy, 98 (90.7%) of patients retained their ovarian function, and 23 (21.3%) achieved clinical pregnancy during the follow-up period. Average FSH after chemotherapy was 11.6 ± 17.9 IU/l 1 year after the treatment resp. 9.0 ± 13.8 at the 2 years interval. There were significantly more patients with chemotherapy induced diminished ovarian reserve (chDOR) among the group receiving escalated BEACOPP chemotherapy in comparison with the other types of treatment (58.1% vs. 87.9% resp. 95.5%). CONCLUSION: The rate of chDOR is significantly higher after EB poly-chemotherapy and there is no tendency for improvement in time. The 2 + 2 chemotherapy with GnRH-a required for more advanced HL retained ovarian function significantly better after 2 years. Another important advantage of GnRH-a co-treatment is the excellent control of patient's menstrual cycle.

[Guidlines for the good laboratory practice in assisted reproduction - clean environment]. / Smernice ke správné laboratorní praxi pri asistované reprodukci - cistota prostredí

The guidlines are the recommendation for good laboratory practice in embryological laboratories. In this first part the requirements of the clean environment from the point of view of the oocytes, sperm, and embryos protection against infection and from point of view of the embryological laboratory staff health protection were described.

[Activities and responsibilities of workers in embryologic and andrologic laboratories in assisted reproduction centers]. / Cinnosti a odpovednosti pracovníku embryologické a andrologické laboratore v centrech asistované reprodukce.

This paper presents the current status and rules for the laboratory staff activities and their competences in the centers of assisted reproduction. The rules were processed by the members of the Association of Reproductive Embryology (ARE) committee under the current legislation. Committee members of the Czech Sterility and Assisted Reproduction Society and Czech Gynecology and Obstetric Society approved these rules as obligatory for assisted reproduction centres in Czech Republic.

[Current issues of assisted reproduction in the Czech Republic]. / Aktuální otázky asistované reprodukce v Ceské republice.

INTRODUCTION: Participants of the lunch table discussion held during the 22nd symposium of assisted reproduction in Brno discussed some current topics of assisted reproduction. DISCUSSED TOPICS: More than 150 participants at round tables discussed 10 topics: 1. IVF in native AR cycle,2. observation of the embryo development dynamics, 3. evaluation and support of endometrial receptivity,4. increased number of elective single embryo transfers (eSET), 5. transport of gametes and embryos in the Czech Republic and between the Czech Republic and abroad, 6. National registry of assisted reproduction,7. new view on sperm pathology, 8. problems with the SAR membership records, 9. surogacy motherhood and 10. preimplantation genetic diagnosis and preimplantation genetic screening. CONCLUSIONS: All findings were presented in the afternoon session. Some of the topics brought concrete results. Some topics could not be clearly concluded and will be the subject of further discussions. A brief summary of those discussion conclusions presents this paper.

[Theoretic and practical content of the clinical embryology]. / Teoretická a praktická nápln oboru klinická embryologie.

Clinical embryology is in whole Europe respected health care specialization. The specialists in this field are certified by the European Society of Human Reproduction and Embryology (ESHRE). In the Czech Republic the specialization has been formally established by the Government Order No 31/ 2010 after 30 years unformal existence. Concurrently with the wide development of new techniques in assisted reproduction, there is necessary to define the content of clinical embryology as a health care, scientific and educational discipline. The definition authorized by the Committee of Association of Reproductive Embryology (ARE) is given in this article..

[Ovarian tissue cryopreservation in cancer patients--six years of clinical experience]. / Kryokonzervace ovariální tkáne u onkologických pacientek--6 let klinických zkuseností.

OBJECTIVE: Presentation of clinical results and experience with this technique during past six years. DESIGN: Original paper. SETTINGS: Gynekologicko-porodnická klinika LF MU a FN Brno, Interní hemato-onkologická klinika LF MU a FN Brno, Department of Obstetrics and Gynecology. Hadassah University Hospital Ein-Karem, Jerusalem, Izrael. INTRODUCTION: Ovarian tissue cryopreservation (OTC) and its future auto-transplantation becomes an alternative for patients to prevent serious damage of ovarian function by oncology treatment. METHODS: Patient is indicated to OTC in case of high risk of ovarian failure due to planned chemotherapy and impossibility to use other oncofertility techniques. Ovarian tissue harvesting is done by laparoscopy in short-term general anesthesia. After tissue processing the samples are cryopreserved in programmable automatic freezer or by vitrification. The auto-transplantation of ovarian tissue is planned after the complete cure of patient's malignancy. Our workplace doesn't have own experience with tissue transplantation - until now cryopreserved tissue has not yet been utilized by the patients. Clinical experience with this technique gained by our team during academic stay in abroad Israeli clinic is presented. RESULTS: During the years of 2005-2011 the OTC was performed in 19 cancer patients before chemotherapy. In majority of cases, patients suffered from blood or lymph node systemic malignancy (84%). Average age of women was 26 years. The patient set consisted of mostly nulliparous women (88%). Patient's average body mass index was 23,9 kg/m2. The length of systemic chemotherapy averaged 7.1 months. Time from fertility preservation counseling to chemotherapy was not exceeding one week (7.2 days on average). Ovarian tissue harvesting was conducted by laparoscopic surgery in all cases. The length of surgery did not exceed 60 minutes and no surgical complications were observed. The case of ovarian tissue transplantation performed on abroad university settings is discussed. CONCLUSION: In the consensus of with international guidelines OTC is offered to patients with high risk of ovarian failure doe to cytotoxic oncology treatment. Research in the field of oncofertility is focused on the techniques of in-vitro folliculogenesis in retrieved ovarian tissue.

[New methods increasing assisted reproduction results]. / Nové metody zvysující úspesnost asistované reprodukce.

OBJECTIVE: The brief review of historical development and application of the assisted reproduction methods at our centre and the recent methods increasing the assisted reproduction results. The new mentioned methods are sperm selection before the intracytoplazmatic sperm injection (PICSI = preselected sperm intracytoplasmic injection) and continuous embryo development monitoring. DESIGN: Review article.

[Heterotopic pregnancy as an complication during pregnancy and labour--the case report]. / Tehotenství a porod zdravého dítete komplikované koincidencí heterotopické gravidity po IVF a placenty perkrety s následnou hysterektomií.

UNLABELLED: Heterotopic gravidity (HG) is a rare co-existence of intrauterine and ectopic pregnancy with higher incidence in pregnancies after in vitro fertilisation (IVF). Diagnosis of HG is demanding. HG is commonly identificated when rupture or hemoperitoneum occurs. CASE REPORT: 35-year-old woman after 3rd cycle of IVF for absolute tubar factor. Two embryos transfered. Ectopic implatation of second embryo in uterine corner occured. Patient hospitalized with acute deteriorating symptoms of hemoperitoneum in 6th week of gestation. Diagnostic laparoscopy performed with ectopic cornual gravidity detection. Decision made to retain the intrauterine gravidity. Induction of the labour in the 40th week of gestation performed. Placenta retained after the delivery of a normal newborn. Lysis manualis performed, uterine hypothonia followed. On suspicion of placenta percreta laparotomy indicated. Acute supracervical abdominal simplex hysterectomy performed. Histologic examination confirmed the diagnosis of placenta percreta. CONCLUSION: The diagnosis of HG is based on the assesment of clinical symptoms, transvaginal ultrasound and endoscopic diagnostics with eventual intervention. In case of hemoperitoneum urgent surgical intervention indicated. Placenta percreta is a rare complication increased in incidence by the presence of myometrial dammage usually from past uterine surgery. Presence of the scar tissue and abnormal placentation might also be a random coincidence. Placenta percreta is a life threatening complication with a great risk of hemmorhage and commonly requires acute surgical intervention.

Combination of fertility preservation strategies in young women with recently diagnosed cancer.

BACKGROUND/AIMS: The study describes clinical management and outcomes of currently available fertility preservation techniques in a set of 154 young female cancer patients. METHODS: Patients in reproductive age with newly diagnosed cancer were offered embryo or oocyte cryopreservation, ovarian tissue cryopreservation and the administration of GnRH analogues during chemotherapy. Particular attention was given to the technical aspects and clinical application of these fertility preservation techniques. RESULTS: During the study period (2004-2009), 154 young female cancer patients were offered fertility preservation counseling. Patient's average age was 29.4 years and average parity was 0.7 children. Administration of GnRH analogues (n = 123, 79.9%) and ovarian tissue cryopreservation (n = 15, 9.7%) were the most commonly used fertility preservation strategies. In 20 cases (16.1%), the combination of several fertility preservation techniques was offered to individually selected patients. CONCLUSIONS: Combination of fertility preservation techniques gives young cancer patients the best chance for future fertility and should be concentrated in specialized centers.