Obravnava žensk v obdobju predmenopavze, ob menopavzi in kasneje se je v novem tisočletju pomembno spremenila. Randomizirane klinične raziskave so bistveno omejile indikacije za uvedbo hormonskega zdravljenja (HZ) in s tem menopavzno medicino postavile pred velik izziv. Na srečo so najnovejša dognanja potrdila, da je ob pravilni uporabi in izbiri HZ korist še vedno bistveno večja od tveganja. Zato smo pripravili posodobljena stališča o menopavzni medicini, ki so v skladu z aktualnimi mednarodnimi priporočili in prilagojena posebnostim slovenskega prostora.

Introduction: This study aimed to evaluate the experience of patients undergoing patient-centered care (PCC) at three fertility clinics in Southeastern Europe in terms of the treatment cost coverage and quality of care provided. Methods: Couples who underwent assisted reproductive technology (ART) were divided into three groups: (i) Slovenian patients covered under the national insurance programme; (ii) self-pay patients receiving treatment in their country of origin/a clinic abroad; and (iii) cross-border self-pay patients receiving treatment in Maribor, Slovenia. A standardized questionnaire, the “Patient Centeredness Questionnaire on Infertility Treatment (PCQ-Infertility)”, was translated into each patient’s native language and administered to 400 couples, of whom 78% completed it. For each of the three patient groups, the mean scores of the PCC domains and indicators were calculated. These data were then used for the score gap analysis. Results: Overall, the domain assessing the competence level of medical staff produced the biggest score domain gap (SDG) in the group of cross-border, self-pay patients, while the smallest score was observed for the health organization domain in the group of domestic patients with reimbursed cycles. In our research, the indicator corresponding to information about any possible side-effects of the prescribed medication produced the smallest score indicator gap (SIG), while the biggest SIG was observed for staff members assigned to be contacted for questions at any time. Conclusion: The results did not show that the financing method of ART treatment affected patient experiences of PCC. Although the competence of the medical staff is rated highly, domestic patients with reimbursed ART cycles continue to experience a lack of PCC in the health organization domain.

Sir, We appreciate your interest in our article detailing the development of performance indicators (PIs) for clinical practice in ART (Vlaisavljevic et al., 2021) and your comments. PIs are objective measures for evaluating critical healthcare domains (patient safety, effectiveness, equity, patientcenteredness, timeliness and efficiency). In the setting of an ART laboratory and clinic, quality indicators are necessary for systematically monitoring and evaluating laboratories’ and clinics’ contributions to patient care (ISO15189-2012) and they represent an important element within the Quality Management System (De los Santos et al., 2016). Very practically, PIs are to be used and monitored within a laboratory/clinic to act upon fluctuations. In case of dropping PIs, the clinic should find the root causes and remediate them. Similarly, in the case of rising PI values, the reasons for the improvement should be investigated and attempts should be made to maintain the improved process. The commentator states that ‘the final goal of ART should be the birth of a healthy baby for a couple who struggles with that’. We could not agree more and, in fact, we stated in our paper that ‘A singleton live full-term healthy baby is the most relevant standard of success in ART’ (Min et al., 2004). There is not much debate regarding the use of live birth rate (LBR) for evaluating interventions. In our paper, however, LBR was investigated as a PI and was found to be non-efficient for monitoring daily clinical practice, whereas clinical pregnancy rate (CPR) was. Following confirmation of clinical pregnancy after ART, any adverse outcome that will not lead to live birth is unlikely to be associated with the performance of the clinician or the lab. On top of that, CPR offers the possibility to react more quickly in performing rootcause analyses. Even if a relevant standard of success and an important parameter in registries, LBR is not the best indicator to be monitored for quality management in an ART centre. The commentator further comments on the definition of a reference population for PI monitoring, defined as female patients <40 years old, using own fresh oocytes, ejaculated spermatozoa (fresh or frozen), any insemination method (i.e. routine IVF and ICSI) and no preimplantation genetic testing. The commentator specifically refers to the exclusion of patients above 40 years old, stresses the importance of female age on the outcomes of ART procedures and cycles and suggests bringing the PI estimation back to the ‘real world’. Referring to the explanation above of the aim and use of PIs, it is reasonable, as for laboratory key performance indicators (KPIs), to limit fluctuations from variability in the population treated and improve the accuracy of the PI estimation by applying a reference population. It is because we acknowledge the impact of female age on the different PIs that a reference population was defined. The same reference population cited in the Vienna consensus was chosen for the Maribor consensus paper as centres would like to relate laboratory PIs to clinical PIs. The group did provide indicators related to poor, normal or high responders making it possible to calculate indicators regardless of age. The age criterion in the definition of the reference population may be revised in the future, possibly aided by algorithms that take into account a lot more variables. Machine learning approaches will be able to analyse a lot more data, follow trends in real-time and alert clinics when deviations are starting to appear. Finally, the commentator refers to the blastocyst transfer and single embryo transfer (SET) rates, stating that both parameters should have been included as PI. Blastocyst transfer is a strategy that a centre may adopt and take into account when evaluating PIs but it cannot be used to evaluate performance within a centre. We do acknowledge the value of SET as a marker of good clinical practice. It is a relevant strategy, but on the other hand, we also consider that it is currently largely dependent local legislation and couple’s decisions. ESHRE is preparing an evidence-based guideline on embryo transfer, which will support SET as embryo transfer strategy and the recommendation that it will be included in a future update of the defined PIs. In conclusion, we would like to emphasize that the proposed list of PIs represents the minimum standard that gives clinics a tool to monitor the clinical performance and to implement if needed, corrective measures. PIs are often related to quality assessments for obtaining a license or quality label, but they are actually there for the benefit of patients and staff. Even without an accreditation or certification label, each clinic should start by looking critically at their clinical practice with the goal to improve clinical care and clinical outcomes. Since most of the papers on indicators in ART focusses on the laboratory aspect, this paper wanted to give clinics a minimal list of indicators to start monitoring their clinical performance. Although challenging and maybe not ambitious enough according to the commentator, we do hope that centres will jump the wagon concerning clinical PIs leading to improvements in clinical practice in ART.

Research question: Does laser-induced artificial blastocoel collapse result in better blastocyst cryopreservation survival and a higher live birth rate (LBR) in comparison with intact counterparts? Design: Half of the supernumerary blastocysts from IVF cycles were randomly selected before vitrification for laser-induced artificial collapsing or vitrification in intact form. A matched case-control study of first transfers of single blastocysts artificially collapsed (case) or intact (control) before vitrification was conducted. Controls were matched to cases on a 1:1 ratio by female age, parity, fresh and vitrified cycle protocol, blastocyst age and quality, resulting in 309 case-control pairs. Results: The two groups were comparable in terms of their characteristics. Survival rates in the case and control groups (97.8% and 95.7%; P = 0.133) were comparable, but the optimal survival rate was higher in the case group (78.2% and 69.3%; P = 0.03). Clinical pregnancy rates (38.2% and 35.3%; P = 0.518), miscarriage rates (15.2% and 22%; P = 0.190), LBR per transfer (32.4% and 27.5%; P = 0.221) and LBR per warmed blastocyst (31.6% and 26.3%; P = 0.137) were not statistically different between the case and control groups. No significant difference in preterm births (11.1% versus 15.7%), birthweights (3333 +/- 723 g versus 3304 +/- 609 g) or sex ratio (49.3% versus 50.7% boys) was observed between the two groups. No major malformations were detected in the study population. Conclusions: Compared with vitrification of intact blastocysts, collapsed blastocysts resulted in a significantly higher optimal survival rate, and although they resulted in a 5% higher LBR, this was not significant for the chosen sample size. Neonatal outcomes were comparable in the two groups.

Abstract STUDY QUESTION Is it possible to define a set of performance indicators (PIs) for clinical work in ART, which can create competency profiles for clinicians and for specific clinical process steps? SUMMARY ANSWER The current paper recommends six PIs to be used for monitoring clinical work in ovarian stimulation for ART, embryo transfer, and pregnancy achievement: cycle cancellation rate (before oocyte pick-up (OPU)) (%CCR), rate of cycles with moderate/severe ovarian hyperstimulation syndrome (OHSS) (%mosOHSS), the proportion of mature (MII) oocytes at ICSI (%MII), complication rate after OPU (%CoOPU), clinical pregnancy rate (%CPR), and multiple pregnancy rate (%MPR). WHAT IS KNOWN ALREADY PIs are objective measures for evaluating critical healthcare domains. In 2017, ART laboratory key PIs (KPIs) were defined. STUDY DESIGN, SIZE, DURATION A list of possible indicators was defined by a working group. The value and limitations of each indicator were confirmed through assessing published data and acceptability was evaluated through an online survey among members of ESHRE, mostly clinicians, of the special interest group Reproductive Endocrinology. PARTICIPANTS/MATERIALS, SETTING, METHODS The online survey was open for 5 weeks and 222 replies were received. Statements (indicators, indicator definitions, or general statements) were considered accepted when ≥70% of the responders agreed (agreed or strongly agreed). There was only one round to seek levels of agreement between the stakeholders. Indicators that were accepted by the survey responders were included in the final list of indicators. Statements reaching less than 70% were not included in the final list but were discussed in the paper. MAIN RESULTS AND THE ROLE OF CHANCE Cycle cancellation rate (before OPU) and the rate of cycles with moderate/severe OHSS, calculated on the number of started cycles, were defined as relevant PIs for monitoring ovarian stimulation. For monitoring ovarian response, trigger and OPU, the proportion of MII oocytes at ICSI and complication rate after OPU were listed as PIs: the latter PI was defined as the number of complications (any) that require an (additional) medical intervention or hospital admission (apart from OHSS) over the number of OPUs performed. Finally, clinical pregnancy rate and multiple pregnancy rate were considered relevant PIs for embryo transfer and pregnancy. The defined PIs should be calculated every 6 months or per 100 cycles, whichever comes first. Clinical pregnancy rate and multiple pregnancy rate should be monitored more frequently (every 3 months or per 50 cycles). Live birth rate (LBR) is a generally accepted and an important parameter for measuring ART success. However, LBR is affected by many factors, even apart from ART, and it cannot be adequately used to monitor clinical practice. In addition to monitoring performance in general, PIs are essential for managing the performance of staff over time, and more specifically the gap between expected performance and actual performance measured. Individual clinics should determine which indicators are key to the success in their organisation based on their patient population, protocols, and procedures, and as such, which are their KPIs. LIMITATIONS, REASONS FOR CAUTION The consensus values are based on data found in the literature and suggestions of experts. When calculated and compared to the competence/benchmark limits, prudent interpretation is necessary taking into account the specific clinical practice of each individual centre. WIDER IMPLICATIONS OF THE FINDINGS The defined PIs complement the earlier defined indicators for the ART laboratory. Together, both sets of indicators aim to enhance the overall quality of the ART practice and are an essential part of the total quality management. PIs are important for education and can be applied during clinical subspecialty. STUDY FUNDING/COMPETING INTEREST(S) This paper was developed and funded by ESHRE, covering expenses associated with meetings, literature searches, and dissemination. The writing group members did not receive payment. Dr G.G. reports personal fees from Merck, MSD, Ferring, Theramex, Finox, Gedeon-Richter, Abbott, Biosilu, ReprodWissen, Obseva, PregLem, and Guerbet, outside the submitted work. Dr A.D. reports personal fees from Cook, outside the submitted work; Dr S.A. reports starting a new employment in May 2020 at Vitrolife. Previously, she has been part of the Nordic Embryology Academic Team, with meetings were sponsored by Gedeon Richter. The other authors have no conflicts of interest to declare. DISCLAIMER This document represents the views of ESHRE, which are the result of consensus between the relevant ESHRE stakeholders and where relevant based on the scientific evidence available at the time of preparation. The recommendations should be used for informational and educational purposes. They should not be interpreted as setting a standard of care, or be deemed inclusive of all proper methods of care nor exclusive of other methods of care reasonably directed to obtaining the same results. They do not replace the need for application of clinical judgment to each individual presentation, nor variations based on locality and facility type. Furthermore, ESHREs recommendations do not constitute or imply the endorsement, recommendation, or favouring of any of the included technologies by ESHRE.

Purpose: The aim of the study was to establish whether follicular fluid (FF) estradiol (E2) and progesterone (P) measurement could be used to predict the outcome of unstimulated IVF/ICSI cycles. Methods: 91 women underwent 150 unstimulated IVF/ICSI cycles (78 IVF and 72 ICSI). Follicular fluid samples were collected at the time of oocyte recovery. Using the ttest, FF E2 and FF P levels and their ratios (FF P/FF E2) were compared between cycles with successful (with oocyte) and unsuccessful (without oocyte) oocyte recovery, between cycles with and without fertilization and between nonconception and conception cycles. Results: The oocyte recovery rate was 86.7% (123/150), the fertilization rate 68.3% (84/123) and the pregnancy rate per oocyte recovery 14.0% (21/150). The average FF E2 level was 3530 ± 1339 nmol/L, average FF P 20649 ± 9489 nmol/L and average FF P/FF E2 ratio 7.18 ± 6.42. There were no statistically significant differences in FF E2, FF P levels and their ratio between cycles with unsuccessful and successful oocyte recovery, between cycles without and with fertilization, and between nonconception and conception cycles. Conclusion: From the FF E2 and P levels and their ratio, it is not possible to make inferences to the likelihood of oocyte recovery, fertilization and conception in unstimulated IVF/ICSI cycles.

Background: Measurements of health-related quality of life (HRQoL) among celiac disease patients using a validated questionnaire have been lacking in Slovenia. This study aims to measure HRQoL in celiac disease (CD) patients using EQ-5D internationally validated questionnaire and comparing it to the HRQoL of the general population. Methods: In this cross sectional analysis all of the approximately 2,000 members of the Slovenian Celiac Society were invited to take part. We used a 3 step approach for recruitment and data collection. HRQoL was evaluated through the EuroQoL EQ-5D-5L instrument (Slovenian version) and analysed using the ordinal logistic regression. Results: Out of 321 patients who gave their consent, 247 celiac patients were included in the study (77%). 68% of the participants were female and 53% of them lived in an urban setting. Most patients originated from North-East Slovenia, whereas approximately 30% of patients came from other Slovenian regions. The EQ-5D respondents’ self-reported health status at the time of the study show that most patients have slight or no problems when living with CD. The duration of the gluten-free diet, academic education and rare (<1x year) doctor visits affect EQ-5D in a positive way. On the other hand, higher age and chronic rheumatic disease were negatively associated with EQ-5D also when compared to the general population. Conclusion: This is the first Slovenian study to measure the HRQoL of Slovenian CD patients, using an internationally validated questionnaire. The results of our study show that HRQoL is slightly impaired among Slovenian patients with CD. Clinical characteristics are better determinants of their HRQoL than socio-demographic factors. Greater awareness of the impact of CD on patients’ HRQoL would improve the holistic management of CD patients.

Background and objective: Automated follicle detection in ovarian ultrasound volumes remains a challenging task. An objective comparison of different follicle-detection approaches is only possible when all are tested on the same data. This paper describes the development and structure of the first publicly accessible USOVA3D database of annotated ultrasound volumes with ovarian follicles. Methods: The ovary and all follicles were annotated in each volume by two medical experts. The USOVA3D database is supplemented by a general verification protocol for unbiased assessment of detection algorithms that can be compared and ranked by scoring according to this protocol. This paper also introduces two baseline automated follicle-detection algorithms, the first based on Directional 3D Wavelet Transform (3D DWT) and the second based on Convolutional Neural Networks (CNN). Results: The USOVA3D testing data set was used to verify the variability and reliability of follicle annotations. The intra-rater overall score yielded around 83 (out of a maximum of 100), while both baseline algorithms pointed out just a slightly lower performance, with the 3D DWT-based algorithm being better, with an overall score around 78. Conclusions: On the other hand, the development of the CNN-based algorithm demonstrated that the USOVA3D database contains sufficient data for successful training without overfitting. The inter-rater reliability analysis and the obtained statistical metrics of effectiveness for both baseline algorithms confirmed that the USOVA3D database is a reliable source for developing new automated detection methods.

ROSUS 2019 – Računalniška obdelava slik in njena uporaba v Sloveniji 2019 je strokovna računalniška konferenca, ki jo od leta 2006 naprej vsako leto organizira Inštitut za računalništvo iz Fakultete za elektrotehniko, računalništvo in informatiko, Univerze v Mariboru. Konferenca povezuje strokovnjake in raziskovalce s področij digitalne obdelave slik in strojnega vida z uporabniki tega znanja, pri čemer uporabniki prihajajo iz raznovrstnih industrijskih okolij, biomedicine, športa, zabavništva in sorodnih področij. Zbornik konference ROSUS 2019 združuje 14 strokovnih prispevkov petdesetih avtorjev, od tega vabljeno predavanje ter 4 demonstracijske prispevke. Prispevki podajajo najnovejše dosežke slovenskih strokovnjakov s področij digitalne obdelave slik in strojnega vida, osvetljujejo pa tudi trende in novosti na omenjenih strokovnih področjih. Velik poudarek prispevkov je na promoviranju ekonomske koristnosti aplikacij računalniške obdelave slik in vida v slovenskem prostoru. Takšne računalniške aplikacije zaradi visoke natančnosti, robustnosti in izjemnih hitrosti pri obdelovanju informacij nudijo namreč nove priložnosti za uveljavitev na trgu visokih tehnologij.

STUDY QUESTION: What is good practice in ultrasound (US), and more specifically during the different stages of transvaginal oocyte retrieval, based on evidence in the literature and expert opinion on US practice in ART? SUMMARY ANSWER: This document provides good practice recommendations covering technical aspects of US-guided transvaginal oocyte retrieval (oocyte pick up: OPU) formulated by a group of experts after considering the published data, and including the preparatory stage of OPU, the actual procedure and post-procedure care. WHAT IS KNOWN ALREADY: US-guided transvaginal OPU is a widely performed procedure, but standards for best practice are not available. STUDY DESIGN, SIZE, DURATION: A working group (WG) collaborated on writing recommendations on the practical aspects of transvaginal OPU. A literature search for evidence of the key aspects of the procedure was carried out. Selected papers (n = 190) relevant to the topic were analyzed by the WG. PARTICIPANTS/MATERIALS, SETTING, METHODS: The WG members considered the following key points in the papers: whether US practice standards were explained; to what extent the OPU technique was described and whether complications or incidents and how to prevent such events were reported. In the end, only 108 papers could be used to support the recommendations in this document, which focused on transvaginal OPU. Laparoscopic OPU, transabdominal OPU and OPU for IVM were outside the scope of the study. MAIN RESULTS AND THE ROLE OF CHANCE: There was a scarcity of studies on the actual procedural OPU technique. The document presents general recommendations for transvaginal OPU, and specific recommendations for its different stages, including prior to, during and after the procedure. Most evidence focussed on comparing different equipment (needles) and on complications and risks, including the risk of infection. For these topics, the recommendations were largely based on the results of the studies. Recommendations are provided on equipment and materials, possible risks and complications, audit and training. One of the major research gaps was training and competence. This paper has also outlined a list of research priorities (including clarification on the value or full blood count, antibiotic prophylaxis and flushing, and the need for training and proficiency). LIMITATIONS, REASONS FOR CAUTION The recommendations of this paper were mostly based on clinical expertise, as at present, only a few clinical trials have focused on the oocyte retrieval techniques, and almost all available data are observational. In addition, studies focusing on OPU were heterogeneous with significant difference in techniques used, which made drafting conclusions and recommendations based on these studies even more challenging. WIDER IMPLICATIONS OF THE FINDINGS: These recommendations complement previous guidelines on the management of good laboratory practice in ART. Some useful troubleshooting/checklist recommendations are given for easy implementation in clinical practice. These recommendations aim to contribute to the standardization of a rather common procedure that is still performed with great heterogeneity.

Objective: Celiac disease is associated with higher risk of infertility, recurrent abortions, and adverse outcomes in pregnancy and in puerperium. The aim of the study was to analyse the association between celiac disease and reproductive disorders in the group of celiac patients and compare these to healthy controls. Methods: A retrospective case-control matched study. The association between celiac disease and menstrual cycle, gyneco-obstetrical complications was assessed with a questionnaire specifically developed for the study. 144 celiac women and 61 celiac men, members of Slovenian Celiac Society, together with 71 healthy women and 31 healthy men participated in the study. Results: A higher percentage of celiac women (27.1%) had difficulties in conception of the first child when compared to healthy controls (12.7%) (p = 0.042). In addition, celiac women experienced more complications than healthy controls during the pregnancy, such as abortions or intrauterine growth retardation (p < 0.005). In our study, the prevalence of reproductive problems was not the same in celiac males and females. Altogether 2 celiac men (3.3%) reported having fertility problems, however, the difference between male cases and controls was not statistically significant (p = 0.548). Conclusion: Physicians should examine women with unexplained infertility, recurrent abortions or intrauterine growth retardation for undiagnosed celiac disease. Compared with healthy women, women with celiac disease have increased risk of spontaneous abortions, preterm delivery and fewer successful pregnancies. (C) 2019 Elsevier B.V. All rights reserved.

Background. It has passed more than 50 years from the developmental phase of ovulation induction. During this period new medications have been introduced, new protocols and dosage established, but the regimen, that would suit all women, has not been designed yet. Methods. The success of ovulation induction in assisted reproduction technologies (ART) does not depend only on medications used, but is influenced by contributing key factors, such as woman’s age, characteristics of the menstrual cycle, body mass index, ovarian reserve and concomitant diseases. The first successful pregnancy followed ART in natural cycle without medications. Because of a relatively low success rate natural cycle was replaced in 70’s by protocols that included clomiphene-citrate or gonadotropins. The introduction of gonadoliberin agonists represented the greatest advantage in this field. The use of human menopausal gonadotropins and recombinants: recombinant FSH, recombinant LH and recombinant HCG in combination with GnRH agonists resulted in significantly higher pregnancy rate (cumulative up to 65 %), but also higher multiple pregnancy rate and ovarian hyperstimulation rate. That is why cheaper, less complicated and patient friendly principles have been renewed, including natural cycle, minimal and mild ovarian stimulation (the use of clomiphene-citrate, letrozole and small doses of HMG or rFSH) that enable ovulation induction and pregnancy in about 30 % of treated women. For a half of the century sophisticated protocols of ovarian stimulation have been developed, but recent European recommendations favour the use of less aggressive, cheaper, effective and patient friendly methods of ovulation induction in ART. There are also protocols for low responding ovaries, which we classify as development of three or less follicles 16 mm in size, only one dominant follicle, or if in past there had been previous cancellations of the cycle because of less than three follicles developed in spite of correct stimulation with gonadotropins. In the literature there are some suggestions how to treat such patients: – long protocol with higher daily doses of gonadotropins, – lowering doses of GnRH agonists or stopping the application soon or immediately after stimulation with gonadotropins has started, – short term use of GnRH agonists in follicular phase, – sequential use of CC and exogene gonadotropins. Ovarian response is monitored by serum estradiol determinations and vaginal ultrasound measurement of follicular size together with echographic estimation of endometrial development. The procedure must comply with each individual and consider her obligations. There should be regular controls, if the dose of gonadotropins is suiting. The application of HCG should be optimized, the hyperstimulation of ovaries should be avoided and the possibility of multiple pregnancies should be lowered. We should also consider the economical side of the use of drugs and the development of the laboratory techniques in reproductive biology. Conclusions. For a half of the century sophisticated protocols of ovarian stimulation have been developed, but recent European recommendations favour the use of less aggressive, effective and patient friendly methods of ovulation induction in ART

Objective Celiac disease (CD) is an autoimmune disorder associated with numerous health problems, including reproductive disorders. This study was performed to analyze the association between CD and the menstrual cycle in a group of patients with CD and compare these patients’ characteristics with those of healthy women. Methods The study included 145 patients with CD (age, 15–51 years) and 162 healthy women (age, 18–55 years). Age at menarche and characteristics of the menstrual cycle were obtained by an anonymous questionnaire developed for the study. Results The age at onset of menarche was 12 to 14 years in 72.9% of the patients with CD and 77.3% of the healthy controls. For most patients (74.2%), the length of the menstrual cycle was around 27 to 28 days with 4 to 5 days of bleeding. Furthermore, 8.4% of patients versus 5.9% of controls experienced bleeding between cycles. Conclusions Our results suggest that in Slovenia, the age at menarche in patients with CD is 12.7 years, which is comparable with that in healthy women. We conclude that CD (treated or untreated) may not be associated with late menarche.

Background. Spontaneous miscarriage isn’t an unexpected complication either in early spontaneous or in ART pregnancy. Previous studies showed that incidence of SM was slightly increased in ART pregnancies in comparison with spontaneous ones after adjusting for maternal age and previous SM. Our objective was to examine the relationship between SM and embryo quality after transfer of one or two blastocysts. Materials and methods. The total of 1433 stimulated IVF and ICSI cycles achieved in our center in the period from 2001 to 2002 after SBT or DBT were retrospectively analyzed. Of these, in the final analysis we included only cases with positive βhCG for which complete data on pregnancy outcome were available – 418 cycles in total. Results. The mean age of patients was 32.2 ± 4.5 years. IVF was performed in 133 cases and ICSI was performed in 285 cases, SBT in 69 and DBT in 349. After SBT, SM rate was 11.6 % and after DBT it was 12.0 %. In SBT group we didn’t find significant relationship between embryo quality and SM (logistic regression: c2 = 0.88; p > 0.05). In this groups, using standard statistical tests, we also couldn’t find significant difference in SM rate between subgroups where optimal or suboptimal quality blastocyst was transferred (Yates corrected c2 test: p > 0.05, Student’s t-test: p > 0.05). In DBT group, we found a strong relationship between embryo quality and SM (logistic regression: c2 = 10.12; p < 0.01). After standard analysis, we confirmed significant difference between subgroups with different combinations of blastocyst quality: after transfer of both optimal blastocysts SM rate was 8.5 %, after transfer of optimal and suboptimal blastocyst SM rate was 10.1 % and after transfer of both suboptimal blastocyst SM rate was 25.4 % (Kruskal-Wallis test: p < 0.001; one-way ANOVA: p < 0.001). We confirmed strong relationship between age of the patients and SM (logistic regression: c2 = 14.57; p < 0.0001). Conclusions. In our study SM rate was 11.9 % which was lower than in previous reports; it was even at the lower limit of expected SM rate in general population. This discrepancy was probably the consequence of longer selection period of blastocysts in in vitro conditions. We didn’t find a significant relationship between SM and blastocyst quality after SBT, which was probably due to the small sample size. A strong relationship between SM and blastocyst quality after DBT was proved. We also confirmed a strong relationship between age of the patients and SM.

Vitrified human blastocysts show varied re-expansion capacity after warming. This prospective observational study compared behaviour of artificially collapsed blastocysts (study group patients, n = 69) to that of blastocysts that were vitrified without artificial collapse (control group patients, n = 72). Warmed blastocysts were monitored by time-lapse microscopy and blastocoel re-expansion speed and growth patterns compared between study and control groups. These parameters were also retrospectively compared between blastocysts that resulted in live birth and those that failed. Artificially collapsed blastocysts re-expanded on average 15.01 µm2/min faster than control blastocysts (P = 0.0013). Warmed blastocysts expressed four different patterns of blastocoel growth. The pattern showing contractions at the end of culture was observed to have a lower prevalence in control blastocysts, which coincided with the lower incidence of hatching in this group. Re-expansion speed and prevalence of growth patterns were comparable between blastocysts that did and did not result in a live birth. This was seen in the study and control groups. Despite faster re-expansion and different growth patterns of artificially collapsed blastocysts, live birth rate did not differ between groups. However, this result should be interpreted with caution due to the small sample size and high risk of bias.

Background. Vitrification is a new, simpler and more rational way of cryopreservation of redundant embryos in in vitro fertilisation. The purpose of this study was to introduce vitrification of blastocysts in clinical practice and compared two commercial cryoprotectants. Methods. We started vitrifying blastocysts in 2008. Two commercial vitrification protocols were used, Vitrification Cooling/Warming (MediCult, Denmark) – (MC), and Vitrification Freeze/Thaw (IrvineScientific, ZDA) – (IS). IS medium includes two cryoprotectants: dimethylsulfoxide (DMSO) and ethylene glycol, meanwhile MC medium contains only ethylene glycol. Among 1030 in vitro fertilizated cycles, we preserved redundant blastocysts in 404 (39.2 %) cycles in liquid nitrogen. At the begining MC vitrification protocol was used in 253 cycles and later IS protocol in 151 cycles. 127 transfers (248 devitrified blastocysts) were performed using MC protocol and 71 transfers (124 blastocysts) using IS protocol. We compared the survival rate of blastocysts, rate of clinical and ongoing pregnancies and miscarriages. Cycles with at least one optimal blastocyst were evaluated separately. Results. Survival rate of vitrified blastocysts by IS protocol was statistically significantly higher than survival rate by MC protocol (88.7 % or 77 %; P = 0.0103). Implantation rate of devitrified blastocysts by IS and MC protocol are comparable (22.7 % or 17.8 %). The rate of miscarriages does not statistically differ between the protocols (18.2 % or 16.7 %). There are no statistically significant differences between IS and MC protocols in clinical (31.8 % or 24 %) and ongoing pregnancies (26.1 % or 20 %). Pregnancy rate in cycles with at least one optimal devitrified blastocyst is 45.7 % by IS protocol and 36.4 % by MC protocol. In the same cycles, the rate of miscarriages is 5.7 % by IS and 4.5 % by MC protocol. Conclusions. Higher values in all parameters were by IS media, even though the differences were not statistically significant because of low number of cycles. The only statistically significant difference between protocols was in survival rate in favor of IS protocol. The study continues.