ART Laboratory Research Articles

Risk factors for mismatch in the ART laboratory: an analysis of 73719 electronic witnessing points

Risk factors for mismatch in the ART laboratory: an analysis of 73719 electronic witnessing points

O-211 Exploring risk factors of mismatch in the ART laboratory: An analysis based on 73719 electronic witnessing points

Abstract Study question What are the risk factors associated with mismatch (MM) and genuine mismatch (GMM) in the ART laboratory? Summary answer MM is associated with individual operators, oocyte denudation (OD) and embryo transfer (ET) processes, while GMM related to OD and extended work hours (>4pm). What is known already Mix-up of gametes or embryos in the ART laboratory causes dramatic consequences to both the patients and clinics. Recent studies using electronic witnessing system (EWS) equipped with radio frequency ID (RFID) tags, have revealed that most MM events only involved empty cell culture vessels, while the true mix-up risk arose from a subgroup of GMM involving culture vessels carrying gametes or embryos belonging to different patients. Exploration of risk factors associated with MM and GMM occurrence, separately, is still currently lacking in the literature. Study design, size, duration A total of 73719 witnessing points were retrospectively extracted from an EWS database, representing 11210 consecutive treatment cycles performed at Fertility North between January 2016 and July 2023. The 11210 treatment cycles comprised 5472 fresh IVF, 3732 frozen embryo transfer, 93 oocyte freeze, and 1913 intrauterine insemination cycles. Participants/materials, setting, methods All laboratory work areas were equipped with RFID readers. A MM was triggered by detection of RFID tagged cell culture vessels (either carrying biological material or not) and/or ID cards that belong to different patients in the same work area. A GMM was defined when a MM had involved biological materials and/or patients during procedure, with a potential mix-up. Risk factors evaluated included time of the day, weekdays, weekend, public holiday, operators, and laboratory processes. Main results and the role of chance A total of 138 (0.19%) MMs and 16 (0.02%) GMMs were identified during the study period, without actual mix-up. MM rate was the highest during 12-2pm (0.28%, p < 0.001) compared to other periods of the day, whilst GMM was the highest after 4pm (0.13%, p = 0.004). Neither MM nor GMM was associated with any specific weekday, weekend or public holiday (p > 0.05, respectively). Amongst 8 laboratory processes investigated, ET (0.36%, p < 0.001) led to the highest MM rate, followed by OD (0.35%, p = 0.021), but no increase was observed in oocyte collection, sperm preparation, insemination, embryo culture, cryopreservation and Assign ID. OD was the only process associated with higher GMM (0.13%, p < 0.001). Furthermore, Two of the 20 assessed operators were identified to produce more MMs (0.59%, p < 0.001, and 0.40%, p = 0.004; respectively) while GMM rates were similar among all operators (p > 0.05). Multivariate logistic regression indicated that the OD (adjusted odds ratio or aOR=2.147, p = 0.011) and ET (aOR=2.118, p < 0.001) processes as well as 2 individual operators (aOR=3.511, p < 0.001; aOR=2.538, p = 0.002; respectively) were independent risk factors to MM, but not the 12-2pm period of the day (aOR=1.341, p > 0.05). Finally, both extended work hours beyond 4pm (aOR=8.651, p = 0.005) and OD (aOR=9.538, p < 0.001) were independent risk factors to GMM. Limitations, reasons for caution Each individual ART laboratory has a unique team of scientists and workflow arrangement so findings in our study may not be universally applicable. GMM occurs at an extremely low frequency and is often calculated based on large numbers, therefore caution ought to be taken at statistical interpretation of data. Wider implications of the findings GMMs should be separated from MMs for analysis considering the very different clinical implications. Sufficient staffing with regular break and streamlined workflow are highlighted in preventing mix-up in the ART laboratory. Using electronically logged data, EWS enables identification of site-specific risk factors, which facilitates the development of corresponding preventative protocols. Trial registration number not applicable

Artificial oocyte activation improves ICSI outcomes following unexplained fertilization abnormalities

Research Question: Is Artificial Oocyte Activation (AOA) effective for patients with unexplained low or no fertilisation following IVF/ICSI? Design: All IVF/ICSI cases resulting in total fertilisation failure or fertilisation rate ≤25% at Ninewells Assisted Conception Unit, Dundee, Scotland January 2014 - December 2021 (n=231) were reviewed contemporaneously. After exclusion of obvious stimulation, egg, sperm and/or ART laboratory factors, patients with ≥1 cycle of IVF/ICSI resulting in apparently unexplained fertilisation abnormalities were offered research investigations, including sperm immunocytochemistry for phospholipase C zeta protein expression (ethical approval 22/SS/0104). This retrospective case-controlled cohort study evaluated laboratory and clinical outcomes for 39 couples (15 attended for sperm studies research) that subsequently undertook ICSI-AOA with Ca2+ ionophore (GM508 Cult-Active 1; Gynemed). Results: Comparing preceding IVF/ICSI and subsequent ICSI-AOA for each patient, number of eggs collected was similar, however ICSI-AOA resulted in a significantly improved fertilisation rate (57.2% vs. 7.1%; p<0.0001). The uplift was even more profound for a subset identified with PLCζ deficiency (66.3% vs. 4.6%; p<0.0001). Overall, ICSI-AOA resulted in a higher number of fresh embryo transfers (94.6% vs. 33.3%; p<0.0001), significantly higher clinical pregnancy rate (CPR) and live birth rate (LBR; 17.9% vs. 2.6%; p=0.025), as well as a significant increase in cycles with surplus embryos suitable for cryostorage (43.6% vs. 0%; p<0.0001) and increased cumulative CPR (30.8% vs. 2.6%; p<0.0001) and LBR (28.8% vs. 2.6%; p=0.0002). Conclusion: AOA is a powerful tool that can transform clinical outcomes for couples experiencing apparently unexplained fertilisation abnormalities. PLCζ assays have the potential to be valuable diagnostic tools to determine patient selection for ICSI-AOA and research efforts should continue to focus on their development.

Candida contamination of embryos in the ART laboratory: a case report and literature review

Candida contamination of embryos in the ART laboratory: a case report and literature review

Embryological Perspectives on Gamete and Embryo Donation

This article addresses the issue of gamete and embryo donation from the perspective of the assisted reproduction laboratory, where the so-called "clinical" embryologists work making all the current variants of in vitro fertilization including the andrology laboratory, oocyte collection, conventional insemination or through ICSI, embryo culture, gamete and embryo quality evaluation, embryo transfer, cryopreservation of gametes and embryos, preimplantation genetic studies, results analyses and quality control, assessment and improvement. To think that the responsibility of an ART laboratory only attains the strictly technical or logistical areas is an error or a simplification that excludes embryologists from other important matters. Clinical embryologists have the obligation to ensure that procedures follow strict ethical rules, highlight deviations or errors from these canons, and refuse to act in cases of violation of said standards. Donating embryos to assist in others´ efforts on building a family is an important option for patients who are considering the disposal of cryopreserved embryos, in excess those needed to meet their own fertility goals.&#x0D; Gamete donation, a more widespread procedure, can help us to better understand certain aspects of embryo donation. Donation must be a free transaction, not influenced by guilt or any other type of pressure or coercion of any kind.&#x0D; We are facing a totally new issue because we are the only species on the planet that can manipulate our own gametes and embryos in a context where assisted reproduction represents a dramatic challenge to the social values that govern our way of being born, without precedent in human history.&#x0D; &#x0D; &#x0D;

Failed Fertilization in ART: What are the Causes and Methods to Improve Fertilization

Introduction of clinical ICSI made a revolution in achieving higher fertilization in ART programs. However, ICSI does not completely eliminate the risk of fertilization failure as total fertilization failure (TFF) occurs in 1-3% of ICSI cycles. TFF leads to unexpected premature termination of the ART treatment, inflicting a substantial psychological and financial burden on patients. The potential reasons for TFF include inability of the oocyte to activate the cascade of fertilization events, suboptimal oocytes or sperm quality, and technical issues in performing ICSI. Specifically, oocyte-related factors include oocyte cytoplasmic immaturity caused by insufficient protein synthesis or aberrant signalling pathways, spindle abnormalities, and defective proteins involved in fertilization. Sperm-related factors are primarily associated with abnormalities in the structure, expression, and localization pattern of PLC[Formula: see text]. Several laboratory approaches involving pharmacological agents help in activation of the oocytes thereby prevent the fertilization failure. However, as of now, there is no conclusive evidence to demonstrate the benefit of artificial oocyte activation hence, TFF following ICSI remains an ongoing and significant challenge in ART laboratory.

Batik Motif Weton Sunday Pon Created by Nurohmad at Omah Kreatif Dongaji in Jakob Sumardjo's Aesthetic Study

This research examines the aesthetics of Nurohmad's Batik Weton Sunday Pon motif. This research is qualitative research using Jakob Sumardjo's aesthetic approach. This research was conducted in Nurohmad's art laboratory, Omah Kreatif Dongaji, in Sawit Hamlet Panggungharjo Sewon Bantul. The research data sources are divided into three. Namely, person data sources (interview results to resource persons, employees, customers, and cultural experts), place data sources (research locations and descriptions of events in them), and paper data sources (data collection in the form of photo documentation, images, books, journals, and others related). This research used three data collection techniques: participatory observation, unstructured interviews, and documents. The research instruments used human instruments, observation guides, interview guides, and document guides, where the researcher is the main instrument. Data validity testing in this study used triangulation of sources and techniques. Data analysis was done in three stages: data reduction, presentation, and conclusion drawing. The results of this study are the aesthetics of the Sunday Pon Batik Weton motif according to Jakob Sumardjo in terms of 1) Referent, the motif of Batik Weton Sunday Pon is the work of Batik Weton Sunday Pon it self. 2) Concept can be interpreted as Weton Sunday Pon in Javanese cultural calculations. 3) Symbols can be studied into two types, namely discursive and presentative; for discursive symbols in Batik Weton Sunday Pon motifs, it can be interpreted that the motif images represent the sun, flowers, clouds, and birds, while for presentative symbols, for example, the sun is interpreted as a lamp and is accepted as a respected person. So on, each symbol has a different meaning.

Esther Fernández. To Embody the Marvelous: The Making of Illusions in Early Modern Spain

Esther Fernández. <i>To Embody the Marvelous: The Making of Illusions in Early Modern Spain</i>

Assuring quality in assisted reproduction laboratories: assessing the performance of ART Compass - a digital art staff management platform.

Staff management is the most cited ART/IVF laboratory inspection deficiency. Small ART/IVF clinics may be challenged to perform these activities by low staff volume; similarly, large ART/IVF networks may be challenged by high staff volume and large datasets. Here, we sought to investigate the performance of an automated, digital platform solution to manage this necessary task. The ART Compass (ARTC) digital staff management platform was used to assess the clinical decision-making of ART laboratory staff. The survey modules presented standardized instructions to technologists and measured inter- and intra-technologist variability for subjective "clinical decision-making" type questions. Internal and external comparisons were achieved by providing technologists two answers: (1) a comparison to their own lab director and (2) to the most popular response collectively provided by all lab director level accounts. The platform is hosted on HIPAA compliant Amazon web servers, accessible via web browser and mobile applications for iOS (Apple) and Android mobile devices. Here, we investigated the performance of a digital staff management platform for single embryologist IVF practices and for three IVF lab networks (sites A, B, C) from 2020 to 2022. Embryology dish preparation survey results show variance among respondents in the following: PPE use, media volume, timing of oil overlay, and timing of moving prepared dishes to incubators. Surveying the perceived Gardner score and terms in use for early blastocysts reveals a lack of standardization of terminology and fair to poor agreement. We observed moderate inter-technologist agreement for ICM and TE grade (0.47 and 0.52, respectively). Lastly, the clinical decision of choice to freeze or discard an embryo revealed that agreement to freeze was highest for the top-quality embryos, and that some embryos can be highly contested, evenly split between choice to freeze or discard. We conclude that a digital platform is a novel and effective tool to automate, routinely monitor, and assure quality for staff-related parameters in ART and IVF laboratories. Use of a digital platform can increase regulatory compliance and provide actionable insight for quality assurance in both single embryologist practices and for large networks. Furthermore, clinical decision-making can be augmented with artificial intelligence integration.

Training and competency assessment of Clinical Embryologists and licensing of the profession in European countries.

How is the acquisition and testing of theoretical and practical knowledge in Clinical Embryology and the licensing of ART laboratory personnel carried out in European countries? Twelve out of 31 European countries have established some kind of verification of laboratory competency and skills in ART: in 7 countries, this was related to licensing, but where organized education for Clinical Embryologists existed, there were vast differences in the way these processes were undertaken. In 2015, a report by the ESHRE Embryology Certification Committee concluded that regardless of the large number of people working in IVF laboratories, Clinical Embryology was only recognized as an official profession in 3 out of 27 European national health systems. In most countries, Clinical Embryologists needed to be officially registered under an alternative profession and there were limited opportunities for organized education in this specialist field. Five years after this report, the ESHRE Working Group on Embryologist Training Analysis conducted a survey to collect detailed information about how Clinical Embryologists from different European countries are acquiring their theoretical knowledge and practical skills in ART, and how their level of education and competence in Clinical Embryology is verified. Two questionnaires about the possibilities for acquiring the education and training needed to work in ART and verification of this knowledge were prepared by the ESHRE Working Group on Embryologist Training Analysis. The first was sent in 2020 to a panel of invited lead European Embryologists who attended an Expert Meeting held in Rome, Italy. In order to have a more comprehensive and updated picture, in 2021 the same survey was also sent to the ESHRE Committee of National Representatives (CNRs). At the end of 2021, the second survey with specific questions, more focused on Clinical Embryologists' training and licencing, was sent to the CNRs who reported on verification of education in Clinical Embryology. The first survey consisted of 17 questions. It was initially submitted to 14 lead Embryologists and then resubmitted to the 34 ESHRE CNRs. Representatives from 31 countries responded. A second survey with 23 questions was sent to the 12 ESHRE CNRs who reported an established national system of verification of education in Clinical Embryology, with specific questions focused on the training of Clinical Embryologists. All 12 CNRs responded. Analysis showed that European national education programmes in Clinical Embryology could be split into 4 categories: non-existent (13 countries), recommended (5 countries), simple compulsory (9 countries), and complex compulsory (4 countries). A national document stating the minimum education requirements for staff to work in an IVF laboratory was reported by 19 respondents. The requirement to follow a prescribed theoretical and laboratory training programme in ART was compulsory in 9 and 10 countries, respectively. Some form of verification of laboratory skills, theoretical knowledge in ART, and continuing professional development was required in 12, 10, and 9 countries, respectively. A national trainee's logbook format was reported by seven respondents and a national tutorial system was available in six countries. Only seven countries had official licensing of ART laboratory staff. The title of Clinical Embryologist was not recognized in 13 countries and in 6 countries, it was used only by professional bodies, while in 12 countries the profession was at least cited in governmental regulations. The ESHRE Clinical Embryologist Certificate was officially recognized in eight countries. The survey took place in two steps and the results were then combined to provide a representative picture for most of the European countries sampled. The vast majority, but not all, of the CNRs answered the request to participate in the survey. The professional recognition of Clinical Embryology within Europe is steadily evolving. However, it remains a concern that many countries continue to not recognize Clinical Embryology as a profession, with a vast difference in the reported organization of educational and training programmes and verification of skills. It is recommended that a training programme for Clinical Embryology and ART in Europe should be standardized and relevant issues should be addressed by competent authorities and European Union institutions. ESHRE is best placed to take a leading role in this educational process. The Working Group members who are the authors of this article did not receive payments for the completion of this study. The authors have no conflicts of interest to declare.

P-150 clinical pregnancy rates of oocyte donor cycles are similar when universal media and/or oocyte-specific media are used for oocyte vitrification and warming

Abstract Study question Did the use of universal vitrification/warming media have an impact on the clinical pregnancy rates in oocyte donation cycles compared with oocyte-specific vitrification/warming media? Summary answer Using universal media for oocyte vitrification and warming yields similar clinical pregnancy rates but higher oocyte survival rates than oocyte-specific media. What is known already Vitrification is now the gold standard for oocyte and embryo cryopreservation. Several commercial kits are available, some are designed for specific cell types (e.g., oocytes, zygotes, cleavage-stage embryos or blastocysts) and others are suitable for several stages, therefore termed “universal”. While the composition and the exposition protocol of stage specific media are optimized for specific cell types, “universal” media display a single composition, therefore exposition protocols need to be adapted to each specific developmental stage to ensure optimal cell survival rates. The potential use of the universal media allows to optimize the management of media in ART laboratories. Study design, size, duration This is a retrospective monocentric study. We compared the results obtained in our center for oocyte vitrification in donor cycles with the successive use of oocyte-specific vitrification/warming media (RapidVit/Warm™ Oocyte, Vitrolife) and afterwards universal vitrification/warming media (RapidVit/Warm™ Omni, Vitrolife). We analyzed 111 oocyte recipient ICSI cycles performed between March 2016 and July 2020 (86 recipient couples) and their 81 corresponding oocyte donation cycles (79 donors) with these media. Participants/materials, setting, methods Oocyte recipient ICSI cycles were divided in three groups depending on the combination of vitrification and warming media used for the oocyte vitrification and warming procedures: “specific/specific” (S/S), “specific/universal” (S/U) and “universal/universal” (U/U). The primary outcome was the clinical pregnancy rate per embryo transfer. Secondary outcomes were the oocyte survival rates, fertilization rates, cleavage and blastocyst rates, live birth and miscarriage rates. Main results and the role of chance The age of the donors (31.7+/-3.6 yo) and the recipients (35.1+/-4.5) was similar for the three groups (p &amp;gt; 0.05). Total FSH dose was similar for the 3 groups (1814.2+/-505UI) (p &amp;gt; 0.05) Clinical pregnancy rates were similar when universal vitrification/warming media were used (25.6% for S/U; 25.8% for U/U) compared with oocyte-specific vitrification/warming media (12.2% for S/S, p = 0.25). We observed higher oocyte survival rates when universal media were used (93.7% for U/U, 85.5% for S/U) compared with the use of the oocyte-specific media (75.6% for S/S, p &amp;lt; 0.0001). Fertilization (74.9% for U/U; 68% for S/U; 68.1% for S/S), cleavage (97.9% for U/U; 94.6% for S/U; 89.3% for S/S), and blastulation rates (46.6% for U/U; 50.3% for S/U; 41.7% for S/S) were not different between the three groups (p &amp;gt; 0.05). There was no difference in miscarriage (6.5% for U/U; 10.2% for S/U; 4.9% for S/S) and live birth rates (19.4% for U/U; 15.4% for S/U; 7.3% for S/S) between the 3 groups (p &amp;gt; 0.05). Limitations, reasons for caution The preliminary results of this retrospective study need to be confirmed by a larger prospective study. Wider implications of the findings The use of vitrification/warming universal media, which improves the management of media used for vitrification of oocytes and embryos in ART laboratory, allows to obtain similar clinical pregnancy rates for oocyte recipient donors. Trial registration number not applicable

O-097 Building the perfect cleanroom IVF lab and maintaining it

Abstract Designing and installing a new ART laboratory is not something one does every day and the challenges one faces are many. Putting together a planning team with all concerned parties and stakeholders already in the very early phase of the project is crucial. The aim of the planning team is to critically consider all wishes and requirements, to weigh all the pros and cons and make realistic choices. The final scenario should comply with regulations and/or legislation. This implicated that IVF laboratory facilities should be constructed according to cleanroom regulations to control air-borne contaminants. The construction costs of laboratory space per square meter is about the same as the cost of an operating room. That's why it's so important to discuss space needs and layout well in advance in the planning team. To avoid future bottlenecks during operation, is might be wise to construct multifunctional facilities i.e. an room dedicated to embryo replacements can be designed without much extra cost in such a way that it is also suitable for performing egg retrieval if necessary. Once the cleanroom is build and the HVAC is working, compliance with the designated classification has to be demonstrated in the formal process of qualification (or validation). Provide enough space for cryostorage. This storage room should be a dedicated, locked room ideally not too far from the laboratory cleanroom and should be easily accessed for liquid nitrogen supply. As an alternative, a centralized liquid nitrogen distribution via a vacuum isolated pressured line (VIP) can be considered. One of the most important choices in terms of space requirements and layout is the type and number of workstations, equipment and incubators. Once the number and exact types are known, their exact position should be drawn on the plans in order to determine the necessary outlet points for medical gasses, electricity plugs and network connections.

Recalculating the staff required to run a modern assisted reproductive technology laboratory

What number of staff is sufficient to perform increasingly complicated processes in today's modern ART laboratories? The adequate number of personnel required for the efficient and safe operation of modern ART laboratories needs to be calculated. In today's modern ART laboratories, the amount of time required to perform increasingly complicated processes has more than doubled, with a downward trend in the amount of work an embryologist can do. Different workload unit values have been used to evaluate each workload task and efficiency in a particular ART laboratory, as well as to occasionally compare one laboratory with another. Seven senior embryologists working at different IVF centers, three public and four private centers, participated in this multicenter study conducted between 2019 and 2020. We prepared a survey to create a calculator for staff using the average (of three attempts) time spent in every laboratory by each embryologist of the center to perform any ART process. Different laboratory processes and activities related to quality control, time spent and conventional human double witnessing were included in the survey. To calculate the number of processes that each embryologist can perform per year, an embryologist was considered to be having a full-time contract and working 7 or 8 h/day. The times included in the calculation of each task were those corresponding to the 95th percentile. For the calculations, Microsoft® Office Excel® Professional Plus 2019 was used. The survey showed that the time needed per embryologist to perform the different processes necessary for a classic IVF cycle without time lapse (TL) was 8.11 h, and with TL, it was 10.27 h. The calculated time also considered the time spent in documentation handling, cycle preparation, database management and conventional human double witnessing verification. An ICSI without TL needed 8.55 h, and with TL, it needed 10.71 h. An ICSI-PGT without a TL cycle needed 11.75 h, and with TL, it needed 13.91 h. Furthermore, 1.81 h should be added for every vitrification support needed. The time needed to control more than 200 critical steps, including equipment control and culture parameters, was 30 min per day plus 3.9 min per device to control.The time spent in semen analysis (including documentation handling, cycle preparation and database management) or intrauterine insemination with a partner sperm was 2.7 h. For donor sperm, an additional hour was required for the management involved. The time required to perform a testicular biopsy and cryopreserve the sample was 4 h. Similarly, the time required to perform seminal cryopreservation was 3.7 h. The study was conducted considering a full-time contract embryologist working 7 or 8 h/day, 5 days a week, with days off according to the Spanish regulations. However, our findings can be adapted to foreign regulations using the developed online calculation platform. A new advanced staff calculator allows any IVF laboratory to estimate the minimum number of embryologists necessary without compromising the security or success of the results. Nevertheless, we recommend a minimum of two qualified embryologists in every laboratory, regardless of the workload. This work was funded by the Asociación para el Estudio de la Biología de la Reproducción (ASEBIR). None of the authors has any conflict of interest to declare. N/A.

Offspring conceived through ART have normal thyroid function in adolescence and as young adults.

STUDY QUESTIONAre there differences in thyroid function between adolescents and young adults conceived with and without ART?SUMMARY ANSWERThis study demonstrated no evidence of clinically relevant differences in thyroid function between adolescents and young adults conceived with and without ART.WHAT IS KNOWN ALREADYStudies to date have reported an increase in subclinical hypothyroidism in offspring conceived after ART. It has been suggested that the increase in maternal estrogen (E2) after fresh embryo transfers could affect thyroid function of the offspring. Suboptimal thyroid function at a young age can cause irreversible damage to the central nervous system, which makes early detection and correct treatment essential.STUDY DESIGN, SIZE, DURATIONThe Growing Up Healthy Study (GUHS) is a prospective cohort study, which aimed to recruit all adolescents born after conception with ART between 1991 and 2001 in the study area. The included participants (n = 303, aged 13–20 years) completed various health assessments. Depending on the age at enrolment, participants completed thyroid assessments at the 14- or 20-year follow-up. The outcomes of these replicated thyroid assessments were compared to those of participants conceived without ART from the Raine Study Generation 2 (Gen2). The Gen2 participants (n = 2868) were born between 1989 and 1992 and have been recognized to be representative of the local population.PARTICIPANTS/MATERIALS, SETTING, METHODSThyroid function assessments were compared between n = 134 GUHS and n = 1359 Gen2 adolescents at age 14 years and between n = 47 GUHS and n = 914 Gen2 young adults at age 20 years. The following mean thyroid hormone concentrations were compared between the cohorts: thyroid-stimulating hormone (TSH), free triiodothyronine (fT3), free thyroxine (fT4) and thyroid peroxidase antibodies (TPOAb). The prevalence of the following thyroid hormone profiles, based on individual thyroid hormone concentrations, was compared: euthyroidism, subclinical and overt hypo- and hyperthyroidism and thyroid autoimmunity. Outcomes were compared between the cohorts, and univariately between fresh embryo transfers (ET) and frozen ET (FET) within the GUHS. The correlation between maternal peak E2 concentrations (pE2) and fT4 was assessed within the GUHS.MAIN RESULTS AND THE ROLE OF CHANCEAll mean thyroid function outcomes fell within the normal range. At both ages, we report no differences in TSH concentrations. At age 14 years, lower fT3 concentrations (4.80 versus 5.35 pmol/L, P < 0.001) and higher fT4 concentrations (12.76 versus 12.19 pmol/L, P < 0.001) were detected in the GUHS adolescents compared to Gen2 adolescents. At age 20 years, higher fT3 and fT4 concentrations were reported in GUHS adolescents (4.91 versus 4.63 pmol/L, P = 0.012; 13.43 versus 12.45 pmol/L, P < 0.001, respectively) compared to Gen2 participants. No differences in the prevalence of subclinical and overt hypo- and hyperthyroidism or thyroid autoimmunity were demonstrated between the cohorts at age 14 and 20 years. Thyroid function did not differ between ET and FET, and no correlation between pE2 and fT4 was reported.LIMITATIONS, REASONS FOR CAUTIONThe observational nature of the study limits the ability to prove causation. Furthermore, the comparison of ET and FET offspring at age 20 years may be lacking power. We were unable to differentiate between different types of ART (e.g. IVF versus ICSI) owing to the low number of ICSI cycles at the time of study. As ART laboratory and clinic data were collected contemporaneously with the time of treatment, no other data pertaining to the ART cycles were sought retrospectively; hence, some factors could not be accounted for.WIDER IMPLICATIONS OF THE FINDINGSThis study does not support previous findings of clinically relevant differences in thyroid function when comparing a cohort of adolescents conceived after ART to counterparts conceived without ART. The minor differences detected in fT3 and fT4 were considered not biologically relevant. Although these findings appear reassuring, they warrant reinvestigation in adulthood.STUDY FUNDING/COMPETING INTERESTSThis project was funded by an NHMRC Grant (Hart et al., ID 1042269). R.J.H. is the Medical Director of Fertility Specialists of Western Australia and a shareholder in Western IVF. He has received educational sponsorship from MSD, Merck-Serono and Ferring Pharmaceuticals. P.B. is the Scientific Director of Concept Fertility Centre, Subiaco, Western Australia. J.L.Y. is the Medical Director and a shareholder of PIVET Medical Centre, Perth, Western Australia.TRIAL REGISTRATION NUMBERN/A.

The Bioethical Discourse in the Development of a Musician-Performer

Involvement of a modern performer in the dialogue of concepts of recompense and biocentrism regulates its experimentation with bio and recorded music by awakening a sense of responsibility at the level of the epigenetic rule. Therefore, the purpose of this article is to substantiate a pedagogical model of bioethical education of a musician-performer - creation of a favourable environment to motivate them to comprehend the value of what serves as confirmation of God-like dignity of a person and the general weal for the glory of the Creator. The essence of the proposed pedagogical model is the concept of musical performance as a workroom of soul. Its leading idea is metaphorization of the intonated sense of emotional revelation as an anthropological discourse. Specifically, the basis of the proposed model appears the connectivism theory of metaphorical interpretation. The direction of such performative activity is determined by the strategy of creative clustering, which involves playing out of music scenarios of the value-based choice on the path to “creative innocence” or “the path of the heart”. The regulatory function in implementation of the mentioned strategy performs the methodology of pedagogical and metaphorical creativity. The technology of performative improvisation and the contextual approach to the game are factors in activating the mental resources of the musician-performer. The effectiveness of implementation of the proposed pedagogical model provides creation of a special event platform in the art laboratory.

Abstract OT2-19-01: Presurgical treatment with ribociclib and letrozole in patients with locally advanced breast cancer: The NEOLETRIB study

Abstract Background: The introduction of CDK4/6 inhibitors has been one of the most pivotal breakthroughs in breast cancer therapy in recent decades. A growing body of evidence now suggests that novel targeting inhibitors of CDK4/6, as well as MAPK and PI3K inhibitors, might have anti-tumor effects beyond tumour cell cytostasis, including immunomodulation. Specifically, studies have demonstrated that CDK4/6 inhibitors can have immunogenic effects, through the induction of a senescent-like phenotype in tumor cells, changes in tumor cell metabolism, and direction modulation of T cell function. Trial design: NEOLETRIB is a multicenter, single-arm, open-label, neoadjuvant, phase II trial. Eligibility criteria: Patients suffering from locally advanced breast cancer defined as either large T2 (&amp;gt; 3 cm), or T3/T4, and/or N2-3 are suitable for inclusion. Both, luminal-A and luminal-B tumors are allowed. HER-2 positive and triple-negative patients are excluded from this trial, as are patients with patients with evidence of distant metastasis. Treatment: All patients receive neoadjuvant therapy for at least 6 months with ribociclib (600 mg daily, 21 days on/7 days off) and letrozole (2.5 mg daily). Premenopausal women will also receive goserelin (3.6 mg s.c. every 28 days) according to clinical practice. Specific aims: The primary objective is to evaluate the intra-tumor immunological effects in individual patients treated with letrozole and ribociclib in the neoadjuvant setting. Secondary objectives include evaluation of clinical and pathological response to the treatment combination, identification of early and late molecular mechanisms of treatment adaptation and resistance, and investigation of the effects of the treatment on the gut microbiota and vice versa. As an exploratory objective, we aim to study the relationship between cancer treatment effects, the immune competent cells in the blood and the gut microbiota. Long-term follow-up will continue after the primary analysis for up to 10 years or until death, withdrawal of consent, loss to follow-up, or trial closure. To allow for the planned laboratory sub-studies, tumor tissue biopsies, blood and stool samples will be collected at baseline, day 21 and at time of surgery, with additional blood and stool samples collected at day 90. The samples will be analyzed to evaluate and measure numerous parameters including levels of cytokines and metabolites, cell free DNA-fragments, and intra-tumor gene expression/regulation using state of the art laboratory methods together with international collaborators. Advanced MRI-techniques are used at baseline and following 3 and 6 months on treatment for clinical efficacy measurements in the breast. Statistical methods: Due to the nature of this single-arm study with longitudinal sampling from individual patients, the statistics will be descriptive in nature, focusing on intra-patient comparisons over time. The inclusion goal of 100 patients is based on the response rate of neoadjuvant trials with ribociclib to allow for a minimum of 30 patients in the responder &amp; non-responder arms, which is sufficient for hypothesis generating results. Present accrual and target accrual: Following 8 weeks after initiation of this trial, the first 10 patients have been enrolled. The target accrual is 100 patients with complete study procedures. Citation Format: Xavier Tekpli, Shom Goel, Barbro Holm, Lars-Egil Fallang, Laurens Cornelus Reitsma, Stephanie Beate Geisler, Hilde Presterud Ødegård, Cathrine Bergquist Fosskaug, Ida Caroline Lindem, Manouchehr Seyedzadeh, Jonn Terje Geitung, Joana Reis, Kjell Inge Gjesdal, Unn-Cathrin Buvarp, Marie Loeng, Kristine Kleivi Sahlberg, Alina Carmen Porojnicu, Helle Kristine Skjerven, Andliena Tahiri, Shakila Jabeen, Torben Lüders, Vahid Bemanian, Jørgen Jahnsen, Marianne Lyngra, Antoni Hurtado, Jason Carroll, Shiuan Chen, Vessela Kristensen, Jürgen Geisler. Presurgical treatment with ribociclib and letrozole in patients with locally advanced breast cancer: The NEOLETRIB study [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr OT2-19-01.

Oxidative Stress Measurement in Frozen/Thawed Human Sperm: The Protective Role of an In Vitro Treatment with Myo-Inositol.

Despite its widespread use, sperm cryopreservation induces serious detrimental alterations in sperm function; indeed, it is commonly associated with decreased sperm viability and motility, and DNA fragmentation. Mechanisms of human sperm cryodamage are thought to be multifactorial, but oxidative stress seems to have a prominent role. A huge amount of data supported the cryoprotective effect of different antioxidants able to minimize the detrimental effects of reactive oxygen species (ROS) and improve the quality of spermatozoa. Among others, myo-inositol is one of the most powerful and has been reported to be effective in improving sperm quality and motility when used both in vivo and in vitro. This study aimed to determine the in vitro impact of myo-inositol in ameliorating sperm oxidative status during sperm cryopreservation. In particular, we demonstrated a significant improvement of sperm parameters (vitality and motility) when myo-inositol was added after sperm thawing (p < 0.05). Moreover, we showed that myo-inositol induces a significant increase in oxygen consumption, the main index of oxidative phosphorylation efficiency and ATP production. Finally, by means of 2D-electrophoresis, we demonstrated a significant decrease in the level of carbonyl groups, the main structural changes occurring in conditions of oxidative stress (p < 0.05). In conclusion, the sperm cryopreservation procedure we developed, assuring the reduction of ROS-induced sperm modifications, may improve the in vitro procedure currently used in ART laboratory for sperm cryostorage.

Online pathology request platform at the Semmelweis University

The 1st Department of Pathology and Experimental Cancer Research, Semmelweis University (Budapest, Hungary) has a broad network of clinical partners, many of which are non-university hospitals. A separate hospital information system and a local laboratory workflow management system is used at the Department. University clinics use the hospital information system for electronic requesting of tests. Non-university partners have no access to the systems, requesting tests is paper-based, registration of the requests at the pathology lab is manual and laborious. Our main objective was to improve the efficiency of the sample registration step of the pathology workflow. Applying the Plan-Do-Check-Act procedure, a quality improvement project has been carried out and an online, subspecialty-based requesting application tool, interfaced with the current laboratory information system, was developed. The average sample registration time improved with 65% among the early user partners. The past years have shown smaller, inefficient pathology labs decreasing in number and integrated into larger regional diagnostic centers. Both issues of efficiency and quality assurance and problems rooted in human resources are drivers of further centralisation. The numbers of test requests and samples from non in-house partners are expected to be increased in the pathology labs in the future. Efficient and safe sample tracking has to start at the site of sample acquisition. State of the art laboratory information systems should support this expansion of competence. Orv Hetil. 2021; 162(49): 1962-1967.

‘The way to improve ART outcomes is to introduce more technologies in the laboratory’

To address the proposition that ‘the way to improve ART outcomes is through the introduction of more technologies in the laboratory’, it is prudent to first define what is considered to be improved outcomes. Evidently, this equates to an increase in the live birth rate but it should also include parameters such as time to pregnancy, cumulative pregnancy per oocyte retrieval and health of the resultant child. Furthermore, being able to maintain clinical results week in, week out through quality management also contributes to the overall success of a clinic, and hence can be considered an improved outcome. With regards to these outcomes, it is offered that not only does the introduction of several new technologies (defined here as instrumentation, techniques and enhanced computer utilization and analysis) have the potential to improve outcomes, but also some of them have the capacity to facilitate automation and standardization in the ART laboratory. Although the automation of procedures can be perceived as a justifiable goal itself, in this contribution the emphasis is on how new technologies could help more patients become parents of healthy children in the shortest possible time.

INCUBATOR Art Lab: Reimagining Biotech Futures through Integrated Laboratory Practices

INCUBATOR Art Lab is an art and science research laboratory at the University of Windsor, Ontario, Canada. This image/text document explores the invisible integrated laboratory practices developed within INCUBATOR Art Lab that reimagining how scientific research is conducted within institutional settings towards more joyful and inclusive biotech futures. The piece describes new modes of engaging with institutional bureaucracy, designing infrastructure, and community-building efforts that are central to how INCUBATOR Art Lab functions as a feminist bioart laboratory.