bluuebunny/crossref_metadata_2025_split huggingface dataset:
Highly efficient CRISPR systems for loss-of-function and gain-of-function research in pear calli
Authors: Ming Meiling , Long Hongjun , Ye Zhicheng , Pan Changtian , Chen Jiali , Tian Rong , Sun Congrui , Xue Yongsong , Zhang Yingxiao , Li Jiaming , Qi Yiping , Wu Jun 
Published: June 2022

Abstract
CRISPR/Cas systems have been widely used for genome engineering in many plant species. However, their potentials have remained largely untapped in fruit crops, particularly in pear, due to the high levels of genomic heterozygosity and difficulties in tissue culture and stable transformation. To date, only a few reports on the application of the CRISPR/Cas9 system in pear have been documented, and have shown very low editing efficiency. Here we report a highly efficient CRISPR toolbox for loss-of-function and gain-of-function research in pear. We compared four different CRISPR/Cas9 expression systems for loss-of-function analysis and identified a potent system that showed nearly 100% editing efficiency for multi-site mutagenesis. To expand the targeting scope, we further tested different CRISPR/Cas12a and Cas12b systems in pear for the first time, albeit with low editing efficiency. In addition, we established a CRISPR activation (CRISPRa) system for multiplexed gene activation in pear calli for gain-of-function analysis. Furthermore, we successfully engineered the anthocyanin and lignin biosynthesis pathways using both CRISPR/Cas9 and CRISPRa systems in pear calli. Taking these results together, we have built a highly efficient CRISPR toolbox for genome editing and gene regulation, paving the way for functional genomics studies as well as molecular breeding in pear.
O-018 Sperm count is increased by diet-induced weight loss and maintained by exercise or GLP-1 analogue treatment: a randomised controlled trial
Authors: Andersen E , Juhl C R , Kjøller E T , Lundgren J R , Janus C , Saupstad M , Ingerslev L R , Jensen S B K , Holst J J , Stallknecht B M , Madsbad S , Torekov S S , Barrès R 
Published: June 2022

Abstract

Study question
Does diet-induced weight loss improve semen parameters, and are these possible improvements maintained with sustained weight loss?

Summary answer
An 8-week low-calorie diet-induced weight loss was associated with improved sperm count, which was maintained after one year in men who maintained weight loss.

What is known already
Obesity is associated with impaired semen parameters. Weight loss improves metabolic health in obesity, but there is a lack of knowledge on the acute and long-term effects of weight loss on semen parameters.

Study design, size, duration
This is a substudy of men with obesity enrolled in a randomised, controlled, double-blinded trial (the S-LITE trial). The trial was conducted between August 2016 and November 2019. A total of 56 men was included in the study and assigned to an initial 8-week low-calorie diet (800 kcal/day) followed by randomisation to 52 weeks of either: placebo, exercise training, and placebo (exercise), the GLP-1 analogue liraglutide (liraglutide) or liraglutide in combination with exercise training (combination).

Participants/materials, setting, methods
Inclusion criteria were men who delivered semen samples, 18 to 65 years of age and a body mass index between 32 to 43 kg/m2, but otherwise healthy. The study was carried out at Hvidovre Hospital and at the University of Copenhagen, and the participants were from the Greater Copenhagen Area. We assessed semen parameters, anthropometrics and collected blood samples before (T0) and after the 8-week low-calorie dietary intervention (T1) and after 52 weeks (T2).

Main results and the role of chance
The men lost on average 16.5 kg (95% CI: 15.2-17.8) bodyweight during the low-calorie diet, which increased sperm concentration 1.49 fold (95% CI: 1.18-1.88, P&lt;0.01) and sperm count 1.41 fold (95% CI: 1.07-1.87, P&lt;0.01). These improvements were maintained for 52 weeks in men who maintained the weight loss but not in men who regained weight. Semen volume, sperm motility and motile sperm count did not change.

Limitations, reasons for caution
The S-LITE trial was a randomised controlled trial of weight loss maintenance. Analysis of semen was preregistered to explore the effects of weight loss and weight loss maintenance on semen parameters. Due to the small sample size, definite inferences cannot be made.

Wider implications of the findings
This study shows that sperm concentration and sperm count were improved after a diet-induced weight loss in men with obesity. Our findings indicate that both liraglutide and exercise as weight maintenance strategies may be used to maintain the improvements in sperm concentration and count.

Trial registration number
H-16027082
O-022 A randomised controlled trial on Recurrent Implantation Failure treatment in a study model performed on women who failed egg donation cycles using GM-CSF (MOLGRAMOSTIM)
Authors: Sbracia M , Scarpellini F 
Published: June 2022

Abstract

Study question
Is the use of GM-CSF (molgramostim) effective in the treatment of recurrent implantation failure in egg donation cycles?

Summary answer
The clinical use of GM-CSF in women experienced implantation failure in egge donation cycles may be useful,

What is known already
The GM-CSF is a cytokine promoting leukocyte growth as well as trophoblast development. Recurrent implantation failure (RIF) is a clinical entity referring when implantation repeatedly failed to reach a stage recognizable by pelvic ultrasound in IVF cycles. There is no universally accepted definition for RIF. Several factors may determine implantation failure, such as maternal age, embyo aneuploidy, uterine anomalies. The efficacy of a specific treatment is difficult to test in this affection since embryos in general cases show an implantation rate of less than 50%.

Study design, size, duration
The study was conducted to the CERM-Hungaria, Rome, Italy, from the January 2020 to May 2021 on women with recurrent implantation failure after egg donation cycles. Inclusion criteria were: women, aged in between 35-49 years old underwent egg donation cycles with at least three previous transfers with good quality blastocysts that failed to reach pregnancy with no uterine defects (included adenomyosis), no systemic diseases.

Participants/materials, setting, methods
50 women with recurrent implantation failure after egg donation cycles. Patients were randomly subdivided in two groups: one (25 women) treated with subcutaneous GM-CSF (Molgramostim) 0.3mg/kg/daily from the day before embryo transfer to the b-hcg day. The treatment was continued until the 8th week of gestation: the control group (25 women) was treated with subcutaneous saline solution infusion in the same way of the study group. Primary outcome was the ongoing pregnancy rate.

Main results and the role of chance
Epidemiological data of the two groups did not show statistically significant differences. The ongoing pregnancy rate in the group treated with GM-CSF was 84.0% (21/25) whereas in the control group was 44.0% (11/25), P &lt; 0.0072. The absolute risk reduction was 40.0% (95%CI: 15.8%-64.2%). The NNT was 3.0 (95%CI: 1.6-6.3).

Limitations, reasons for caution
This study has a limited number of patients and more studies are needed to confirm these findings. Furthermore, recurrent implantation failure is a clinical entity difficult to classify and define, consequently to extrapolate a common indication for all patients with this should be carefully done.

Wider implications of the findings
The clinical use of GM-CSF in women experienced implantation failure may be an interesting treatment, if these data will confirmed, this treatment can be extended to all patients with recurrent implantation failure. Furthermore, the study model used may ba an interesting way to study recurrent implantation failure.

Trial registration number
NCT01718210
O-039 Novel sperm centrosome function, structure, and composition during fertilization
Authors: Avidor-Reiss T 
Published: June 2022

Abstract

Sperm centrioles play a critical role in sperm differentiation and embryo development, and they are implicated in infertility and miscarriages. However, the precise way centrioles function and are inherited during fertilization in mammals remains unresolved. A key reason for this lack of understanding is that many proteins are only transiently present in the sperm centriole because of a process known as Centrosome Reduction and that sperm centrioles exhibit dramatic species-specific adaptions. It was long widely accepted that structural degeneration and functional inactivation occur at one or two of the centrioles in mammals, resulting in a vestigial remnant. This led to the belief that human spermatozoa have only one functional centriole (the Proximal Centriole, PC). While Centrosome Reduction happens in mice, our studies suggest that two centrioles are present, and both are essential in non-murine mammals. However, the Distal Centriole (DC) has a highly atypical structure and composition, while the PC is only modestly modified. These findings led us to a new model: Centriole Remodeling, which states that preexisting spermatid centrioles, along with their pericentriolar material (PCM), go through a structural and compositional transformation during differentiation resulting in remodeled centrioles and PCM that functions as a dynamic basal complex (DBC) in the spermatozoon and as a precursor for the embryonic centrioles. A crucial mystery remaining is: what are the centrioles' essential functions? To answer this question, we study the sperm centrioles' function and remodeling process in infertile men and rabbit mutants with partially disabled sperm centrioles.

Trial registration number
-
O-040 Investigating why early embryonic mitoses are highly error prone by real time imaging
Authors: Ellenberg J 
Published: June 2022

Abstract
Maintenance of genomic integrity during early embryogenesis is essential for faithful embryonic development. However, mitotic errors during preimplantation development are surprisingly common and despite the fundamental importance of the cell divisions at the beginning of life, the mechanisms that control the correct progression through mitosis during pre-implantation development are not well characterized. The sensitivity of mammalian embryos to light and its demanding in vitro culture requirements have limited real-time imaging during the first embryonic divisions in the past. Today, advances in light-sheet microscopy allow in toto imaging of preimplantation mouse development from zygote to blastocyst at single chromosome resolution and enable us to study spindle assembly, chromosome restructuring and segregation. This presentation will discuss the insights we have gained by real-time imaging of the first cell divisions in mammalian life, and the role of the key molecular mechanisms that regulate mitosis during the critical first stages of embryo development.
O-042 ESHRE good practice recommendations on chromosomal mosaicism
Authors: De Rycke M , Capalbo A , Coonen E , Coticchio G , Fiorentino F , Goossens V , MCheik S , Rubio C , Sermon K , Sfontouris I , Spits C , Vermeesch J , Vermeulen N , Wells D , Zambelli F , Kakourou G 
Published: June 2022

Abstract
The implementation of high-resolution genome-wide methods, usually next-generation sequencing-based, in preimplantation genetic testing (PGT), has led to the frequent detection of embryos with chromosomal mosaicism (whole chromosome and/or segmental aberrations). Despite a growing series of papers showing the birth of healthy babies following the transfer of embryos indicating mosaicism on PGT-A analysis - albeit with lower implantation rates and higher miscarriage rates in comparison with euploid embryo transfer - many questions remain, making it difficult to decide on how to handle chromosomally mosaic embryos in the clinic.
A dedicated ESHRE working group developed good practice recommendations on how to manage chromosomal mosaicism in clinical practice. The recommendations were formulated based on the expert opinion of the working group while taking into consideration the published data and outcomes of a survey on current practices in 239 PGT laboratories and ART clinics, mostly within Europe, Asia and America. The recommendations with regards to detection and management of chromosomal mosaicism were developed following the manual for development of ESHRE good practice recommendations with a stakeholder review of the paper on the ESHRE website. In addition to the recommendations, the working group identified missing information and scientific questions, which should guide further research in PGT, with relevance to the occurrence, detection and impact of chromosomal mosaicism.
Trial registration number:
O-051 The male reproductive health initiative - global collaboration and progress update
Authors: De Jonge C 
Published: June 2022

Abstract
The Male Reproductive Health Initiative (MRHI) was launched in 2018 and is a global collaboration of more than a dozen key opinion leaders in research, medicine, and patient support groups, from the Americas, Australia, the United Kingdom, and Europe.
The mission of the Male Reproductive Health Initiative (MRHI) is to advance the science and practice of male reproductive medicine. The MRHI accomplishes its mission by promoting and contributing to high quality research in both basic and clinical science, forging collaborative relationships with professional societies to contribute to a mission of education, and through advocacy of male reproductive health to all stakeholders including patient support groups, funding agencies, non-for-profit organizations, industry, and the public.
Since its inception, interest in MRHI is expanding, as evidenced by invitations from a dozen or more professional societies from around the world requesting presentations to educate their memberships about the initiative. Helping to spread the scientific, medical, and social importance of male reproductive health are six publications and an additional three manuscripts in press, submitted and or being finalized for submission. These papers, each building on the next, highlight gaps in knowledge that exist regarding diagnosis and treatment of male infertility, how lifestyle and the environment impacts the father and his offspring, and the connection between male reproductive health and somatic health. These gaps collectively raise a present and growing global alarm for the man, his partner and family, and society.
O-052 Male fertility testing - new horizons, ideas and research
Authors: Salas-Huetos A 
Published: June 2022

Abstract
The diagnosis of male infertility is primarily based on the macroscopic analysis of semen and microscopic analysis of sperm (mainly concentration and count, motility, and morphology) through the seminogram. Although six different editions of the “WHO Manual for the Laboratory Examination and Processing of Human Semen” have been published to date, the diagnostic potential of this approach has been called into question. Additionally, recent studies report a significant decline in sperm counts and fecundability in general. Therefore, the need for alternative diagnostic approaches that effectively predict fertility in men is urgently needed.
In this talk we will cover what the limitations of the current semen analysis are, what novel tests are available and more important the new horizons, ideas and research in male fertility testing. We will focus in: 1) the usefulness of sperm functionality analyses (e.g., vitality, plasma membrane and acrosome integrity, plasma membrane lipid disorder, mitochondrial membrane potential, mitochondrial peroxide and superoxide levels, intracellular levels of ROS and Ca2+, and sperm chromatin condensation, among others), 2) the clinical implications of sperm DNA fragmentation and the newest recommendations for assisted reproduction clinics, 3) the newly described monogenic causes of human male infertility, and 4) how comorbidities are related with infertility.
O-112 Fertility awareness: Educating the teenagers in europe
Authors: Özdemir E 
Published: June 2022

Abstract
The term fertility awareness has been long associated with specific methods of family planning. Within the context of reproductive health and fertility issues, fertility awareness is a level of knowledge of the society on the topics of human reproduction system, fertility life cycle, possible threats to reproductive health and the ways of its protection. Moreover, as the birth rates have been continuously falling throughout Europe and the number of European citizens with fertility issues has increased during the last 50 years, fertility and infertility have been put into the spotlight for scientists, policy-makers, non-government organizations and other stakeholders. Therefore, the level of fertility awareness should be increased among all society groups. The main objective of the Fertility Awareness Project is to digitally reach teenagers aged 15-18 in various European countries in order to check their knowledge of fertility and to raise their awareness level. Fertility Awareness Project will follow a knowledge-based, holistic, systemic approach built on two pillars: collecting information and providing information. For this purpose, a digital tool will be developed to measure/increase fertility awareness and this tool will be disseminated in 4 selected European countries among teenagers aged 15-18, aiming to reach as many respondents as possible. After the data collection, a large effort will be devoted to evaluate the results based on the application usage by the target group. Thus, the Fertility Awareness Project will provide reliable information on the levels of fertility awareness among young people in Europe and the outcome of the project will help us to advocate for actions to promote fertility education on EU and national levels.
O-114 Preimplantation genetic testing for monogenic diseases without necessary familial members
Authors: Li R 
Published: June 2022

Abstract
Preimplantation genetic testing for monogenic diseases (PGT-M) is available for any single gene defects theoretically, as long as the disease-causing locus has been unequivocally identified. However, PGT-M has faced great challenges for couples with more than one genetic mutation, de novo mutations or without essential family members for linkage analysis. In order to overcome these challenges, we developed a comprehensive strategy which includes preimplantation genetic testing for aneuploidies (PGT-A) and chromosomal structural rearrangements (PGT-SR) study, as well as monogenic diagnosis study for couples bearing de novo mutations or without necessary familial members. The innovation of our strategy is to use the gamete (polar body or single sperm) or embryo as proband for linkage analyses (GEPLA) to detect an embryo’s carrier status. GEPLA strategy uses affected embryo as proband or gametes carrying mutant alleles as proband for SNP linkage analyses, that were suitable for all types of patients mentioned above. Using this novel developed method, nine autosomal dominant polycystic kidney disease (ADPKD) couples with either de novo mutation or without a positive family history were recruited. Moreover, other two couples with single gene disorders (SMA and NF2) which lack of essential family members for linkage analysis including de novo mutation, and also combined with reciprocal translocation were recruited too. For nine ADPKD couples, a total of 34 embryos from 13 PGT-M cycles were examined and two couples were successfully delivered healthy babies. For the other two couples, 15 embryos were screened, and two embryos were determined as free of the monogenic disease and specific chromosomal abnormalities created by reciprocal translocations. This study provides a creative approach for embryo diagnosis of patients with de novo mutations or patients lacking of essential family members for linkage analysis, and even for reciprocal chromosome translocation concurrently.
O-152 The role of South-South and Triangular Cooperation in the establishment of national and regional registries
Authors: Zegers F , Dyer S , De Mouzon J , Kupka M , Banker M , Ishihara O , Chambers G.M , Adamson G.D 
Published: June 2022

Abstract
The United Nations refers to South-South cooperation as a broad framework of collaboration between developing countries in the Global South. It can take place on a bilateral, regional, or interregional basis. Its main purpose is to share knowledge, skills, resources and successful initiatives to meet development goals through concerted partnerships. Triangular cooperation implies the presence of traditional donor countries and organizations which facilitate South-South initiatives. Both forms of cooperation carry distinct benefits and differ from the more traditional North-South partnerships.
This presentation will describe how, under the umbrella and support of ICMART, regional and interregional ART registries in the developing South, specifically the Latin American registry (REDLARA) and the African registry (ANARA), are contributing to the development, sustainability and cohesion in ART monitoring among the many countries they represent. With an innovative and standardized ART registry as a backbone, 35 countries in these two continents are now organized in regional reproductive networks increasing their recognition, visibility and impact,
We shall demonstrate that this south-south and triangular cooperation, built upon a spirit of community and regional belonging is a strong facilitator of ART monitoring and transfer of technology to the developing South, and its ongoing expansion and reach. In Latin America, where REDLARA was established 30 years ago, local and expedited channels of communication have furthermore underpinned the training of embryologists, clinicians and other health care professionals through regional networks of professionals and certified training centers. This has resulted in enhanced ART self-reliance including the capability to implement new ART technologies and also monitor their availability, utilization, effectiveness and safety. Similar initiatives, strongly supported by REDLARA and ICMART, are starting to be implemented among African institutions via ANARA.
Lastly, we shall demonstrate that our regional as well as global ART registries, together with other international organizations, have contributed to the dissemination of knowledge and serve as monitors and external quality controls now for 35 countries and almost 300 institutions in Latin America and Africa. Furthermore, given that both continents are progressively implementing the same cycle-based reporting system, new knowledge will be gained by examining regional and country similarities and differences.