7 research outputs found

    Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

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    #172 : The Comparison of Intra-Cytoplasmic Sperm Injection (ICSI) Treatment Outcome for Patient Stimulated with Progestine-Primed Ovarian Stimulation (PPOS) and Gonadotrophin-Releasing Hormone Agonist (GnRHa) Long Protocol

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    Background and Aims: GnRHa long protocol is a traditional method with benefits such as synchronisation of follicular development, higher oocytes recovery, and improved endometrial thickness but it possesses higher fees with long injection timeline. However, PPOS is an ovarian stimulation protocol that can block the surge of luteinizing hormone (LH) through progesterone instead of the traditional method. This prospective randomized trial is to investigate whether PPOS can be as effective as GnRHa long protocol in ICSI treatment for patients. Method: A total of 55 patients with mean age 36.67 who underwent ICSI cycle in TMC Fertility Centre Ipoh, Malaysia from Jan 2015 to December 2022 was analysed between PPOS and GnRHa long protocol. The embryological and clinical outcomes were measured using statistical analysis T-test with a significant value at p<0.05. Patients above 42-years-old was excluded in this study. Results: Basic characteristics such as infertility duration, age, anti-mullerian hormone (AMH), antra-follicle count (AFC) and body mass index (BMI) were comparable in both groups. There was no significant difference found between PPOS and GnRHa long protocol in the number (mean± SD) oocytes retrieved [9.33±7.38 vs 8.55±7.44, p=0.352]; fertilized oocytes [5.79±4.92 vs 4.03±4.12, p=0.081]; number of blastocysts formed [4.35±2.82 vs 3.11±3.23, p=0.147] and utilized blastocysts [2.79±1.42 vs 2.35±2.66, p=0.300]. There was also no significant difference in clinical pregnancy rate (p=0.456) in both groups. No patient from either group experienced cycle cancellation due to the low number of oocytes expected. No ovarian hyperstimulation (OHSS) syndrome was reported from both groups as well. Conclusion: PPOS in combination with embryo cryopreservation as an ovarian stimulation regimen was as effective as GnRHa long protocol during controlled ovarian stimulation (COH) under different endocrinal mechanisms as it can achieve comparable embryological and clinical outcome. Hence, it can be an alternative treatment for infertile patients in ART in terms of cost and treatment timeline

    #42 : Comparison of Fertilization, Blastocyst Utilization, Clinical Pregnancy Rate (cPR), Implantation Rate (IR) and On-Going Pregnancy Rate Using Fresh and Frozen-Thawed Micro-Epididymal Sperm Aspiration (MESA) Sperm

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    Background and Aim: To compare the fertilization, blastocyst utilization, clinical pregnancy rate (cPR), implantation rate (IR) and on-going clinical pregnancy rate between fresh versus frozen-thawed micro-epididymal aspirated sperm (MESA) in intra-cytoplasmic sperm injection (ICSI) cycle. Methods: A retrospective study of a total of 27 MESA-ICSI cycles done from the year 2019 to 2022 was carried out. Patients were categorized into fresh MESA-ICSI group (n=16) and frozen-thawed MESA-ICSI group (n=11). The maternal mean age of both groups was 33.0 and 32.5 respectively. For fresh MESA-ICSI group, retrieved sperm were being injected on the same day while frozen-thawed MESA-ICSI group, sperm sample were being frozen after the MESA procedure and utilized in the future. Sample was processed using sperm medium (COOK MEDICALⓇ). All embryos were cultured in time-lapse monitoring incubator and assessed according to modified Gardner’s grading system. On day 5/6, blastocyst range from grade A to C will be vitrified for frozen embryo transfer (FET). A total of 22 FET procedures were done, and statistical analysis was performed using Fisher’s exact test, p<0.05. The mean number of embryos transferred for MESA-ICSI group and frozen-thawed MESA-ICSI group are 1.43 and 1.13 respectively. Results: There is no significant difference in the fertilization rate between fresh MESA-ICSI, 71.1% (106/149) and frozen-thawed MESA-ICSI group, 65.4% (70/107). The blastocyst utilization rate between fresh and frozen-thawed MESA-ICSI group are also similar, 45.3% (48/106) and 51.4% (36/70) respectively. Besides, the cPR shown higher in the fresh MESA-ICSI group, 64.3% (9/14) compared to frozen-thawed MESA-ICSI group, 37.5% (3/8). The IR, 50.0% (10/20) and on-going pregnancy rate, 57.1% (8/14) were higher in the fresh MESA-ICSI group compared to frozen-thawed MESA-ICSI group, 33.3% (3/9) and 12.5% (1/8) respectively, p<0.05. Conclusion: The results show the trend of higher cPR, IR and on-going pregnancy rate in fresh MESA-ICSI group compared to frozen-thawed MESA-ICSI group. However, further study is required to conduct in a larger population in order to obtain a significant result

    #361 : What Does My Embryo Score Mean? The Relationship Between KIDScore, Life Whisperer AI Score, and Embryo Euploidy

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    Background and Aims: Many models have been developed for predicting an embryo’s euploidy or implantation potential. These models assign continuous scores to embryos, based on annotation performed by embryologists according to morphokinetic parameters, or based on deep learning analysis of embryo images or video, with higher scores indicating higher probability of implantation and/or euploidy. However, the actual probability of implantation or euploidy at a given score is not always clear. We aimed to investigate the relationship between scores assigned by an annotation model (KIDScoreD5 v3.1, Vitrolife), an image analysis model (Life Whisperer, Presagen), and embryo euploidy. Method: From January 2019 to May 2021, PGT-A by NGS performed on blastocysts in Thomson Hospital Kota Damansara in Malaysia identified 273 euploid and 212 aneuploid blastocysts. Since blastocyst culture in a timelapse incubator (Embryoscope+, Vitrolife) is routine in our centre, the KIDScore for each blastocyst was known. Still image data of the same embryos was uploaded to the Life Whisperer (LW) platform. Embryos were grouped based on their KIDScore and LW score, and the percentage of euploid and aneuploid embryos in each group was calculated. Results: In both models, there were more embryos in higher score groups, as only embryos with good morphology underwent PGT-A (Figures 1 and 2). In KIDScore groups up to score 8, approximately 50% of embryos were euploid, while in KIDScore groups >8-9 and >9-10, 76.6% and 75.5% of embryos were euploid respectively (Figure 1). In LW groups up to score 7, approximately 50% of embryos were euploid, while in LW groups >7-8, >8-9, and >9-10, the percentage increased to 56.1%, 58.6%, and 66.7% respectively (Figure 2). Conclusion: Increasing KIDScores and LW scores correspond with increased percentages of euploid embryos. However, the increase is not constant, especially for KIDScores. These results can be used to estimate the probability of euploidy at a given embryo score

    Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

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    In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

    Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

    Get PDF
    International audienceIn 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field
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