Animal Studies Research Articles

Efficacy of MAGE-A4 long peptide as a universal immunoprevention cancer vaccine

BackgroundThe clinical application of peptide vaccines in tumor immunotherapy holds significant promise. Peptide-based tumor vaccines are currently subject to certain limitations in clinical trials, including the challenge of inducing a sustained response from CD4+ T helper cells and cytotoxic T lymphocytes (CTL), as well as human leukocyte antigen (HLA) restrictions.MethodsThrough the utilization of biological information methodology, a screening process was conducted to identify three potential long peptides that are specifically targeted by the MAGE-A4 antigen. The candidate long peptides were subjected to in vitro testing using human peripheral blood lymphocytes as samples to evaluate their immunogenicity and immune function. The antitumor properties and preliminary mechanism of the long peptide vaccine were investigated through the use of a mouse model designed for the prevention of triple negative breast cancer (TNBC).ResultsThree predicted multi-epitope long peptides targeting MAGE-A4 have shown to have a strong immunogenicity, with a total positive rate of 72% across different HLA subtypes in Chinese populations. they can also increase the levels of the costimulatory factor CD137 and tumor necrosis factor-alpha (TNF-α), activate T cells, and boost the cytotoxic activity. Results from an animal study have revealed that the long-peptide vaccine, both on its own and in combination with R848, has displayed impressive anti-tumor and target-specific capabilities. Moreover, it has the ability to increase the expression of effector memory T cells and central memory T cells.ConclusionsThis study was the first to screen three multi-epitope long peptides targeting MAGE-A4 and assess their immunogenicity, immune function, and potential as adjuvant peptides. The results showed that the MAGE-A4 long peptide vaccine can be used as a novel immunoprophylaxis method to prevent TNBC. Moreover, the proposed development model is capable of screening multiple target antigens, which lead to its clinical application.

P-336 Efficacy of Gonadotrophin (GnRH) antagonists for endometriosis-associated pelvic pain - A systematic review and meta-analysis

Abstract Study question Are Gonadotropin-releasing hormone (GnRH) antagonists effective and safe in managing pain associated with endometriosis? Summary answer GnRH antagonists seem effective and safe in managing endometriosis-associated pain for at least 12 weeks of use. There is uncertainty with the evidence. What is known already GnRH agonists are commonly used as effective second-line treatment for endometriosis-associated pain. However, these agonists have drawbacks and are not well-tolerated by many women. GnRH antagonists, offer potential advantages over the agonists while providing similar efficacy. They lack the initial stimulatory effects thereby avoiding the initial flare-up of symptoms, which is common with agonists. Furthermore, unlike agonists, which are mainly available as injections, the antagonist are also available as oral preparations, which allow titration of doses. In-vitro and animal studies have shown antagonists to induce regression of endometriotic lesions by apoptosis. However, their efficacy in relieving endometriosis-related pains remains uncertain. Study design, size, duration Extensive electronic search was conducted by the Cochrane Gynaecology and Fertility group, up to October 2023 to identify randomised controlled trials (RCTs) evaluating efficacy of GnRH antagonists in managing endometriosis-related pains compared with placebo and/or GnRH agonists. The main searched databases included CENTRAL, MEDLINE and EMBASE. Screening and selection of suitable studies were undertaken. Study eligibility criteria included treatment for ≥12 weeks, laparoscopically confirmed endometriosis and assessment of pelvic pain scores as the primary endpoint. Participants/materials, setting, methods Primary end-point was a change in endometriosis-related pain score after 12-week treatment with GnRH antagonists compared with placebo and/or GnRH agonists. Changes in pain and quality of life (QoL) scores and adverse-events of GnRH-antagonists were compared between the antagonist versus the agonist/placebo groups. Data were analysed using RevMan software (V5.4) to calculate the standard mean difference (SMD) for continuous data, and the risk ratios (RRs) with 95% confident intervals (95% CI) for categorical data. Main results and the role of chance Seven eligible RCTs (n = 3043) were included, all of which compared GnRH-antagonist versus placebo, and only three also compared versus GnRH-agonist (leuprorelin). The GnRH-antagonists included ASP1707, Elagolix, Linzagolix and Relugolix. Meta-analysis of overall pelvic pain data from seven RCTs (n = 1684) showed significantly greater reduction in pain scores after 12 weeks of GnRH-antagonist versus placebo at low (SMD -0.29 95%CI -0.39 to -0.19; I2=0) and high doses (SMD-0.53 95%CI -0.63 to -0.43, I2=38%). Similarly, meta-analysis of other pains (dysmenorrhoea, non-menstrual pain, and dyspareunia) showed significantly greater pain score reduction with GnRH-antagonists versus placebo. Meta-analysis of overall pelvic pain from two RCTs (n = 263) comparing GnRH-antagonist versus leuprorelin showed comparable reduction in pain scores after 12 weeks at low (SMD 0.33 95%CI 0.09 to 0.58, I2=44%) and high doses (SMD 0.06 95%CI -0.19 to 0.30, I2=0%). Similarly, meta-analysis of other forms of pain revealed comparable reductions of pain scores between both drugs. Meta-analysis of QoL data (2RCTs, n = 259) revealed significantly greater improvements in scores with leuprorelin versus antagonists at low (SMD 0.74 95% CI 0.46 to 1.01; I2=98%) and high doses (SMD 0.35, 95%CI, 0.09 to 0.60, I2=97%). The most frequently reported treatment-related side-effects were hot-flushes, headaches and nasopharyngitis. No life-threatening side-effects were reported. Limitations, reasons for caution There was high heterogeneity between studies (e.g. differences in antagonists and pain scoring systems). No information was provided on prior endometriosis surgery, which is a confounding factor. Additionally, declared COI by authors of all trials could introduce high risk of bias. Evidence certainty was therefore deemed low to very low. Wider implications of the findings GnRH-antagonists seem to offer a promising treatment for endometriosis-related pain, with comparable efficacy to the agonists, but with several potential advantages. However, given the low quality and uncertainty of the evidence in this meta-analysis, high quality RCTs with robust designs are needed before making any firm recommendations for clinical practice. Trial registration number not applicable

Blood–brain barrier integrity is linked to cognitive function, but not to cerebral arterial pulsatility, among elderly

Blood–brain barrier (BBB) disruption may contribute to cognitive decline, but questions remain whether this association is more pronounced for certain brain regions, such as the hippocampus, or represents a whole-brain mechanism. Further, whether human BBB leakage is triggered by excessive vascular pulsatility, as suggested by animal studies, remains unknown. In a prospective cohort (N = 50; 68–84 years), we used contrast-enhanced MRI to estimate the permeability-surface area product (PS) and fractional plasma volume (vp\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$${v}_{p}$$\\end{document}), and 4D flow MRI to assess cerebral arterial pulsatility. Cognition was assessed by the Montreal Cognitive Assessment (MoCA) score. We hypothesized that high PS would be associated with high arterial pulsatility, and that links to cognition would be specific to hippocampal PS. For 15 brain regions, PS ranged from 0.38 to 0.85 (·10−3 min−1) and vp\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$${v}_{p}$$\\end{document} from 0.79 to 1.78%. Cognition was related to PS (·10−3 min−1) in hippocampus (β = – 2.9; p = 0.006), basal ganglia (β = – 2.3; p = 0.04), white matter (β = – 2.6; p = 0.04), whole-brain (β = – 2.7; p = 0.04) and borderline-related for cortex (β = – 2.7; p = 0.076). Pulsatility was unrelated to PS for all regions (p > 0.19). Our findings suggest PS–cognition links mainly reflect a whole-brain phenomenon with only slightly more pronounced links for the hippocampus, and provide no evidence of excessive pulsatility as a trigger of BBB disruption.

P-253 Development of embryos from aged mothers transferred to the uterus of young mice

Abstract Study question Is the development of embryos from advanced maternal age (AMA) mice, similar to those from young maternal age (YMA) mice when transferred to young uteri? Summary answer AMA mouse embryos are characterized by altered organ morphology, particularly of the brain, which may predispose to neurodevelopmental disorders. What is known already Human and animal studies have linked AMA pregnancy to elevated risk of neurodevelopmental disorders and behavioural alterations in offspring, primarily attributed to compromised uterine (maternal) conditions. In human, it was also suggested that postnatal care of older mother may influence offspring behaviour. Animal studies revealed that behavioural alterations of AMA offspring are programmed during pregnancy and so they persist in litters fostered by young mouse. It remains unclear whether an embryo of older mother will undergo normal development after embryo transfer to younger female’s uterus. Study design, size, duration Embryos for Advanced Maternal Age, AMA (n = 105) and control Young Maternal Age, YMA (n = 44) groups were collected from thirty females 8-14 months old and six females 3-5 months old, respectively and transferred to pseudo-pregnant mice for further development. At embryonic day 16 (E16) foetuses were collected, fixed for histology and evaluated using Theiler and Kaufman’s classification of mouse development, where E16 corresponds to Theiler stage 24 (TS24) Participants/materials, setting, methods CBA x C57BL6 hybrid mice were used throughout the experiment. At E16, embryo survival rate (number of foetuses collected / number of transferred embryos) was scored for both groups. Foetuses were assessed for visible abnormalities and underwent histological evaluation of organs accordingly to TS 24. Parameters such as size/diameter, signs of vacuolization, cell density and layers of cerebral cortex in brain of foetuses were analysed. Values were considered different when p < 0.05, Mean±SD; Mann-Whitney test. Main results and the role of chance At E16, the survival of AMA embryos did not differ significantly from YMA control (39/105; 37% vs. 20/44; 45%, respectively) however, 5% of AMA foetuses (2/39) displayed oedema or cerebral haemorrhage. Histological analysis revealed alterations in the head/brain of AMA in comparison to YMA foetuses, such as smaller head size (head area in mid-sagittal section: 25.25±4.87 vs. 31.39±3.79 mm2, p < 0.05), thinner cerebral cortex (435.78±12.95 vs. 515.22±17.78µm, p < 0.005), reduced ratio of cortical plate (CP) to cerebral cortex thickness (0.20±0.03 vs. 0.26±0.009, p < 0.05) and an increased ratio of ventricular zone (VZ) to cerebral cortex thickness (0.29±0.002 vs. 0.24±0.002, p < 0.05). Moreover, decreased cellular density in CP layer (10378.86 vs. 12066.40 cells/mm2, P < 0.05) was noted. AMA brains were characterised by high degree of cellular vacuolisation, particularly in medulla oblongata, suggestive for spongiosis (1550.27 vs. 571.92 vacuoles/mm2, p < 0.05). These brain alterations may cause neurodevelopmental disorders in offspring. AMA foetuses had: smaller bronchial diameter (31.52 vs. 36.62µm, p < 0.05), no ossification centres and reduced skin wrinkling, corresponding to TS23 (that is previous developmental stage). These observations may suggest a delayed development of AMA foetuses. Limitations, reasons for caution The results of controlled animal experiments play a key role in developing conceptual models and formulating hypothesis about target species. However, extrapolation of these results to humans requires caution because species-specific developmental features may differ. Wider implications of the findings Our study demonstrates that advanced maternal age (AMA) impacts oocyte/embryo programming in mice. Morphological changes in AMA-conceived foetuses suggest potential risks for neurodevelopmental and other health issues in offspring, even when carried in the uterus of a young female, emphasizing the far-reaching effects of AMA on progeny health. Trial registration number not applicable

Antiglycation Effect of Jabuticaba (Plinia cauliflora) and Its Potential Role in Delaying Cataract Formation in Streptozotocin-Induced Diabetic Rats

Jabuticaba fruit (Plinia cauliflora) is widely consumed in various forms such as juice, jam, wine, and liquors; however, its potential therapeutic effects on diabetic complications remain inadequately explored. We aimed to investigate the potential antiglycation activity of Jabuticaba, identify the active compounds through bioassay-guided fractionation, and assess its effects on cataract formation in a Streptozotocin-induced diabetic type 1 rat model. Through bioassay-guided fractionation, we identified gallic acid (IC50: 24.7 µg/mL), protocatechuic acid (IC50: 1.22 µg/mL), and an ellagitannin, Repandinin B (IC50: 0.55 µg/mL), as active compounds contributing to antiglycation effects. In the animal study, the addition of Jabuticaba juice extract to the drinking water at a concentration of 0.5% (w/v) for 12 weeks demonstrated an amelioration in cataract progression. These results suggest that Jabuticaba has high antiglycation effects leading to the delaying of cataract formation in type 1 diabetes.

Resveratrol as an Anti-inflammatory Agent in Coronary Artery Disease: A Systematic Review, Meta-Analysis and Meta-Regression.

Resveratrol is a non-flavonoid polyphenol that shows promise in reducing pro-inflammatory factors and maintaining endothelial function, which hints at its potential role in slowing atherosclerosis and preventing acute coronary events. To study the cardioprotective effects of resveratrol on inflammatory mediators and endothelial function in patients with coronary artery disease (CAD). A thorough search was conducted in databases (Cochrane Library, ProQuest, PubMed, LILACS, ScienceDirect, Springer, Taylor&Francis, CNKI, Wanfang, and Weipu) until September 24, 2023. The vasopro-inflammatory mediators, endothelial function and outcomes related to cardiovascular events were observed. Titles and abstracts were assessed, and bias was evaluated with Cochrane RoB 2.0. Heterogeneity of results was explored by meta-regression, certainty of evidence was assessed by the GRADE system, and conclusive evidence was enhanced by trial sequence analysis. Ten randomized controlled trials and 3 animal studies investigated resveratrol's impact on inflammatory mediators and endothelial function. In primary prevention studies, meta-analysis showed a significant reduction (95% CI: -0.73 to -0.20; P=0.0005) in tumor necrosis factor-α (TNF-α) expression with resveratrol, demonstrating a dose-dependent relationship. No significant difference was observed in interleukin-6 (IL-6) expression with P=0.58 for primary prevention and P=0.57 for secondary prevention. Vascular endothelial nitric oxide synthase (eNOS) expression was significantly increased after resveratrol pre-treatment following CAD events. Secondary prevention studies yielded no significant results; however, meta-regression identified associations between age, hypertension, and lower doses with the extent of TNF-α alterations. High certainty of evidence supported TNF-α reduction, while evidence for IL-6 reduction and eNOS elevation was deemed low. Resveratrol reduces TNF-α in individuals at risk for CAD, specifically 15 mg per day. However, its usefulness in patients with confirmed CAD is limited due to factors such as age, high blood pressure, and insufficient dosage. Due to the small sample size, the reduction of IL-6 is inconclusive. Animal studies suggest that resveratrol enhances endothelial function by increasing eNOS. (PROSPERO registration No. CRD42023465234).

P-074 The effect of sperm from male patients who use electronic cigarettes on the euploidy ratio of embryos during in vitro fertilization procedures

Abstract Study question What is the effect of electronic cigarettes as a substitute for regular cigarettes on sperm and subsequent embryo development in IVF patients? Summary answer E-cigarettes are just as harmful to male fertility as regular cigarettes, reduce the euploidy rate of the embryo, and have negative effects on pregnancy. What is known already Many harmful substances from smoking are known to negatively affect sperm development and function. It causes high elevations in sperm DNA fragmentation (SDF), which are associated with embryo euploidy rates. Recently, the use of e-cigarettes is increasing as an alternative to regular cigarettes because they are less harmful and safer. However, several animal studies have shown that e-cigarettes are just as harmful as regular cigarettes and can have negative effects on sperm and fertility. Preliminary data showed that SDF increased with e-cigarettes compared to regular cigarettes. Therefore, we would like to find out whether e-cigarettes affect embryo euploidy. Study design, size, duration In this study, from January 2022 to December 2023, 1,725 cycles of PGT-A (PGT for aneuploidy) were analyzed among the patients of IVF. The average ages of men and women were 40.0±4.6 and 38.7±4.0. A total of 4,796 embryos were analyzed (2.8 ± 2.0 per case) for euploidy rates. Participants/materials, setting, methods Blastocyst biopsy and PGT-A were performed, and embryo euploidy rates was confirmed. In male, the number of no smoking IVF cycles was 1061 and the number of embryo biopsies was 2.9 ± 2.1. The number of IVF cycles using regular cigarettes is 370, and the number of embryo biopsies is 2.8 ± 2.1. The number of IVF cycles using e-cigarettes was 294, and the number of embryo biopsies was 2.5 ± 1.6. Main results and the role of chance Based on the total female age, the embryo euploidy rate was nonsmoking (27.3%), regular cigarettes (23.6%), and e-cigarettes (18.8%). Among women under 38 years of age, e-cigarettes (29.9%) had a lower euploidy rate than regular cigarettes (37.4%), and nonsmoking (38.2%) had the highest rate. Among women over 38, the euploidy rate dropped significantly compared to those under 38, including nonsmoking (18.9%), but it was also lower for e-cigarettes (11.6%) than regular cigarettes (17.7%). Based on female age under 38 years old, the embryo euploidy rate among male age under 39 years old was nonsmoking (39.0%), regular cigarettes (38.3%), and e-cigarettes (31.7%). And among men over 40, the group with the lowest embryo euploidy rate was those who used e-cigarettes (no smoking (35.0%), regular cigarettes (32.6%), e-cigarettes (27.7%)). Based on the age of women over 38 years old, the euploidy rate among men under 39 years of age is nonsmoking (23.9%), regular cigarettes (21.6%), and e-cigarettes (12.7%). And the euploidy rate among men over 40 years of age is nonsmoking (16.8%), regular cigarettes (16.5%), and e-cigarettes (11.8%). nonsmoking is essential to maintain a high euploidy rate in embryos, and e-cigarettes may have a more adverse effect on pregnancy than regular cigarettes. Limitations, reasons for caution Because there are various types of e-cigarettes, it is expected that the degree of harmfulness will vary, but the number of patients was not sufficient to conduct research on each type of e-cigarette. Therefore, it is believed that the above problems should be solved through additional research. Wider implications of the findings This study is a study that links the harmful effects of cigarettes and the polyploidy of embryos. Electronic cigarettes, used as an alternative to regular cigarettes, may reduce embryo euploid production and cause recurrent pregnancy failure. Therefore, it is believed that electronic cigarettes should also be managed appropriately. Trial registration number not applicable

Termination of Resuscitation Rules and Survival Among Patients With Out-of-Hospital Cardiac Arrest

Termination of resuscitation (TOR) rules may help guide prehospital decisions to stop resuscitation, with potential effects on patient outcomes and health resource use. Rules with high sensitivity risk increasing inappropriate transport of nonsurvivors, while rules without excellent specificity risk missed survivors. Further examination of the performance of TOR rules in estimating survival of out-of-hospital cardiac arrest (OHCA) is needed. To determine whether TOR rules can accurately identify patients who will not survive an OHCA. For this systematic review and meta-analysis, the MEDLINE, Embase, CINAHL, Cochrane Library, and Web of Science databases were searched from database inception up to January 11, 2024. There were no restrictions on language, publication date, or time frame of the study. Two reviewers independently screened records, first by title and abstract and then by full text. Randomized clinical trials, case-control studies, cohort studies, cross-sectional studies, retrospective analyses, and modeling studies were included. Systematic reviews and meta-analyses were reviewed to identify primary studies. Studies predicting outcomes other than death, in-hospital studies, animal studies, and non-peer-reviewed studies were excluded. Data were extracted by one reviewer and checked by a second. Two reviewers assessed risk of bias using the Revised Quality Assessment Tool for Diagnostic Accuracy Studies. Cochrane Screening and Diagnostic Tests Methods Group recommendations were followed when conducting a bivariate random-effects meta-analysis. This review followed the Preferred Reporting Items for a Systematic Review and Meta-Analysis of Diagnostic Test Accuracy Studies (PRISMA-DTA) statement and is registered with the International Prospective Register of Systematic Reviews (CRD42019131010). Sensitivity and specificity tables with 95% CIs and bivariate summary receiver operating characteristic (SROC) curves were produced. Estimates of effects at different prevalence levels were calculated. These estimates were used to evaluate the practical implications of TOR rule use at different prevalence levels. This review included 43 nonrandomized studies published between 1993 and 2023, addressing 29 TOR rules and involving 1 125 587 cases. Fifteen studies reported the derivation of 20 TOR rules. Thirty-three studies reported external data validations of 17 TOR rules. Seven TOR rules had data to facilitate meta-analysis. One clinical study was identified. The universal termination of resuscitation rule had the best performance, with pooled sensitivity of 0.62 (95% CI, 0.54-0.71), pooled specificity of 0.88 (95% CI, 0.82-0.94), and a diagnostic odds ratio of 20.45 (95% CI, 13.15-31.83). In this review, there was insufficient robust evidence to support widespread implementation of TOR rules in clinical practice. These findings suggest that adoption of TOR rules may lead to missed survivors and increased resource utilization.

From Lab Bench to Hope: Emerging Gene Therapies in Clinical Trials for Alzheimer's Disease.

Alzheimer's disease is a progressive neurodegenerative disorder that affects memory and cognitive abilities, affecting millions of people around the world. Current treatments focus on the management of symptoms, as no effective therapy has been approved to modify the underlying disease process. Gene therapy is a promising approach that can offer disease-modifying treatment for AD, targeting various aspects of the pathophysiology of the disease. This review presents a comprehensive overview of the current state of gene therapy research for AD, with a specific focus on clinical trials and preclinical studies that have used nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), apolipoprotein E2 (APOE2), and human telomerase reverse transcriptase (hTERT) as therapeutic gene therapy approaches. These gene targets have shown potential to alleviate the neuropathology of AD in animal studies and have demonstrated feasibility and safety in non-human primates. Despite the failure of the NGF gene therapy approach in clinical trials, we have reviewed and highlighted the reported findings and evaluations from the trials. Furthermore, the review included the conclusions of postmortem brain tissue analysis of AD patients who received NGF gene therapy. The goal is to learn from the failed trials and improve the approach in the future. Although gene therapy shows promise, it faces several challenges and limitations, including optimizing gene delivery methods, enhancing safety and efficacy profiles, and determining long-term results. This review contributes to the growing body of literature on innovative treatments for AD and highlights the need for more research and development to advance gene therapy as a viable treatment option for AD.

P-779 GLP-1 receptor agonist treatment in early pregnancy and risk of pregnancy complications - A nationwide cohort study

Abstract Study question Does periconceptionally exposure to GLP-1 receptor agonist (GLP-1 RA) treatment increase the risk of pregnancy complications? Summary answer Perconceptionally exposure to GLP-1 RA had no association with pregnancy complications when compared to a control group with balanced covariates. What is known already The prevalence of obesity continues to increase globally, affecting half the adult population in Western countries, increasing the risk of infertility and obstetric and neonatal complications. Weight loss might improve risks, leading to an interest in effective weight loss methods such as GLP-1 RAs. However, none of the GLP-1 RAs are approved for use during pregnancy and at least two months before to avoid fetal toxicity. The current recommendations are based on a fetal safety principle and data from animal studies, clearly underscoring the need for more human evidence about the effect of GLP-1 RA use in the periconceptional period. Study design, size, duration A Danish nationwide observational cohort study of 522,209 pregnancies from October 2009 until June 2019 with data from health and socioeconomic registries at Statistics Denmark: the National Prescription Register, National Patient Register, Birth Register, Residential Register, and the National Diabetes Registry. All registries had complete information until June 2019, except the National Prescription Register which was until December 2018. The unique personal identification number used in Denmark enables direct linkage of individual-level information across registries. Participants/materials, setting, methods Exposure was defined as the redemption of GLP-1 RA prescription, liraglutide, +/- eight weeks from last menstruation. Primary outcome: any complication (pre-eclampsia, gestational diabetes (GDM), preterm birth (<37 + 0), low-birth-weight (<2500g), small-for-gestational-age, or large-for-gestational-age (LGA)). Evaluated risk of individual outcomes, and effect on gestational age, birth weight, and placenta weight. Outcomes were compared to an unexposed control group, weighted using fast generalized matching on propensity score estimated from relevant confounders. Risk ratio, 95% confidence interval (CI). Main results and the role of chance Within the study population of 522,209 pregnant women, we identified 93 pregnancies exposed to liraglutide in early pregnancy. The exposed women were on average older, had a higher BMI (on average 40% higher), and had a sixty times higher frequency of diabetes (DM1 or DM2) compared to unexposed women giving birth in the same period (p < 0.001). We found that liraglutide exposure was associated with a higher risk of obstetrical complications, pre-eclampsia, GDM, and preterm birth. There was no difference in the birth weight, but those exposed were more likely to give birth to an LGA child (liraglutide, p < 0.01). However, we did not find any evidence of increased risk when utilizing a propensity score matching method (liraglutid, p > 0.1). This indicates that liraglutide, in itself, is not driving the increased risk, but the indication for a GLP-1 RA prescription (e.g. diabetes or obesity) is the causal factor. The finding did not depend on the choice of method for propensity score matching. Findings pertaining to liraglutide and semaglutide, encompassing the period from 2009 to 2023, will be showcased at the primary conference. Limitations, reasons for caution Weight loss treatment was off-label for most of the study time. Wider implications of the findings The study is important for counseling women periconceptionally exposed to GLP-1 RA, that there is no evidence of an increased risk of pregnancy complications. It calls for studies on the safety profile for GLP-1 RA treatment in women, enabling early weight management for the benefit of the mother and child. Trial registration number not applicable

Oral administration of oligo fucoidan improves the survival rate, quality of life, and immunity in patients with lung cancer.

Lung cancer, the most commonly diagnosed cancer globally, has the highest incidence and mortality rates in Taiwan. It can be divided into two types. Non-small cell lung cancer (NSCLC) accounts for approximately 85% of lung cancers, which is further divided into adenocarcinoma, squamous cell carcinoma, and large cell lung cancer accounting for approximately 40%, 25%, and 15% of NSCLC cases, respectively. Small cell lung cancer accounts for approximately 15% of lung cancers. Early systemic therapy NSCLC was based on chemotherapy, and immunotherapy is currently under development. Fucoidan, from brown seaweed extracts, shows promise in mitigating radiation-induced lung fibrosis in animal studies, suggesting its potential as an adjuvant for radiation therapy-related lung fibrosis in lung cancer patients. However, the clinical utility of such adjuvant therapy in lung cancer treatment remains uncertain. The purpose of this study was to investigate the effects of oral administration of oligo-fucoidan on the survival rate, quality of life, and immunity of patients with lung cancer. Subjects with Non-small cell lung cancer aged between 20 and 80 were collected from outpatient clinics, divided into control group (n = 7): conventional therapy and fucoidan group (n = 13): received conventional therapy+ oral supplementation of oligo-fucoidan (550 mg × 4 tablets). Data were collected before the study, at weeks 4, 12, and 24 during the study, and to collect 20 ml of peripheral blood, for analysis biochemical data, liver and kidney function, lymphocyte population, inflammation cytokines, and using EORTC QLQ-C30 questionnaire to assess quality of life. The survival rates of the subjects in the control and fucoidan groups were 20% and 28.6%, respectively. During the study, patients in the fucoidan group experienced a better quality of life than those in the control group, but this difference lacked statistical significance. Oligo-fucoidan increases the CD19 lymphocyte population. The patients in the fucoidan group also had Lower inflammatory cytokine. Oligo-fucoidan holds promise as an adjuvant therapy to enhance the survival rate, quality of life, and immune function in patients with lung cancer.

P-169 Oocyte-to-embryo transition is regulated by availability of exogenous aminoacids in culture medium

Abstract Study question Is de novo protein synthesis during the conversion of oocytes into embryos coupled with aminoacid availability more than with gene transcription? Summary answer The oocyte-to-embryo transition is modulated post-transcriptionally by exogenous aminoacids, including translational reactivation of otherwise transcriptionally silent maternal genes. What is known already Countless studies documented how genes’ mRNAs are regulated up or down during the oocyte-to-embryo transition, with maternal genes being switched off and embryonic genes being switched on. This switching off and on has been poorly explored at the protein level, because it required to administer radioactive or non-canonical aminoacids to embryos in vitro or in the genital tract of the mother, in animal models. However, radioactive aminoacids could not be applied for too long as they harm cells, while non-canonical aminoacids are recognized by cells as unnatural and are metabolized differently. A promising solution is provided by non-radioactive isotopic aminoacids. Study design, size, duration In vivo-fertilized mouse oocytes (zygotes) were cultured to blastocysts in KSOM medium, which was adapted for live-cell proteomic labeling by replacing 2 of the 12 Eagle’s essential aminoacids (EAAs) with isotopic but non-radioactive counterparts (Arginine and Lysine, 13C 15N). The 7 non-essential Eagle’s aminoacids (NEAAs) were left untouched. Labeled blastocysts were sampled at 96 hours of culture in vitro. Controls were cultured in parallel under non-labeling conditions in conventional KSOM. Participants/materials, setting, methods B6C3F1 females were stimulated with eCG+hCG and mated to CD1 males. Zygotes were labeled in vitro by culture in KSOM containing polyvinylpyrrolidone in place of albumin, isotopic Arginine and Lysine in place of non-isotopic counterparts, with or without NEAAs and with or without the remaining 10 EAAs, at 37 °C under 5% CO2. In vitro-labeled blastocysts were transplanted to uterus to ascertain viability, or compared to non-labeled blastocysts by cell counting, mass spectrometry and RNA sequencing. Main results and the role of chance A total of 5426 mouse blastocysts were generated. Isotopically labeled embryos formed blastocysts with total cell numbers and full-term ability similar to unlabeled controls (blastocyst rate ≥68%, p ≥ 0.11; cell count ≥58, p ≥ 0.62; birth rate ≥25%, p ≥ 0.33; Wilcoxon test). When isotopic Arginine and Lysine were present as the sole aminoacids, the blastocyst proteome consisted of 54% unlabeled proteins, 30% semi-labeled proteins and 16% completely labeled proteins. The balance changed to 12%-51%-37% and 6%-56%-38% when NEAAs and NEAAs+EAAs, respectively, were added in culture on top of isotopic Arginine and Lysine (p = 1.9E-09; chi test). This rebalancing did not mirror in the transcriptomes, which were conserved in the three groups (p > 0.08, Wilcoxon test). The proteins of 42 maternal-effect genes (PMID 35047854), which are downregulated during the oocyte-to-embryo transition, were unlabeled when isotopic Arginine and Lysine were present as the sole aminoacids, but became increasingly more labeled (49%, 66%) when NEAAs and NEAAs+EAAs were provided on top of isotopic Arginine and Lysine. Collectively, this information supports that the oocyte-to-embryo transition is not a cell-autonomous process, but features a dynamic post-translational response to the composition of culture medium, including an altered pace of degradation and de novo synthesis of maternal proteins. Limitations, reasons for caution This is an animal study. Mature and fertilized mouse oocytes were collected from oviduct, which is not how human embryos are produced in medically assisted reproduction. The role of autophagy as internal source of aminoacids was not investigated. The oocyte-to-embryo transition takes 2-3 days longer in humans than in mice. Wider implications of the findings As seen in mice, so also in humans the oocyte-to-embryo transition could be modulated by aminoacids, considering that human media are mouse embryo-tested and the requirement of aminoacids for development is conserved across species. To recognize that culture media actively change the embryos, also in medically assisted reproduction, is overdue. Trial registration number not applicable

O-075 Particulate matter (PM2.5 and PM10) exposure prior to oocyte collection is associated with decreased live birth rates in subsequent frozen embryo transfers

Abstract Study question Does exposure to particulate matter air pollution prior to oocyte collection (OPU) or frozen embryo transfer (FET) impact FET live birth rates? Summary answer Increasing PM10 and PM2.5 exposure prior to OPU is associated with decreased odds of live birth in subsequent FET, independent of the conditions at FET. What is known already Ambient air pollution is increasing, and exposure is associated with adverse reproductive health outcomes, including infertility, miscarriage, preterm birth and low birth weight. Animal studies have shown that air pollutants can impair spermatogenesis, oogenesis, ovarian reserve and embryogenesis. These findings are corroborated by human epidemiological studies. The impact of ambient air pollution on IVF outcomes is uncertain. Additionally, whether pollutants exert a greater impact during oocyte development or early pregnancy is uncertain. Therefore, studies of FETs offer an opportunity to delineate different phases of reproduction that may be adversely affected by pollution. Study design, size, duration A retrospective analysis of 3,657 FETs in a single Australian centre from January 2013 – December 2021. All FETs during the study period were included; data were collected from the clinic database. Air pollutant concentrations for each day during the study were obtained from the Governmental monitoring body, including SO2, CO, O3, NO2, PM10 and PM2.5. Evaluation of FETs allowed separation of environmental effects during oocyte development and during early pregnancy. Participants/materials, setting, methods Quartiles were created for pollutants averaged over 24 hours, 2 weeks, 4 weeks and 3 months prior to OPU and prior to FET. Multi-pollutant models were created to account for co-exposures and were adjusted for variables at the time of OPU and FET. Bonferroni correction was made to account for the 4 timepoints tested on each outcome. Linear trends were assessed for each model, and pairwise comparisons against the first quartile were performed. Main results and the role of chance Overall, air quality was excellent, with pollutant levels within published safe limits on > 99% of days during the study period. Increasing PM10 exposure in the 2 weeks prior to OPU was associated with decreased odds of livebirth (linear trend p = 0.009); the odds of livebirth decreased by 38% when PM10 concentrations were in the highest quartile compared with the lowest (OR 0.62, p = 0.010). Increasing PM2.5 exposure in the 3 months prior to OPU was also associated with decreased odds of livebirth (linear trend p = 0.011). No significant linear trends were observed between livebirths and other pollutants. The summer seasonal association with livebirth that we have previously demonstrated consistently held in the multi-pollutant models. No significant linear trends were observed between air pollutants and biochemical pregnancy, clinical pregnancy, miscarriage, gestation at birth or embryological outcomes. Limitations, reasons for caution The retrospective design is a limitation. Whilst corrections were made for multiple pollutant co-exposures, other environmental conditions, patient factors and repeated cycles, we were unable to account for all potential covariates. To reduce the risk of type 1 error due to the multiple timeframes, we utilised a Bonferroni correction. Wider implications of the findings Even when PM2.5 and PM10 exposures are within so-called “safe” ranges, increasing exposure in the weeks and months prior to OPU are associated with decreasing livebirth rates, independent of the conditions at the time of FET. Trial registration number 2022/ET000980

Biomechanical effects of hemin and sildenafil treatments on the aortic wall of chronic-hypoxic lambs.

Introduction: Gestation under chronic hypoxia causes pulmonary hypertension, cardiovascular remodeling, and increased aortic stiffness in the offspring. To mitigate the neonatal cardiovascular risk, pharmacological treatments (such as hemin and sildenafil) have been proposed to improve pulmonary vasodilation. However, little is known about the effects of these treatments on the aorta. Therefore, we studied the effect of hemin and sildenafil treatments in the aorta of lambs gestated and raised at highlands, thereby subjected to chronic hypoxia. Methods: Several biomechanical tests were conducted in the descending thoracic aorta (DTA) and the distal abdominal aorta (DAA), assessing 3 groups of study of hypoxic animals: non-treated (Control) and treated either with hemin or sildenafil. Based on them, the stiffness level has been quantified in both zones, along with the physiological strain in the unloaded aortic duct. Furthermore, a morphological study by histology was conducted in the DTA. Results: Biomechanical results indicate that treatments trigger an increment of axial pre-stress and circumferential residual stress levels in DTA and DAA of lambs exposed to high-altitude chronic hypoxia, which reveals a vasodilatation improvement along with an anti-hypertensive response under this characteristic environmental condition. In addition, histological findings do not reveal significant differences in either structure or microstructural content. Discussion: The biomechanics approach emerges as a valuable study perspective, providing insights to explain the physiological mechanisms of vascular function. According to established results, alterations in the function of the aortic wall may not necessarily be explained by morphostructural changes, but rather by the characteristic mechanical state of the microstructural components that are part of the studied tissue. In this sense, the reported biomechanical changes are beneficial in mitigating the adverse effects of hypobaric hypoxia exposure during gestation and early postnatal life.

Renal denervation: a key approach to hypertension and cardiovascular disease.

Sympathetic activation plays a critical role in the development of hypertension and cardiovascular disease, including heart failure and arrhythmias. Renal nerves contribute to the regulation of blood pressure and fluid volume through renal sympathetic efferent nerves, and to the modulation of sympathetic outflow through renal sensory afferent nerves. Previous studies including ours suggest that selective afferent renal denervation with preservation of efferent renal nerves can significantly decrease central sympathetic outflow in animal models of hypertension with renal damage. In Dahl salt-sensitive rats fed high salt diet from an early age, a model of hypertensive heart failure, this central sympathoinhibition by afferent renal denervation may attenuate the development of heart failure without significant blood pressure reduction. Accumulating clinical evidence supports the efficacy of renal denervation as an antihypertensive treatment. However, it remains important to clarify the appropriate indications and predictors of responders to renal denervation in the treatment of hypertension. Several clinical studies suggest beneficial effects of renal denervation in patients with heart disease, with or without hypertension, although most were not sham-controlled. In particular, some clinical studies have demonstrated that renal denervation reduces the incidence of atrial fibrillation or cardiovascular events even without a significant antihypertensive effect. It is essential to accumulate more insightful data in patients undergoing renal denervation, to establish the efficacy of renal denervation in patients with cardiovascular disease in the clinical setting, and to elucidate the therapeutic mechanisms of renal denervation and the renal nerves-linked pathophysiology of cardiovascular disease in basic research. This review outlines the effects of renal denervation on sympathetic activity and organ damage in animal models of hypertension and hypertensive heart failure, including our own data. Beyond the antihypertensive effects, the beneficial effects of renal denervation on cardiovascular disease are also discussed based on clinical studies. Several animal and clinical studies suggest the cardioprotective effects of renal denervation even in the absence of significant blood pressure reduction, probably due to its sympathoinhibitory effects.

Dyslipidemia and hyperglycemia induce overexpression of Syndecan-3 in erythrocytes and modulate erythrocyte adhesion.

Erythrocytes are important vascular components that play vital roles in maintaining vascular homeostasis, in addition to carrying oxygen. Previously, we reported that the changes in the internal milieu (e.g., hyperglycemia or hypercholesterolemia) increase erythrocyte adhesion to various ECM components, potentially through altering glycosaminoglycans (GAGs). In this study, we have investigated the expression of syndecan (Sdc) family members that could be involved in mediating cytoadherence under conditions of dyslipidemia and hyperglycemia. Among the Sdc family members analyzed, we found significant overexpression of Sdc-3 in erythrocyte membranes harvested from high-fat-fed control and diabetic animals. Animal studies revealed a positive correlation between Sdc-3 expression, blood sugar levels, and erythrocyte adhesion. In the human study, diabetic cohorts with BMI >24.9 showed significantly increased expression of Sdc-3. Interestingly, blocking the Sdc-3 moiety with an anti-Sdc-3 antibody revealed that the core protein might not be directly involved in erythrocyte adhesion to fibronectin despite the GAGs bringing about adhesion. Lastly, Nano LC-MS/MS verified the presence of Sdc-3 in erythrocyte membranes. In conclusion, the high-fat diet and diabetes modulated Sdc-3 expression in the erythrocyte membrane, which may alter its adhesive properties and promote vascular complications.

P-629 In vivo follicular activation (IVFA) through repeated ovarian punction: results of an experimental study in a premature ovarian insufficiency rodent model

Abstract Study question Can the new IVFA therapy effectively promote ovarian follicle development and increase oocyte number in a premature ovarian insufficiency (POI) animal model. Summary answer The IVFA technique reveals favourable reproductive outcomes in the animal model, demonstrating increased oocyte retrieval without compromising quality, making a promising advancement in fertility restoration. What is known already POI is characterized by the loss of the primordial follicular pool before the age of 40. Ovarian fragmentation for in vitro follicular activation (IVA) is a technique based on the disruption of the ovarian extracellular matrix, subsequently promoting actin polymerization and interfering with the evolutionarily conserved tumour suppressor Salvador-Hippo-Warts (SHW) signalling pathway. This has been proven to increase expression of downstream growth factors, thus promoting the primordial follicle growth and the generation of mature oocytes. Yet, this procedure is invasive and expensive. Consequently, a demand exists for less invasive in vivo therapies. Study design, size, duration Experimental animal study. Sixty 6-week-old C57BL/6 female mice were divided into control (CT) and POI groups. Each group, was further divided into three subsets (n = 10/group): without ovarian punctures (CT-0 and POI-0), 15 punctures (CT-15 and POI-15), and 22 punctures (CT-22 and POI-22). Twenty-one days after POI induction, the new IVFA technique was performed on the anaesthetized animals. Subsequently, controlled ovarian stimulation (COS) was initiated, followed by euthanasia for the oocyte retrieval and histological ovarian evaluation. Participants/materials, setting, methods POI condition was induced with intraperitoneally injections of 120 mg/kg of cyclophosphamide and 12 mg/kg of busulfan. The IVFA technique involved ovarian punctures using a 30-gauge needle. COS was induced with 10 IU of PMSG and 10 IU of hCG 48 hours later, leading to euthanasia for oocytes and ovaries collection. Ovaries (n = 6/group) were fixed for follicular counts and immunohistochemistry. Oocyte quality assessment included reactive oxygen species (ROS) levels, mitochondrial fluorescence (Mitotracker), and spindle analysis. Main results and the role of chance The control groups subjected to the IVFA technique (CT-15 and CT-22) demonstrated significantly higher mean total oocyte counts compared to the non-intervention group (CT-0) (30.0±9.8 and 27.7±7.9 vs 18.1±11.7, respectively; p-value<0.05 in both comparisons). Similarly, within the POI model, interventions (POI-15 and POI-22) showed significantly elevated mean total oocyte counts than the non-intervention group (POI-0) (8.1±4.4 and 7.2±2.8 vs 3.8±3.6, respectively; p-value<0.05). The intensity of ROS fluorescence, Mitotracker fluorescence, and spindle analysis in retrieved oocytes were not affected by the IVFA therapy, thereby preserving oocyte quality. Notably, the percentage of growing follicles was significantly increased in the intervention groups compared to the non-intervention groups in the control (60±5.7% and 58.1±4.7% vs 41.5±4.5%; p-value<0.001). In the POI model, statistically significant differences were observed for POI-15 vs POI-0 (57.8±9% vs 41.6±13.3%; p-value<0.05), but not for POI-22 vs POI-0 (51.5±14.2% vs 41.6±13.3%, p=n.s.) Moreover, the global Ki67 proliferation marker demonstrated a significant increase in the percentage of proliferative cells in the CT-15 intervention group compared to the CT-0 group (53±11.2% vs 46.1±15.1; p < 0.01). Finally, in the POI model, the percentage of Ki-67 positive cells was also significantly higher in both IVFA groups (39.4±18.5% and 38.1±17.3% vs 32±17.5%, respectively; p < 0.05). Limitations, reasons for caution Since this is an animal model of POI, further validation of the new therapy with human ovarian samples would be required. The magnitude of the effect could vary due to inter-species variability. Wider implications of the findings The positive results of the new therapy in promoting follicle development and increasing oocyte retrieval, without compromising quality, have significant implications for developing new fertility treatments. If validated in humans, this approach could offer a promising solution for women with POI. Trial registration number not applicable

P-636 Effects of maternal circadian rhythm changes: intergenerational influences on testicular steroidogenesis in offspring

Abstract Study question Do alterations in the maternal circadian rhythm during pregnancy and lactation result in changes to the male offspring’s testicular steroidogenesis intergenerationally? Summary answer The expression of proteins associated with steroidogenesis is altered due to maternal circadian rhythm changes during pregnancy and lactation periods. What is known already Circadian rhythms impact diverse physiological processes, including reproduction. Several clock genes have been linked to reproductive function. However, there is a complex interaction between hormones, fertility, and the circadian clock. It has been shown that disruption of the circadian clock machinery has negative effects on reproductive biology. Current literature indicates a potential relationship between the circadian clock, testicular steroidogenic-related gene expression profiles, and testosterone synthesis. Steroidogenesis involves a complex cascade of enzymatic reactions. An increasing body of evidence from animal studies suggests a relationship between maternal chronodisruption and long-term offspring health and fertility. Study design, size, duration Male offspring from pregnant Wistar rats (n = 60) were exposed to control (12 hours light/12 hours dark), short day (SD) (8 hours light/16 hours dark), or long day (LD) periods (16 hours light/8 hours dark) during only pregnancy, only lactation, or both periods (n = 6/group) categorized as control, SD pregnancy (SD-P), SD lactation (SD-L), SD pregnancy and lactation (SD-P+L), LD pregnancy (LD-P), LD lactation (LD-L), LD pregnancy and lactation (LD-P+L) groups. Participants/materials, setting, methods The expression of critical components of testicular steroidogenesis, including steroidogenic acute regulatory protein (StAR), cholesterol side-chain cleavage enzyme (cytochrome p450scc), 17-beta hydroxysteroid dehydrogenase 3 (HSD17B3), androgen receptor (AR), and luteinizing hormone receptor (LHR), was analyzed in the testis by counting the percentage of positively-stained Leydig cells and ImageJ analysis of immunohistochemistry and western blot. Serum FSH, LH, and testosterone hormones were measured at four zeitgeber time (ZT) points (ZT0, ZT6, ZT12, and ZT18) using ELISA. Main results and the role of chance All of these testicular steroidogenic enzymes were localized in the Leydig cell, and AR was predominantly observed in the Sertoli cell. Although there are no significant differences between the groups in terms of immunolocalization patterns, the expressions of p450scc and AR proteins were significantly decreased in the SD-P and SD-L groups compared to the control group (p < 0.001). In addition, western blot data indicated a decreased expression of p450scc in the SD-P+L group. However, an increased expression of HSD17B3 was detected in both SD-P+L and LD-P+L groups compared to the control group (p = 0.003, p < 0.001, respectively). Among the steroidogenesis-related proteins evaluated, the protein expression levels of StAR and LHR proteins were not altered. FSH showed a statistically significant increase in the LD-L (ZT0), SD-P+L (ZT12), LD-L (ZT12), and LD-P+L (ZT18) groups when compared to the corresponding control groups at the respective time points (p < 0.05). In contrast, LH levels decreased significantly in the LD-P+L (ZT0) group (p < 0.05). Nevertheless, no statistical difference was determined for testosterone levels between groups. Limitations, reasons for caution Most studies on circadian rhythm have used rodent models. Results from animal models are often informative. However, there is a limitation in the appropriate translation of findings in rodents to human studies. Wider implications of the findings The effects triggered by maternal chronodisruption can be intergenerationally transmitted, potentially affecting offspring health. Our results indicate alterations in the expression of proteins involved in testicular steroidogenesis and time-point-dependent changes in the FSH and LH hormone levels due to maternal circadian rhythm changes, further supporting the potential implications for offspring. Trial registration number This work was supported by research grants from the Scientific and Technological Research Council of Turkey (TUBITAK) (SBAG 219S636) and Akdeniz University, The Scientific Research Projects Coordination Unit (TYL-2018-3580).

P-178 Putrescine supplementation enhances embryonic development in patients with Recurrent Implantation Failure (RIF)

Abstract Study question Does putrescine, a type of polyamine, affect embryonic development in patients with recurrent implantation failure (RIF)? Summary answer Putrescine supplementation on cumulus-oocyte complexes (COCs) was associated with better embryologic development from fertilization to blastulation in patients with RIF. What is known already Putrescine, a type of polyamine, plays a role in cell growth and proliferation. Several animal studies suggest the potential impacts of putrescine on embryo development, but its effects on human IVF have yet to be extensively studied. Study design, size, duration This retrospective study analyzed 442 patients who have undergone IVF cycles three or more times from January to December 2023. The average IVF cycle number of RIF patients was 7.4 times. A total of 1356 oocytes from the putrescine group and 1391 oocytes from the control group underwent aspiration and ICSI. Further analysis was conducted by classifying RIF patients into two groups: patients under 40 years of age and patients over 40 years of age. Participants/materials, setting, methods COCs were cultured for 4 hours in a culture medium supplemented with 0.5mM putrescine. 2 hours after oocyte collection, cumulus cells were denuded, and ICSI was performed. Embryonic development was evaluated based on fertilized oocytes (2PN), good quality embryos (GQE) on day 3, and blastulation rates on day 5/6. Group differences were analyzed using t-tests and chi-square tests (p < 0.05). Main results and the role of chance When maternal age was ≥40, the fertilization rate in the putrescine group was significantly higher than that in the control group (86.6% vs. 80.5%, p=0.002). The putrescine group had a numerically higher proportion of embryos with top-grade morphology on day 3 (47.8%, compared with 53.3% for control; p=0.057), without statistical significance. Additionally, the rate of blastulation on day 5 indicated a significant difference between the two groups (14.5% vs. 6.1%, p=0.0001). Further analysis was conducted in younger patients (<40 years old), and it also showed a significantly higher fertilization rate (89.7% vs. 71.1 p=0.0001) and blastocyst development rates on the day 5 in the putrescine group (22.3% vs. 11.1%, p=0.0001), respectively. This study suggests that putrescine supplementation can be utilized to improve embryo quality and facilitate blastocyst formation in IVF procedures for patients with RIF, irrespective of age. Limitations, reasons for caution Putrescine supplementation in IVF showed improved fertilization and blastulation rates, but caution is needed due to the small sample size used. Therefore, a larger sample size will help improve the robustness of the conclusions drawn with respect to the embryologic development in patients with RIF. Wider implications of the findings The study’s findings suggest that putrescine supplementation in IVF may enhance outcomes for patients with RIF, potentially leading to personalized treatments. Trial registration number not applicable

Dietary and circulating branched chain amino acids are unfavorably associated with body fat measures among Chinese adults

Animal studies showed a detrimental effect of dietary branched chain amino acids (BCAAs) on metabolic health, while epidemiological evidence on dietary BCAAs and obesity is limited and inconclusive. We hypothesized that high dietary and circulating BCAAs are unfavorably associated with obesity in community-dwelling adults. We evaluated the 1-year longitudinal associations of dietary BCAA intake and circulating BCAAs with body fat measures. Body weight, height, and circumferences of the waist (WC) and hip (HC) were measured at baseline and again after 1-year. Body composition and liver fat [indicated by controlled attenuation parameter (CAP)] were also assessed after 1-year. Serum BCAA concentrations at baseline were quantified by liquid chromatography mass spectrometry. Diet was collected using 4 quarterly 3-day recalls during the 1-year. The correlation coefficients between dietary and serum BCAAs were 0.12 (P = .035) for total dietary BCAAs, and ranged from -0.02 (soy foods, P = .749) to 0.18 (poultry, P = .001). Total dietary BCAA intake was associated with increase in body weight (β = 0.044, P = .022) and body mass index (BMI, β = 0.047, P = .043). BCAAs from animal foods were associated with increase in HC, while BCAAs from soy foods were associated with weight gain and higher CAP (all P < .05). Serum BCAAs were associated with higher WC, HC, BMI, body fat mass, visceral fat level, and CAP (all P < .05). These results support that dietary and circulating BCAAs are positively associated with the risk of obesity. More cohort studies with validated dietary assessment tools and long-term follow-up among diverse populations are needed to confirm our findings.