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Simple Summary: The naturally occurring derivative of estrogen, 2-methoxyestradiol (2-ME), has been shown to have good anti-cancer properties. However, it is broken down too quickly within the blood to be clinically useful. We designed 2-ME analogs with modifications which could avoid rapid metabolism, would preferably stay in the tumor, and were more toxic to cancer cells. Here, we looked more closely at how these compounds work within cancer cells, and how they communicate within themselves and with their environment. ESE-15-one and ESE-16 interfere with the functioning of the intracellular cytoskeleton (actin and microtubules), sending messages that stop cell division, intracellular transport of proteins, and cell migration and invasion, eventually inducing cell suicide. They also inhibited the movement of cells that make blood vessels that would support tumor growth. Using eggs with chicken embryos, we could show that the compounds reduced the tumor size and diminished the spread of cancer cells in a living system. The microtubule-disrupting agent 2-methoxyestradiol (2-ME) displays anti-tumor and anti-angiogenic properties, but its clinical development is halted due to poor pharmacokinetics. We therefore designed two 2-ME analogs in silico—an ESE-15-one and an ESE-16 one—with improved pharmacological properties. We investigated the effects of these compounds on the cytoskeleton in vitro, and their anti-angiogenic and anti-metastatic properties in ovo. Time-lapse fluorescent microscopy revealed that sub-lethal doses of the compounds disrupted microtubule dynamics. Phalloidin fluorescent staining of treated cervical (HeLa), metastatic breast (MDA-MB-231) cancer, and human umbilical vein endothelial cells (HUVECs) displayed thickened, stabilized actin stress fibers after 2 h, which rearranged into a peripheral radial pattern by 24 h. Cofilin phosphorylation and phosphorylated ezrin/radixin/moesin complexes appeared to regulate this actin response. These signaling pathways overlap with anti-angiogenic, extra-cellular communication and adhesion pathways. Sub-lethal concentrations of the compounds retarded both cellular migration and invasion. Anti-angiogenic and extra-cellular matrix signaling was evident with TIMP2 and P-VEGF receptor-2 upregulation. ESE-15-one and ESE-16 exhibited anti-tumor and anti-metastatic properties in vivo, using the chick chorioallantoic membrane assay. In conclusion, the sulfamoylated 2-ME analogs displayed promising anti-tumor, anti-metastatic, and anti-angiogenic properties. Future studies will assess the compounds for myeloproliferative effects, as seen in clinical applications of other drugs in this class. [ABSTRACT FROM AUTHOR]

Simple Summary: Superovulation is a hormonal treatment used to produce multiple oocytes in cows simultaneously, which is essential for embryo transfer. Its success depends on various factors such as hormone levels, protocols, timing of insemination, weather, and nutrition. However, the impact of feed changes, specifically the energy levels in the concentrate, on embryo production is not well understood. This study explores how changes in concentrate intake, with unlimited access to hay, affect embryo production in indigenous Korean (Hanwoo) cows. We compared embryo production and physiological changes between cows with altered concentrate intake and those with consistent intake. The results offer new insights for managing superovulation in relation to feed concentrate. Superovulation is a technique used to increase the number of oocytes released for fertilization. This study investigated the effects of short-term differences in concentrate feed intake on in vivo embryo production through superovulation in indigenous Korean (Hanwoo) cows. The cows were given fresh water and hay ad libitum and randomly divided into three groups (control (CON, n = 9): 2.0 kg/day (unchanged diet); low concentrate (LC, n = 10): 0 kg/day; and high concentrate (HC, n = 8): 4.0 kg/day) according to the amount of formula they were fed. This feeding treatment began seven days before the start of the hormonal treatment for superovulation. From the results, the LC group had the greatest weight change and the lowest body condition score at harvest, followed by the CON and HC groups (p < 0.05). The LC group had the highest number of harvesting embryos, followed by the HC and CON groups (p < 0.05). Estradiol, progesterone, glucose, total cholesterol, high-density lipoprotein, low-density lipoprotein, total protein, and blood urea nitrogen concentrations did not differ between the groups, except for a temporary increase in the HC group on day 0. These findings suggest that more embryos may be harvested when short-term changes in concentrate intake are made during superovulatory responses in Hanwoo cows. [ABSTRACT FROM AUTHOR]

Simple Summary: Embryos release microRNAs (miRNAs) into their surrounding spent media during early development. Gene expression is expected to vary between embryos of good and poor developmental potential; therefore, identifying miRNAs unique to good-quality embryos can aid in embryo selection. This study aimed to characterize the miRNA expression in the spent media of embryos identified as slow- versus fast-growing, as developmental timing may be indicative of embryo quality. Distinct miRNA populations were detected in the spent media conditioned with bovine embryos growing at different developmental rates at the two-cell, eight-cell, and blastocyst stage in vitro. The results highlight the novel and non-invasive miRNA biomarkers of early embryo development. In bovine embryos, the microRNA (miRNA) expression has been profiled at each stage of early development in vitro. The miRNAomic analysis of spent media has the potential to reveal characteristics of embryo health; however, applications are limited without categorizing miRNA profiles by embryo quality. Time-lapse imaging has shown the timing of embryo development in vitro may be indicative of their developmental potential. The study aimed to characterize miRNAs in the spent media of bovine embryos with different growth rates during the pre-implantation phase. Bovine cumulus–oocyte complexes were aspirated from ovaries, fertilized, and cultured to blastocyst stage of development. At the 2-cell, 8-cell, and blastocyst stage, each microdrop of 30 presumptive zygotes were classified as slow- or fast-growing based on the percentage of embryos that had reached the desired morphological stage. A comparative analysis was performed on the spent media of slow- and fast-growing embryos using the results of a GeneChip miRNA 4.0 array hybridization. In total, 34 differentially expressed miRNAs were identified between the comparison groups: 14 miRNAs were found in the 2-cell samples, 7 in the 8-cell samples, and 12 in the blastocyst samples. The results demonstrate distinct miRNAs populations can be identified between slow- and fast-growing embryos, highlighting the novel biomarkers of developmental potential at each stage of pre-implantation development. [ABSTRACT FROM AUTHOR]

Promyelocytic leukemia (PML) nuclear bodies (PML-NBs) are core–shell-type membrane-less organelles typically found in the nucleus of mammalian somatic cells but are absent in mouse oocytes. Here, we deliberately induced the assembly of PML-NBs by injecting mRNA encoding human PML protein (hPML VI -sfGFP) into oocytes and investigated their impact on fertilization in which oocyte/embryos undergo multiple types of stresses. Following nuclear membrane breakdown, preassembled hPML VI -sfGFP mRNA-derived PML-NBs (hmdPML-NBs) persisted in the cytoplasm of oocytes, forming less-soluble debris, particularly under stress. Parthenogenetic embryos that successfully formed pronuclei were capable of removing preassembled hmdPML-NBs from the cytoplasm while forming new hmdPML-NBs in the pronucleus. These observations highlight the beneficial aspect of the PML-NB-free nucleoplasmic environment and suggest that the ability to eliminate unnecessary materials in the cytoplasm of metaphase oocytes serves as a potential indicator of the oocyte quality. [ABSTRACT FROM AUTHOR]

The research is aimed to elucidate the role of plant hormones in regulating the development of hybrid embryos in Hydrangea macrophylla. Fruits from the intraspecific cross of H. macrophylla 'Otaksa' × 'Coerulea' were selected at the globular, heart, and torpedo stages of embryo development. Transcriptome sequencing and differential gene expression analysis were conducted. The results showed that fruit growth followed a single "S-shaped growth curve, with globular, heart, and torpedo embryos appearing at 30, 40, and 50 d post-pollination, respectively, and the embryo maintaining the torpedo shape from 60 to 90 d. A total of 12,933 genes was quantified across the three developmental stages, with 3359, 3803, and 3106 DEGs in the S1_vs_S2, S1_vs_S3, and S2_vs_S3 comparisons, respectively. Among these, 133 genes related to plant hormone biosynthesis and metabolism were differentially expressed, regulating the synthesis and metabolism of eight types of plant hormones, including cytokinin, auxin, gibberellin, abscisic acid, and jasmonic acid. The pathways with the most differentially expressed genes were cytokinin, auxin, and gibberellin, suggesting these hormones may play crucial roles in embryo development. In the cytokinin pathway, CKX (Hma1.2p1_0579F.1_g182670.gene, Hma1.2p1_1194F.1_g265700.gene, and NewGene_12164) genes were highly expressed during the globular embryo stage, promoting rapid cell division in the embryo. In the auxin pathway, YUC (Hma1.2p1_0271F.1_g109005.gene and Hma1.2p1_0271F.1_g109020.gene) genes were progressively up-regulated during embryo growth; the early response factor AUX/IAA (Hma1.2p1_0760F.1_g214260.gene) was down-regulated, while the later transcriptional activator ARF (NewGene_21460, NewGene_21461, and Hma1.2p1_0209F.1_g089090.gene) was up-regulated, sustaining auxin synthesis and possibly preventing the embryo from transitioning to maturity. In the gibberellin pathway, GA3ox (Hma1.2p1_0129F.1_g060100.gene) expression peaked during the heart embryo stage and then declined, while the negative regulator GA2ox (Hma1.2p1_0020F.1_g013915.gene) showed the opposite trend; and the gibberellin signaling repressor DELLA (Hma1.2p1_1054F.1_g252590.gene) increased over time, potentially inhibiting embryo development and maintaining the torpedo shape until fruit maturity. These findings preliminarily uncover the factors affecting the development of hybrid H. macrophylla embryos, laying a foundation for further research into the regulatory mechanisms of H. macrophylla hybrid embryo development. [ABSTRACT FROM AUTHOR]

Acute myeloid leukemia (AML) is characterized by the abnormal proliferation and differentiation arrest of myeloid progenitor cells. The clinical treatment of AML remains challenging. Promoting AML cell differentiation is a valid strategy, but effective differentiation drugs are lacking for most types of AML. In this study, we generated Tg(drl:hoxa9) zebrafish, in which hoxa9 overexpression was driven in hematopoietic cells and myeloid differentiation arrest was exhibited. Using Tg(drl:hoxa9) embryos, we performed chemical screening and identified four FDA-approved drugs, ethacrynic acid, khellin, oxcarbazepine, and alendronate, that efficiently restored myeloid differentiation. The four drugs also induced AML cell differentiation, with ethacrynic acid being the most effective. By an RNA-seq analysis, we found that during differentiation, ethacrynic acid activated the IL-17 and MAPK signaling pathways, which are known to promote granulopoiesis. Furthermore, we found that ethacrynic acid enhanced all-trans retinoic acid (ATRA)-induced differentiation, and both types of signaling converged on the IL-17/MAPK pathways. Inhibiting the IL-17/MAPK pathways impaired ethacrynic acid and ATRA-induced differentiation. In addition, we showed that ethacrynic acid is less toxic to embryogenesis and less disruptive to normal hematopoiesis than ATRA. Thus, the combination of ethacrynic acid and ATRA may have broader clinical applications. In conclusion, through zebrafish-aided screening, our study identified four drugs that can be repurposed to induce AML differentiation, thus providing new agents for AML therapy. [ABSTRACT FROM AUTHOR]

Simple Summary: Gonadal teratomas are tumours that can be benign or malignant and affect the gonads, specifically the ovaries in women and the testicles in men. They are predominantly young tumours that affect children and adolescents; therefore, early diagnosis is essential to apply the most suitable therapy to preserve fertility. In this article, based on articles found in the literature, we analysed the past, present and future of gonadal teratomas, starting from the description of the possible causes, the distinction between benign and malignant, and consequent diversification of the diagnostic and, therefore, therapeutic procedure. Furthermore, we concluded on the importance of a team that involves various specialists, including the pathologist, for the correct diagnosis and therapy to guarantee the best possible quality of life for the patient. Teratomas are neoplasms arising from germ cells and encompass tissues derived from two or more embryonic germ layers, including ectoderm, mesoderm, and endoderm. These tumours typically localize along the midline or in paramedian positions and can manifest as gonadal (20%) or extragonadal (80%) entities. Although gonadal teratomas are uncommon, they represent the predominant type of gonadal tumour in the paediatric population. They comprise approximately 20–25% of all ovarian tumours in females and about 3–5% of all testicular tumours in males. Ovarian teratomas exhibit a higher incidence in early childhood and adolescence, whereas testicular teratomas are more prevalent during the first three months of life and between the ages of 15 and 19. While the majority of paediatric gonadal teratomas are benign, malignant or mixed variants may also arise, necessitating more aggressive therapeutic interventions. [ABSTRACT FROM AUTHOR]

In recent years, the scientific community has increasingly recognized the complex multi-scale competency architecture (MCA) of biology, comprising nested layers of active homeostatic agents, each forming the self-orchestrated substrate for the layer above, and, in turn, relying on the structural and functional plasticity of the layer(s) below. The question of how natural selection could give rise to this MCA has been the focus of intense research. Here, we instead investigate the effects of such decision-making competencies of MCA agential components on the process of evolution itself, using in silico neuroevolution experiments of simulated, minimal developmental biology. We specifically model the process of morphogenesis with neural cellular automata (NCAs) and utilize an evolutionary algorithm to optimize the corresponding model parameters with the objective of collectively self-assembling a two-dimensional spatial target pattern (reliable morphogenesis). Furthermore, we systematically vary the accuracy with which the uni-cellular agents of an NCA can regulate their cell states (simulating stochastic processes and noise during development). This allows us to continuously scale the agents' competency levels from a direct encoding scheme (no competency) to an MCA (with perfect reliability in cell decision executions). We demonstrate that an evolutionary process proceeds much more rapidly when evolving the functional parameters of an MCA compared to evolving the target pattern directly. Moreover, the evolved MCAs generalize well toward system parameter changes and even modified objective functions of the evolutionary process. Thus, the adaptive problem-solving competencies of the agential parts in our NCA-based in silico morphogenesis model strongly affect the evolutionary process, suggesting significant functional implications of the near-ubiquitous competency seen in living matter. [ABSTRACT FROM AUTHOR]

In order to comprehensively describe and evaluate Chinese olive seeds, this study examined 33 varieties (strains) of Chinese olive seeds to address the limitations of previous research on quantitative trait variation and grading among Chinese olive seed varieties (strains). The research specifically focused on evaluating the morphological characteristics, seed locule structure, embryo composition, and phenotypic quantitative traits of Chinese olive seeds. The results indicated that Chinese olive seeds consisted of a core, seed coat, and embryo. Typically, the core contained two to four locules, with each locule containing zero to one embryos. Eight distinct structural variations were identified, with the number of locules per seed ranging from two to four and the number of embryos ranging from one to four. The most common structural types were '3-locule 1-embryo' and '3-locule 2-embryo', each occurring in 100% of the varieties (strains) studied. These two structural types also had a high average proportion within each variety (line), accounting for 50.17% and 42.06%, respectively. The average dimensions of a seed were 31.20 mm in length and 10.89 mm in width, with a shape index of 2.89 and weighing 1.55 g. These quantitative traits displayed significant variation, with the coefficient of variation being highest for single seed weight (19.35%) and lowest for seed length (9.39%). Normality tests revealed that seed width, seed length, and single seed weight followed a normal distribution. These traits were categorized into five levels based on specific points, with probabilities of occurrence approximately 10%, 20%, 40%, 20%, and 10%, respectively. The findings of this study are crucial for understanding and utilizing Chinese olive seed traits and provide valuable insights for the establishment of seed trait standards and data normalization. [ABSTRACT FROM AUTHOR]

Simple Summary: Congenital defects have been described in almost every vertebrate group. In crocodiles, teratology alterations have been described in captive animals (pets, zoos, farms) such as Crocodylus niloticus and Gavialis gangeticus. The present study aimed to characterize congenital malformations of C. acutus from a farm in Lomas de Matunilla, Ballestas, Bolívar, Colombia. The analyzed eggs presented macroscopic malformations, with 42 different types of anomalies observed. Limb and tail malformations (29%) were the most common changes observed. The American crocodile (Crocodylus acutus, Cuvier, 1807) (Class Reptilia, Family Crocodylidae) is a crocodile species inhabiting the Neotropics. Congenital defects have been described in almost every vertebrate group. In crocodiles, teratology alterations have been described in captive animals (pets, zoos, farms) such as Crocodylus niloticus or Gavialis gangeticus. The present study aimed to characterize congenital malformations of C. acutus from a farm in Lomas de Matunilla, Ballestas, Bolívar, Colombia. A total of 550 unhatched eggs were examined after embryo death. A total of 61 embryos presented malformations, with 42 different types of anomalies observed. Limb and tail malformations (29%) were the most common malformations observed. Several malformations, such as cephalothoracopagus, thoracopagus, sternopagus, xiphopagus twins, campylorrachis scoliosa, and acrania, were documented in crocodiles for the first time. Research in teratology enhances our understanding of crocodile biology. It plays a role in their conservation and management, thus helping to ensure the long-term viability of these species in their natural habitats. [ABSTRACT FROM AUTHOR]

Neurogenesis is the process by which new brain cells are formed. This crucial event emerges during embryonic life and proceeds in adulthood, and it could be influenced by environmental pollution. Non-combustion-derived magnetite represents a portion of the coarse particulate matter (PM) contributing to air and water pollution in urban settings. Studies on humans have reported that magnetite and other iron oxides have significant damaging effects at a central level, where these particles accumulate and promote oxidative stress. Similarly, magnetite nanoparticles can cross the placenta and damage the embryo brain during development, but the impact on neurogenesis is still unknown. Furthermore, an abnormal Fe cation concentration in cells and tissues might promote reactive oxygen species (ROS) generation and has been associated with multiple neurodegenerative conditions. In the present study, we used zebrafish as an in vivo system to analyze the specific effects of magnetite on embryonic neurogenesis. First, we characterized magnetite using mineralogical and spectroscopic analyses. Embryos treated with magnetite at sub-lethal concentrations showed a dose–response increase in ROS in the brain, which was accompanied by a massive decrease in antioxidant genes (sod2, cat, gsr, and nrf2). In addition, a higher number of apoptotic cells was observed in embryos treated with magnetite. Next, interestingly, embryos exposed to magnetite displayed a decrease in neural staminal progenitors (nestin, sox2, and pcna markers) and a neuronal marker (elavl3). Finally, we observed significative increases in apoeb (specific microglia marker) and interleukin-1b (il1b), confirming a status of inflammation in the brain embryos treated with magnetite. Our study represents the very first in vivo evidence concerning the effects of magnetite on brain development. [ABSTRACT FROM AUTHOR]

The benzophenone (BP) family, including oxybenzone (BP-3), a prevalent sunscreen ingredient and environmental contaminant, has raised concerns since the year 2005. This study investigated oxybenzone toxicity in zebrafish (Danio rerio) eleutheroembryos and brine shrimp (Artemia salina) nauplii, focusing on the LC50 and developmental impacts. Zebrafish embryos (0.100–1.50 mg/L BP-3, 96 h) and A. salina (0.100–5.00 mg/L BP-3, 48 h) were tested with ultrasound-assisted emulsified liquid-phase microextraction (UA-ELPME) used for zebrafish tissue analysis. HPLC-DAD determined BP-3 concentrations (highest: 0.74 ± 0.13 mg/L). Although no significant zebrafish embryo mortality or hatching changes occurred, developmental effects were evident. Lethal concentrations were determined (A. salina LC50 at 24 h = 3.19 ± 2.02 mg/L; D. rerio embryos LC50 at 24 h = 4.19 ± 3.60 mg/L), with malformations indicating potential teratogenic effects. A. salina displayed intestinal tract alterations and D. rerio embryos exhibited pericardial edema and spinal deformities. These findings highlight oxybenzone's environmental risks, posing threats to species and ecosystem health. [ABSTRACT FROM AUTHOR]

Worldwide meat consumption and production have nearly quintupled in the last 60 years. In this context, research and the application of new technologies related to animal reproduction have evolved in an accelerated way. The objective of the present study was to apply nanoemulsions (NEs) as carriers of lipids to feed bovine embryos in culture media and verify their impact on the development of embryos produced in vitro. The NEs were characterized by particle size, polydispersity, size distribution, physical stability, morphology using atomic force microscopy (AFM), surface tension, density, pH, and rheological behavior. The NEs were prepared by the emulsification/evaporation technique. A central composite rotatable design (CCRD) was used to optimize the NE fabrication parameters. The three optimized formulations used in the embryo application showed an emulsion stability index (ESI) between 0.046 and 0.086, which reflects high stability. The mean droplet diameter analyzed by laser diffraction was approximately 70–80 nm, suggesting a possible transit across the embryonic zona pellucida with pores of an average 90 nm in diameter. AFM images clearly confirm the morphology of spherical droplets with a mean droplet diameter of less than 100 nm. The optimized formulations added during the higher embryonic genome activation phase in bovine embryos enhanced early embryonic development. [ABSTRACT FROM AUTHOR]

Day 3 embryo quality is a predictor of in vitro fertilization (IVF) success rates in cleavage-stage embryo transfer. However, the association between day 3 embryo quality and clinical outcomes in blastocyst transfer policy is largely unknown. This retrospective study included 1074 frozen–thawed single day 5/6 blastocyst transfers between January 2019 and December 2022. Three groups were assessed depending on whether the transferred blastocyst derived from a top-quality, good-quality, or poor-quality embryo at day 3. The analysis was conducted independently for each blastocyst quality group (top, good, and poor) using multivariable logistic regression. We applied a Factorial Analysis of Mixed Data (FAMD) to reduce the potential collinearity between the covariates used in the model. All the blastocysts included in this study were obtained from the first ICSI freeze-all cycles. The cleavage and blastocysts stages were assessed between 67 ± 0.5 (day 3), 115 ± 0.5 (day 5), and 139 ± 0.5 (day 6) hours post-insemination (hpi), respectively. After adjusting for the day of transfer (day 5 or day 6) and FAMD dimensions, no statistical differences in a β-HCG, clinical pregnancy, and live birth were observed among the same-quality blastocysts derived from different day 3 embryo quality groups (top = A, good = B, and poor = C). Our findings showed that a day 3 embryo quality assessment may be unnecessary in planned freeze-all blastocyst cycles. [ABSTRACT FROM AUTHOR]

Simple Summary: Simple Summary: Embryo reinsertion into the same donor mare after embryo flushing would be useful for some reproductive techniques. However, this procedure has been unsuccessful in the past. This study envisages to gain insights into the mechanisms by which embryos fail after embryo flushing and reinsertion in the same mare. The results of this study showed that embryo mortality after embryo reinsertion is not due to luteal deficiency. In contrast, it seems that changes in the endometrial environment due to bacterial contamination and/or inflammation during the uterine lavage are more likely to be responsible for the destruction of most reinserted embryos. The effect of embryo reinsertion immediately after embryo flushing was studied. In Experiment 1, eight mares were used during 32 cycles (8 cycles in each group). For the first two groups, inseminated mares were flushed 8 days after ovulation and prostaglandin F2α was not administered: in group EF-ET (embryo flushing and embryo transfer) the embryo was reinserted in the same donor mare, while in the EF group, no further procedure was performed. In the third group (ET), non-inseminated mares (recipients) received a Day 8 embryo. Progesterone concentration was measured before EF/ET and 72 h after in the three groups. In Experiment 2, twelve mares were used during 17 cycles in two groups, EF-ET (n = 11) and ET (n = 6), as in Experiment 1, except that every mare was flushed 24 h after embryo transfer to retrieve the embryo. Fewer pregnancies resulted after transfer in EF-ET cycles (0/8, 0%) than in the ET group (6/8, 75%). Progesterone concentration decreased significantly (p = 0.05) 72 h after EF-ET but not in EF or ET cycles (p > 0.1). Three mares from the EF-ET showed full luteolysis and signs of endometritis. In Experiment 2, more (5/6; p = 0.08) grade 1 embryos were recovered in the ET compared to the EF-ET group (3/7); 4 embryos were graded 3–4 (were broken or had signs of degeneration) in the EF-ET group but none in the ET group. In both groups, capsule fragments were obtained as indicative of the presence of a recently destroyed embryo in the EF-ET (n = 3) and ET (n = 1) groups. Positive bacterial cultures were obtained in 2/11 and 1/6 embryo flushes from the EF-ET and ET groups, respectively. [ABSTRACT FROM AUTHOR]

Global methylation levels differ in in vitro- and in vivo-developed embryos. Follicular fluid (FF) contains extracellular vesicles (EVs) containing miRNAs that affect embryonic development. Here, we examined our hypothesis that components in FF affect global DNA methylation and embryonic development. Oocytes and FF were collected from bovine ovaries. Treatment of zygotes with a low concentration of FF induced global DNA demethylation, improved embryonic development, and reduced DNMT1/3A levels. We show that embryos take up EVs containing labeled miRNA secreted from granulosa cells and the treatment of zygotes with EVs derived from FF reduces global DNA methylation in embryos. Furthermore, the methylation levels of in vitro-developed blastocysts were higher than those of in their vivo counterparts. Based on small RNA-sequencing and in silico analysis, we predicted miR-29b, -199a-3p, and -148a to target DNMTs and to induce DNA demethylation, thereby improving embryonic development. Moreover, among FF from 30 cows, FF with a high content of these miRNAs demethylated more DNA in the embryos than FF with a lower miRNA content. Thus, miRNAs in FF play a role in early embryonic development. [ABSTRACT FROM AUTHOR]

Reduced oxygen during embryo culture in human ART prevents embryo oxidative stress. Oxidative stress is also the major mechanism by which maternal diabetes impairs embryonic development. This study employed induced hyperglycemia prepubertal mice to mimic childhood diabetes to understand the effects of varying oxygen tension during in vitro embryonic development. The oocytes were fertilized and cultured at low (≈5%) oxygen (LOT) or atmospheric (≈20%) oxygen tension (HOT) for up to 96 h. Embryo development, apoptosis in blastocysts, inner cell mass (ICM) outgrowth proliferation, and Hif1α expression were assessed. Though the oocyte quality and meiotic spindle were not affected, the fertilization rate (94.86 ± 1.18 vs. 85.17 ± 2.81), blastocyst rate (80.92 ± 2.92 vs. 69.32 ± 2.54), and ICM proliferation ability (51.04 ± 9.22 vs. 17.08 ± 3.05) of the hyperglycemic embryos were significantly higher in the LOT compared to the HOT group. On the other hand, blastocysts from the hyperglycemic group, cultured at HOT, had a 1.5-fold increase in apoptotic cells compared to the control and lower Hif1α transcripts in ICM outgrowths compared to the LOT. Increased susceptibility of embryos from hyperglycemic mice to higher oxygen tension warrants the need to individualize the conditions for embryo culture systems in ART clinics, particularly when an endogenous maternal pathology affects the ovarian environment. [ABSTRACT FROM AUTHOR]

Simple Summary: Early embryonic mortality is one of the main causes of poor reproductive health and is influenced by many factors, including mycotoxins and their metabolites in feed. When several mycotoxins are present in feed, they can have different effects and cause significant animal health problems; however, their synergistic effects have not yet been explored. We found that the blastocyst rate was affected by acetonitrile as a vehicle for the mycotoxin. When the desired mycotoxin concentrations were diluted with the lowest possible amount of acetonitrile, we found no statistical difference between blastocyst rates, although there was a trend suggesting that the combination caused poorer blastocyst development. However, a less invasive vehicle should be used to see if the combination of mycotoxins can affect blastocyst rates. The quality of animal feed is increasingly affected by weather conditions, high humidity, and damage to grains, which have led to various mycotoxin-producing moulds. The aim of this study was to determine the effects of the combination of deepoxy-deoxynivalenol and beta-zearalenol on the development of preimplantation bovine embryos, the extent to which the presence of both mycotoxin metabolites affects the development of in vitro cultured bovine embryos, or whether the effect of both toxins enhances embryotoxicity. Ovaries were transported from the abattoir to the laboratory and, after maturation and fertilisation, zygotes were placed in an embryo culture medium (IVC) with different mycotoxin metabolite concentrations diluted in acetonitrile. It was found that the blastocyst rate of cleaved embryos was affected by 1 μL acetonitrile in 400 μL medium (0.25%) compared to the group without acetonitrile. For this reason, it was decided to use acetonitrile as a control group, and the desired mycotoxin metabolite concentrations were diluted in the lowest possible amount of acetonitrile (0.5 μL) that could be accurately added to the study groups. There was no statistical difference when the higher mycotoxin metabolite concentrations were added. [ABSTRACT FROM AUTHOR]

Leptospirosis is an infectious disease that affects domestic animals, wild animals, and humans. It represents a public health problem and has an important economic impact on livestock. This study aims to investigate the importance of genital and transplacental infection in the epidemiology of leptospirosis in cows maintained in Caatinga biome conditions, Northeastern Brazil, as well as reporting organs colonized by Leptospira spp. in embryos and fetuses. Blood, urinary tract (urine, bladder, and kidney), and reproductive tract (vaginal fluid, uterus, uterine tube, ovary, and placenta) samples were collected from 15 slaughtered pregnant cows. Two embryos and 13 fetuses were sampled. Central nervous system and choroid ovoid samples were collected from embryos. Blood, central nervous system, lung, peritoneal liquid, abomasal content, liver, spleen, urine, bladder, kidney, and reproductive system samples were collected from fetuses. Diagnostic methods included the microscopic agglutination test (MAT) using a collection of 24 serovars belonging to 17 different pathogenic serogroups of five species as antigens, as well as polymerase chain reaction (PCR). Anti-Leptospira spp. antibodies were found in 9 cows (60%), while 13 cows (86.67%) had at least one organ or urine with leptospiral DNA. No fetus was seroreactive. Among the embryos and fetuses, 13 (86.67%) presented leptospiral DNA, proving a high frequency of transplacental infection (100%). For cows, the most frequent biological materials regarding Leptospira spp. DNA detection were placenta (13 out of 15 samples; 86.7%), uterus (10 out of 15 samples; 66.7%), and vaginal fluid (5 out of 15 samples; 33.3%), while, for fetuses/embryos, the most frequent PCR-positive samples were choroid ovoid (1/2; 50%), spleen (6/13; 46.2%), kidney (5/13; 38.5%), and central nervous system (5/15; 33.3%). Sequenced samples based on the LipL32 gene presented 99% similarity with L. borgpetersenii. The results indicate that transplacental infection is an efficient way of spreading Leptospira spp. in cows maintained in Caatinga biome conditions. Therefore, prevention and control strategies must include actions that interrupt transmission through this alternative route. [ABSTRACT FROM AUTHOR]

Polyethylene glycol can abrogate plant seed dormancy and alleviate salt–alkali stress damage to plants, but its role in embryonic dormancy abrogation and germination in Sorbus pohuashanensis is not yet clear. The mechanism by which polyethylene glycol promotes the release of embryonic dormancy may be related to the synthesis and metabolism of endogenous hormones, reactive oxygen species and reactive nitrogen. In this article, germination in indoor culture dishes was used, and the most suitable conditions for treating S. pohuashanensis embryos, with polyethylene glycol (PEG) and sodium carbonate (Na2CO3), were selected. Germination was observed and recorded, and related physiological indicators such as endogenous hormones, reactive oxygen species and reactive nitrogen were measured and analyzed to elucidate the mechanism of polyethylene glycol in alleviating salt–alkali stress in S. pohuashanensis embryos. The results showed that soaking seeds in 5% PEG for 5 days is the best condition to promote germination, which can increase the germination rate of embryos under salt–alkali stress by 1–2 times and improve indicators such as germination speed and the germination index. Polyethylene glycol led to an increase in gibberellin (GA), indole-3-acetic acid (IAA), ethylene (ETH), cytokinin (CTK), nitric oxide (NO), soluble protein and soluble sugar in the embryos under salt–alkali stress; increased activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), nitrate reductase (NR) and nitric oxide synthase (NOS) in the embryos; a reduction in the accumulation of abscisic acid (ABA), hydrogen peroxide (H2O2) and malondialdehyde (MDA). Therefore, it is suggested that the inhibitory effect of polyethylene glycol on the salt–alkali-stress-induced germination of S. pohuashanensis embryos is closely related to the response of endogenous hormones, reactive oxygen species and nitric oxide signalling. [ABSTRACT FROM AUTHOR]

The possibility of detecting the developmental competence of individually cultured embryos through analysis of spent media is a major current trend in an ART setting. However, individual embryo culture is detrimental compared with high-density group culture due to the reduced concentration of putative embryotropins. The main aim of this study was to identify an individual culture system that is not detrimental over high-density group culture in the bovine model. Blastocyst rates and competence were investigated in a conventional (GC) group, semi-confined group (MG), and individual culture (MS) in a commercial microwell device. Main findings showed that: (1) individual embryos can be continuously cultured for 7 days in ~70 nL microwells (MS) without detrimental effects compared with the GC and MG; (2) MS and MG blastocysts had a reduced number of TUNEL-positive cells compared to GC blastocysts; (3) though blastocyst mean cell numbers, mitochondrial activity, and lipid content were not different among the three culture conditions, MS blastocysts had a higher frequency of small-sized lipid droplets and a reduced mean droplet diameter compared with GC and MG blastocysts. Overall, findings open the way to optimize the development and competence of single embryos in an ART setting. [ABSTRACT FROM AUTHOR]

Huntington's disease (HD) arises from expanded CAG repeats in exon 1 of the Huntingtin (HTT) gene. The resultant misfolded HTT protein accumulates within neuronal cells, negatively impacting their function and survival. Ultimately, HTT accumulation results in cell death, causing the development of HD. A nonhuman primate (NHP) HD model would provide important insight into disease development and the generation of novel therapies due to their genetic and physiological similarity to humans. For this purpose, we tested CRISPR/Cas9 and a single-stranded DNA (ssDNA) containing expanded CAG repeats in introducing an expanded CAG repeat into the HTT gene in rhesus macaque embryos. Analyses were conducted on arrested embryos and trophectoderm (TE) cells biopsied from blastocysts to assess the insertion of the ssDNA into the HTT gene. Genotyping results demonstrated that 15% of the embryos carried an expanded CAG repeat. The integration of an expanded CAG repeat region was successfully identified in five blastocysts, which were cryopreserved for NHP HD animal production. Some off-target events were observed in biopsies from the cryopreserved blastocysts. NHP embryos were successfully produced, which will help to establish an NHP HD model and, ultimately, may serve as a vital tool for better understanding HD's pathology and developing novel treatments. [ABSTRACT FROM AUTHOR]

Brown algae are multicellular organisms that have evolved independently from plants and animals. Knowledge of the mechanisms involved in their embryogenesis is available only for the Fucus, Dictyota, and Ectocarpus, which are brown algae belonging to three different orders. Here, we address the control of cell growth and cell division orientation in the embryo of Saccharina latissima, a brown alga belonging to the order Laminariales, which grows as a stack of cells through transverse cell divisions until growth is initiated along the perpendicular axis. Using laser ablation, we show that apical and basal cells have different functions in the embryogenesis of this alga, with the apical cell being involved mainly in growth and basal cells controlling the orientation of cell division by inhibiting longitudinal cell division and thereby the widening of the embryo. These functions were observed in the very early development before the embryo reached the 8-cell stage. In addition, the growth of the apical and basal regions appears to be cell-autonomous, because there was no compensation for the loss of a significant part of the embryo upon laser ablation, resulting in smaller and less elongated embryos compared with intact embryos. In contrast, the orientation of cell division in the apical region of the embryo appears to be controlled by the basal cell only, which suggests a polarised, non-cell-autonomous mechanism. Altogether, our results shed light on the early mechanisms of growth rate and growth orientation at the onset of the embryogenesis of Saccharina, in which non-cell-specific cell-autonomous and cell-specific non-cell-autonomous processes are involved. This complex control differs from the mechanisms described in the other brown algal embryos, in which the establishment of embryo polarity depends on environmental cues. [ABSTRACT FROM AUTHOR]

Embryo rescue is a vital technique in cucurbit breeding and propagation, addressing challenges such as embryo abortion, poor seed viability, and incompatibility barriers. This method involves the excision of immature embryos from seeds followed by their in vitro culture on a nutrient medium, providing an environment conducive to their growth and development. In cucurbits, embryo rescue has been extensively utilized to overcome barriers to hybridization, enabling the production of interspecific and intergeneric hybrids with desired traits. Various factors, including genotype, developmental stage of embryos, and culture conditions, influence the success of embryo rescue in cucurbits. Optimal nutrient formulations, growth regulators, and culture techniques are critical for promoting embryo germination, shoot elongation, and subsequent plantlet establishment. Additionally, embryo rescue facilitates the recovery of valuable genetic material from wild and exotic cucurbit species, expanding genetic diversity and developing novel cultivars with improved traits such as disease resistance, yield, and quality. This review highlights the principles, applications, and advancements in embryo rescue technology in cucurbits, emphasizing its significance in cucurbit breeding programs and crop improvement efforts. [ABSTRACT FROM AUTHOR]

Hox gene clusters are crucial in embryogenesis. It was observed that some Hox genes are located in order along the telomeric to centromeric direction of the DNA sequence: Hox1, Hox2, Hox3.... These genes are expressed in the same order in the ontogenetic units of the Drosophila embryo along the anterior–posterior axis. The two entities (genome and embryo) differ significantly in linear size and in-between distance. This strange phenomenon was named spatial collinearity (SP). Later, it was observed that, particularly in the vertebrates, a temporal collinearity (TC) coexists: first Hox1 is expressed, later Hox2, and later on Hox3.... According to a biophysical model (BM), pulling forces act at the anterior end of the cluster while a cluster fastening applies at the posterior end. Hox clusters are irreversibly elongated along the force direction. During evolution, the elongated Hox clusters are broken at variable lengths, thus split clusters may be created. An empirical rule was formulated, distinguishing development due to a complete Hox cluster from development due to split Hox clusters. BM can explain this empirical rule. In a spontaneous mutation, where the cluster fastening is dismantled, a weak pulling force automatically shifts the cluster inside the Hox activation domain. This cluster translocation can probably explain the absence of temporal collinearity in Drosophila. [ABSTRACT FROM AUTHOR]

Simple Summary: The low developmental efficiency of cloned pig embryos limits the application of the pig cloning technique. In this study, a small molecule drug called CPI was added to the culture medium of nuclear donor cells to enhance the developmental capacity of subsequent cloned pig embryos. CPI treatment of nuclear donor cells changed the cellular energy metabolism status from oxidative phosphorylation to glycolysis, and significantly improved the developmental competence of subsequent cloned pig embryos. This study provides a simple approach to facilitate the development and application of pig cloning technology. Somatic cell nuclear transfer (SCNT) technology holds great promise for livestock industry, life science and human biomedicine. However, the development and application of this technology is limited by the low developmental potential of SCNT embryos. The developmental competence of cloned embryos is influenced by the energy metabolic status of donor cells. The purpose of this study was to investigate the effects of CPI, an oxidative phosphorylation inhibitor, on the energy metabolism pathways of pig fibroblasts and the development of subsequent SCNT embryos. The results showed that treatment of porcine fibroblasts with CPI changed the cellular energy metabolic pathways from oxidative phosphorylation to glycolysis and enhanced the developmental ability of subsequent SCNT embryos. The present study establishes a simple, new way to improve pig cloning efficiency, helping to promote the development and application of pig SCNT technology. [ABSTRACT FROM AUTHOR]

Numerous post-translational modifications are involved in oocyte maturation and embryo development. Recently, lactylation has emerged as a novel epigenetic modification implicated in the regulation of diverse cellular processes. However, it remains unclear whether lactylation occurs during oocyte maturation and embryo development processes. Herein, the lysine lactylation (Kla) modifications were determined during mouse oocyte maturation and early embryo development by immunofluorescence staining. Exogenous lactate was supplemented to explore the consequences of modulating histone lactylation levels on oocyte maturation and embryo development processes by transcriptomics. Results demonstrated that lactylated proteins are widely present in mice with tissue- and cell-specific distribution. During mouse oocyte maturation, immunofluorescence for H3K9la, H3K14la, H4K8la, and H4K12la was most intense at the germinal vesicle (GV) stage and subsequently weakened or disappeared. Further, supplementing the culture medium with 10 mM sodium lactate elevated both the oocyte maturation rate and the histone Kla levels in GV oocytes, and there were substantial increases in Kla levels in metaphase II (MII) oocytes. It altered the transcription of molecules involved in oxidative phosphorylation. Moreover, histone lactylation levels changed dynamically during mouse early embryogenesis. Sodium lactate at 10 mM enhanced early embryo development and significantly increased lactylation, while impacting glycolytic gene transcription. This study reveals the roles of lactylation during oocyte maturation and embryo development, providing new insights to improving oocyte maturation and embryo quality. [ABSTRACT FROM AUTHOR]

Coconut palms (Cocos nucifera L.) are globally significant palms with both economic and cultural value. Despite the increasing demand for coconut products, production is decreasing globally due to palm senility, pests, and diseases. It has been estimated that over half of the world's coconut palms need to be replaced immediately. The coconut industry has acknowledged that conventional propagation methods are unlikely to yield sufficient high-quality planting material. Therefore, coconut tissue culture is considered a potential solution to this problem. By using coconut tissue culture, a large number of plantlets can be obtained in a short period of time. In this study, the quality of explants and the development stage (visible shoot/non-visible shoot) of coconut used for micropropagation were examined. To our knowledge, little research has been undertaken on this aspect of coconut micropropagation. Our results indicated that tender coconut fruit exhibited an advantage over mature fruits. In addition, coconut plumule explants subjected to an extended storage of 15 days demonstrated enhanced development compared to those without storage. Notably, smaller embryos utilized as explants displayed superior callus formation compared to their larger counterparts. Finally, embryos possessing shoots exhibited improved callus initiation, albeit accompanied by a more pronounced browning effect. Further investigations are required to obtain more knowledge about the most suitable conditions for plumule explants that lead to optimal callus initiation. [ABSTRACT FROM AUTHOR]

Cancer poses significant emotional challenges for children and adolescents, despite improvements in survival rates due to new therapies. However, there is growing concern about the long-term effects, including fertility issues. This review examines recent advancements and future directions in fertility preservation within a pediatric population subjected to oncological therapies. Worldwide, there is variability in the availability of fertility preservation methods, influenced by factors like development status and governmental support. The decision to pursue preservation depends on the risk of gonadotoxicity, alongside factors such as diagnosis, treatment, clinical status, and prognosis. Currently, options for preserving fertility in prepubertal boys are limited compared to girls, who increasingly have access to ovarian tissue preservation. Adolescents and adults have more options available, but ethical considerations remain complex and diverse. [ABSTRACT FROM AUTHOR]

The present study compares sugarcane-wax purified policosanols sourced from Cuba (Raydel®) and China (BOC Sciences) and utilized following the synthesis of reconstituted high-density lipoproteins (rHDL). The two policosanols exhibited distinctly different ingredient ratios of long-chain aliphatic alcohols, particularly 1-octacosanol (C28) and 1-tetratriacotanol (C34). After synthesizing rHDL with apolipoprotein A-I (apoA-I), the two policosanols bound well with phospholipid and apoA-I to form the discoidal rHDL. Notably, rHDL-1, containing Cuban policosanol, displayed the largest particle diameter at approximately 78 ± 3 nm. In contrast, both control rHDL (rHDL-0) and rHDL containing Chinese policosanol (rHDL-2) exhibited smaller particles, with diameters of approximately 58 ± 3 nm and 61 ± 2 nm, respectively. Furthermore, rHDL-1 demonstrated enhanced anti-glycation activity, safeguarding apoA-I from degradation within HDL, and displayed the antioxidant ability to inhibit LDL oxidation. A microinjection of each rHDL into zebrafish embryos in the presence of carboxymethyllysine (CML) revealed rHDL-1 to have the strongest antioxidant activity with the highest embryo survivability and normal developmental morphology. Dermal application to recover the wound revealed rHDL-1 to have the highest wound-healing activity (75%) and survivability (92%) in the cutaneous wound area in the presence of CML. In adult zebrafish, injecting CML (250 μg) caused acute death and hyperinflammation, marked by heightened neutrophil infiltration and interleukin (IL)-6 production in liver. However, co-administering rHDL-1 notably increased survival (85%) and exhibited strong anti-inflammatory properties, reducing IL-6 production while improving the blood lipid profile. However, a co-injection of rHDL-2 resulted in the lowest survivability (47%) with more hepatic inflammation. In conclusion, Cuban policosanol (Raydel®) has more desirable properties for the in vitro synthesis of rHDL with stronger anti-glycation and antioxidant activities than those of Chinese policosanol (BOC Sciences). Moreover, Raydel-policosanol-integrated rHDL demonstrates a noteworthy effect on accelerated wound healing and robust anti-inflammatory properties, leading to increased survivability in zebrafish embryos and adults by effectively suppressing CML-induced hyperinflammation. [ABSTRACT FROM AUTHOR]

In vitro-generated blastocyst-like structures are of great importance since they recapitulate specific features or processes of early embryogenesis, thus avoiding ethical concerns as well as increasing scalability and accessibility compared to the use of natural embryos. Here, we combine cell reprogramming and mechanical stimuli to create 3D spherical aggregates that are phenotypically similar to those of natural embryos. Specifically, dermal fibroblasts are reprogrammed, exploiting the miR-200 family property to induce a high plasticity state in somatic cells. Subsequently, miR-200-reprogrammed cells are either driven towards the trophectoderm (TR) lineage using an ad hoc induction protocol or encapsulated into polytetrafluoroethylene micro-bioreactors to maintain and promote pluripotency, generating inner cell mass (ICM)-like spheroids. The obtained TR-like cells and ICM-like spheroids are then co-cultured in the same micro-bioreactor and, subsequently, transferred to microwells to encourage blastoid formation. Notably, the above protocol was applied to fibroblasts obtained from young as well as aged donors, with results that highlighted miR-200′s ability to successfully reprogram young and aged cells with comparable blastoid rates, regardless of the donor's cell age. Overall, the approach here described represents a novel strategy for the creation of artificial blastoids to be used in the field of assisted reproduction technologies for the study of peri- and early post-implantation mechanisms. [ABSTRACT FROM AUTHOR]

Aspartame, a widely used artificial sweetener, is present in many food products and beverages worldwide. It has been linked to potential neurotoxicity and developmental defects. However, its teratogenic effect on embryonic development and the underlying potential mechanisms need to be elucidated. We investigated the concentration- and time-dependent effects of aspartame on zebrafish development and teratogenicity. We focused on the role of sirtuin 1 (SIRT1) and Forkhead-box transcription factor (FOXO), two proteins that play key roles in neurodevelopment. It was found that aspartame exposure reduced the formation of larvae and the development of cartilage in zebrafish. It also delayed post-fertilization development by altering the head length and locomotor behavior of zebrafish. RNA-sequencing-based DEG analysis showed that SIRT1 and FOXO3a are involved in neurodevelopment. In silico and in vitro analyses showed that aspartame could target and reduce the expression of SIRT1 and FOXO3a proteins in neuron cells. Additionally, aspartame triggered the reduction of autophagy flux by inhibiting the nuclear translocation of SIRT1 in neuronal cells. The findings suggest that aspartame can cause developmental defects and teratogenicity in zebrafish embryos and reduce autophagy by impairing the SIRT1/FOXO3a axis in neuron cells. [ABSTRACT FROM AUTHOR]

Background: Does the Time-lapse Incubator (TLI) add value to reproductive outcomes when its two components, undisturbed culturing and morphokinetic embryo grading, are separated. Methods: A prospective pilot, randomized, controlled, double-blinded, single-center study was conducted during the years 2016–2020. In total, 102 patients were randomized into three groups: (1) conventional incubation with morphological evaluation only (n = 34), (2) TLI with both morphological and morphokinetic evaluations (n = 32), and (3) TLI with morphological evaluation only (n = 36). All arms were cultured in ESCO-MIRI® incubators. A total of 1061 injected mature oocytes were evaluated (420 in arm 1, 285 in arm 2, and 356 in arm 3). The primary outcome was live birth rates. Secondary outcomes included clinical and cumulative pregnancy rates as well as embryo quality. Embryos in arm 3 were retrospectively evaluated for their morphokinetic score. Results: No significant difference was found in the live birth rate for single embryo transfer cycles (SET) (35% vs. 31.6% vs. 24%, p = 0.708) or double embryo transfer (DET) cycles (41.7% vs. 38.5% vs. 36.4%, p = 0.966). Comparable pregnancy rates, clinical pregnancy rates, and cumulative pregnancy were found for similar top-quality embryos for days 2, 3, and blastocyst stages across groups. A similar number of embryos were suitable for either transfer or cryopreservation within the different groups. For 62.8% of the embryos in arm 3, the morphokinetic and morphologic evaluations were similar. In only 2/36 (5.6%) treatment cycles, the use of morphokinetic scoring may have helped the patient avoid undergoing an additional treatment cycle. In the other cases, morphokinetic scoring would not have changed the end point of pregnancy. Conclusions: The two components of the TLI system—undisturbed culturing and morphokinetic embryo grading—do not appear to have a significant additional value in reproductive outcome, although these results should be validated by an RCT. [ABSTRACT FROM AUTHOR]

Simple Summary: The ingestion of excessive nitrate can affect the thyroid gland and cause thyroid dysfunction in humans. In the present study, amphibian embryos were exposed to nitrate, thyroxine and methimazole (a thyroid peroxidase inhibitor) during embryonic development to further explore the effects of nitrate on the thyroid. The results showed that nitrate, thyroxine and methimazole inhibited embryo growth and development. Additionally, methimazole and high concentrations of nitrate downregulated the genes related to thyroid morphogenesis and cholesterol metabolism, while upregulating the genes related to inflammation and apoptosis. These suggested that nitrate not only damaged the thyroid gland, but also affected the formation of the thyroid, thus affecting embryonic development. In recent years, nitrate (NO3-N) pollution in water bodies has been increasing due to the excessive use of nitrogen-based fertilizers. Exposure to NO3-N during the development of amphibian embryos may have lasting effects on the growth and development of individuals and even threaten their survival, but the toxicity mechanism of NO3-N in amphibian embryos prior to thyroid morphogenesis remains unclear. In the present study, Bufo gargarizans was selected as the model organism to investigate the toxic effects of 10 mg/L and 100 mg/L NO3-N exposure (N10 and N100) on amphibian embryos using methimazole (MMI) and exogenous thyroxine (T4) as the reference groups. We found that T4, MMI, N10 and N100 inhibited B. gargarizans embryo growth and development, with MMI and N100 showing the earliest and strongest effects. Transcriptome analysis revealed that MMI and NO3-N (especially N100) significantly downregulated genes related to thyroid morphogenesis and cholesterol metabolism, while upregulating genes related to inflammation and apoptosis. Together, these results contribute to a deeper understanding of the complex mechanisms by which NO3-N disrupts B. gargarizans embryonic development, reveal the potential risks of NO3-N pollution to other aquatic organisms, and provide insights into the conservation of a broader ecosystem. [ABSTRACT FROM AUTHOR]

Neuromesodermal progenitors (NMPs), serving as the common origin of neural and paraxial mesodermal development in a large part of the trunk, have recently gained significant attention because of their critical importance in the understanding of embryonic organogenesis and the design of in vitro models of organogenesis. However, the nature of NMPs at many essential points remains only vaguely understood or even incorrectly assumed. Here, we discuss the nature of NMPs, focusing on their dynamic migratory behavior during embryogenesis and the mechanisms underlying their neural vs. mesodermal fate choice. The discussion points include the following: (1) How the sinus rhomboidals is organized; the tissue where the neural or mesodermal fate choice of NMPs occurs. (2) NMPs originating from the broad posterior epiblast are associated with Sox2 N1 enhancer activity. (3) Tbx6-dependent Sox2 repression occurs during NMP-derived paraxial mesoderm development. (4) The nephric mesenchyme, a component of the intermediate mesoderm, was newly identified as an NMP derivative. (5) The transition of embryonic tissue development from tissue-specific progenitors in the anterior part to that from NMPs occurs at the forelimb bud axial level. (6) The coexpression of Sox2 and Bra in NMPs is conditional and is not a hallmark of NMPs. (7) The ability of the NMP pool to sustain axial embryo growth depends on Wnt3a signaling in the NMP population. Current in vitro models of NMPs are also critically reviewed. [ABSTRACT FROM AUTHOR]

Simple Summary: The study of the earliest stages of pig embryo development has several potential uses: feeding a growing population, medical applications that require the use of genetically modified organisms, rare breed conservation, and the use of pig embryos as a research model for progressing IVF technology in both humans and other (e.g., endangered) species. Despite this, barriers to the widespread adoption of pig embryo production include fat-rich cells that are difficult to see down a microscope, low success rates, and slow adoption of new technologies such as artificial intelligence. Here, we fully characterise pig early embryo development in an undisturbed system, using videos derived from an incubator equipped with in-built cameras. We find that a rapid first cell division, from one to two cells, correlates with ongoing embryo success and that this early-stage measure alone can be an accurate predictor of future development. Conversely, we find that re-fusion of cells following division is associated with poor rates of embryo development. Such predictors may allow for the early commercial selection of high-quality embryos and upscaling of production so that the true potential of pig embryology can be realised. The study of pig preimplantation embryo development has several potential uses: from agriculture to the production of medically relevant genetically modified organisms and from rare breed conservation to acting as a physiologically relevant model for progressing human and other (e.g., endangered) species' in vitro fertilisation technology. Despite this, barriers to the widespread adoption of pig embryo in vitro production include lipid-laden cells that are hard to visualise, slow adoption of contemporary technologies such as the use of time-lapse incubators or artificial intelligence, poor blastulation and high polyspermy rates. Here, we employ a commercially available time-lapse incubator to provide a comprehensive overview of the morphokinetics of pig preimplantation development for the first time. We tested the hypotheses that (a) there are differences in developmental timings between blastulating and non-blastulating embryos and (b) embryo developmental morphokinetic features can be used to predict the likelihood of blastulation. The abattoir-derived oocytes fertilised by commercial extended semen produced presumptive zygotes were split into two groups: cavitating/blastulating 144 h post gamete co-incubation and those that were not. The blastulating group reached the 2-cell and morula stages significantly earlier, and the time taken to reach the 2-cell stage was identified to be a predictive marker for blastocyst formation. Reverse cleavage was also associated with poor blastulation. These data demonstrate the potential of morphokinetic analysis in automating and upscaling pig in vitro production through effective embryo selection. [ABSTRACT FROM AUTHOR]

As more and more of the available genomic data have been published, several databases have been developed for deciphering early mammalian embryogenesis; however, less research has been conducted on the regulation of the expression of natural immunity genes during early embryonic development in dairy cows. To this end, we explored the regulatory mechanism of innate immunity genes at the whole-genome level. Based on comparative genomics, 1473 innate immunity genes in cattle were obtained by collecting the latest reports on human innate immunity genes and updated bovine genome data for comparison, and a preliminary database of bovine innate immunity genes was constructed. In order to determine the regulatory mechanism of innate immune genes in dairy cattle early embryos, we conducted weighted co-expression network analysis of the innate immune genes at different developmental stages of dairy cattle early embryos. The results showed that specific module-related genes were significantly enriched in the MAPK signaling pathway. Protein–protein interaction (PPI) analysis showed gene interactions in each specific module, and 10 of the highest connectivity genes were chosen as potential hub genes. Finally, combined with the results for differential expressed genes (DEGs), ATF3, IL6, CD8A, CD69, CD86, HCK, ERBB3, LCK, ITGB2, LYN, and ERBB2 were identified as the key genes of innate immunity in dairy cattle early embryos. In conclusion, the bovine innate immunity gene set was determined and the co-expression network of innate immunity genes in the early embryonic stage of dairy cattle was constructed by comparing and analyzing the whole genome of bovines and humans. The findings in this study provide the basis for exploring the involvement and regulation of innate immune genes in the early embryonic development of dairy cattle. [ABSTRACT FROM AUTHOR]

Mitochondria undergo a myriad of changes during pre-implantation embryo development, including shifts in activity levels and mitochondrial DNA (mtDNA) replication. However, how these distinct aspects of mitochondrial function are linked and their responsiveness to diverse stressors is not well understood. Here, we show that mtDNA content increased between 8-cell embryos and the blastocyst stage, with similar copy numbers per cell in the inner cell mass (ICM) and trophectoderm (TE). In contrast, mitochondrial membrane potential (MMP) was higher in TE than ICM. Culture in ambient oxygen (20% O2) altered both aspects of mitochondrial function: the mtDNA copy number was upregulated in ICM, while MMP was diminished in TE. Embryos cultured in 20% O2 also exhibited delayed development kinetics, impaired implantation, and reduced mtDNA levels in E18 fetal liver. A model of oocyte mitochondrial stress using rotenone showed only a modest effect on on-time development and did not alter the mtDNA copy number in ICM; however, following embryo transfer, mtDNA was higher in the fetal heart. Lastly, endogenous mitochondrial dysfunction, induced by maternal age and obesity, altered the blastocyst mtDNA copy number, but not within the ICM. These results demonstrate that mitochondrial activity and mtDNA content exhibit cell-specific changes and are differentially responsive to diverse types of oxidative stress during pre-implantation embryogenesis. [ABSTRACT FROM AUTHOR]

Three new species of the genus Caridina are described from the northernmost part of Australia. Caridina darwin n. sp. resembles Caridina temasek Choy and Ng, 1991 but differs in the armature of the rostrum, the development of epipods on the pereiopods and the absence of an appendix interna on the male first pleopods. Caridina magnovis n. sp. resembles Caridina serratirostris de Man, 1892 but differs in the armature of the ventral margin of the rostrum, a shorter stylocerite, a stouter carpus of the first pereiopod, the number and size of spiniform setae on the third and fifth pereiopods, the shape of the preanal carina and the size of the embryos (referred to as "eggs" in most previous publications). Caridina wilsoni n. sp. resembles Caridina gracilirostris de Man, 1892 but differs in the size of the embryos and in some length to width ratios of the segments of the pereiopods. Detailed morphological descriptions of all three new species are given. A molecular phylogeny (mt DNA 16S) supports the morphospecies hypothesis and illustrates the phylogenetic relationship with morphologically similar species from outside Australia. [ABSTRACT FROM AUTHOR]

Seeds are one of the most important characteristics of plant evolution. Within a seed, the embryo, which will grow into a plant, can survive harsh environments. When the seeds are mature, the mother plant will disperse them from its body, allowing them to be taken away to grow in a new place. Otherwise, if the young generation grows alongside the mother plants in the same place, they will compete for sunlight and nutrition. The mother plants use different strategies to send away their seeds. One of these strategies is endozoochory, which means that the seeds disperse via ingestion by animals. There is a conflict between the seeds' abilities to attract animals and protect the embryo within the digestion systems of animals. Magnolia seeds exhibit typical endozoochory. The seed coats of Magnolia feature sarcotestas and sclerotestas. The sarcotesta, which is fleshy, bright-colored, and edible, attracts animals. The sclerotesta is hard and woody, protecting the embryo from the digestive systems of animals. In this study, we used scanning electron and light microscopes to examine the development of the sarcotesta and sclerotesta of Magnolia stellata seed coats. The results showed that the sarcotesta and sclerotesta come from the outer integument. This result confirms the hypothesis of Asa Gray from 1848. The dependence of the seed dispersal strategy on structural development is discussed. [ABSTRACT FROM AUTHOR]

Nitrogen-based fertilizers represent the most common fertilization tools, particularly used in crop food agriculture, despite the low cost-efficiency and the high negative environmental impact. At present, there is still inadequate information available about the effects of urea on human health; nevertheless, previous studies in animals observed that high urea concentration exposure can damage different tissues, including the brain. In several vertebrates, a crucial factor involved in neuronal cell formation is represented by the gas molecule, nitric oxide (NO), derived from the conversion of arginine to citrulline through the enzymatic activity of nitric oxide synthases (NOS). In zebrafish, three different isoforms of the NOS gene are known: nos1, nos2a, and nos2b. In the present study we show that nos1 represents the unique isoform with a stable high expression in the brain and spinal cord during all the embryonic stages of zebrafish development. Then, by using a specific transgenic zebrafish line, Tg(HuC:GFP), to mark neuronal cells, we observed nos1 to be specifically expressed in neurons. Interestingly, we observed that urea exposure at sub-lethal doses affected cell proliferation and the number of nos1-expressing cells, inducing apoptosis. Consistently, brain NO levels were observed to be reduced in urea-treated animals compared to untreated ones. This finding represents the first evidence that urea exposure affects the expression of a key gene involved in neuronal cell formation during embryonic development. [ABSTRACT FROM AUTHOR]

Sapium sebiferum is a tree with socioeconomic, environmental, and medicinal value. S. sebiferum seeds possess physiological dormancy, which is induced by endogenous inhibitors and can be broken by cold stratification. However, the putative spatiotemporal changes in inhibitor activity are currently unknown, which can hinder the propagation of S. sebiferum seeds. The objective of this study was to investigate the spatiotemporal dynamics of inhibitor activity and its effect on germination during the cold stratification of S. sebiferum seeds. An extractant consisting of 80% methanol was used to extract the inhibitory substance from the seed coat and endosperm at different stages of cold stratification. The extract was then applied to both the Chinese cabbage seeds and excised embryos of S. sebiferum. The germination percentage and germination index were used to assess the inhibitor activity of S. sebiferum seeds. The germination of non-stratified S. sebiferum seeds was completely inhibited (0% germination). As the stratification duration was extended, the germination percentage of S. sebiferum seeds gradually increased. However, dormancy persisted until the stratification duration reached 120 d; at this point, the mean germination was 81.3%. The germination test on Chinese cabbage seeds revealed a significant increase from 10.0% (stratified for 0 d) to 91.2% (stratified for 120 d) when treated with endosperm extracts. The germination indexes also increased from 0.0 to 40.3, indicating a decrease in the inhibitory activity of endosperm extracts. The seed coat extracting solution showed varying dynamic changes. The lowest germination was observed after 60 d of stratification, with no significant differences among the results of 0 d, 30 d, and 60 d. However, after stratification for 90 d, the germination percentage of Chinese cabbage seeds increased. The germination percentage of excised embryos exhibited similar changes to those of Chinese cabbage seeds. This study discovered that endogenous inhibitors were present in both the seed coat and endosperm of S. sebiferum seeds, and the inhibitor activity was higher in the endosperm. The spatiotemporal patterns of inhibitor activity suggest that the endogenous inhibitors move from the endosperm to the seed coat during early cold stratification stages, aiding in the physiological dormancy release of S. sebiferum seeds. These findings enhance our understanding of seed biology in S. sebiferum and will facilitate high-efficiency seed propagation. [ABSTRACT FROM AUTHOR]

The presence of per- and polyfluoroalkyl substances (PFASs) in aquatic environments is often persistent and widespread. Understanding the potential adverse effects from this group of chemicals on aquatic communities allows for better hazard characterization. This study examines impacts on zebrafish (Danio rerio) embryo physiology, behavior, and lipid levels from exposure to perfluorooctanoic acid (PFOA), perfluorohexane sulfonate (PFHxS), and heptadecafluorooctanesulfonic acid (PFOS). Embryos were exposed to lethal and sublethal levels of each chemical and monitored for alterations in physiological malformations, mortality, lipid levels, and behavior (only PFOA and PFHxS). The predicted 50% lethal concentrations for 120 hpf embryos were 528.6 ppm PFOA, 14.28 ppm PFHxS, and 2.14 ppm PFOS. Spine curvature and the inability of the 120 hpf embryos to maintain a dorsal-up orientation was significantly increased at 10.2 ppm PFHxS and 1.9 ppm PFOS exposure. All measured 120 hpf embryo behaviors were significantly altered starting at the lowest levels tested, 188 ppm PFOA and 6.4 ppm PFHxS. Lipid levels decreased at the highest PFAS levels tested (375 PFOA ppm, 14.4 PFHxS ppm, 2.42 ppm PFOS). In general, the PFAS chemicals, at the levels examined in this study, increased morphological deformities, embryo activity, and startle response time, as well as decreased lipid levels in 120 hpf zebrafish embryos. [ABSTRACT FROM AUTHOR]

Chromosome segregation in female germ cells and early embryonic blastomeres is known to be highly prone to errors. The resulting aneuploidy is therefore the most frequent cause of termination of early development and embryo loss in mammals. And in specific cases, when the aneuploidy is actually compatible with embryonic and fetal development, it leads to severe developmental disorders. The main surveillance mechanism, which is essential for the fidelity of chromosome segregation, is the Spindle Assembly Checkpoint (SAC). And although all eukaryotic cells carry genes required for SAC, it is not clear whether this pathway is active in all cell types, including blastomeres of early embryos. In this review, we will summarize and discuss the recent progress in our understanding of the mechanisms controlling chromosome segregation and how they might work in embryos and mammalian embryos in particular. Our conclusion from the current literature is that the early mammalian embryos show limited capabilities to react to chromosome segregation defects, which might, at least partially, explain the widespread problem of aneuploidy during the early development in mammals. [ABSTRACT FROM AUTHOR]

Camelina is an oil seed crop that is enjoying increasing interest because it has a particularly valuable fatty acid profile, is modest regarding its water and nutrient requirements, and is comparatively resilient to abiotic and biotic stress factors. The regeneration of plants from cells accessible to genetic manipulation is an essential prerequisite for the generation of genetically engineered plants, be it by transgenesis or genome editing. Here, immature embryos were used on the assumption that their incomplete differentiation was associated with totipotency. In culture, regenerative structures appeared adventitiously at the embryos' hypocotyls. For this, the application of auxin- or cytokinin-type growth regulators was essential. The formation of regenerative structures was most efficient when indole-3-acetic acid was added to the induction medium at 1 mg/L, zygotic embryos of the medium walking stick stage were used, and their hypocotyls were stimulated by pricking to a wound response. Histological examinations revealed that the formation of adventitious shoots was initiated by locally activated cell division and proliferation in the epidermis and the outer cortex of the hypocotyl. While the regeneration of plants was established in principle using the experimental line Cam139, the method proved to be similarly applicable to the current cultivar Ligena, and hence it constitutes a vital basis for future genetic engineering approaches. [ABSTRACT FROM AUTHOR]

Simple Summary: This study investigated the developmental differences between in vivo-developed (IVV) and in vitro embryos (in vitro fertilization (IVF) and parthenogenetic activation (PA)) in pigs using single-cell RNA sequencing data from the oocyte to the 8-cell stages. Findings show that in vitro embryos have more similar developmental trajectories and lower gene diversity, especially PA, compared to IVV. Significant variations were observed in maternal mRNA, influencing mRNA splicing, energy metabolism, and chromatin remodeling, where key genes like SMARCB1 and HDAC1 (in vivo) and SIRT1 and EZH2 (in vitro) might be playing crucial roles. Despite similar ZGA timings across embryo types, IVV embryos exhibited more upregulated genes, particularly during major ZGA, and global epigenetic modification differences were evident from the oocyte stage and continued to expand during minor ZGA. They also uniquely upregulated genes linked to mitochondrial function, ATP synthesis, and oxidative phosphorylation. A notable difference in mRNA degradation between in vivo and in vitro embryos was observed during major ZGA, and the timely decay of maternal mRna in IVV except in in vitro embryos was related to phosphatase activity and cell junctions. The study also highlighted the higher expression of mitochondrially encoded genes in in vitro embryos, though nucleosome occupancy and ATP8 expression were higher in IVV. In vitro-fertilized (IVF) and parthenogenetically activated (PA) embryos, key to genetic engineering, face more developmental challenges than in vivo-developed embryos (IVV). We analyzed single-cell RNA-seq data from the oocyte to eight-cell stages in IVV, IVF, and PA porcine embryos, focusing on developmental differences during early zygotic genome activation (ZGA), a vital stage for embryonic development. (1) Our findings reveal that in vitro embryos (IVF and PA) exhibit more similar developmental trajectories compared to IVV embryos, with PA embryos showing the least gene diversity at each stage. (2) Significant differences in maternal mRNA, particularly affecting mRNA splicing, energy metabolism, and chromatin remodeling, were observed. Key genes like SMARCB1 (in vivo) and SIRT1 (in vitro) played major roles, with HDAC1 (in vivo) and EZH2 (in vitro) likely central in their complexes. (3) Across different types of embryos, there was minimal overlap in gene upregulation during ZGA, with IVV embryos demonstrating more pronounced upregulation. During minor ZGA, global epigenetic modification patterns diverged and expanded further. Specifically, in IVV, genes, especially those linked to H4 acetylation and H2 ubiquitination, were more actively regulated compared to PA embryos, which showed an increase in H3 methylation. Additionally, both types displayed a distinction in DNA methylation. During major ZGA, IVV distinctively upregulated genes related to mitochondrial regulation, ATP synthesis, and oxidative phosphorylation. (4) Furthermore, disparities in mRNA degradation-related genes between in vivo and in vitro embryos were more pronounced during major ZGA. In IVV, there was significant maternal mRNA degradation. Maternal genes regulating phosphatase activity and cell junctions, highly expressed in both in vivo and in vitro embryos, were degraded in IVV in a timely manner but not in in vitro embryos. (5) Our analysis also highlighted a higher expression of many mitochondrially encoded genes in in vitro embryos, yet their nucleosome occupancy and the ATP8 expression were notably higher in IVV. [ABSTRACT FROM AUTHOR]

This study investigated whether metabolomic fingerprints of bovine embryo growth media improve the prediction of successful embryo implantation. In this prospective cohort study, the metabolome from in vitro-produced day 7 blastocysts with successful implantation (n = 11), blastocysts with failed implantation (n = 10), and plain culture media without embryos (n = 5) were included. Samples were analyzed using an AbsoluteIDQ® p180 Targeted Metabolomics Kit with LC-MS/MS, and a total of 189 metabolites were analyzed from each sample. Blastocysts that resulted in successful embryo implantation had significantly higher levels of methionine sulfoxide (p < 0.001), DOPA (p < 0.05), spermidine (p < 0.001), acetylcarnitine-to-free-carnitine ratio (p < 0.05), C2 + C3-to-free-carnitine ratio (p < 0.05), and lower levels of threonine (nep < 0.001) and phosphatidylcholine PC ae C30:0 (p < 0.001) compared to control media. However, when compared to embryos that failed to implant, only DOPA, spermidine, C2/C0, (C2 + C3)/C0, and PC ae C30:0 levels differentiated significantly. In summary, our study identifies a panel of differential metabolites in the culture media of bovine blastocysts that could act as potential biomarkers for the selection of viable blastocysts before embryo transfer. [ABSTRACT FROM AUTHOR]

Generating synthetic time series data, such as videos, presents a formidable challenge as complexity increases when it is necessary to maintain a specific distribution of shown stages. One such case is embryonic development, where prediction and categorization are crucial for anticipating future outcomes. To address this challenge, we propose a Siamese architecture based on diffusion models to generate predictive long-duration embryonic development videos and an evaluation method to select the most realistic video in a non-supervised manner. We validated this model using standard metrics, such as Fréchet inception distance (FID), Fréchet video distance (FVD), structural similarity (SSIM), peak signal-to-noise ratio (PSNR), and mean squared error (MSE). The proposed model generates videos of up to 197 frames with a size of 128 × 128 , considering real input images. Regarding the quality of the videos, all results showed improvements over the default model (FID = 129.18, FVD = 802.46, SSIM = 0.39, PSNR = 28.63, and MSE = 97.46). On the coherence of the stages, a global stage mean squared error of 9.00 was achieved versus the results of 13.31 and 59.3 for the default methods. The proposed technique produces more accurate videos and successfully removes cases that display sudden movements or changes. [ABSTRACT FROM AUTHOR]

Embryo morphology and morphokinetics have been studied for their association with euploid embryos. However, the results are controversial, especially in the advanced-aged women group, when the risk of aneuploidy increases significantly. This prospective cohort study evaluated the association between embryo development between day-3 cleavage and day-5 blastocyst stages and euploidy rates, determined using preimplantation genetic testing for aneuploidy (PGT-A). Embryos from women aged 35 years and above who underwent intracytoplasmic sperm injections and PGT-A were studied. Day-3 cleavage-stage embryos were evaluated for their cell number, and day-5 blastocyst-stage embryos were evaluated for their morphological grade. Embryo development from day 3 to day 5 was categorized as either good or poor development and evaluated for its association with the PGT-A results. We evaluated 325 embryos from 101 infertile couples. It was found that 55.17% of blastocysts with good development and 29.83% with poor development were euploid. A significant association was found between embryo development and euploidy rates in advanced-aged women (p < 0.001). Also, there were significantly higher rates of euploid embryos with good blastocyst morphological grades, especially blastocyst expansion grades and trophectoderm grades. In conclusion, embryo morphokinetics shows promising results in predicting euploidy in advanced female age. [ABSTRACT FROM AUTHOR]

Simple Summary: The in vitro production (IVP) of bovine embryos has increased in popularity in the past few decades with improvements in our ability to harvest oocytes from genetically elite heifers and cows, fertilize them in vitro, and successfully culture embryos to the stage where they can be transferred. One issue with IVP is that the efficiency of embryo production is less than ideal. One likely reason for the poor development of IVP embryos is the excessive exposure of oocytes and embryos to reactive oxygen species (ROS). These molecules are produced as a normal by-product of energy generation; however, excess accumulation of ROS during the IVP of oocyte and embryo culture will damage nucleic acids, lipids, and proteins in ways that compromise development potential and post-transfer embryo survival. Molecules that react with and block ROS-induced damage are commonly referred to as antioxidants. This review explores the use of five common antioxidants to limit ROS-induced damage and promote the IVP of embryos. The in vitro production (IVP) of bovine embryos has gained popularity worldwide and in recent years and its use for producing embryos from genetically elite heifers and cows has surpassed the use of conventional superovulation-based embryo production schemes. There are, however, several issues with the IVP of embryos that remain unresolved. One limitation of special concern is the low efficiency of the IVP of embryos. Exposure to reactive oxygen species (ROS) is one reason why the production of embryos with IVP is diminished. These highly reactive molecules are generated in small amounts through normal cellular metabolism, but their abundances increase in embryo culture because of oocyte and embryo exposure to temperature fluctuations, light exposure, pH changes, atmospheric oxygen tension, suboptimal culture media formulations, and cryopreservation. When uncontrolled, ROS produce detrimental effects on the structure and function of genomic and mitochondrial DNA, alter DNA methylation, increase lipid membrane damage, and modify protein activity. Several intrinsic enzymatic pathways control ROS abundance and damage, and antioxidants react with and reduce the reactive potential of ROS. This review will focus on exploring the efficiency of supplementing several of these antioxidant molecules on oocyte maturation, sperm viability, fertilization, and embryo culture. [ABSTRACT FROM AUTHOR]