Search Results (5,316)

Maternal metabolic dysfunction adversely influences embryonic muscle oxidative capacity and mitochondrial biogenesis, increasing the child’s long-term risks of developing obesity and metabolic syndrome in later life. This pilot study explored the mechanistic basis of embryonic muscle metabolic programming, employing non-invasive magnetic field exposures. Brief (10 min) exposure to low-energy (1.5 milliTesla at 50 Hertz) pulsing electromagnetic fields (PEMFs) has been shown in mammals to promote oxidative muscle development, associated with enhanced muscular mitochondriogenesis, augmented lipid metabolism, and attenuated inflammatory status. In this study, quail eggs were used as a model system to investigate the potential of analogous PEMF therapy to modulate embryonic muscle oxidative capacity independently of maternal influence. Quail eggs were administered five 10-min PEMF exposures to either upward-directed or downward-directed magnetic fields over 13 days. Embryos receiving magnetic treatment exhibited increased embryo weight, size, and survival compared to non-exposed controls. Upward exposure was associated with larger embryos, redder breast musculature, and upregulated levels of PPAR-α and PGC-1α, transcriptional regulators promoting oxidative muscle development, mitochondriogenesis, and angiogenesis, whereas downward exposure augmented collagen levels and reduced angiogenesis. Exposure to upward PEMFs may hence serve as a method to promote embryonic growth and oxidative muscle development and improve embryonic mortality. Full article

Fat deposition plays a key role in determining porcine meat quality traits, with lipid droplets serving as critical organelles for lipid storage in adipose tissue. Inhibiting lipid droplet biogenesis disrupts the lipid storage capacity of adipocytes. The Stearoyl-CoA Desaturase (SCD) family is crucial in regulating polyunsaturated fatty acid/monounsaturated fatty acid (PUFA/MUFA) composition, while its role in lipid droplet formation remains unclear. This study employed CRISPR/Cas9 to create SCD1-deficient porcine renal epithelial cells (PK15), enabling an investigation into SCD1’s role in fatty acid composition and lipid droplet regulation. RNA-seq analysis was conducted to elucidate the mechanisms underlying SCD1’s impact on lipid droplet numbers. Results showed that SCD1 deletion significantly decreased triacylglycerols (TAG) content, altered fatty acid composition, and decreased lipid droplet numbers. Conversely, SCD1 overexpression increased lipid droplet numbers, confirming SCD1’s role in regulating lipid droplet abundance. RNA-seq analysis revealed that SCD1 regulates lipid metabolism via Calsyntenin 3β (CLSTN3B). Experimental validation confirmed the SCD1-CLSTN3B regulation of lipid droplet numbers. In summary, we discovered the role of SCD1 in regulating the number of lipid droplets, highlighting its potential impact on lipid metabolism and adipocyte function in pigs. Full article

This study investigated the effects of dietary supplementation with the probiotic Pediococcus acidilactici on the reproductive performance and offspring viability of male Atlantic salmon (Salmo salar). A total of 48 mature males were divided into three groups—Group A (control, no probiotic), Group B (60-day supplementation), and Group C (120-day supplementation). The fish were kept in 10 m³ tanks at a constant temperature of 9 °C under a natural photoperiod and evaluated for multiple reproductive and physiological parameters. The results demonstrate that Group C exhibited significantly higher gonad weight, gonadosomatic index (GSI), and sperm concentration than Groups A and B. The blood biochemistry revealed reduced bilirubin and alanine aminotransferase (ALT) levels in Group C; however, values were within normal reference ranges for the species in all groups. While sperm quality parameters such as membrane integrity and motility showed no significant differences, fertility and embryo viability were significantly higher in Group C across all three developmental stages (180, 300, and 380 accumulated thermal units, ATUs). Additionally, Group C showed lower rates of embryonic malformation and mortality at later stages of development. Statistical analyses confirmed a significant interaction between treatment duration and embryo viability, with the 120-day probiotic treatment (Group C) producing the best outcomes. These findings support the hypothesis that P. acidilactici supplementation positively influences male reproductive parameters and improves embryo quality and survival in Atlantic salmon. Full article

MicroRNAs (miRs) are epigenetic regulators of several metabolic diseases, including gestational diabetes mellitus (GDM). Objectives: Following a systematic review, we propose a pattern of key circulating miRs associated with placental changes and their potential role in the fetus. Methods: A systematic investigation of studies published between January 2011 and July 2024 was conducted in the PubMed, ScienceDirect, Trip Database, and Wiley databases. A total of 90 articles were analyzed. Results: Two hundred twenty-six circulating microRNAs were identified in women with GDM, and fifty miRs were validated by PCR, with miRs-16-5p, -29a-5p, and -195-5p being the most frequently reported. Interestingly, miR-16-5p was also expressed in the placenta but not in umbilical cord blood or amniotic fluid. Conversely, miR-126-3p was expressed in circulation, the placenta, umbilical cord blood, and amniotic fluid. Several reports describe high expression levels of miR-518d in maternal circulation, umbilical cord blood, and placenta. Controversial results regarding the expression of miR-29a-3p, -137, and -148a-3p were identified when comparing umbilical cord blood and the placenta. Conclusions: In silico analyses suggest that the miR-29 family, as well as miRs-16-5p, -126-3p, -195-5p, and -518b, may be involved in alterations in the heart, brain, and kidneys in the embryo when exposed to a hyperglycemic environment. Full article

Nile tilapia (Oreochromis niloticus) is the most widely farmed tilapia species globally, making it one of the most important aquaculture species. To meet increasing demand, hatcheries occasionally use artificial breeding techniques such as hormonal induction to synchronize breeding. Despite the common use of human chorionic gonadotropin (hCG) in fish breeding, no detailed protocol has been established specifically for Nile tilapia. The objective of this study is to establish an effective hCG-induced artificial breeding protocol for gene editing and aquaculture production, optimizing fertilization, hatching, and survival rates. We employed a single intramuscular injection of 2 IU/g hCG to induce ovulation. The protocol achieved an average fertilization rate of 88.3% and a larval survival rate of 90.5%, demonstrating its potential for obtaining high-quality embryos for functional studies and enhancing reproductive performance on a commercial scale. Full article

Copper-based nanoparticles (as Cu₂O) are a key component in marine antifouling paints and, as coatings degrade, release nanoparticles that can affect a wide range of non-target organisms. This study investigates the impact of Cu₂O nanoparticles on the early development of urchins Arbacia lixula, Paracentrotus lividus and Sphaerechinus granularis, and benchmarks their toxicity against similarly sized Cu and CuO nanoparticles and ionic copper. Concentration-dependent toxicity was noted for all forms of copper at concentrations in the 1 to 5000 µg L⁻¹ range. EC₅₀ values after Cu₂O exposure indicated that A. lixula (99 µg L⁻¹) was generally more sensitive than the other two species, with EC₅₀ values of 371 µg L⁻¹ and 606 µg L⁻¹ noted for S. granularis and P. lividus, respectively. The same trend across species was noted for both Cu and CuO, although these nanoparticles generally showed higher EC₅₀ values, indicating lower toxicity compared to Cu₂O. LC₅₀ values qualitatively parallel the corresponding EC₅₀ values, with Cu₂O consistently the most toxic, while Cu was less harmful, and CuO did not reach LC₅₀ at any concentration. Again, greatest lethality was noted in A. lixula. While copper ion release from Cu was much greater than from CuO and Cu₂O, the latter showed similar or greater toxicity to developing embryos compared to Cu. This indicates that copper ions are not the sole driver of toxicity of Cu₂O, but there may also be a contribution derived from Cu₂O redox activity within cells or at membranes that negatively impact oxidative stress defence mechanisms and metabolic pathways. Full article

Milk fat is an important nutritional component and flavor substance in dairy products. Its content and composition directly affect the nutritional value, processing characteristics, and economic benefits of dairy products. Therefore, an in-depth exploration of the molecular mechanisms that influence milk protein synthesis holds profound significance for dairy farming and dairy production. Molecular biology techniques were used to construct CDKN1A overexpression and interference vectors. Using BMECs (bovine mammary epithelial cells) as the experimental model, the vectors were transfected into the cells via liposome mediation to investigate the effect of CDKN1A on the expression of genes related to milk protein synthesis. The results showed that the CDKN1A overexpression and interference vectors were successfully constructed, and the overexpression of CDKN1A reduced milk protein synthesis, and the interference of CDKN1A enhanced milk protein synthesis. This finding provides an important theoretical basis for dairy farming and dairy production. By regulating the expression level of CDKN1A, it is possible to achieve precise control of milk protein yield in dairy cows. It also offers a potential target for the development of new feed additives or drugs. These additives or drugs can promote milk protein synthesis by regulating the activity of CDKN1A, providing new strategies and methods for the sustainable development of the dairy industry. Full article

Preimplantation genetic testing (PGT) has been used in various forms over the last two decades. PGT involves testing early embryos following in vitro fertilization and has now become an accepted part of genetic testing. Nowadays, PGT serves as a resource for couples who have a family history of monogenic disorders, wherein the fetus is at high risk of inheriting the condition. PGT is also used to improve pregnancy outcomes in IVF patients in cases of recurrent IVF implantation failure, recurrent miscarriages, as well as male factor. It is also used in screening for sex-linked disorders and sourcing stem cells for therapy. The latest PGT direction is polygenic embryo screening (PES, PGT-P), which allows the identification of embryos that are at elevated risk for significant diseases in adulthood, such as coronary artery disease (CAD), diabetes, hypertension, and breast cancer. As the prevalence and the potential for the use of PES grow, fundamental ethical issues have been underlined, raising concerns about the broader implications of genetic testing. This narrative review summarizes indications, methods, applications, and limitations for PGT, with a particular focus on PES. Full article

Male fertility is strongly influenced by environmental exposures, lifestyle, and advancing age. While advanced paternal age (APA) has been linked with a progressive decline in male fertility, poor reproductive outcomes, and decreased offspring health, the molecular mechanisms underlying these alterations remain unclear. In this work, we investigated the impact of men’s age on human sperm protein expression and phosphorylation to identify molecular alterations possibly responsible for the age-associated decline in male fertility. Semen samples from volunteers attending fertility consultations at the Hospital of Aveiro were collected, analyzed according to WHO’s guidelines, and processed by the density gradient technique. The proteome and phosphoproteome of 19 normozoospermic human sperm samples divided into four age groups were evaluated by mass spectrometry: ≤30 years old; 31–35 years old; 36–40 years old; and >40 years old. Proteomic analysis revealed 46 differentially expressed proteins (DEPs) between groups, some of them associated with infertility-related phenotypes. Gene ontology (GO) analysis, performed using the DAVID database, revealed that DEPs in older men were enriched in pathways related to stress response, metabolism, and embryo implantation. Additionally, 94 differentially phosphorylated sites corresponding to 76 differentially expressed phosphorylated proteins between the groups were identified, related to key reproductive processes such as sperm motility, spermatogenesis, and sperm binding to zona pellucida, and involved in metabolic and stress response pathways, like HSF1 activation. The set of proteins and phosphorylated residues altered in the sperm fraction usually used in assisted reproductive technology (ART) highlights the need to consider the age of the male partner during fertility assessment and treatment planning. These markers can also be used to explain cases of idiopathic infertility, failure in ART, or repeated abortion associated with APA, overcoming the subjectivity of the conventional semen analysis. Full article

Preimplantation genetic testing for aneuploidy (PGT-A) is widely used to select euploid embryos for in vitro fertilization (IVF), but its accuracy in predicting the implantation potential for full segmental aneuploid (Seg-A) embryos remains unclear. In this study, we investigated chromosomal concordance between clinically biopsied trophectoderm (TE) and inner cell mass (ICM) in 175 donated blastocysts, which comprised those clinically diagnosed as euploid (13), Seg-A (36), segmental mosaicism (Seg-M) (60), whole-chromosome aneuploid (Who-A) (52), and whole-chromosome mosaicism (14). Using next-generation sequencing (NGS), we found that TE–ICM concordance rates were higher for euploid (85%) and Who-A (94%) embryos but significantly lower for Seg-A (25%) and Seg-M embryos (33%). For Seg-A, the euploidy rate in the ICM was 19% and the euploidy rate in the ICM was 63% for Seg-M. These low concordance rates may be due to technical and biological artifacts of PGT-A for Seg-A. Despite the significant discordance between TE and ICM, a subset of Seg-A embryos demonstrated euploidy. While clinically diagnosed euploid embryos remain the preferred choice, Seg-A embryos should be considered as having implantation potential. In particular, Seg-A results should be clearly distinguished from Who-A results and not routinely categorically discarded. Further research is required to refine the selection criteria, aided by parental karyotyping or re-biopsy, and to develop more reliable embryo assessment methods to ensure the accurate evaluation of reproductive potential and support shared decision making between doctors and patients. Full article

Background: Given the common occurrence of mosaicism and aneuploidy in IVF embryos, our study aimed to retrospectively identify whether specific chromosomal regions or individual chromosomes are predominantly affected in our clinic. Understanding these patterns can improve embryo selection, reduce miscarriage risks, and enhance genetic counseling. At the same time, due to the limited data on potential comorbidities in affected children, our findings aim to support both clinicians and patients in making informed decisions. Methods: The retrospective clinical study included 461 PGT-A biopsies from our clinic database (September 2023–December 2024) to determine whether specific chromosome regions or individual chromosomes (C) are more likely to be mosaic or aneuploid. Results: Among the 461 embryos analyzed in our clinic, the incidence rate of mosaicism was 16.70% whereas the aneuploidy rate was 32.10%. Our results showed that mosaicism tends to target a specific chromosomal region in embryos, namely the chromosome 1 to 9 region, in particular chromosomes 7, 1, 9. On the other hand, aneuploidy targets the chromosomal region chromosome 16 to 22, particularly chromosomes 16, 19, and 22. Conclusions: Our data suggest that mosaicism and aneuploidy affect the genome in an uneven manner and are often concentrated in specific chromosomal regions, with mosaicism primarily affecting the C1–C9 region and aneuploidy targeting the C16–C22 region. These data highlight the need for further research to understand these patterns and the impact of IVF methods on chromosomal targeting. Comparative studies could also be helpful in genetic counseling by clarifying the implications of the levels of mosaicism in the newborn. Full article

A previous study demonstrated that a 19-nucleotide edit, encoding a six amino acid substitution in the bovine CD46 gene, dramatically reduced bovine viral diarrhea virus (BVDV) susceptibility in a cloned Gir (Bos indicus) heifer. The present study aimed to replicate this result in American Angus (Bos taurus) using genetically matched controls and larger sample sizes. CRISPR/Cas9-mediated homology-directed repair introduced the identical CD46 edit, encoding the A₈₂LPTFS amino acid sequence, into exon 2 of CD46 in primary Angus fibroblasts. Thirty-three cloned embryos (22 CD46-edited and 11 unedited) were transferred to recipient cows. However, all pregnancies resulted in pre- and perinatal losses due to cloning-related abnormalities, preventing in vivo BVDV challenge. Consequently, ex vivo BVDV susceptibility assays were performed on primary fibroblast cell lines rescued from deceased cloned Angus calves. Infection studies revealed significantly reduced susceptibility in the edited lines, comparable to the resistance previously observed from the edited Gir heifer. These studies extend the applicability of this finding from Gir to the most common US beef breed, Angus, suggesting the potential for broad application of CD46 editing in BVDV control. Continued advancements in cloning technology will enhance the potential of gene-editing for producing disease-resistant livestock. Full article

Bisphenol A (BPA) is a widespread environmental endocrine disruptor with significant neurodevelopmental and behavioral risks. The present study explored the role of the circadian clock protein NR1D1 in mediating BPA-induced anxiety-like behavior and brain inflammation early in life. Zebrafish embryos exposed to BPA exhibited anxiety-like behavior characterized by altered motor activity patterns. Notably, BPA exposure suppressed the expression of the circadian clock gene nr1d1, accompanied by increased transcriptional and protein levels of pro-inflammatory cytokines, including IL-6, IL-1β, and TNF-α. These changes created a pro-inflammatory microenvironment that disrupted dopamine system homeostasis, contributing to the observed behavioral abnormalities. Activation of NR1D1 using GSK effectively reversed BPA-induced inflammatory responses and restored normal dopamine levels and behavioral phenotypes. These findings highlight NR1D1 as a critical regulator linking circadian rhythm disruption, neuroinflammation, and dopaminergic dysfunction to anxiety-like behavior. This study provides novel insights into the mechanisms underlying BPA-induced neurotoxicity and identifies NR1D1 as a potential therapeutic target for mitigating the adverse effects of early-life BPA exposure. Full article

Food colourants are widely used additives classified as either synthetic or natural. In recent years, consumers have increasingly favoured natural options, considering them safer and potentially beneficial due to their nutritional properties. This study examined the effects of a natural food colourant, commercially known as Vegan Red (RVEG), on zebrafish embryonic development. Its impact was compared with cochineal red E120, of animal origin, and the synthetic dye E124, which are associated with hyperactivity in children and allergies. Shield stage embryos were exposed for 72 h and then examined using a multidisciplinary approach to assess the effects on conventional toxicity endpoints, such as survival, hatching rate, heart rate, genotoxicity, and behavioural interferences, including the impact on muscle ultrastructure. The results demonstrated that RVEG, as well as E120, do not affect hatching, heart rate, and motility parameters. However, RVEG moderately alters skeletal muscle organisation and, more relevant, the expression of the gfap, chchd2, and notch1a genes. Based on standard toxicity parameters, the findings indicated that RVEG is less toxic than E124 and E120, but that the alterations induced in gene expression and muscle anatomy raise safety concerns. Full article

As the primary active component of Astragalus membranaceus, Astragaloside IV (AS-IV) is widely recognized in pharmacological research for its multifaceted therapeutic potential, particularly its antioxidative, immunostimulatory, and cardioprotective properties. Oxidative stress is an important mechanism in the induction of many diseases. The present study investigates the antioxidative mechanism of Astragaloside IV in zebrafish, using menaquinone exposure to induce oxidative stress conditions. The findings revealed that AS-IV effectively attenuated oxidative stress-induced mortality and morphological abnormalities in zebrafish. AS-IV exhibited a concentration-dependent protective effect against developmental abnormalities, with progressive reduction in pericardial effusion, body curvature, and growth retardation observed at higher doses. Moreover, AS-IV treatment not only effectively reduced reactive oxygen species (ROS) accumulation and attenuated oxidative DNA damage but also significantly decreased apoptosis in the cardiac region of zebrafish embryos under oxidative stress conditions. Western blot analysis revealed that AS-IV treatment significantly reduced the protein levels of both Cleaved Caspase-3 and γ-H2AX, indicating its ability to inhibit DNA damage-induced apoptosis. AS-IV mediates its antioxidant defense mechanisms through the activation of the nuclear factor erythroid 2-related factor 2 (NRF2) signaling pathway, inducing the significant upregulation of cytoprotective enzymes. This molecular mechanism underlies the observed phenotypic improvements in oxidative stress-related damage. Upstream analysis demonstrated that AS-IV activates NRF2 primarily through protein kinase B (AKT/PKB) pathway modulation, independent of KEAP1 regulation. Comprehensive mechanistic analysis reveals that Astragaloside IV mitigates oxidative stress-induced apoptosis in zebrafish through coordinated regulation of the AKT/NRF2/HO-1/Caspase-3 signaling axis. Full article