Search Results (1,913)

The maturation of oocytes is a critical step in mammalian reproduction, involving dynamic regulation of gene expression. Therefore, investigating how gene expression varies during different stages of oocyte maturation is highly important. This study employed single-cell RNA sequencing (scRNA-seq) to analyze bovine oocytes at the germinal vesicle (GV) and metaphase II (MII) stages. The results identified 1787 differentially expressed genes (DEGs) between the two stages, with 1556 genes upregulated and 231 downregulated in the GV stage. Further investigation using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses indicated that the upregulated genes are mainly involved in mitochondrial functions and energy metabolism, whereas the downregulated genes are primarily associated with signaling pathways. Validation through RT-qPCR confirmed that COA4, TKT and GPX4 were significantly higher in GV-stage oocytes, while ISG15, MAP1LC3C and ZEB2 were notably downregulated. This study highlights significant gene expression differences between GV and MII bovine oocytes, underscoring the vital roles of genes related to energy metabolism and signaling during oocyte maturation. The expression patterns of these genes provide important molecular markers for further elucidating the mechanisms underlying oocyte maturation. Full article

Background/Objectives: The emergence of methicillin-resistant Staphylococcus aureus (MRSA) necessitates innovative strategies to overcome conventional resistance mechanisms. This study investigated the potential of the natural flavonolignan silibinin (SIL) as an antivirulence agent against S. aureus, with a particular emphasis on its putative multi-target antibacterial activity and its capacity to potentiate the effects of ciprofloxacin (CIP). Methods: The antibacterial and antivirulence properties of SIL were assessed using both in vitro and in silico approaches. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined, and its synergistic interaction with CIP was evaluated using checkerboard assays. Inhibition of biofilm formation, as well as disruption of established biofilms, was assessed using an MTT-based viability assay. Staphyloxanthin (STX) inhibition was examined through pigment extraction and spectrophotometric quantification of pathway intermediates. Molecular docking studies were conducted to predict the binding affinities of the compounds to key bacterial targets, while safety was evaluated through haemolytic and cytotoxicity assays. Results: SIL exhibited weak to moderate direct antibacterial activity (MICs of 256–512 µg/mL), which is characteristic of many natural product scaffolds. Notably, SIL potentiated the activity of CIP, reducing its MIC by up to fourfold against selected resistant strains of S. aureus. SIL significantly inhibited biofilm formation and disrupted established mature biofilms in a strain-dependent manner. In vitro metabolic profiling and in silico analyses provided mechanistic insights into the effects of SIL on STX biosynthesis. Precursor accumulation data suggest inhibition at the diapophytoene desaturase (CrtN) catalytic step, representing a potential mechanism not previously reported for flavonolignans. Molecular docking studies further predicted favourable binding affinities for CrtM and other key targets. Importantly, SIL exhibited no haemolytic activity and low cytotoxicity in macrophages at synergistic concentrations. Conclusions: This study provides evidence that SIL functions as a dual-action agent, potentiating ciprofloxacin efficacy while reducing STX production and inhibiting biofilm formation, thereby impairing key virulence mechanisms of S. aureus. These findings, together with its favourable safety profile, provide a strong rationale for the development of silibinin-based topical adjuvants to combat drug-resistant Staphylococcus infections in humans. Full article

Background/Objectives: To address the limitations of natural curcumin, this study focuses on the functional evaluation of structurally optimized derivatives. We aimed to elucidate structure–activity relationships (SAR) and the stage-specific molecular mechanisms of adipogenesis inhibition using an in vitro cellular assay. Methods: Four novel curcuminoids were synthesized and evaluated in 3T3-L1 preadipocytes against natural curcumin (Curcuminoid I). Efficacy and mechanisms were assessed via cell viability assays, quantitative Oil Red O staining, and time-dependent transcriptional profiling (qPCR/Western blotting) of the KLF family and master regulators. Results: SAR analysis identified Curcuminoid III (symmetric 3,5-dimethoxy-4-hydroxy) as the most potent and safe candidate, whereas Curcuminoid IV exhibited cytotoxicity. Time-course analysis revealed a distinct step-wise inhibition mechanism wherein Curcuminoid III significantly upregulated the differentiation repressor KLF2 at the immediate-early phase. This rapid modulation effectively prevented the subsequent induction of pro-adipogenic factors, including KLF9, KLF15, PPARγ, and C/EBPα, in the mid-stage (3–5 d). Consequently, the expression of the maturation marker aP2 was robustly suppressed by the late stage (5–7 d). Conclusions: The symmetric 3,5-dimethoxy-4-hydroxy substitution pattern appears to confer strong anti-adipogenic activity to Curcuminoid III. Early modulation of the KLF2–PPARγ axis at the onset of differentiation may initiate a cascading inhibitory effect throughout the adipogenic program. These findings highlight the potential of structurally optimized plant-derived bioactive compounds as regulators of metabolic cell fate. Full article

Due to its high efficiency and safety, oocyte vitrification finds broad application in many fields of life sciences, such as clinical assisted reproduction and conservation of animal genetic resources. However, vitrification may cause cellular damage and reduce the quality of oocytes and their cumulus cells (CCs), which could be closely related to disorders in lipid metabolism. At present, the impact of vitrification upon the lipid profile of oocytes and CCs has not been systematically elucidated. In this study, we used porcine germinal vesicle cumulus–oocyte complexes (COCs) as a model to analyze their lipid characteristics after vitrification and in vitro maturation (IVM), utilizing untargeted lipid metabolomics. Our results showed that an overall count of 37 down-regulated and 8 up-regulated differential lipids was identified in the vitrified oocytes. Pathway analysis confirmed the enrichment in glycerophospholipid metabolism and fat digestion and absorption, etc. Combined with transcriptomic analysis, three enriched pathways were revealed, including the AMPK signaling pathway, metabolic pathways, and fatty acid elongation. On the other hand, a total of four down-regulated and eight up-regulated differential lipids were detected in the vitrified CCs. Pathway enrichment implicated autophagy, glycerophospholipid metabolism, etc. A joint analysis of metabolomic and transcriptomic data revealed four enrichment pathways, including cholesterol metabolism, fat digestion and absorption, regulation of lipolysis in adipocytes, and metabolic pathways. Notably, the supplementation of lysophosphatidylcholine during IVM attenuated oxidative stress, enhanced mitochondrial activity, and enhanced the viability and embryonic development of cryopreserved porcine oocytes. The results indicate that vitrification alters lipids in oocytes and CCs, and the supplementation of lipids plays a role in improving the quality of vitrified oocytes. Full article

Background: Dodder, the dried mature seed of Cuscuta chinensis Lam. (CCL), has demonstrated anti-tumor activity, but its molecular and immunological mechanisms in clear cell renal cell carcinoma (ccRCC) remain unclear. Objective: To identify potential targets and elucidate the immune mechanisms by which CCL exerts therapeutic effects against ccRCC. Methods: A network pharmacology approach was employed to predict CCL’s bioactive components and their putative targets in ccRCC. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were used to explore relevant pathways. Molecular docking validated the binding of key compounds to hub proteins. In vitro assays—including cell viability, colony formation, invasion, and apoptosis measurements—assessed the effects of quercetin, a principal CCL constituent, on 786-O renal carcinoma cells. Flow cytometry was performed to determine the percentage of CD163⁺ cells. An in vivo xenograft model evaluated CCL’s anti-tumor efficacy. Western blotting, flow cytometry, and multiplex immunohistochemistry (mIHC) examined the modulation of signaling pathways and immune cell markers. Results: Network pharmacology identified IL-6, EGFR, TLR4, MMP9, CD44, and IFN-γ as core targets of CCL in ccRCC. Enrichment analyses implicated immune regulation, inflammation modulation, and PI3K/AKT signaling inhibition. Molecular docking revealed strong quercetin–MMP9 binding affinity. Immuno-correlation analyses indicated that high MMP9 levels positively correlated with macrophage infiltration and M2 polarization, suggesting a role in tumor immune escape. Quercetin significantly reduced the viability of 786-O cells in a dose-dependent manner, showing approximately 45% inhibition at 80 μM (p < 0.01). In addition, quercetin decreased MMP9 expression and reduced the proportion of CD163-positive macrophages. These effects were reversed by FSL-1 TFA (Toll-like receptor 2/6 agonist), which is the agonist of MMP-9. In the xenograft model, tumor volume in the quercetin-treated group was reduced by approximately 50% compared with the control group. Conclusions: CCL, particularly its active component quercetin, may inhibit ccRCC progression via inhibiting MMP9-mediated M2 macrophage polarization. Full article

Polysialic acid (PSA), a highly negatively charged glycan attached mainly to the neural cell adhesion molecule (NCAM), is emerging as a critical but underrecognized extracellular regulator of glutamatergic neurotransmission. While previous literature has focused on PSA’s developmental roles, increasing evidence indicates that PSA–NCAM also contributes to synaptic plasticity mechanisms in the mature brain. This review integrates evidence from structural biophysics, single-channel electrophysiology, and disease models to explain how PSA modulates glutamate receptor gating to control learning and memory. We synthesize findings from biochemical reconstitution, electrophysiological recordings, and in vivo studies to show that PSA can modulate α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor open probability, burst duration, and cooperative gating without affecting conductance, thereby promoting long-term potentiation. Conversely, PSA selectively suppresses GluN2B-containing extrasynaptic N-methyl D-Aspartate (NMDA) receptor activity by lowering open probability and calcium influx, maintaining an optimal balance between potentiation and depression while providing neuroprotection. Disruption of PSA–NCAM signaling in developmental and disease models, including prenatal cannabinoid exposure and neurodegeneration, produces cognitive deficits reversible by PSA restoration. Notably, much of the current evidence derives from in vitro systems, with relatively few studies conducted in vivo, and studies employing PSA mimetics mostly, which should be considered when interpreting physiological relevance. Collectively, the available evidence suggests that PSA functions as an extracellular modulator linking synaptic glycans to glutamate receptor regulation and plasticity related signaling pathways, highlighting the potential importance of extracellular glycan mechanisms in the control of synaptic function. Full article

To overcome the seasonal constraints of explant availability and facilitate genetic improvement in Catalpa ovata, this study established a dual-pathway in vitro regeneration system (encompassing adventitious shoot organogenesis and somatic embryogenesis) using mature zygotic embryos. We systematically evaluated the synergistic effects of maternal genotypes, plant growth regulators (PGRs), basal media, and the histone deacetylase inhibitor Trichostatin A (TSA). Genotype screening revealed significant divergence in regenerative potential, with the half-sib family 32F17 exhibiting superior responsiveness (84.7% callus induction). A high cytokinin-to-auxin ratio (ZA3 medium) optimally drove direct shoot organogenesis. For adventitious shoot proliferation, the addition of TDZ significantly improved the multiplication coefficient (up to 2.99 on ZB4 medium), although a physiological trade-off with shoot elongation was observed. In parallel, the application of 10 µM TSA significantly enhanced somatic embryogenesis from embryogenic calli, effectively alleviating the inhibitory constraints of exogenous PGRs. For rhizogenesis, the DKW basal medium proved superior to half-strength MS, with the ZE3 treatment (0.1 mg·L⁻¹ NAA + 0.1 mg·L⁻¹ IBA) yielding the highest rooting frequency (69.6%) and robust root architecture. Notably, while somatic embryo conversion remained recalcitrant, plantlets derived exclusively from the adventitious shoot organogenesis pathway were successfully acclimatized ex vitro. These transplanted plantlets exhibited consistently high survival rates (83.1–84.4%) across all tested genotypes, effectively overcoming the initial genotype-dependent recalcitrance. Collectively, this optimized protocol provides a reliable technical platform for the large-scale clonal propagation and biotechnological breeding of C. ovata. Full article

Objectives: This study aimed to investigated the role of Giardia extracellular vesicles (EVs) in intercellular communication and to evaluated their potential as vaccine candidates. Methods: The immunomodulatory effects of Giardia EVs were assessed in mouse macrophages and human monocyte-derived dendritic cells (Mo-DCs), with a particular focus on key inflammatory signaling pathways. In vivo immunogenicity was evaluated following EV administration, and the antigenic composition of EV cargo was characterized by proteomic analysis. Results: Giardia EVs activated pro-inflammatory signaling pathways in mouse macrphages, including SAPK/JNK, ERK1/2, and NF-κB. This activation was associated with IκB-α degradation and nuclear translocation of p65. Furthermore, EV stimulation significantly upregulated the expression of pro-inflammatory genes, including Il1β, Il6, Il4, Ptgs2, Nos2, and Tnf, with log₂ fold changes ranging from 3.9 to 15.8. Consistently, EVs increased iNOS protein expression (28–45%) and nitrite production (9.6–12.3-fold). In human Mo-DCs, Giardia EVs promoted cellular maturation, as evidenced by increased expression of MHC-II, CD80, and CD86, and enhanced T-cell proliferation with a Th1-skewed profile. In vivo immunization induced antigen-specific antibody responses, with IgG subclass distribution indicative of a balanced Th1/Th2 response. Proteomic analysis identified immunoreactive EV-associated proteins, including elongation factor 1-alpha, α-7.3 giardin, tubulin, and variant surface proteins (VSPs), which are well-established antigens in Giardia infection, with prominent bands observed at approximately 22 kDa and 50 kDa. Conclusions: Collectively, these findings demonstrate that Giardia EVs modulate innate immune responses in vitro, elicit antigen-specific humoral immunity in vivo, and contain conserved immunogenic proteins. These properties support their potential as a promising cell-free vaccine platform against giardiasis. Full article

The SH-SY5Y cell line is widely used as an in vitro model for pharmacological and molecular investigations of Parkinson’s disease (PD). The use of SH-SY5Y cells in PD research critically relies on their ability to differentiate into a mature, post-mitotic, dopaminergic (DAergic) neuronal phenotype. However, SH-SY5Y cells are inherently heterogeneous since they are firstly catecholaminergic cells and may express diverse phenotypic markers besides the DAergic ones. These properties seem to be determined by the differentiation protocol that is employed, thus meaning it is crucial to obtain proper cell types. This systematic review aims to discuss the main differentiation protocols used in PD research over the last 30 years. They include inducers such as retinoic acid (RA), the phorbol ester TPA, and the BDNF. Among the 514 studies that were screened, 249 employed these inducers. Then, we quantitatively report the ability of these protocols to differentiate SH-SY5Y cells in mature DAergic neurons, evaluating morphology, differentiation markers, and DAergic markers among the studies that specifically compared differentiated to undifferentiated SH-SY5Y cells (61 studies over 249). As our research shows, despite the highest usage of the RA differentiation protocol, the combination of RA with the BDNF inducer seems to increase the expression and the acquisition of a DAergic phenotype. Nevertheless, during this analysis, some limitations emerged, highlighting the intrinsic phenotypic heterogeneity of these cells, thereby limiting their suitability according to the specific biological question under investigation. A deep investigation into the literature about the molecular phenotypic features of differentiated SH-SY5Y cells may eventually help us to understand the advantages and disadvantages of each protocol that was employed, and adequately set experiments around the PD research. Full article

This scoping review mapped the clinical readiness of directly additively manufactured (AM) dental ceramics for single-unit definitive restorations (crowns, veneers, and partial-coverage restorations) using a predefined review-specific five-tier readiness framework (R1–R5) designed to organize evidence maturity from restoration-relevant foundational studies to comparative clinical evidence. MEDLINE (PubMed), Scopus, Web of Science Core Collection, EBSCO (Dentistry and Oral Sciences Sources), and ClinicalTrials.gov were searched from inception to February 2026, with citation tracking. Thirty-five sources were included: 31 in vitro studies and 4 clinical studies. Evidence clustered in preclinical tiers, with most studies classified as restoration-level in vitro investigations (R2, 22/35) or foundational specimen-level studies explicitly linked to restorative performance (R1, 9/35); only one feasibility study reached R3 (1/35), three studies provided comparative clinical evidence (R4, 3/35), and no R5-level evidence was identified. The additively manufactured definitive restorations evaluated were zirconia-based. Most restoration-level studies addressed zirconia crowns (18/35), with fewer studies focusing on veneers/laminates (5/35) and occlusal veneers/tabletops (2/35). Across AM routes (most commonly vat photopolymerization ceramic workflows and nanoparticle jetting) outcomes focused on fit/adaptation, manufacturing accuracy, mechanical performance, and aging simulations; clinical studies reported short- to mid-term performance using standardized evaluation criteria. Overall, the evidence suggests technical feasibility and increasing restoration-level evaluation under controlled conditions, but clinical applicability remains preliminary because higher-readiness clinical evidence is still limited. Future work should prioritize standardized reporting, clinically relevant aging/fatigue paradigms, and longer-term comparative clinical studies. Full article

Serotonin is a critical morphogen in early development, yet the mechanisms regulating its homeostasis in the preimplantation embryo remain unclear, particularly under conditions of maternal antidepressant exposure. Here, we investigated embryonic serotonergic autonomy using mouse models of pharmacological transport blockade (maternal fluoxetine treatment) and in vitro treatment with the monoamine oxidase inhibitor pargyline. We employed immunofluorescence, RT-qPCR, and live-cell imaging to assess metabolic flux, gene expression, and physiological health. We demonstrate that monoamine oxidase functions as a metabolic firewall, progressively maturing from zygote to blastocyst to degrade excess amines. Paradoxically, maternal serotonin transporter blockade triggered significant intracellular serotonin hyper-accumulation in blastocysts, associated with a trend toward a compensatory upregulation of the biosynthetic gene Ddc. While this serotonin overload did not compromise morphology, mitochondrial function, or pluripotency marker expression, it induced a robust epigenetic response. Excess serotonin promoted elevated H3Q5ser immunoreactivity in both nuclear and cytoplasmic compartments via a transglutaminase-dependent mechanism. These findings reveal that the preimplantation embryo possesses a resilient, autonomous serotonergic system capable of compensatory synthesis. However, environmental fluctuations are chemically recorded via transglutaminase-mediated serotonylation, representing an epigenetic mark that warrants further long-term study within the Developmental Origins of Health and Disease (DOHaD) framework. Full article

This study aimed to evaluate the impact of coenzyme Q10 (CoQ10) supplementation during in vitro maturation (IVM) and/or vitrification of immature dromedary camel oocytes on their subsequent in vitro developmental competence. Additionally, total antioxidant capacity (TAC) and malondialdehyde (MDA) concentrations were assessed in the IVM spent medium. In experiment 1, cumulus–oocyte complexes (COCs, n = 808) collected from slaughtered dromedary camel ovaries were cultured in IVM media supplemented with either 0, 25, 50, or 100 μM CoQ10 for 36 h. Matured oocytes were then fertilized in vitro with epididymal camel spermatozoa. Eighteen hours post-insemination (pi), presumptive zygotes were cultured in vitro for 7 days. In experiment 2, a total of 875 COCs were randomly allocated to one of four experimental groups, namely (a) Vit−/IVM− (control) group, where COCs were vitrified in vitrification solution (VS; 25% DMSO + 25% EG) and matured in IVM media without CoQ10 supplementation; (b) Vit+/IVM− group, in which COCs were vitrified in a VS supplemented with 50 µM CoQ10 and matured in IVM media without CoQ10 supplementation; (c) Vit−/IVM+ group, where COCs were vitrified in VS without CoQ10 supplementation and matured in IVM media supplemented with 50 µM CoQ10; and (d) Vit+/IVM+ group, where COCs were vitrified in VS and matured in IVM media, both supplemented with 50 µM CoQ10. Following vitrification and warming, oocyte viability was evaluated morphologically and by trypan blue staining. Viable oocytes were then matured, fertilized, and cultured in vitro. In experiment 3, TAC and MDA concentrations in the IVM spent media were analyzed. Results showed that 50 µM CoQ10 supplementation to IVM media enhanced cumulus expansion, nuclear maturation, cleavage, and blastocyst rates (experiment 1). Adding 50 µM CoQ10 to the VS enhanced (p ≤ 0.05) oocyte viability compared to those vitrified in CoQ10-free media. Cumulus cell expansion and nuclear maturation rates were higher (p ≤ 0.05) in Vit−/IVM+ than in Vit+/IVM+ and Vit−/IVM− groups. Furthermore, cleavage and blastocyst rates were the highest (p ≤ 0.05) in Vit−/IVM+ group (experiment 2). The concentrations of TCA were higher, and the concentrations of MDA were lower (p ≤ 0.05) in Vit−/IVM+ than in other groups (experiment 3). In conclusion, supplementation of CoQ10 in the maturation medium of vitrified–warmed immature dromedary camel oocytes may enhance their in vitro developmental competence. Full article

Background/Objectives: Metabolic dysfunction-associated steatotic liver disease (MASLD) involves the interplay of hepatic lipid accumulation and immune-mediated inflammatory signaling, yet human-relevant in vitro systems that capture both processes simultaneously in a scalable format remain limited. The objective of this study was to develop and characterize a matrix-free 3D hepatocyte–macrophage co-culture model enabling simultaneous assessment of lipid accumulation and NF-κB-mediated inflammatory activation under glucolipotoxic stress. Methods: A 3D liver co-culture model was established by combining HepG2 hepatocyte-like cells with phorbol 12-myristate 13-acetate (PMA)-differentiated THP-1 macrophage-like cells stably expressing a NF-κB–Luc2 reporter. Spheroids were generated using a hanging-drop method in standard 96-well plates and matured for 8–10 days. Mature spheroids were subjected to acute 24 h glucolipotoxic challenge combining high glucose and palmitic acid and assessed for neutral lipid accumulation, NF-κB reporter activation (luciferase), and macrophage marker expression (qPCR). Results: Time-course characterization demonstrated progressive hepatocyte marker remodeling (albumin, alpha-fetoprotein, CYP3A4) and dynamic macrophage phenotype shifts (CD14, CD206, MARCO, TREM2). Acute glucolipotoxic challenge induced dose-dependent increases in neutral lipid accumulation and NF-κB reporter activation, accompanied by coordinated macrophage-associated transcriptional changes consistent with lipid-handling and tissue-remodeling programs. Post-challenge metabolic activity was retained under the selected stress conditions. As a proof-of-concept demonstration, three botanical extracts showed distinct attenuation profiles across the lipid and inflammatory endpoints. Conclusions: This 3D hepatocyte–macrophage co-culture model provides orthogonal readouts of steatosis and NF-κB-mediated inflammatory activation under glucolipotoxic stress, offering a reproducible, fit-for-purpose screening tool for investigating early glucolipotoxic hepatic responses and evaluating candidate compounds in a defined in vitro setting. Full article

Objectives: To quantitatively evaluate the inhibitory effects of commercially available probiotic formulations (Probien, Enterogermina, Reflor) applied as intracanal medicaments against mature dual-species biofilms of Enterococcus faecalis (E. faecalis) and Candida albicans (C. albicans) using a qPCR-based in vitro root canal model, with calcium hydroxide included as the reference intracanal medicament for comparison. Materials and Methods: Root canal specimens containing mature dual-species biofilms were medicated with probiotic–poloxamer gel formulations (Probien, Enterogermina, or Reflor) or calcium hydroxide (reference inhibitory control); infected but untreated canals served as the non-inhibitory control, and sterile non-inoculated specimens were included to confirm procedural sterility. After a 7-day intracanal application period, microbial loads were quantified at baseline and post-treatment by qPCR, and results were expressed as delta cycle threshold (ΔCt), colony-forming equivalents (CFE/mL), and percentage reduction values. Results: A total of 78 specimens (n = 13 per group) were analyzed. No significant intergroup differences were found in E. faecalis ΔCt or reduction percentages (p > 0.05), indicating its persistence despite intracanal medication. For C. albicans, differences among groups were significant (p < 0.001). Calcium hydroxide showed the strongest antifungal effect, producing marked ΔCt and CFE reductions versus probiotic and positive control groups, whereas probiotic formulations displayed only limited antifungal activity and no measurable inhibition against E. faecalis. Conclusions: Under the conditions of this in vitro model, the tested commercially available probiotic formulations—originally developed for gastrointestinal use—did not demonstrate significant antimicrobial effects against mature E. faecalis–C. albicans biofilms. These findings should be interpreted in the context of the absence of probiotic formulations specifically designed for intracanal use and the distinct ecological characteristics of the root canal system, which represents a closed, low-oxygen environment dominated by hard-tissue surfaces. Rather than excluding the potential of probiotics in endodontics, the present results highlight the need for root canal–adapted probiotic strains and delivery strategies tailored to intracanal conditions. Clinical Relevance: This in vitro study provides experimental insight into the limitations of directly applying commercially available gastrointestinal probiotic formulations within the root canal system. The findings highlight the importance of developing root canal–specific probiotic strains and delivery strategies tailored to the unique ecological conditions of the intracanal environment, thereby informing future translational and experimental research in biological endodontics. Full article

Ovarian reserve and reproductive life are closely linked concepts in female reproductive biology. The ovarian reserve consists of primordial follicles and refers to the number and quality of oocytes (eggs) remaining in the ovaries at any given time. Follicular dynamics shape a woman’s reproductive lifespan, ultimately leading to menopause. Elucidating the underlying genetic and molecular pathways of follicle maturation and depletion is thus crucial for understanding menopausal onset and progression, both in normal and pathophysiological contexts, such as primary ovarian insufficiency, defined as menopause before the age of 40. A key factor in ovarian differentiation and fertility maintenance is FOXL2, a forkhead family transcription factor that plays a crucial role in follicle formation and development, ovarian maintenance, and sex determination. By employing a ChIP-Seq approach in mice, we identified a previously unreported binding of FOXL2 to a Tsc1 regulatory region. Our data, along with a thorough literature review, support the hypothesis that FOXL2-mediated activation of Tsc1 in granulosa cells can help maintain primordial follicles in a dormant state by suppressing mTORC1 signalling. Understanding the mechanisms behind ovarian reserve may lay the foundation for developing novel fertility preservation strategies, improving fertility treatment protocols and promoting in vitro activation of cryopreserved ovarian tissue to support folliculogenesis. Full article