Microplastics (MPs) is an escalating aquatic environmental crisis that poses significant threats to marine organisms, especially mussels. Here, we compare the cumulative toxic effects of the two most abundant morphotypes of MPs in the environment, microspheres, and microfibers, on the gill and digestive gland (DG) of Mytilus galloprovincialis in a dose-dependent (1, 10, and 100 mg/L) and time-dependent (1, 4, 7, 14, 21 days exposure) manner. DNA fragmentation assessment through TUNEL assay revealed consistency in the pattern of morphological disturbance degree and cell apoptosis proportions indicated by histopathological analysis. Upon the acute phase of exposure (day 1–4), gill and DG treated with low MPs concentration exhibited preserved morphology and low proportion of TUNEL+ cells. At higher concentrations, spherical and fibrous MP-induced structural impairments and DNA breakage occurred at distinct levels. 100 mg/L microfibers was lethal to all mussels on day 21, indicating the higher toxicity of the fibrous particles. During the chronic phase, both morphological abnormalities degree and DNA fragmentation level increased over time and with increasing concentration, but the differentials between the spherical and fibrous group was gradually reduced, particularly diminished in 10 and 100 mg/L in the last 2 weeks. Furthermore, analysis of transcriptional activities of key genes for apoptosis of 100 mg/L–day 14 groups revealed the upregulation of both intrinsic and extrinsic apoptotic induction pathway and increment in gene transcripts involving genotoxic stress and energy metabolism according to MP morphotypes. Overall, microfibers exert higher genotoxic effects on mussel. In response, mussels trigger more intense apoptotic responses together with enhanced energy metabolism to tolerate the adverse effects in a way related to the accumulation of stimuli.

Glioblastoma multiforme (GBM) heterogeneity causes a greater number of deaths than any other brain tumor, despite the availability of alkylating chemotherapy. GBM stem-like cells (GSCs) contribute to GBM complexity and chemoresistance, but it remains challenging to identify and target GSCs or factors that control their activity. Here, we identified a specific GSC subset and show that activity of these cells is positively regulated by stabilization of methyl CpG binding domain 3 (MBD3) protein. MBD3 binds to CK1A and to BTRCP E3 ubiquitin ligase, triggering MBD3 degradation, suggesting that modulating this circuit could antagonize GBM recurrence. Accordingly, xenograft mice treated with the CK1A activator pyrvinium pamoate (Pyr-Pam) showed enhanced MBD3 degradation in cells expressing high levels of O⁶-methylguanine-DNA methyltransferase (MGMT) and in GSCs, overcoming temozolomide chemoresistance. Pyr-Pam blocked recruitment of MBD3 and the repressive nucleosome remodeling and deacetylase (NuRD) complex to neurogenesis-associated gene loci and increased acetyl–histone H3 activity and GSC differentiation. We conclude that CK1A/BTRCP/MBD3/NuRD signaling modulates GSC activation and malignancy, and that targeting this signaling could suppress GSC proliferation and GBM recurrence.

This study aims to investigate the effects of polystyrene microplastics (PS-MPs) on the entire female reproductive cycle and to elucidate the molecular mechanisms underlying their adverse impact on female ovaries. Additionally, it develops an in vitro follicle culture system as a novel methodological approach to evaluate reproductive toxicity, mimicking in vivo reproductive outcomes. First, PS-MPs were characterized using FTIR spectroscopy, TEM, and fluorescence microscopy. To assess reproductive toxicity, female mice were exposed to polystyrene microplastics (PS-MPs) at a dose of 30 mg/kg with an average particle size of 1 μm for 35 days. As a result, PS-MPs accumulated in the ovaries, leading to increased follicular atresia and apoptosis of granulosa cells. TEM revealed abnormal mitochondrial morphology in granulosa cells. Post-superovulation treatment, significant differences were noted in the number of ovulated metaphase II (MII) oocytes, spindle chromosome integrity, mitochondrial patterns, and ROS levels compared to controls. Mating with PS-MPs-exposed females led to fewer offspring. The in vitro follicle culture system proved promising for assessing PS-MPs reproductive toxicity. Immunohistochemistry showed increased Cleaved Caspase 3 and decreased Bcl2 levels in PS-MPs-treated groups, indicating apoptosis in granulosa cells. PS-MPs activate JNK and ERK pathways to mediate cell death, while impairing AKT signaling, reducing granulosa cell survival and ovarian function. This study highlights PS-MPs adverse reproductive effects and aids in developing strategies to protect female reproductive health.

Nuclear architecture underlies the transcriptional programs within the cell to establish cell identity. As previously demonstrated, long-range chromatin interactions of the Oct4 distal enhancer (DE) are correlated with active transcription in naïve state embryonic stem cells. Here, we identify and characterize extreme long-range interactions of the Oct4 DE through a novel CRISPR labeling technique we developed and chromosome conformation capture to identify lethal giant larvae 2 ( Llgl2 ) and growth factor receptor-bound protein 7 ( Grb7 ) as putative functional interacting target genes in different chromosomes. We show that the Oct4 DE directly regulates expression of Llgl2 and Grb7 in addition to Oct4 . Expression of Llgl2 and Grb7 closely correlates with the pluripotent state, where knock down of either result in loss of pluripotency, and overexpression enhances somatic cell reprogramming. We demonstrated that biologically important interactions of the Oct4 DE can occur at extreme distances that are necessary for the maintenance of the pluripotent state.

This study aimed to derive mature oocytes from murine preantral follicles cultured in a biomimetic ovary with a porcine scaffold using decellularization technology. We evaluated the DNA content and the presence of cell and extracellular matrix (ECM) components, including collagen, elastin, and glycosaminoglycans (GAGs), in decellularized (decell) porcine ovaries. The DNA content inthe decell ovarian tissues was approximately 94 % less than that in native tissues (66 ± 9.8 ng/mg vs. 1139 ± 269 ng/mg). Furthermore, the ECM component integrity was maintained in the decell ovarian tissue. The soluble collagen concentration of native ovarian tissue (native) was 195.34 ± 15.13 μg/mg (dry wt.), which was less than 878.6 ± 8.24 μg/mg for the decell ovarian tissue due to the loss of cellular mass. Hydrogels derived from decell porcine ovaries were prepared to develop an in vitro biomimetic ovary with appropriate ECM concentration (2−6 mg/mL). Scanning electron microscope (SEM) imagining revealed that the complex fiber network and porous structure were maintained in all groups treated with varying ECM concentration (2–6 mg/mL). Furthermore, rheometer analysis indicated that mechanical strength increased with ECM concentration in a dose-dependently. The preantral follicles cultured with 4 mg/mL ECM showed high rates of antral follicle (66 %) and mature oocyte (metaphase II) development (47 %). The preantral follicles cultured in a biomimetic ovary with a decell porcine scaffold showed a higher rate of antral follicle and mature oocytes than those cultured in other biomaterials such as collagen and Matrigel. In mature oocytes derived from antral follicles, meiotic spindles and nuclei were stained using a tubulin antibody and Hoechst, respectively. Two-cell embryos were developed from MII oocytes following parthenogenetic activation. Preantral follicles were cultured in a biomimetic ovary derived from the ECM of a decell porcine ovary, and embryos were generated from MII oocytes. This biomimetic ovary could contribute to restoring fertility in infertile women with reduced ovarian function, benefit mating efforts for endangered species, and maintain animals with valuable genetic traits.