Search Results (735)

Objectives: Alzheimer’s disease (AD) is associated with impaired adult hippocampal neurogenesis (AHN). This study aimed to establish an in vitro model of Aβ_1–42 oligomer-damaged neural stem cells (NSCs) and to employ the 5×FAD mouse model of AD in vivo, and to evaluate the therapeutic effects of brain-derived neurotrophic factor-loaded hyaluronic acid hydrogel (BDNF-HA gel) on AHN. Methods: In vitro, BDNF-HA gel was co-cultured with Aβ_1–42 oligomer-impaired NSC spheres and evaluate NSC proliferation, migration, and differentiation. In vivo, BDNF-HA gel was infused intracerebroventricularly into 5×FAD mice. Using BrdU labeling, immunofluorescence, anterograde transsynaptic viral tracing, and behavioral tests, we assessed the effects of BDNF-HA gel on adult neurogenesis, newborn neuron integration into memory circuits, and cognitive function. Results: In vitro, BDNF-HA gel attenuated Aβ_1–42-induced NSC apoptosis, restored proliferation and migration, promoted differentiation into neuroblasts, newborn neurons, and oligodendrocytes, and alleviated mitochondrial depolarization and loss of mitochondrial mass. In vivo, despite the absence of significant Aβ plaques reduction in 5×FAD mice, BDNF-HA gel markedly enhanced NSC proliferation and neurogenesis in the subventricular zone (SVZ) and subgranular zone (SGZ). Behavioral tests further revealed significant improvements in object recognition, spatial working memory, and spatial reference memory. Conclusions: BDNF-HA gel can effectively counteract the toxic microenvironment induced by Aβ oligomers, promoting NSC proliferation, migration, and differentiation into neurons. Without altering the Aβ burden, it significantly enhances adult neurogenesis and rescues cognitive deficits in AD mice. Full article

Scientific knowledge-based fishery management is essential to ensure the sustainability of marine resources, particularly in regions where fisheries are data-limited. This study assessed the stock status of six shallow-water demersal fish species (Bagre marinus, Cathorops mapale, Diapterus rhombeus, Eucinostomus argenteus, Haemulopsis corvinaeformis, and Lutjanus synagris) in the Colombian Caribbean Sea using three complementary length-based models: length-based indicators (LBIs), length-based spawning potential ratio (LBSPR), and the Length-Based Bayesian Biomass estimator (LBB). The integrated results demonstrated that five species (C. mapale, D. rhombeus, E. argenteus, H. corvinaeformis, and L. synagris) are currently overexploited (F/M > 1 and B/B_MSY < 1), while B. marinus is experiencing overfishing (F/M > 1 and B/B_MSY > 1), with a high risk of surpassing its maximum sustainable yield. These outcomes confirm that demersal fish populations in the Colombian Caribbean are being exploited beyond sustainable biological limits. With the aim of promoting stock recovery and long-term sustainability, this study recommends the implementation of recently evaluated management measures focused on (i) the implementation and enforcement of Bycatch Reduction Devices (BRDs); (ii) the regulation and monitoring of trawl net mesh sizes to improve selectivity patterns; (iii) the establishment of spatial and temporal closures in critical spawning areas for demersal fish species; and (iv) the strengthening of fishery monitoring and data collection systems. The findings provide critical baseline information and a methodological framework to support evidence-based fishery management and conservation strategies in tropical multispecies fisheries under data-limited conditions. Full article

This scoping review examines bovine respiratory disease (BRD) risk factors across beef production systems to clarify their contribution to disease occurrence and outcomes and to identify gaps that limit effective prevention. Following the PRISMA-ScR guidelines, eight databases (EBSCOhost, Google Scholar, MDPI, PubMed, ScienceDirect, Journal of Animal Science, Journal of Dairy Science, and Web of Science) were searched for peer-reviewed studies published between 2004 and 2024, yielding 91 eligible studies from 133 screened articles. Data were charted to assess how BRD risk factors have been investigated across cow–calf, stocker/backgrounding, and feedlot systems and categorized into biological, operational, and environmental domains. Twenty-three major risk factors were identified, with transportation stress and commingling most frequently studied in stocker and feedlot systems, while management-related factors such as vaccination were more commonly emphasized in cow–calf operations. The evidence base was heavily skewed toward feedlot populations (62%), with limited focus on cow–calf (13%) and stocker (7%) systems and few longitudinal studies spanning multiple production stages. Overall, the findings reveal substantial heterogeneity in research emphasis and highlight critical gaps in early-life and cross-stage risk assessment. These findings underscore an urgent need for longitudinal, cross-stage cohort studies and standardized BRD surveillance frameworks to address the early-life evidence gap and provide a foundation for more effective, data-driven, integrated disease prevention strategies. Full article

Retinitis pigmentosa (RP) is a genetically heterogeneous group of inherited retinal degenerations with primary degeneration of rod photoreceptors followed by secondary cone loss. We investigated whether downregulating Nrl (neural retina leucine zipper), a key transcription factor specifying rod fate, can reprogram rods into a more resilient state. In a transgenic Nrl^N/N mouse in which Nrl was markedly downregulated, the rod phenotype became more like a rod precursor, particularly in the inferior retina. Crossing Nrl^N/N mice with two rod degeneration models, rd1 (Pde6b^rd1/rd1) and rhodopsin P23H knock-in (Rho^P23H/P23H) mice, showed significantly improved photoreceptor survival in double-mutant mice. In addition, AAV-mediated delivery of shRNA targeting Nrl mRNA substantially enhanced photoreceptor survival in rd10 (Pde6b^rd10/rd10) mice. These findings demonstrate that downregulation of Nrl reprograms rods and confers broad resistance to degeneration across multiple RP models. AAV-mediated Nrl knockdown represents a promising mutation-independent therapeutic strategy for autosomal recessive and dominant forms of RP. Full article

Background and Objectives: Potential anti-neoplastic effects of resveratrol, which has antioxidant features combined with clomipramine, which has antineoplastic features, or with gemcitabine, used as a nucleoside analog widely used in chemotherapy, were evaluated together and individually on the HL-60 leukemia cells in this in vitro screening study. Materials and Methods: HL-60 cells were treated with gemcitabine, clomipramine, resveratrol, or their combinations at concentrations ranging from 1 to 200 µM. Cell viability was assessed at 24, 48, and 72 h using the trypan blue exclusion method, and results are expressed as a percentage of time-matched untreated controls. Cell proliferation was further evaluated by bromodeoxyuridine (BrdU) immunohistochemical labeling. All experiments were performed in triplicate, and statistical analyses were conducted using one-way analysis of variance (ANOVA) with post hoc comparisons. Results: Gemcitabine markedly reduced HL-60 cell viability at all concentrations and time points (p < 0.001), indicating strong time-dependent cytotoxicity, with a significant drop in BrdU proliferation index at 48 h (p < 0.001). Clomipramine exhibited a biphasic response: high concentrations decreased viability (p < 0.05), while low concentrations allowed partial recovery by 72 h. Resveratrol showed concentration-dependent cytotoxicity, with reduced viability at high concentration and near-control levels at low concentration by 72 h; BrdU indices remained significantly lower than control (p < 0.001). Combination treatments with gemcitabine showed no additive cytotoxic or antiproliferative effects (p > 0.05). A transient enhanced effect was observed in the clomipramine + resveratrol group at 24 h (p < 0.01 vs. clomipramine; p < 0.05 vs. gemcitabine). Conclusions: Gemcitabine, clomipramine, and resveratrol all exhibited inhibitory effects on cell proliferation in HL-60 cell cultures. However, the combination treatments did not show additional cytotoxicity or additive effects. These findings suggest that while each of these compounds individually has the potential to inhibit cell growth, their combined application does not enhance the cytotoxic effects beyond those observed with single treatments. These findings highlight the necessity of a rational approach when considering novel drug combinations. Full article

Background/Objectives: Immune evasion remains a critical barrier to effective hepatocellular carcinoma (HCC) therapy. Lactate dehydrogenase A (LDHA) drives lactate accumulation and histone lysine lactylation (Kla), reshaping the immunosuppressive microenvironment, while bromodomain-containing protein 4 (BRD4) sustains B7-H3 transcription via super-enhancer occupancy. Despite their synergistic roles in the lactate–Kla–B7-H3 immunosuppressive axis, no dual-target inhibitor simultaneously engaging both proteins has been reported. This study aimed to discover dual LDHA/BRD4 inhibitors from natural product libraries using an integrated AI-driven computational pipeline. Methods: We established a multi-tier virtual screening cascade comprising Lipinski/QED drug-likeness filtration, DiffDock-based AI docking, QuickVina binding energy validation, PLIP interaction profiling, 200 ns all-atom molecular dynamics simulations, MM-GBSA binding free energy calculations, and density functional theory analysis. Natural product libraries from COCONUT and CMNPD databases (84,730 compounds post-filtration) were screened against both targets. Results: High-throughput DiffDock screening identified 11 dual-target hits, from which CNP0038114.1 and CMNPD16582 emerged as prioritized lead candidates. All four protein–ligand complexes maintained structural stability throughout MD simulations, with MM-GBSA binding free energies ranging from −27.24 to −32.45 kcal/mol, predominantly driven by van der Waals interactions. DFT calculations revealed distinct electronic profiles: CNP0038114.1 exhibited a narrow HOMO–LUMO gap (2.718 eV) favoring charge-transfer reactivity, whereas CMNPD16582 displayed a larger gap (4.822 eV), suggesting superior chemical stability. Conclusions: This computational study furnishes two novel natural product leads for targeting the lactate–Kla–B7-H3 immunosuppressive axis in HCC, establishing a generalizable AI-driven workflow for dual-target inhibitor discovery. Full article

Background and Objectives: The objective of this study is to evaluate whether deferoxamine modulates cell biological properties, such as proliferation and wound closure of porcine corneal endothelial cells (CECs) in vitro, and whether the treatment of CECs with deferoxamine results in an enhanced expression of vascular endothelial growth factor (VEGF). Materials and Methods: Corneal endothelial cells were extracted from porcine globes within 24 h postmortem. Immunohistochemistry for the endothelial Na⁺/K⁺-ATPase was performed to confirm the cells’ endothelial origin. To assess CEC viability and proliferation, a water-soluble tetrazolium salt (WST-1) and 5-bromo-2′-deoxyuridine (BrdU) assay were performed. Corneal endothelial wound closure was evaluated using a wound closure assay. VEGF mRNA expression was evaluated using real-time polymerase chain reaction (rt-PCR). Results: The extracted corneal endothelial cells showed a typical hexagonal morphology with Na⁺/K⁺-ATPase staining of the cell membrane. The treatment with 200 µM deferoxamine significantly increased CEC viability to 121 ± 24% compared to the control group (p = 0.0024). Corneal endothelial cell proliferation did not show any significant changes under the treatment with deferoxamine (p > 0.05). Both 100 µM and 200 µM deferoxamine led to a significantly smaller remaining wound area of 82.4 ± 6.7% and 78.7 ± 6.2% (p < 0.0001) in comparison to the control group after 24 h of treatment in the wound closure assay. Treatment with 200 µM deferoxamine significantly induced VEGF mRNA expression to 1.67- ± 0.57-fold from 1.00- ± 0.03-fold in the control group (p = 0.0006). Conclusions: Deferoxamine effectively enhances corneal endothelial cell viability and wound healing associated with an overexpression of VEGF. Thus, deferoxamine is a potent modulator of cell biological properties of corneal endothelial cells and maintains their integrity in vitro. Full article

Resistance to apoptosis represents a major therapeutic challenge in advanced and recurrent endometrial cancer. This study investigated whether vinorelbine, alone or in combination with lithium chloride (LiCl) or medroxyprogesterone acetate (MPA), suppresses tumor growth through non-apoptotic mechanisms in a three-dimensional spheroid model of Ishikawa endometrial cancer cells. Proliferation was assessed by BrdU incorporation, cell cycle distribution by flow cytometry, and apoptosis by Annexin V/propidium iodide staining. Vinorelbine-based treatments significantly reduced DNA synthesis and were associated with decreased S-phase fractions and altered cell cycle distribution. These effects were sustained over time and were more pronounced in combination treatments. Importantly, no consistent increase in apoptotic cell death was observed, and most cells remained viable despite strong suppression of proliferation. These findings indicate that vinorelbine-based regimens inhibit tumor spheroid growth predominantly through cytostatic mechanisms rather than induction of apoptosis. This non-apoptotic growth-control strategy may be relevant for the management of apoptosis-resistant endometrial cancer. Full article

Background/Objectives: Adult neural stem cells retain the capacity to generate immature neuronal lineages; however, pharmacological approaches that robustly enhance neurogenic activity remain limited. To identify compounds with intrinsic activity under physiologically restrictive conditions, we aimed to screen for small molecules that promote neural stem cell proliferation in the absence of exogenous growth factors and are compatible with central nervous system drug discovery. Methods: We developed a human neural stem cell–based phenotypic screening cascade performed under growth factor–free conditions. Compound activity was evaluated in vitro by ATP-based proliferation assays, BrdU incorporation, and assessment of neurogenic marker analysis. In vivo neurogenic effects were assessed in adult rats by BrdU labeling and immunohistochemical analysis of BrdU/Nestin- and BrdU/DCX-positive cells in the subventricular zone and hippocampal subgranular zone, together with pharmacokinetic analysis to assess brain exposure. Results: Using this platform, we identified Lead-238 as a small-molecule that enhanced neural stem cell proliferation and neurogenic output in vitro. In vivo, Lead-238 increased neurogenic activity in the subventricular zone, as evidenced by elevated numbers of BrdU-positive, BrdU/DCX-positive, and BrdU/Nestin-positive cells, whereas no detectable effects were observed in the hippocampal subgranular zone. Lead-238 achieved sufficient brain exposure, and its neurogenic effects were not readily explained by established neurogenic pathways. Conclusions: These findings demonstrate that growth factor–free phenotypic screening using human neural stem cells is an effective approach for identifying compounds that enhance adult neurogenic activity and identify Lead-238 as a small molecule that increases neurogenic activity in the subventricular zone without detectable effects in the hippocampal subgranular zone. Full article

Breast cancer (BC) remains the most diagnosed malignancy among women worldwide, with approximately 2.3 million new cases and over 670,000 deaths reported annually. Resistance to conventional chemotherapeutic agents and treatment-related toxicity remain major challenges in BC management. Resveratrol, a naturally occurring polyphenol, has been proposed as a potential modulator of chemotherapy response; however, comparative evidence regarding its interaction with different classes of chemotherapeutic agents is limited. This study aimed to comparatively assess the effects of resveratrol on the cytotoxic, antiproliferative, and apoptotic responses induced by adriamycin, taxol, and cisplatin in MCF-7 BC cells. MCF-7 cells were treated with adriamycin, taxol, cisplatin, and resveratrol, either alone or in combination, across multiple concentrations for 24, 48, 72, and 96 h. Cell viability was evaluated using the trypan blue exclusion assay. Cellular proliferation was assessed via BrdU incorporation, while apoptosis and cell death profiles were analyzed using Annexin V staining and flow cytometry. Exposure to individual chemotherapeutic agents induced a significant time- and dose-dependent reduction in MCF-7 cell viability (p < 0.001). Resveratrol co-treatment further modulated chemotherapy-induced cytotoxicity in an agent- and time-dependent manner. Combination treatments markedly suppressed DNA synthesis compared with single-agent exposure (p < 0.01) and significantly increased apoptotic cell populations. Flow cytometric Annexin V/PI analysis demonstrated that early apoptotic cells ranged from 3.2–11.3% in single-agent treatments and increased to 0.04–50.4% in resveratrol-based combination groups. Similarly, late apoptotic/secondary necrotic cell fractions increased from 4.1–4.4% in single-agent treatments to 2.1–69.9% following combination therapy, indicating a substantially higher overall cell death response in selected treatment conditions. This study demonstrates that resveratrol modulates the cytotoxic and apoptotic responses of MCF-7 BC cells to adriamycin, taxol, and cisplatin in an agent- and time-dependent manner. The findings indicate that resveratrol acts as a context-dependent modulator of chemotherapy response. Although resveratrol generally enhanced cytotoxic and apoptotic responses in combination treatments, the magnitude of these effects varied depending on the chemotherapeutic agent and exposure conditions. Further preclinical and clinical studies are warranted to define their therapeutic relevance. Full article

Background/Objectives: Endometrial cancer frequently develops resistance to therapy, partly due to the ability of tumor cells to adapt to cellular stress through non-apoptotic mechanisms. Mitochondrial dysfunction and cytoskeletal remodeling are increasingly recognized as key components of stress adaptation; however, their structural relationship under pharmacological stress in three-dimensional (3D) tumor models remains poorly characterized. The present study aimed to investigate the ultrastructural and phenotypic effects of lithium chloride (LiCl)-induced stress in 3D endometrial cancer spheroids, with a particular focus on mitochondrial alterations and intermediate filament organization. Methods: Three-dimensional spheroids generated from Ishikawa endometrial cancer cells were exposed to lithium chloride at concentrations of 1, 10, or 50 mM for defined time periods. Cell viability, proliferative activity, and clonogenic capacity were assessed using Trypan Blue exclusion, BrdU incorporation, and soft agar assays. Ultrastructural changes were examined by transmission electron microscopy to evaluate mitochondrial morphology, cytoplasmic organization, and intermediate filament distribution. Results: LiCl exposure resulted in a dose- and time-dependent reduction in cell viability, proliferation, and clonogenic potential in 3D spheroids. Ultrastructural analysis revealed pronounced mitochondrial swelling, cristae disorganization, and membrane-associated mitochondrial alterations. These changes were consistently accompanied by conspicuous accumulation and reorganization of intermediate filaments in close spatial proximity to damaged mitochondria, suggesting a structural association between cytoskeletal remodeling and mitochondrial injury. Across all experimental conditions, classical apoptotic ultrastructural features, including chromatin condensation and apoptotic body formation, were not observed. Conclusions: Together, these observations indicate that lithium chloride elicits a stress phenotype in 3D endometrial cancer spheroids that primarily manifests at the organelle and cytoskeletal levels, rather than through classical apoptotic execution. Although descriptive in nature, the present study highlights intermediate filament accumulation as a prominent structural feature of lithium-induced mitochondrial stress and establishes a structural reference point for future studies aimed at further investigating mitochondrial–cytoskeletal relationships during pharmacological stress in endometrial cancer. Full article

Gastric cancer (GC) is a highly prevalent and rapidly progressing cancer with a poor prognosis, primarily due to chemoresistance and treatment-related toxicity. Moracin D (MD), a benzofuran extracted from Morus alba L., has shown potential antitumor effects in various malignancies, although its impact on GC remains limited. The aim of this study was to assess the anticancer potential of MD in human gastric cancer cell lines and subcutaneous xenograft models. We examined cell proliferation, clonogenic ability, cell cycle progression, and apoptosis using MTT, BrdU, colony formation assays, flow cytometry, Western blotting, and immunohistochemistry. Our findings suggest that MD selectively inhibited GC cell proliferation and reduced DNA synthesis in vitro. It also inhibited colony formation and tumor growth in vivo, affecting GC cell clonogenicity without affecting body weight or vital organs, and without overt toxicity under the experimental conditions tested. Mechanistically, MD was found to induce G₂/M cell-cycle arrest, potentially through modulation of cyclin B1 and CDK1, and to trigger apoptosis in GC cells, which may involve the mitochondrial pathway as suggested by changes in Bcl-2 and pro-apoptotic protein levels. While Bcl-2 overexpression partially reversed MD-induced inhibition of proliferation and apoptosis, further studies are required to confirm its role as a mediator. Additionally, MD enhances the anticancer effects of 5-fluorouracil (5-FU) through synergistic mechanism. This study highlights the observed antiproliferative and proapoptotic effects of MD in preclinical models and suggests its potential as monotherapy or in combination with 5-FU as a promising therapeutic approach in the treatment of gastric cancer. Full article

Hydrogen peroxide (H₂O₂) is a key extracellular redox signaling molecule that regulates diverse physiological processes, including immune cell activation and proliferation. However, its role in maintaining extracellular redox balance and mediating intercellular signaling remains underexplored. In this study, we investigated how extracellular depletion of H₂O₂ by catalase modulates intracellular signaling pathways in macrophages. Catalase treatment effectively depleted extracellular H₂O₂ in a concentration- and time-dependent manner, leading to activation of mitogen-activated protein kinase (MAPK) pathways, including extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38, as well as nuclear translocation of the nuclear factor κB (NF-κB) p65 subunit. Perturbation of extracellular redox status resulted in robust upregulation of inflammatory and oxidative stress–related genes, including cyclooxygenase-2 (COX-2), C-C motif chemokine ligand 5 (CCL5), inducible nitric oxide synthase (iNOS), and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. This transcriptional response was accompanied by increased nitric oxide (NO) production and enhanced nuclear translocation and DNA-binding activity of nuclear factor erythroid 2–related factor 2 (Nrf2). Mechanistically, our data suggest that NO-mediated S-nitrosylation contributes to activation of the cellular antioxidant response. In addition, catalase-mediated depletion of extracellular H₂O₂ significantly (p < 0.05) suppressed 5-bromo-2′-deoxyuridine (BrdU) incorporation, indicating inhibition of macrophage proliferation. Together, these findings demonstrate that extracellular H₂O₂ functions as a physiological redox signal that maintains cellular homeostasis, and that its removal triggers a coordinated intracellular response involving both inflammatory activation and antioxidant defense. This study highlights the critical role of extracellular redox balance in shaping macrophage function and provides mechanistic insight into how changes in the oxidative environment regulate downstream immune signaling pathways. Full article

Glioblastoma is one of the most aggressive tumours with a poor prognosis and a modest survival rate after diagnosis. Several trials for a more targeted and effective treatment are in progress. Protein kinase CK2 is upregulated in glioblastoma and creates a favourable environment for cell proliferation by supporting several survival pathways. Inhibitors of CK2 kinase activity were shown to restrict growth rate or to induce apoptosis in different cell culture and animal models. Recently, we described the selective CK2 inhibitor 6,7-dichloro-1,4-dihydro-8-hydroxy-4(4 methylphenylamino)methylen]dibenzo [b,d]furan 3(2H)-one (TF). In this study, we found that TF effectively reduces the proliferation of A1207 glioblastoma cells with an EC₅₀ value of 13.7 µM, which is equal to the EC₅₀ value of CX-4945, which was the first CK2 inhibitor in clinical phase II trials (13.9 µM). We investigated the effect of TF and temozolomide (TMZ) as a single or combination treatment in two glioblastoma cell lines, A1207 and U87. The treatment was carried out over 48 or 72 h, and, subsequently, the biological effects were evaluated. The proliferation of both cell lines was significantly impaired by the application of the drugs, and combination treatment with TF and TMZ proved superior to the individual treatments. Not only proliferation, as determined by cell confluence assays and BrdU incorporation, but also viability in terms of metabolic activity and cytotoxicity were affected by the treatment. The decrease in proliferation and viability is partly due to the induction of apoptosis, with both cell lines differing in terms of the pattern of apoptotic caspases. Taken together, TF in combination with TMZ may be a promising candidate for the treatment of glioblastoma in the future. Full article

Sildenafil, a selective phosphodiesterase-5 (PDE5) inhibitor, supports vascular remodeling, but its effects on angiogenesis and regeneration of ischemic muscle tissue are not fully understood. We investigated the function of sildenafil by employing a murine hindlimb model of ischemia, in which ischemia and angiogenesis is induced by femoral artery ligation (FAL) in the lower leg of mice. Then, 7 days after FAL or sham operation, gastrocnemius muscles of sildenafil-treated and control mice were isolated and processed for histological and immunofluorescence analyses. Sildenafil treatment led to reduced apoptotic areas within the ischemic tissue (ascertained via TUNEL assay) and increased angiogenesis, evidenced by a higher capillary-to-muscle fiber ratio and an augmented number of proliferating capillary cells (CD31⁺/CD45⁻/BrdU⁺), compared to controls. We observed a decrease in the total count of leukocytes (CD45⁺) in sildenafil-treated mice. Regarding macrophage infiltration, we found a reduced total number of macrophages (CD68⁺), along with a shift in macrophage polarization toward the pro-angiogenic and anti-inflammatory M2-like phenotype (CD68⁺/MRC1⁺). In summary, we show that sildenafil treatment contributes to angiogenesis and the regeneration of ischemic muscle tissue, most likely by attenuating inflammatory responses and influencing macrophage polarization in direction to regenerative M2-like polarized macrophages. Full article