Background/Objectives: Osteotoxicity is a severe complication of Methotrexate (MTX) chemotherapy, characterized by oxidative stress and disrupted bone remodeling. The primary objective of this study was to investigate the cytoprotective mechanisms of the antioxidant Alpha-Lipoic Acid (ALA) against MTX-induced osteotoxicity, specifically focusing on its modulation of oxidative stress, apoptosis, and Mitogen-Activated Protein Kinase (MAPK) signaling pathways. Methods: Murine osteocyte-like MLO-Y4 cells were cultured and exposed to a fixed dose of MTX (10−5 M), either alone or concurrently with ALA (50 μmol/L) for 48 h. Biochemical profiling was performed using specific enzyme-linked immunosorbent assays (ELISA) and colorimetric kits to evaluate pro- and anti-apoptotic proteins (Caspase-3, Bax, Bcl-2, Wee1, GRP78, GADD153, AIF), active MAPK components (p-JNK, p-ERK), and standard oxidative stress parameters (TAS, TOS, SOD, GPx). Results: MTX treatment induced significant cellular stress, evidenced by elevated Caspase-3, Bax, p-JNK, and p-ERK levels, alongside a critical reduction in Bcl-2 expression. MTX also markedly increased TOS while depleting TAS, SOD, and GPx levels. Conversely, co-treatment with ALA significantly mitigated these cytotoxic responses. ALA restored the Bax/Bcl-2 balance, effectively downregulated both p-JNK and p-ERK activation, and substantially reinforced the cellular antioxidant defense system by enhancing TAS, SOD, and GPx activities, although recovery to baseline control levels was partial. Conclusions: ALA exerts robust in vitro cytoprotective effects against MTX-induced osteotoxicity in MLO-Y4 cells by counteracting oxidative stress and inhibiting aberrant apoptotic and MAPK signaling. These findings establish a mechanistic baseline, underscoring the need for subsequent in vivo dose–response studies to validate ALA’s therapeutic potential in chemotherapy management.

Background: The emergence of multidrug-resistant (MDR) Gram-negative pathogens, namely Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii and Helicobacter pylori, necessitates urgent therapeutic alternatives. This scoping review aimed to summarize the current evidence on the efficacy of lytic bacteriophages against these critical MDR pathogens, and to identify existing research gaps and implementation challenges. Methods: The literature search was conducted by searching PubMed, Web of Science, and Scopus AI for studies published from 2015 to 2025. The inclusion criteria focused on experimental and human studies evaluating phage therapy against MDR, extensively drug-resistant (XDR), or pan-drug-resistant (PDR) strains in the four target species. A total of 172 articles were included. Results: A number of studies showed an increasing trend (2015–2025), focusing mainly on K. pneumoniae (n = 65), P. aeruginosa (n = 55), and A. baumannii (n = 48). No eligible studies for MDR H. pylori were found. All 172 studies confirmed lytic activity, with phage cocktails showing superior antibacterial activity than single phages in four studies. Phages also demonstrated antibiofilm activity (n = 44). Most animal studies reported successful bacterial reduction in animals treated with phages, and 87.5% of 23 human case studies reported patient improvement or infection clearance. However, heterogeneity in the types of animal models used and in dosage and administration routes in human studies was notable. Conclusions: Lytic bacteriophages exhibit strong potential as a new therapeutic option. Key challenges include the lack of data for MDR H. pylori, heterogeneity in animal models, and a paucity of large-scale human clinical trials. Future research must prioritize standardization, mechanistic studies, and conducting robust human trials to enable clinical translation and regulatory acceptance.

Background: Spasmolytic polypeptide-expressing metaplasia (SPEM) is an injury-induced gastric epithelial reprogramming state with limited therapeutic options. Although mitochondrial dysfunction has been implicated in epithelial stress responses, its contribution to SPEM development remains incompletely understood. Traditional herbal decoctions have shown potential in alleviating gastric epithelial injury, yet their underlying mechanisms remain largely unclear. Purpose: This study aimed to investigate whether refined Jianpi Huayu Jiedu Decoction attenuates tamoxifen (TAM)-induced SPEM, with a focus on LCN2-associated mitochondrial dysfunction. Methods: TAM-induced SPEM models were established in mice and gastric epithelial cells. Histological, molecular, and mitochondrial analyses were performed to evaluate SPEM features and epithelial stress responses. UPLC–MS/MS-based chemical profiling, network pharmacology, transcriptomic analysis, and LCN2 knockdown experiments were integrated to explore the underlying regulatory mechanisms. Results: Refined Jianpi Huayu Jiedu Decoction significantly alleviated TAM-induced gastric mucosal injury and suppressed the expression of SPEM-associated markers in vivo and in vitro. JHJD treatment improved mitochondrial function, reduced oxidative stress, and normalized mitochondrial dynamics, accompanied by downregulation of LCN2 expression. Chemical profiling identified multiple bioactive components of JHJD, and integrative analyses combining transcriptomics, network pharmacology, and molecular docking suggested that these components are associated with LCN2-related epithelial stress and mitochondrial regulatory networks. Functional validation further demonstrated that LCN2 knockdown partially recapitulated the protective effects of JHJD on mitochondrial homeostasis and epithelial reprogramming. Conclusions: These findings indicate that refined Jianpi Huayu Jiedu Decoction attenuates TAM-induced SPEM in association with restoration of mitochondrial homeostasis and suppression of LCN2-related stress signaling, providing mechanistic insight into early gastric epithelial reprogramming.

Background/Objectives: Major depressive disorder (MDD) remains a leading cause of disability worldwide and exhibits substantial biological heterogeneity that is not adequately captured by current symptom-based diagnostic systems. While the classical monoamine hypothesis has historically guided antidepressant development, it does not fully account for variability in treatment response, delayed therapeutic onset, or the persistence of cognitive and anhedonic symptoms. Converging evidence from molecular, neuroimaging, and translational studies increasingly implicates glutamatergic dysregulation and impaired neuroplasticity as key mechanisms in depressive pathology. This narrative review aims to integrate monoaminergic and glutamatergic perspectives within a dimensional framework that may help explain clinical heterogeneity and inform mechanism-based treatment strategies. Methods: A narrative synthesis of the literature was conducted using major biomedical databases including PubMed, Scopus, and Web of Science. Preclinical studies, neuroimaging investigations, biomarker research, randomized clinical trials, and meta-analyses examining monoaminergic dysfunction, glutamatergic signaling, neuroplasticity pathways, and rapid-acting antidepressants were reviewed and thematically integrated. Results: Evidence indicates that depressive syndromes may reflect varying contributions of monoaminergic dysregulation and glutamatergic–neuroplastic impairment. Monoaminergic disturbances interact with inflammatory and neuroendocrine processes, including cytokine-driven activation of the kynurenine pathway. In parallel, alterations in glutamatergic signaling, glial function, and BDNF–TrkB–mTOR pathways contribute to synaptic atrophy and network dysfunction. Rapid-acting antidepressants such as ketamine, esketamine, and dextromethorphan–bupropion provide clinical proof-of-concept that direct engagement of synaptic plasticity mechanisms can accelerate symptom improvement, particularly in treatment-resistant depression. Conclusions: Integrating monoaminergic and glutamatergic mechanisms within a “monoamine–glutamate continuum” offers a conceptual framework for understanding depressive heterogeneity and treatment response. Multimodal approaches combining clinical phenotyping with inflammatory, neuroimaging, and molecular markers may ultimately support mechanism-informed precision psychiatry strategies in major depressive disorder.

Background/Objectives: Endophytic fungi represent an alternative source for resveratrol (RES) production. The present study aims to utilize mycoendophytic-derived resveratrol as a reducing agent for the synthesis of silver nanoparticles (AgNPs), in addition to further assay the cytotoxic activity of a RES-conjugated nanocarrier system toward human epidermoid carcinoma A-431 cells. Methods: Alternaria alternata AUMC 16209 was isolated from the stem of grapevine Vitis vinifera L. cultivar prime. Strain identification was achieved through morphological and molecular characterization using ITS sequencing. A. alternata AUMC 16209 exhibited RES production capability upon cultivation on PDB medium for seven days with a total of 8.25 mg/L as determined by HPLC. The crude RES was purified using flash chromatography followed by structure elucidation through 1H and 13C NMR analyses. The purified RES was used for green synthesis of nanoparticles, acting as a reducing agent for silver ions. Results: Stable RES-AgNPs were fabricated at particle sizes ranging from 25 to 47 nm. RES-AgNPs observed a plasmon resonance absorption band at 415 nm with a negative zeta potential value of −38.5 mV. The crystalline structure of RES-AgNPs was addressed through X-ray diffraction analysis. FT-IR spectroscopy confirms the involvement of the functional –OH group and the aromatic C=C bond in the reduction and stabilization process. RES-AgNPs was more efficient to inhibit the cellular proliferation of human epidermoid carcinoma A-431 cells compared to RES alone. Conclusions: This report introduces for the first time an endophytic A. alternata as a sustainable source for RES production and emphasizes its potential for green synthesis of stable AgNPs with promising cytotoxic activity.

Background/Objectives: Epigenetic modifications, particularly DNA methylation, contribute to tumor progression and therapy resistance. Guadecitabine, a hypomethylating agent (HMA), has shown promising clinical activity when combined with carboplatin in preclinical models. We evaluated the combination of guadecitabine with carboplatin as a second-line treatment for extensive-stage small-cell lung cancer (SCLC; NCT03913455), one of the deadliest malignancies. Here, we report methylome changes in peripheral blood mononuclear cells (PBMCs) collected at baseline and during treatment from patients on the trial. Methods: PMBC DNA was analyzed using Infinium HumanMethylationEPIC v1.0 bead chips. Data were processed, and differentially methylated positions (DMPs) were identified and analyzed for pathway enrichment using bioinformatic approaches, and immune deconvolution analyses were conducted to investigate the impact on immune cell composition. Results: Direct comparison of PBMCs between cycle 2 day 5 (C2D5; post-treatment) vs. cycle 1 day 1 (C1D1; pre-treatment) revealed a greater number of hypomethylated DMPs (380 DMPs in C2D5 vs. C1D1 PBMCs; p < 0.05, |β| > 20%). Moreover, when first compared with normal PBMCs from cancer-free controls, the number of hypomethylated DMPs was even greater in C2D5 than in C1D1 (1771 vs. 237 DMPs, respectively; p < 0.05, |β| > 20%). Long interspersed nucleotide elements-1 (LINE-1) were significantly hypomethylated in PBMCs after HMA treatment (C2D5 vs. C1D1). Pathway analysis of hypomethylated DMPs revealed significant alterations in key signaling pathways, including NF-κB, Rho GTPase, and pulmonary fibrosis in C1D1 vs. C2D5. Normal PBMCs to C1D1 PBMCs revealed changes in IL-3 signaling, Fcγ receptor-mediated phagocytosis, and molecular mechanisms of cancer. Deconvolution analysis revealed a greater percentage of monocytes in C1D1 vs. normal PBMCs; after HMA treatment, percentages of monocytes and B cells decreased, while the eosinophil percentage increased in C1D1 vs. C2D5. Conclusions: HMA treatment has a global impact on PBMC methylomes in cancer patients. DNA methylation changes were associated with biological pathways related to PBMC function, and shifts in distinct immune cell populations were observed.

Bovine mastitis remains a major challenge in dairy production despite extensive antimicrobial use, with therapeutic failure increasingly attributed to factors beyond classical antimicrobial resistance (AMR). Growing evidence indicates that treatment inefficacy arises from the combined effects of pharmacokinetic/pharmacodynamic (PK/PD) constraints, biofilm-mediated tolerance, intracellular persistence, and the adaptive capacity of mastitis pathogens. Intramammary therapy is particularly limited by poor tissue penetration, episodic drug elimination via milk flow, and inactivation by milk components, frequently resulting in subtherapeutic exposure at the site of infection. These limitations are amplified in chronic and subclinical mastitis, where biofilms and intracellular reservoirs reduce antimicrobial susceptibility and promote relapse and resistance selection. This narrative review integrates current knowledge on pharmacokinetic and pharmacodynamic (PK/PD) barriers, microbial survival strategies, and antimicrobial resistance (AMR) mechanisms that underlie treatment failure in bovine mastitis. It critically evaluates conventional antimicrobial therapies alongside emerging approaches, including antimicrobial peptides, bacteriophages and endolysins, nanoparticle-based delivery systems, immunomodulators, CRISPR-guided antimicrobials, and drug repurposing strategies. Overall, available evidence highlights the potential of these approaches to enhance therapeutic durability, particularly in settings where biofilm formation, intracellular persistence, and resistance limit conventional treatment efficacy. By mapping research coverage across mastitis phenotypes and therapeutic outcomes, this review identifies key gaps in long-term efficacy and resistance mitigation and underscores the need for PK/PD-guided, biofilm-aware, and resistance-conscious strategies to support durable next-generation mastitis management.

Background/Objectives: Cancer persists as a leading concern in the current medical field, and current therapies are limited by toxicity, cost, and resistance. Targeted inhibition of tubulin polymerization is considered as a promising therapeutic strategy for cancer treatment. Methods: Thirty-one new tubulin polymerization inhibitors were designed via molecular hybridization techniques, and BLI technology was employed to quantitatively investigate their interactions with tubulin. Antiproliferative activities against MCF-7, MDA-MB-231, A549, and HeLa cell lines was evaluated using the CCK8 assay. Apoptosis induction and cell cycle arrest were analyzed by flow cytometry. The anti-tumor activity of compound B6 was validated in a mouse melanoma tumor model. Results: Compounds exhibited varying degrees of antiproliferative activity against four tumor cell lines. Among them, compound B6 was the most promising candidate and displayed strong broad-spectrum anticancer activity with an average IC50 value of 2 μM. The mechanism studies revealed that compound B6 inhibited tubulin polymerization in vitro, disrupted cell microtubule networks, and arrested the cell cycle at G2/M phase. Furthermore, B6 displayed significant in vivo antitumor efficacy in a melanoma tumor model with tumor growth inhibition rates of 70.21% (50 mg/kg). Conclusions: This work shows that B6 is a promising lead compound deserving further investigation as a potential anticancer agent.

Background: Saffron (Crocus sativus L.) corms, often discarded as agricultural by-products, are a promising and sustainable source of bioactive metabolites with potential therapeutic relevance. However, their anticancer potential remains largely underinvestigated. Objectives: This study aimed to compare the phytochemical composition of hydroethanolic extracts from fresh (HEEF) and stored (HEES) saffron corms and to evaluate their anticancer effectiveness against colorectal cancer cells. Methods: Phytochemical profiling was performed using HPLC-ESI-QTOF-MS/MS. Cytotoxicity against T84 and SW480 colorectal cancer cell lines was determined by the crystal violet assay. Apoptosis-related protein modulation was assessed by Western blotting. Additionally, molecular docking, molecular dynamics simulations, and MM/GBSA calculations were used to investigate ligand–target binding affinities and stability. Results: Both extracts contained diverse primary and secondary metabolites, including phenolic acids, flavonoids, triterpenoids, lignans, anthraquinones, carotenoids, sugars, and fatty acids. HEES showed higher relative abundance of key bioactive metabolites than HEEF, which was enriched mainly in primary metabolites. HEES showed significantly greater dose-dependent cytotoxicity, particularly against SW480 cells after 24 h (IC50 = 34.85 ± 3.35). Apoptosis induction was confirmed through increased expression of caspase-9 and p53 in T84 cells. In silico studies revealed strong and stable interactions of major metabolites, especially 3,8-dihydroxy-1-methylanthraquinone-2-carboxylic acid with COX2 and crocetin with VEGFR2. Conclusions: Stored saffron corms possess a richer bioactive profile and show enhanced anticancer effects in vitro compared with fresh saffron corms, suggesting that they may represent a promising source of compounds for the future development of colorectal cancer therapeutics.

Creatine monohydrate is known for its moderate solubility (13 g/L at 25 °C), which limits the feasibility of producing its fast-dissolving forms. Overcoming this limitation is possible through the application of technological approaches, the overview of which is presented in this work, including chemical modification, micronization, granulation, amorphization, formation of solid dispersions, and encapsulation. The results showed the predominance of chemical methods (about 60% of the analyzed patents). At the same time, the use of physical methods and the combination of several technologies can increase both the dissolution rate and the solubility of creatine monohydrate while maintaining its stability. This makes these approaches the most promising for the development of production technology for fast-dissolving forms.