Search Results (717)

Dicarbadodecaboranes (12) (carboranes) are versatile molecular building blocks with unique properties, which allow the expansion of classical medicinal-chemical space. To enable single-photon emission computed tomography (SPECT) imaging of cyclooxygenase-2 (COX-2), we investigated the feasibility of introducing iodine-123 into nido-indoborin 1, a nido-carborane analog of indomethacin with potent and selective cyclooxygenase-2 inhibitory activity. An electrophilic iodination strategy afforded two regioisomers, 2a and 2b, bearing the iodine at the carborane cluster. Compared to nido-indoborin, a reduced COX-2 inhibition potency and selectivity were observed, with 2b exhibiting the more favorable inhibition profile. Radiosynthesis of [¹²³I]2b was achieved by N-chlorosuccinimide–mediated electrophilic substitution of 1, and conditions were optimized, leading to an isolated radiochemical yield of 4%. While the radiotracer displayed high stability in phosphate buffer, ester hydrolysis was observed in human plasma and murine liver microsomes with no significant deiodination in vitro. Cell uptake studies indicated partial COX-2–dependent accumulation but also revealed substantial non-specific uptake and unexpected enhancement of radiotracer uptake in the presence of carborane-based blocking agents. In vivo pilot imaging studies in mice bearing U87 xenografts showed renal and hepatobiliary clearance without measurable tumor accumulation but evidence of deiodination over time. Overall, iodination was feasible, but the resulting compounds lacked the required COX-2-selective tumor accumulation for further radiotracer development. Full article

Natural pigment lycopene (LYC), a carotenoid, possesses antioxidant, anti-apoptotic, anticancer, and immunoenhancing properties. During in vitro culture, this substance protects oocytes and early embryos from damage caused by reactive oxygen species (ROS), thereby enhancing the in vitro maturation (IVM) rate of oocytes and the developmental competence of early embryos. This study aimed to investigate the effects of supplementing different concentrations of LYC (0, 5, 10, and 15 μM) during in vitro culture of sheep oocytes and early embryos on their developmental competence. In contrast to the control group, the 5 μM LYC treatment group displayed a marked increase in the first polar body extrusion rate and the extent of cumulus cell expansion, as well as a significantly higher proportion of normal spindle assembly in sheep oocytes, but 15 μM LYC appeared to negatively affect oocyte maturation. Relative to all other experimental groups, the 5 μM LYC treatment group displayed significantly elevated rates of cleavage and blastocyst rate during early in vitro embryonic development. The levels of ROS in mature oocytes and early embryos were significantly decreased, whereas the GSH level was significantly elevated. Furthermore, LYC treatment significantly enhanced mitochondrial activity and markedly elevated the mitochondrial membrane potential (MMP) in mature oocytes and early embryos. Moreover, the total cell number of blastocysts was significantly increased. Moreover, in early embryos, the transcript levels of genes associated with both oxidative stress and apoptosis were favorably regulated. In conclusion, LYC supplementation boosted the rates of oocyte maturation and blastocyst formation in sheep, while elevating the developmental capacity of early embryos. Full article

Primula nutans Georgi, a medicinal herb used in Mongolian and Tibetan medicine for treating respiratory ailments, is a natural agent with antiobesity potential. We investigated the antiobesity and insulin-sensitizing effects of P. nutans Georgi extract (PGE) using in vitro and in vivo models. In 3T3-L1 preadipocytes, PGE inhibited adipocyte differentiation and lipid accumulation without cytotoxicity, accompanied by the reduced expression of adipogenic transcription factors (PPARG, C/EBPA, and adiponectin) and lipogenic genes (FASN, SCD1, and ACC), particularly during the early stages of adipogenesis. Similar effects were observed in primary stromal vascular cells derived from mouse inguinal white adipose tissue. PGE upregulated C/EBP homologous protein and C/EBPB and was associated with altered cell cycle progression, increased G2/M phase distribution, and the potential disruption of mitotic clonal expansion during early adipogenesis. In HFD-induced obese mice, intraperitoneal administration of PGE (10 or 30 mg/kg) significantly reduced body weight gain, white adipose tissue mass, and hepatic steatosis, independent of food intake. PGE downregulated lipogenic and proinflammatory gene expression in adipose and hepatic tissues and increased AMPK phosphorylation in white adipose tissue. PGE improved glucose tolerance and was associated with enhanced insulin sensitivity, as evidenced by reduced areas under the curve in the glucose tolerance and insulin tolerance tests and increased circulating adiponectin levels. Feature-based molecular networking identified 61 compounds from PGE. Network pharmacology analysis revealed several antiobesity targets, including PPARG and AKT1. Molecular docking analyses suggested favorable binding affinities between major compounds and metabolic regulators. Collectively, these findings suggest that PGE may suppress adipogenesis and improve metabolic parameters in obese mice, supporting its potential as a natural candidate for obesity and related metabolic disorders. Full article

Iris sanguinea features upright, stiff leaves, making it an excellent cut-foliage material, with its tall leaf architecture greatly enhancing ornamental value in landscaping. However, during the leaf expansion phase, plants frequently exhibit loose foliage arrangement, excessive spreading, and compromised mechanical strength, culminating in lodging and a concomitant decline in ornamental quality. Plant height in I. sanguinea is strongly regulated by phytohormones. This study showed that exogenous GA at concentrations of 50 mg·L⁻¹, 100 mg·L⁻¹, and 200 mg·L⁻¹ increased seedling height by 5.7%, 8.8%, and 12.7%, respectively, through foliar spraying on I. sanguinea seedlings grown ex vitro in a greenhouse; conversely, PAC treatment at equivalent concentrations suppressed growth by 19.3%, 21.0%, and 22.2%, respectively. Two pivotal GA signaling components, GAI and GID1a, were isolated from I. sanguinea. Subcellular localization confirmed that both IsGAI and IsGID1a proteins localize to the nucleus. Overexpression vectors pCAMBIA1300-IsGAI-GFP and pCAMBIA1300-IsGID1a-GFP were constructed and expressed in Arabidopsis thaliana. Transgenic lines overexpressing IsGAI showed significantly reduced plant height, hypocotyl elongation, and bolting, whereas IsGID1a overexpression promoted these traits. Exogenous GA application partially reversed the dwarf phenotype induced by IsGAI overexpression and further potentiated the height enhancement observed in IsGID1a-overexpressing lines. This study identifies two key genes controlling plant height and provides a theoretical basis and genetic resources for precisely engineering plant architecture in I. sanguinea. This is especially important for developing dwarf varieties with enhanced ornamental and agronomic traits, offering significant potential in the landscaping and cut flower industries. Full article

Influenza A virus (IAV) remains a major global health threat despite available vaccines and antiviral agents, while current therapies are limited by drug resistance and safety concerns. Curcuminoids exhibit antiviral and anti-inflammatory activities but are constrained by poor water solubility and low bioavailability. To address these limitations, we investigated the antiviral and immunomodulatory properties of a water-solubilized curcuminoid nanoparticle formulation (C–S/M) in both in vitro and in vivo models of IAV infection. To evaluate the potential antiviral and anti-inflammatory effects of C–S/M, we performed a cytopathic effect (CPE) reduction assay in triplicate at 0.001 MOI and quantitative real-time PCR (qRT-PCR) targeting viral NS1 transcripts in MDCK cells. C–S/M suppressed viral NS1 vRNA levels in MDCK cells at lower curcuminoid-equivalent concentrations than native curcuminoids and attenuated IAV-induced TNF-α, IL-6, and IL-8 production. Furthermore, in vivo antiviral efficacy was evaluated in female C57BL/6 mice intranasally infected with IAV and treated orally with C–S/M. Survival, lung viral loads, pulmonary cytokine levels, and splenic immune cell phenotypes were analyzed. In IAV-infected mice, oral administration of C–S/M modestly improved survival and significantly reduced lung viral burden and pulmonary proinflammatory cytokine levels. In addition, in vivo C–S/M treatment was associated with recovery of virus-suppressed T-cell immune responses, including increased Th1 and activated CD8⁺ T cells, reduced regulatory T-cell expansion, and restoration of multifunctional CD4⁺ and CD8⁺ T cells. These findings suggest that C–S/M exerts antiviral and immunomodulatory effects in experimental IAV infection and may serve as a potential adjunctive candidate for further investigation against influenza-associated inflammation. Full article

Quinazolinone derivatives are well-known anticancer agents; anticancer properties are also part of the broad spectrum of biological activity of coumarins. Conjugates containing quinazolin-4(3H)-one and coumarin fragments linked by polymethylene bridges of varying lengths were designed to improve properties of both parental compounds and create new anticancer or antibacterial agents. 3-{3-[(4-Methyl-2-oxo-2H-chromen-7-yl)oxy]propyl}quinazolin-4(3H)-one was synthesized as the base compound. It demonstrated moderate cytotoxicity against leukemia (K562 and HL60) and neuroblastoma (SH-SY5Y) cells in vitro, combined with relatively low acute, subacute, and chronic toxicity in vivo. Conjugates with various substituents and linkers were then synthesized to evaluate the structure–activity relationship. A study of the synthesized compounds on cell cultures showed that the introduction of a methyl substituent into the benzene ring of the coumarin fragment led to both an increase in cytotoxicity and expansion of its spectrum of action. Testing of the hybrids against Gram-positive and Gram-negative bacteria revealed that the introduction of halogens into the quinazoline fragment in the compounds or the elongation of the linker led to the emergence of pronounced antibacterial properties, which were most clearly manifested against Acinetobacter baumanii. The possibility of directing activity of quinazoline-4(3H)-one–coumarin hybrids by varying the substituents and the length of the linker was shown. Full article

Background: Advanced glycation end products (AGEs) and oxidative stress (OS) have been linked to bone complications related to type 2 diabetes mellitus (T2DM). However, the effects of AGEs and OS on bone marrow mesenchymal stromal cells (BMMSCs), which play a key role in bone homeostasis and repair, remain unclear. Objectives: This study aimed to investigate the effects of AGEs on BMMSCs function and the ability of N-acetylcysteine (NAC) to alleviate AGE-induced OS in a T2DM context. Methods: Bone marrow (BM) and BMMSCs were isolated from Zucker diabetic fatty (ZDF) rats, which serve as a T2DM model, and their lean littermates (ZL, controls) at 24 weeks of age. Results: The results show that long-standing T2DM leads to changes in the BM’s cellular composition and BMMSCs function that are distinct from age-related changes. In vitro, AGEs decreased BMMSCs viability, proliferation, and migration. The effects of AGEs were stronger in BMMSCs derived from a T2DM microenvironment. In both T2DM- and ZL-BMMSCs, AGEs induced cytoplasmic ROS, which was differentially reduced by NAC. The effect of NAC on T2DM-BMMSCs was greater when the cells were pre-treated with NAC 24 h before exposure to AGEs, whereas simultaneous exposure to both resulted in a smaller effect. Conclusions: These results show that AGEs impair BMMSCs expansion and functionality. AGE-induced ROS generation may be a critical factor in this impairment, while NAC was able to reduce OS in BMMSCs from a T2DM context. These findings highlight the vicious negative effects of the T2DM microenvironment on BMMSCs and underscore the need for further studies to better understand the underlying mechanisms and to explore strategies aimed at mitigating OS in the T2DM context. Full article

Limbal stem cell (LSC) transplantation is an important treatment for limbal stem cell deficiency (LSCD), but low efficacy in maintaining LSC stemness during in vitro expansion greatly affects its wider application. The primary contributing factors include a low proportion of stem cells and the lack of a stable, supportive microenvironment over prolonged culture. Rabbit corneal tissues preserved under deep cryogenic conditions for more than six months retain viable limbal stem cells (LSCs), and primary LSCs isolated from these tissues exhibit robust stem cell characteristics. It is noteworthy that the NLRP3/Caspase-1/IL-1β signaling axis was activated in corneal epithelial cells, and outer limbal layers preserved for one or two years. Based on these findings, a combined strategy integrating deep cryopreservation with IL-1β induction was established for the processing of limbal tissues. The combined cryogenic and IL-1β preconditioning yielded primary LSCs with maintained p63⁺ cell proportions, a reduction in K3⁺ differentiated cells from approximately 80% to 60%, and a 6.25-fold increase in colony-forming efficiency. In addition, an increased proportion of cells in the G2/M phase and enhanced proliferative capacity were observed. The enriched LSC population also exhibited improved stratified epithelial reconstruction potential. These findings identify an effective strategy for preserving and enriching LSCs from limbal tissue, providing a practical and efficient approach for LSC preparation prior to transplantation. Further in vivo studies will be important to validate the functional performance of these cells in ocular surface reconstruction. Full article

Hair follicles (HFs) are highly accessible mini-organs that house diverse somatic and stem cell populations with broad therapeutic potential. In this study, we investigate the untapped utility of plucked HFs as a non-invasive tissue source for regenerative medicine. We demonstrate the successful isolation and expansion of keratinocytes and mesenchymal stem cells (MSCs) from plucked follicles using an enzyme-free explant culture method. HF-derived keratinocytes retained their epithelial identity and were efficiently reprogrammed into induced pluripotent stem cells (iPSCs). These iPSCs were further directed toward definitive endoderm and pancreatic progenitor fates, confirming their robust autologous regenerative capacity. Flow cytometric analysis of HF-MSCs validated a characteristic mesenchymal profile, and these cells exhibited classical trilineage plasticity alongside the ability to differentiate into dopaminergic neural progenitors. Furthermore, proteomic and vesicular characterization of the autologous HF secretome (aHFS) revealed a rich enrichment of regenerative cytokines and exosomes. The aHFS demonstrated potent wound-healing bioactivity in vitro. Collectively, these findings establish the plucked hair follicle as a highly practical, scalable source for both cell-based and cell-free therapies, highlighting the clinical value of early-stage follicular biobanking for personalized medicine. Full article

Polysaccharide-based hydrogels for gastric retention face the inherent challenge of achieving effective retention through swelling while avoiding mechanical failure. Here, we introduce a strategy by incorporating curdlan into chitosan/sodium polyacrylate interpenetrating networks to reinforce the hydrogel and regulate swelling-induced transport behavior. Curdlan-reinforced chitosan/polyacrylate (CS/CUR/PAAS) hydrogels with varying curdlan content (0–4 wt.%) were synthesized and characterized. Optimal reinforcement was achieved with 2 wt.% curdlan, yielding an indentation hardness of ~80 kPa and an elastic modulus of ~63 kPa without compromising swelling capacity. Under acidic conditions (pH 1.2), the hydrogel swelled rapidly (~50-fold at 3 h; ~140-fold at 8 h) while maintaining structural integrity. Using a dynamic in vitro human stomach simulator (DHSI-IV), the optimized hydrogel demonstrated gastric retention for up to 5 h, with ~60% of the initial mass retained at 6 h. Metformin hydrochloride release followed diffusion-controlled kinetics (~69% over 8 h), governed primarily by pH with secondary shear modulation. Microstructural and rheological analyses revealed that acidic conditions regulated network expansion, viscoelastic relaxation, and pore formation, which in turn controlled transport pathways and drug release. The findings highlight that curdlan reinforcement stabilizes swelling behavior under acidic conditions, offering a robust and pH-responsive strategy for designing mechanically stable, gastric-retentive hydrogels. Full article

Background/Objectives: The rapid expansion of New Approach Methodologies (NAMs) is transforming oral biopharmaceutics by offering mechanistically rich, human-relevant tools that can reduce reliance on animal testing while improving translational confidence. Regulatory agencies, including the Food and Drug Administration (FDA) and the European Medicines Agency (EMA), are increasingly open to NAM-generated evidence, provided that methods are fit-for-purpose and scientifically justified. This review synthesizes current advances and evaluates how NAMs can be integrated across drug-development stages to enhance the prediction of oral absorption, formulation performance, and regulatory decision-making. Methods: A comprehensive literature review was conducted across classical and emerging methodologies, including in vitro permeability and solubility models, organoids, organ-on-a-chip (OoC) systems, machine learning frameworks, and mechanistic approaches such as the physiologically based pharmacokinetic (PBPK) and biopharmaceutics (PBBM) models. Emphasis was placed on physiological relevance, predictive performance, validation status, and regulatory applicability. Results: Classical tools remain essential for the Biopharmaceutics Classification System (BCS)-based biowaivers and risk-based assessments, yet they often lack physiological fidelity. NAMs provide enhanced representation of intestinal architecture, hydrodynamics, transporter activity, and metabolism. Organoids and microphysiological systems generate high-quality permeability and metabolic data, while computational NAMs enable scalable prediction of ADME properties and formulation behavior. When integrated into PBPK/PBBM models, these methods have great potential in predicting in vivo performance in humans. Evidence demonstrates that NAMs can refine, reduce, and, in specific contexts, replace animal studies without compromising scientific rigor. Conclusions: NAMs complement, rather than displace, classical biopharmaceutic tools, enabling a more mechanistic, human-centered, and ethically responsible framework for drug development. Their effective implementation will depend on continued validation, standardization, and regulatory harmonization as the field transitions toward fully NAM-supported biopharmaceutical assessment. Full article

Silver nanoparticles (AgNPs) have gained significant interest across various areas arising from their multifunctional mechanisms. Biomedical applications are one of the areas where the therapeutic and diagnostic potential of AgNPs are highlighted. Considering the expansion of biomedical use of AgNPs, nervous system-based applications, including neuroimaging, neural implant coatings and development of neural tissue-targeted drug delivery systems are some of the potential applications of AgNPs in the current research. However, growing interest in these nervous system related applications and the limited regenerative capacity of neural tissues make it essential to carefully evaluate the potential neurotoxic effects of AgNPs. AgNP-induced responses in neural tissues may differ according to key physicochemical and exposure-related parameters, specifically particle size, shape, surface chemistry, coating properties, protein corona formation, exposure route, dose, and duration. Among the possible mechanisms that may contribute to these responses are blood–brain barrier (BBB) disruption, mitochondrial dysfunction and oxidative stress, neuroinflammation and glial activation, and cell death processes such as apoptosis, autophagy, and ferroptosis. In this review, in the context of the potential neurotoxic effects of AgNPs on the nervous system, the main parameters that determine AgNP neurotoxicity and the possible mechanisms involved are examined in detail, where recent scientific developments in this field are evaluated based on current in vitro and in vivo studies. Full article

(1) Background: Posterior crossbite associated with maxillary transverse deficiency is commonly managed with maxillary expansion, yet the correspondence between laboratory activation behavior and the clinical response of nickel–titanium leaf-spring expanders remains insufficiently defined; therefore, this study aimed to compare in vitro and in vivo performance of the Leaf Expander^® and to assess their agreement. (2) Methods: A retrospective sample of 15 mixed-dentition patients (7–10 years) treated at two university centers with a Leaf Expander^® (6 mm screw; 900 g) was evaluated; interpremolar (E–E), intermolar (6–6), and intercanine (C–C) distances were recorded at baseline (T0, digital models) and at follow-up visits (T1–T5, caliper measurements), while mechanical compression testing (Instron 3365) quantified force release across the activation sequence; normality (Shapiro–Wilk), parametric analyses, and Pearson correlation were used. (3) Results Posterior crossbite correction was achieved in all completed cases, with mean total increases (T0–T5) of 5.4 mm (E–E), 4.4 mm (6–6), and 6.0 mm (C–C); early expansion (T1–T0) averaged 2.5 mm at E–E, and laboratory curves showed an activation peak followed by sustained force release (~6.5–9 N) and a residual-load phase. Agreement between declared activation and clinical response was higher for E–E and 6–6 than for C–C, which showed greater variability. (4) Conclusions: These findings support the Leaf Expander^® as an effective compliance-free slow expansion device and indicate that laboratory force behavior can help interpret the clinical expansion timeline, including delayed expression after activation. Full article

Background: Cardiolipin (CL) is a phospholipid composed of a glycerol linked with two phosphatidate moieties that constitutes an integral part of the human inner mitochondrial membrane under physiological conditions. It is also vital for bacterial membrane transport and key bacterial functions associated with cell division and infection. CL is released in the cytosol or into the extracellular milieu upon cell death and during inflammation. We therefore tested the ability of CL to activate and expand tumor infiltrating lymphocytes (TIL) from patients with epithelial cancer. Methods: TIL were isolated from gastrointestinal tumor tissues and expanded in vitro in the presence of CL. The role of the NLRP3 inflammasome was evaluated using the specific inhibitor MCC950 and siRNA-mediated silencing of NLRP3. Phenotypic changes and T-cell potency were assessed via CXCL9/10 expression levels. To characterize the immune repertoire, deep TCR sequencing was performed to compare the TCR Vα and Vβ CDR3 regions between TIL and the corresponding tumor tissue. Recognition of autologous tumor cells and tumor-specific mutations, including mutations in KRAS and mitochondrial UQCRFS1 (D145V), was assessed using MHC class I and II restriction assays. Results: CL-expanded TIL exhibited increased CXCL9/10 expression, which is associated with increased potency of tissue invasion. CL-TIL exhibited broader recognition of frequently occurring KRAS mutations, and this effect could be blocked with an inhibitor (MCC950) of the NLRP3 pathway, a multiprotein inflammatory complex associated with danger signaling. TIL exhibited an enriched TCR Vα and Vβ CDR3 repertoire compared to tumor tissue, as defined by deep TCR sequencing. TCR αβ+ TIL recognized autologous tumor tissue in an MHC class I– and class II–restricted fashion, including the mutant HLA-DP–restricted mitochondrial protein associated with the electron respiratory chain complex III (UQCRFS1 D145V) presented by autologous tumor cells. Conclusions: CL activates the NLRP3 inflammasome pathway in TIL from patients with GI cancer and increases CXCL9/CXCL10 expression in TIL, resulting in enhanced recognition of mutant cancer–associated target epitopes, including a mitochondrial protein. CL may provide a danger signal: that facilitates TIL expansion via CL-activated pathways. Full article

Adipogenesis is a critical factor in causing obesity, which is a global health problem associated with metabolic disorders, such as insulin resistance and cardiovascular diseases. Natural compounds with anti-adipogenic activity may represent potential approaches for modulating adipocyte function. However, despite increasing interest in natural products, the anti-adipogenic potential of acridone alkaloids, particularly prenylated derivatives, remains largely unexplored. This study examined the effects of N-methylatalaphylline (NMA), a prenylated acridone alkaloid, on adipocyte differentiation, lipid accumulation, and glucose uptake. NMA exhibited anti-adipogenesis, particularly toward preadipocytes, and significantly reduced lipid accumulation in murine 3T3-L1 and human PCS-210-010 adipocytes at nontoxic doses (1.5–6 µM). At 3–6 µM, NMA downregulated adipogenic regulators, including PPARγ, C/EBPα, and SREBP1, along with adipogenic effectors, such as FABP4, adiponectin, LPL, PLIN1, and FAS. Mechanistic studies indicated that NMA treatment was associated with reduced phosphorylation of AKT, ERK, and p38, accompanied by cell cycle arrest and inhibition of mitotic clonal expansion. Meanwhile, activation of AMPK-ACC signaling, which may contribute to suppression of adipogenesis and reduced glucose uptake, was observed in differentiated 3T3-L1 cells after treatment with 6 µM NMA for 48 h. Additionally, molecular docking and molecular dynamics simulations suggested potential interaction between NMA and ERK1, supported by hydrogen bonding and hydrophobic contacts. Overall, these findings suggest that NMA exerts anti-adipogenic effects in vitro by modulating adipocyte proliferation, differentiation, and lipid metabolism. These findings highlight NMA as a promising acridone alkaloid scaffold for anti-adiposity applications, warranting further in vivo validation. Full article