Search Results (1,304)

Nuclear Magnetic Resonance (NMR) spectroscopy has long been a cornerstone in the structural elucidation of molecules, offering unique insights into atomic-level connectivity, conformation, and dynamics. Over the past decades, methodological and technological advances have significantly expanded its capabilities and applications. This manuscript charts the evolution of NMR from classical 1D/2D experiments to modern methods empowered by ultrahigh magnetic fields, cryogenic probes, non-uniform sampling, new methodologies, and hyperpolarization. We emphasize the growing synergy between experiment and computation, where automated analysis, quantum chemical calculations, and machine learning are dramatically enhancing the accuracy and efficiency of structure determination. We also highlight NMR’s broadening scope in areas ranging from complex mixtures and natural products to biomolecular and materials science. Full article

Naphthyridine derivatives have emerged as privileged scaffolds with diverse pharmacological activities, particularly in anticancer and antiviral drug discovery. In this study, a series of naphthyridine-based derivatives (1–10b) was designed, synthesized, and structurally characterized using IR, ¹H/¹³C NMR, and mass spectrometry, and evaluated as dual-function antiproliferative and anti-fowlpox virus agents supported by integrated computational analyses. The synthesized compounds were screened for in vitro antiproliferative activity against HeLa, HCT-116, and MCF-7 cancer cell lines, as well as normal WI-38 lung fibroblasts. Several derivatives exhibited potent cytotoxic activity with enhanced selectivity toward cancer cells. Compound 5b showed the highest activity against HeLa cells, compound 1 was most effective against HCT-116 cells, while compounds 7 and 8 displayed remarkable activity against MCF-7 cells, with compound 7 surpassing doxorubicin and compound 8 demonstrating excellent selectivity toward normal cells. Mechanistic investigations revealed that compounds 7 and 8 acted as dual topoisomerase I/IIβ inhibitors, inducing G2/M cell cycle arrest and intrinsic apoptosis associated with caspase-9 activation and downregulation of topoisomerase II protein expression. Selected derivatives were further evaluated for antiviral activity against fowlpox virus using in ovo and in vivo SPF embryonated chicken egg models, where compounds 2 and 9a exhibited the highest therapeutic indices, comparable to ribavirin, and compound 9a markedly suppressed viral replication and titers in vivo. ADMET profiling, molecular docking, molecular dynamics simulations, and DFT calculations supported the experimental findings and identified compound 10a as the most favorable theoretical candidate. Overall, this integrated experimental–computational approach establishes naphthyridine derivatives as a rationally designed multifunctional chemotype for simultaneous anticancer and antiviral drug development. Full article

Background: There is currently increasing interest in atypical opioid compounds capable of expanding their clinical applications beyond pain management, including the treatment of psychiatric disorders and substance abuse. In this context, the cyclotetrapeptide c[Trp-Phe-D-Pro-Phe] (CJ-15,208, 1) and its derivatives represent an unusual class of opioid peptides. This compound was found to be a mixed KOR/MOR antagonist in vitro, but it acted as an agonist in vivo. For its diverse analogues, it appeared that receptors’ affinity, selectivity, and agonist/antagonist activity greatly varied upon modifications to backbone geometry and the 3D display of pharmacophores. Methods: We utilized NMR, molecular dynamics, and molecular docking to analyze 3D structures and pharmacodynamic properties of selected representative cyclopeptide analogues of 1. Results: The simulations support that, despite its contradictory functional activity in vitro and in vivo, 1 can bind to the active conformation of receptors in an agonist-like fashion. In general, Trp appeared to be the fundamental pharmacophore in the ligand–receptor complexes. In particular, agonists showed a direct interaction between the indole ring and the carboxylate of the conserved Asp(3:32). Conclusions: These studies support a distinctive pharmacodynamic model for this class of compounds, potentially useful for the design of opioid compounds with novel binding/activity profiles and improved therapeutic effects. Full article

Mitochondria play a crucial role in cellular bioenergetics, signaling, and metabolism; yet, many fundamental mechanisms such as the proton transfer along the membranes, the link between membrane curvature and oxidative phosphorylation, and the nanoscale organization of enzyme supercomplexes remain poorly understood due to the limitations of classical biochemical approaches. This review addresses this gap by systematically analyzing the contemporary physical methods used to investigate the mitochondrial structure and function from the micro to nano scale. It covers advanced fluorescence and super-resolution microscopy, electron and volume electron microscopy, and scanning probe techniques, as well as cryo-electron tomography for resolving supramolecular assemblies in near-native conditions. The review highlights the applications of the modern fluorescent probes, expansion and phase microscopy, and machine-learning-based image analysis for a quantitative assessment of the mitochondrial morphology, membrane potential, and dynamics in living cells and tissues. Complementary spectroscopic and scattering methods, including Raman spectroscopy, NMR, and X-ray and neutron scattering, are discussed as tools for probing the redox state, metabolite composition, and membrane organization. Emphasis is placed on integrating high-resolution experimental data with advanced computational frameworks to test competing models of mitochondrial function and pathology, and to guide the development of biomimetic and biomedical technologies. Full article

Natural gas hydrate (NGH) deposits represent a vast and clean energy source. However, sustainable gas production from these resources remains an unsolved technical problem due to potential geohazards and climate challenges. A critical issue in this regard is the difficulty of obtaining in situ samples, which are essential for detailed laboratory studies of NGH’s geomechanical and chemical behavior for safe and green gas production after hydrate dissociation. Currently, the retrieval of representative samples from NGH reservoirs is hindered by significant technological limitations and high costs. Consequently, laboratory-synthesized gas hydrate-bearing sediment (HBS) samples are crucial for controlled research purposes and validating numerical simulation models and are used in the majority of research studies. With this in mind and considering the complexity of synthesizing HBS samples, this study comprehensively reviews different methods of synthesizing gas hydrates in porous media, including excess-gas, excess-water, dissolved-gas, spray, bubble injection, and hybrid techniques. Each method produces distinct hydrate morphologies (e.g., pore-filling, cementing, grain-coating, etc.) and saturation levels, with trade-offs in speed, uniformity, reproducibility, and ease of control. Furthermore, the current review details the synergic application of non-invasive characterization techniques, i.e., X-ray Computed Tomography (CT) and Nuclear Magnetic Resonance (NMR), in studying gas hydrates. CT provides high-resolution three-dimensional (3D) structural images of pore geometry and hydrate distribution, while NMR/MRI (Magnetic Resonance Imaging) quantifies fluid saturations and tracks hydrate formation/dissociation dynamics in real time. The synergistic use of CT and NMR offers a powerful multimodal approach, overcoming individual limitations such as CT’s poor hydrate–water contrast detection and NMR’s indirect hydrate inference, which could help in the sustainable synthesis of particular hydrate morphologies. Finally, the critical analysis of current technological challenges or gaps and also the emerging trends and future directions in the study of HBS, including advanced imaging techniques, AI-assisted analysis, and standardization efforts, etc., are discussed. It was found that the selection of the most appropriate method for natural gas hydrate synthesis is mostly task-specific, and the emerging technologies have facilitated the synthesis of HBS samples with more precise control of morphology, saturation, etc. This review provides the required insights for sustainable synthesis and characterization of hydrate-bearing sediments samples and serves sustainable gas production from natural gas hydrate reservoirs. Full article

Tubulin is a heterodimeric protein composed of α- and β-subunits, which polymerize to form the cell’s microtubules. The latter are key components in mitotic spindle formation and essential targets in anticancer therapy. Compounds such as paclitaxel, tubulysins, dolastatins and synthetic analogues of these latter compounds, including cemadotin, exert their cytotoxic effects by disrupting microtubule dynamics. Previously, we reported the production and anticancer activity of a library of cemadotin analogues featuring a C-terminal tertiary amide functionalized with a variety of N-substituents, thus resulting in compounds occurring as a mixture of amide rotamers. Here we describe a comprehensive NMR and conformational study that provides new insights into the effect of the conformational equilibrium on the binding mode of the novel cemadotin analogues to the tubulin target. The conformational behavior of the isomer equilibrium of cemadotin’s terminal amide bond was investigated by TOCSY and ROESY NMR experiments, which allowed the identification and quantification of individual rotamer populations. A slow interconversion between the s-cis and s-trans amide rotamers was observed under standard NMR conditions (25 °C), indicating a significant energy barrier and conformational rigidity. Molecular docking and saturation transfer difference (STD) NMR experiments were performed with a representative analogue and tubulin to assess the binding mode. The results revealed that the s-trans rotamer is the predominant conformer in solution and exhibits a more favorable interaction with tubulin compared to the s-cis isomer, thus helping to understand the conformational requirements for an improved tubulin binding and the inhibition of the polymerization process. Full article

Unlocking the potential of polymer blends requires innovative strategies that transcend simple mixing. This study presents a novel approach by creating hybrid blends of epoxy and structurally compatible in situ synthesized epoxy acrylate (vinyl ester) resins, further reinforced with halloysite nanotubes (HNTs). We went beyond simple blending by synthesizing the epoxy acrylate (EA) component from the base epoxy resin, ensuring molecular-level compatibility. The epoxy acrylate was successfully synthesized via a ring-opening reaction, as confirmed by FTIR and ¹H-NMR. A series of blends at varying weight ratios of epoxy/epoxy acrylate (75/25, 50/50, and 25/75) was prepared and optimized using dynamic mechanical analysis (DMA) for the best viscoelastic performance and subsequently reinforced with 2 wt% HNTs. Our findings reveal that this unique approach fosters highly interpenetrated polymer networks (IPNs), as evidenced by thermal and viscoelastic behavior. The hybrid epoxy nanocomposite with a 75/25 blend ratio exhibits a superior balance of properties, demonstrating a synergistic enhancement in both thermal and thermomechanical properties compared to the neat epoxy and epoxy acrylate networks. The optimized hybrid epoxy composite exhibits a 147% increase in storage modulus (E′) and a 180% increase in loss modulus (E″) over the neat epoxy composite while enhancing thermal stability. This study not only presents HNT-reinforced epoxy/epoxy acrylate as a new family of robust hybrid nanocomposites but also provides a fundamental blueprint for compatibilizing and reinforcing thermoset blends for advanced applications. Full article

Understanding Li⁺ solvation structure is critical for the rational design of high- and localized high-concentration electrolytes. Here, we present a systematic investigation of tetrahydrofuran (THF)-based electrolytes with lithium bis(trifluoromethanesulfonyl)imide (LiTFSI) using Raman spectroscopy and ⁷Li nuclear magnetic resonance to investigate the local solvation structures. By varying the THF:LiTFSI molar ratio, we observed a transition of Li⁺ solvation from solvent-separated ion pairs to contact ion pairs and aggregates, accompanied by increased structural heterogeneity and constrained local dynamics. Raman spectroscopy captures the evolution of Li⁺–anion coordination with increasing salt concentration, while ⁷Li NMR chemical shifts, line widths, and relaxation times provide complementary insight into changes in the electronic environment and symmetry of Li⁺ coordination. Electrolyte structure is further examined by introducing a hydrofluoroether co-solvent into a concentrated (THF)₂–LiTFSI electrolyte. Raman results show that the local Li⁺–TFSI⁻ coordination structure is preserved upon 1,1,2,2-tetrafluoroethyl-2,2,3,3-tetrafluoropropyl ether (TTE) addition, whereas NMR reveals subtle modifications of the ion-rich solvation clusters. These results provide fundamental insight into Li⁺ solvation and electrolyte localization, offering general design principles for advanced electrolyte systems. Full article

Liquid–liquid phase separation (LLPS) underlies the formation of membraneless cellular compartments, yet experimental strategies that directly connect quantitative LLPS behavior with residue-level structural information remain limited. Here, we present an integrated protocol that combines quantitative LLPS assays with nuclear magnetic resonance (NMR) spectroscopy and structure-guided mutagenesis to regulate protein phase separation. Using the VAPB MSP domain as a representative example, this workflow links residue-specific structural features to macroscopic LLPS behavior and enables suppression or enhancement of phase separation through targeted amino acid substitutions. This protocol provides a generalizable framework for systematic, residue-level regulation of protein LLPS. Full article

Background: Pentamidine isethionate (PTM) and miltefosine (MF) are clinically relevant antiparasitic agents whose use is limited by toxicity, emerging resistance, and the lack of effective co-delivery strategies. Tetronic^® 1307 (T1307), an amphiphilic and thermoresponsive block copolymer, was investigated as a carrier to enable their combination therapy. Methods: PTM and MF were formulated in T1307-based micelles and thermoresponsive gels. The systems were characterized by small-angle neutron scattering (SANS), dynamic light scattering (DLS), and nuclear magnetic resonance spectroscopy (NMR). Antiparasitic activity was evaluated against Leishmania major promastigotes. Results: MF formed stable micelles that efficiently incorporated PTM, generating a “drug-in-drug” architecture. While T1307 alone showed limited PTM loading, MF promoted mixed micelle formation and enhanced PTM incorporation. At physiological temperature and adequate copolymer concentrations, drug-loaded micelles formed thermoreversible gels suitable for topical application. The combined formulations preserved drug activity and exhibited synergistic effects against L. major. Conclusions: T1307 is a promising platform for the co-delivery of PTM and MF, enabling synergistic combination therapy and thermoresponsive gel formation with potential to reduce systemic toxicity and improve treatment administration. Full article

Hydroxyl-terminated polybutadiene (HTPB) propellants are widely used in aerospace applications owing to their excellent mechanical performance and storage stability, which are primarily governed by the crosslinked network formed during curing. Understanding the evolution of this network is therefore essential for optimizing propellant formulations and curing parameters. In this work, the curing behaviors of HTPB-based propellant slurries employing two representative curing agents, toluene diisocyanate (TDI) and isophorone diisocyanate (IPDI), were systematically investigated under isothermal conditions at 60 °C using low-field nuclear magnetic resonance (LF-NMR), combined with infrared spectroscopy, dynamic mechanical analysis, and macroscopic mechanical testing. The curing time and crosslink density of both propellant systems were quantitatively determined by LF-NMR crosslink densitometry, while transverse relaxation time measurements were used to monitor the mobility evolution of different molecular segments during curing. The results show that with increasing curing time, the crosslink density and crosslinked chain content progressively increased, whereas the free chain content decreased, accompanied by a transient increase and subsequent decrease in dangling chains. The curing endpoints of the HTPB/TDI and HTPB/IPDI propellants were determined to be approximately 1.25 days and 5.5 days, with corresponding final crosslink densities of 2.438 × 10⁻⁴ and 2.007 × 10⁻⁴ mol mL⁻¹, respectively. Excellent agreement between LF-NMR results and complementary characterization techniques confirms LF-NMR as an effective tool for studying curing reaction and network evolution in complex solid propellant systems. Full article

Thymosin β4 (Tβ4) is a highly acidic, intrinsically disordered 43-amino-acid peptide with diverse biological functions, yet its interactions with metal ions remain poorly understood. In this study, we provide the first experimental demonstration that Tβ4 forms discrete Zn²⁺-bound adducts and undergoes Zn²⁺-induced aggregation under physiological pH conditions. Combining zeta potential analysis, dynamic light scattering (DLS), electrospray ionization mass spectrometry (ESI-MS), nuclear magnetic resonance (NMR) spectroscopy, and scanning electron microscopy with elemental mapping (SEM/EDS), we show that Zn(II) binding progressively neutralizes Tβ4’s negative surface charge and triggers a sharp aggregation transition. ESI-MS unambiguously identifies Tβ4/Zn(II) complexes of peptide-to-zinc molar ratio 1:3, while DLS and SEM reveal the formation of compact, low-solubility supramolecular assemblies. NMR measurements support a metal-induced aggregation, confirming the absence of folding upon Zn(II) binding. By quantitatively comparing the experimentally determined critical aggregation concentration with physiologically observed extracellular Zn(II) ranges, we demonstrate that aggregation is unlikely in plasma or basal interstitial environments but may become feasible in Zn-rich microdomains, such as the synaptic cleft, where transient Zn(II) levels can exceed 1 μM. These findings introduce a previously unrecognized dimension of Tβ4 chemistry and suggest that a Zn(II)-mediated supramolecular assembly of Tβ4 could influence peptide behavior in neurological or inflammatory conditions characterized by elevated extracellular Zn(II). This work establishes a foundational biochemical framework for future studies aimed at elucidating the biological implications of Tβ4/Zn(II) complexation and aggregation in vivo. Full article

Tight conglomerate reservoirs exhibit strong pore-scale heterogeneity and extremely low permeability, in which spontaneous imbibition is primarily governed by capillary and viscoelastic effects. In this study, the imbibition dynamics of four representative fracturing fluid systems, including slickwater, 3% potassium chloride (KCl) brine, hydrolyzed polyacrylamide (HPAM) viscoelastic fluid, and a nanoemulsion (NE), were investigated using a temperature-controlled nuclear magnetic resonance (NMR) monitoring system. This approach enables real-time quantification of fluid uptake and pore-scale redistribution through time-resolved T₂ spectral analysis. The experimental results reveal a three-stage imbibition process consisting of rapid capillary-driven uptake, viscoelastic-retarded transition, and final equilibrium. Among the four fracturing fluid systems, the nanoemulsion exhibits the lowest interfacial tension (1.72 mN/m), the strongest wettability alteration, and the highest equilibrium recovery (0.76), which is nearly 80% greater than that of slickwater. Based on these observations, a multiscale capillary–viscoelastic coupling model was developed by extending the Lucas–Washburn framework to incorporate pore-size distribution, time-dependent wettability evolution, and viscoelastic damping. The model fits the experimental data well (R² > 0.90) and identifies viscosity as the most influential parameter controlling the imbibition rate (sensitivity = 0.78). Energy analysis further indicates that capillary energy dominates the early stage, whereas viscoelastic energy storage sustains fluid transport during the later stage. SEM observations were further used to qualitatively corroborate pore heterogeneity and pore–mineral associations, supporting the NMR-based pore-scale interpretation. This study provides a quantitative framework for describing non-Newtonian capillary flow in tight conglomerate rocks and enhances the understanding of capillary–viscoelastic interactions relevant to multiphase fluid migration. Full article

Pomegranate peel, accounting for 35–50% of the fruit weight, is an underutilized agri-food by-product. This study applied, for the first time, fermentation with Saccharomyces cerevisiae as a simple and sustainable strategy to simultaneously obtain tannin-rich extracts and polysaccharide fractions with potential prebiotic activity. Peels from two cultivars, Wonderful and G1, differing in peel thickness, were subjected to three fermentation protocols (air- and not air-exposed) and monitored at 25 °C over 48 and 72 h. HPLC-DAD analysis showed that yeast-inoculated fermentation increased total tannin concentration in dry extracts (up to 70%) without inducing chemical modifications to tannin profiles. As determined by Dynamic Light Scattering, fermentation promoted significant depolymerization of native polysaccharides, while DOSY-¹H-NMR analyses revealed the presence of reduced molecular weight fractions down to 26 kDa. In vitro growth assays confirmed that fermented polysaccharides were more efficiently utilized as a carbon source by Bifidobacterium breve and Lactiplantibacillus plantarum compared to non-fermented controls, likely thanks to polysaccharide depolymerization induced by fermentation. The study demonstrated that air-exposed S. cerevisiae fermentation was an effective process alternative to chemical or enzymatic hydrolysis for modifying pomegranate peel pectin directly within a complex matrix, while simultaneously enhancing tannin recovery. This approach represents a possible sustainable strategy for pomegranate peel valorization into functional ingredients. Full article

Soil organic matter (SOM) molecular composition governs its stability and ecological functions in forest ecosystems. Nevertheless, how land-use changes (LUCs) regulate the SOM molecular composition remains poorly understood, particularly the underlying mechanisms mediated by soil properties. This study investigated the effects of LUCs on SOM molecular composition in a subtropical coastal region and examined the driving roles of soil nutrient availability and enzyme activities. The research was conducted in Huanghai National Forest Park, Jiangsu Province, China, focusing on four land-use types converted from historical wheat cropland (W, as control): monoculture plantations of Ginkgo biloba (G) and Metasequoia glyptostroboides (M), a ginkgo–metasequoia mixed forest (GM), and a ginkgo–wheat agroforestry system (GW). Soil samples were collected from 0 to 20 cm and 20–40 cm layers and analyzed for SOM molecular compositions using solid-state ¹³C nuclear magnetic resonance (NMR) spectroscopy. Soil chemical properties and enzyme activity activities were also determined, with redundancy analysis (RDA) and correlation analysis applied to identify key influencing factors. Results demonstrated that LUCs significantly altered SOM molecular composition. The GW system exhibited the highest proportion of labile O-alkyl carbon (42.65%), while the M plantation accumulated greatest levels of stable aromatic carbon (up to 49.25%). During the initial decades following afforestation, soil nutrient availability and enzyme activities were confirmed as pivotal drivers of SOM molecular variation. Specifically, available potassium (AK), ammonium nitrogen (AN), and the carbon/phosphorus (C/P) ratio were significantly correlated with specific SOM components (p < 0.05). The elevated O-alkyl carbon proportion in GW was closely associated with its higher invertase activity. Notably, vertical differentiation in SOM stability was observed across land-use types, with the agroforestry system achieving the highest carbon pool management index in surface soil but showing a weakened capacity for subsoil C stabilization. RDA further confirmed that AK and AN were dominant factors shaping SOM molecular composition. In conclusion, LUCs modulate SOM chemical composition and stability primarily through altering soil nutrient availability and associated enzyme activities. Agroforestry system facilitates labile C accumulation in surface soil, whereas monoculture plantations are more conducive to stable C sequestration, especially in subsoil layers. These findings provide novel mechanistic insights into SOM dynamics following LUCs and offer a theoretical basis for formulating tailored management strategies to enhance C sequestration efficiency in subtropical coastal ecosystems. Full article