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Journal of Structural Biology: X Jun 2024NMR spectroscopy has played a pivotal role in fragment-based drug discovery by coupling detection of weak ligand-target binding with structural mapping of the binding...
NMR spectroscopy has played a pivotal role in fragment-based drug discovery by coupling detection of weak ligand-target binding with structural mapping of the binding site. Fragment-based screening by NMR has been successfully applied to many soluble protein targets, but only to a limited number of membrane proteins, despite the fact that many drug targets are membrane proteins. This is partly because of difficulties preparing membrane proteins for NMR-especially human membrane proteins-and because of the inherent complexity associated with solution NMR spectroscopy on membrane protein samples, which require the inclusion of membrane-mimetic agents such as micelles, nanodiscs, or bicelles. Here, we developed a generalizable protocol for fragment-based screening of membrane proteins using NMR. We employed two human membrane protein targets, both in fully protonated detergent micelles: the single-pass C-terminal domain of the amyloid precursor protein, C99, and the tetraspan peripheral myelin protein 22 (PMP22). For both we determined the optimal NMR acquisition parameters, protein concentration, protein-to-micelle ratio, and upper limit to the concentration of D-DMSO in screening samples. Furthermore, we conducted preliminary screens of a plate-format molecular fragment mixture library using our optimized conditions and were able to identify hit compounds that selectively bound to the respective target proteins. It is hoped that the approaches presented here will be useful in complementing existing methods for discovering lead compounds that target membrane proteins.
PubMed: 38883400
DOI: 10.1016/j.yjsbx.2024.100100 -
International Journal of Nanomedicine 2024This review article discusses the potential of nanomaterials in targeted therapy and immunomodulation for stroke-induced immunosuppression. Although nanomaterials have... (Review)
Review
This review article discusses the potential of nanomaterials in targeted therapy and immunomodulation for stroke-induced immunosuppression. Although nanomaterials have been extensively studied in various biomedical applications, their specific use in studying and addressing immunosuppression after stroke remains limited. Stroke-induced neuroinflammation is characterized by T-cell-mediated immunodepression, which leads to increased morbidity and mortality. Key observations related to immunodepression after stroke, including lymphopenia, T-cell dysfunction, regulatory T-cell imbalance, and cytokine dysregulation, are discussed. Nanomaterials, such as liposomes, micelles, polymeric nanoparticles, and dendrimers, offer advantages in the precise delivery of drugs to T cells, enabling enhanced targeting and controlled release of immunomodulatory agents. These nanomaterials have the potential to modulate T-cell function, promote neuroregeneration, and restore immune responses, providing new avenues for stroke treatment. However, challenges related to biocompatibility, stability, scalability, and clinical translation need to be addressed. Future research efforts should focus on comprehensive studies to validate the efficacy and safety of nanomaterial-based interventions targeting T cells in stroke-induced immunosuppression. Collaborative interdisciplinary approaches are necessary to advance the field and translate these innovative strategies into clinical practice, ultimately improving stroke outcomes and patient care.
Topics: Animals; Humans; Cytokines; Nanomedicine; Nanoparticles; Nanostructures; Stroke; T-Lymphocytes
PubMed: 38882535
DOI: 10.2147/IJN.S456632 -
RSC Advances Jun 20241,2,3-Triazole-based gemini bis-quaternary ammonium surfactants with varying hydrophobic chain length and ethylene or butylene spacers were synthesized and...
1,2,3-Triazole-based gemini bis-quaternary ammonium surfactants with varying hydrophobic chain length and ethylene or butylene spacers were synthesized and characterized. Their surface and aggregation properties were analyzed using tensiometry and conductimetry. The gemini surfactants showed significantly lower CMC values compared to their single-tail counterparts and conventional gemini surfactants described in the literature. The micellization and surface adsorption processes in water can be tailored according to the hydrophobic chain and the spacer length and were substantially improved by the presence of the heterocycle. These surfactants are active against various Gram-positive and Gram-negative bacterial stains, as well as fungi. The gemini surfactant with tetradecyl chain and ethylene spacer (Bis 14-2-14) exhibited the highest activity against all investigated microbial strains.
PubMed: 38882484
DOI: 10.1039/d4ra02177k -
ACS Polymers Au Jun 2024The precise sequence of a protein's primary structure is essential in determining its folding pathways. To emulate the complexity of these biomolecules, functional block...
The precise sequence of a protein's primary structure is essential in determining its folding pathways. To emulate the complexity of these biomolecules, functional block copolymers consisting of segmented triblocks with distinct functionalities positioned in a sequence-specific manner are designed to control the polymer chain compaction. Triblock polymers and and random diblock copolymer consist of a hydrophilic poly(ethylene oxide) (PEO) block and a hydrophobic block with coumarin () and ferrocene () moieties that are grafted in a sequence-specific or random manner onto the hydrophilic block. External stimuli such as UV light, redox, and chemical cues influence the functional hydrophobic block to alter the packing parameters that are monitored with spectroscopic and scattering techniques. Interestingly, the positioning of the stimuli-responsive moiety within the hydrophobic block of , , and affects the extent of the hydrophobic-hydrophilic balance in block copolymers that renders orthogonal control in stimuli-responsive transformation of self-assembled vesicles to micelles.
PubMed: 38882035
DOI: 10.1021/acspolymersau.4c00009 -
Journal of Pharmaceutical and... Jun 2024Omeprazole (OME) is a proton pump inhibitor used to treat gastroesophageal reflux disease associated conditions. The current study presents an Analytical Quality by...
Omeprazole (OME) is a proton pump inhibitor used to treat gastroesophageal reflux disease associated conditions. The current study presents an Analytical Quality by Design-based approach for the development of a CE method for OME impurity profiling. The scouting experiments suggested the selection of solvent modified Micellar ElectroKinetic Chromatography operative mode using a pseudostationary phase composed of sodium dodecyl sulfate (SDS) micelles and n-butanol as organic modifier in borate buffer. A symmetric three-level screening matrix 3//16 was used to evaluate the effect of Critical Method Parameters, including Background Electrolyte composition and instrumental settings, on Critical Method Attributes (critical resolution values, OME peak width and analysis time). The analytical procedure was optimized using Response Surface Methodology through a Central Composite Orthogonal Design. Risk of failure maps made it possible to define the Method Operable Design Region, within which the following optimized conditions were selected: 72 mM borate buffer pH 10.0, 96 mM SDS, 1.45 %v/v n-butanol, capillary temperature 21 °C, applied voltage 25 kV. The method was validated according to ICH guidelines and robustness was evaluated using a Plackett-Burman design. The developed procedure enables the simultaneous determination of OME and seven related impurities, and has been successfully applied to the analysis of pharmaceutical formulations.
PubMed: 38879949
DOI: 10.1016/j.jpba.2024.116295 -
Scientific Reports Jun 2024Acute promyelocytic leukemia (APL) is characterized by rearrangements of the retinoic acid receptor, RARα, which makes all-trans retinoic acid (ATRA) highly effective...
Acute promyelocytic leukemia (APL) is characterized by rearrangements of the retinoic acid receptor, RARα, which makes all-trans retinoic acid (ATRA) highly effective in the treatment of this disease, inducing promyelocytes differentiation. Current therapy, based on ATRA in combination with arsenic trioxide, with or without chemotherapy, provides high rates of event-free survival and overall survival. However, a decline in the drug activity, due to increased ATRA metabolism and RARα mutations, is often observed over long-term treatments. Furthermore, dedifferentiation can occur providing relapse of the disease. In this study we evaluated fenretinide, a semisynthetic ATRA derivative, encapsulated in nanomicelles (nano-fenretinide) as an alternative treatment to ATRA in APL. Nano-fenretinide was prepared by fenretinide encapsulation in a self-assembling phospholipid mixture. Physico-chemical characterization was carried out by dinamic light scattering and spectrophotometry. The biological activity was evaluated by MTT assay, flow cytometry and confocal laser-scanning fluorescence microscopy. Nano-fenretinide induced apoptosis in acute promyelocytic leukemia cells (HL60) by an early increase of reactive oxygen species and a mitochondrial potential decrease. The fenretinide concentration that induced 90-100% decrease in cell viability was about 2.0 µM at 24 h, a concentration easily achievable in vivo when nano-fenretinide is administered by oral or intravenous route, as demonstrated in previous studies. Nano-fenretinide was effective, albeit at slightly higher concentrations, also in doxorubicin-resistant HL60 cells, while a comparison with TK6 lymphoblasts indicated a lack of toxicity on normal cells. The results indicate that nano-fenretinide can be considered an alternative therapy to ATRA in acute promyelocytic leukemia when decreased efficacy, resistance or recurrence of disease emerge after protracted treatments with ATRA.
Topics: Humans; Fenretinide; Leukemia, Promyelocytic, Acute; HL-60 Cells; Apoptosis; Reactive Oxygen Species; Antineoplastic Agents; Nanoparticles; Cell Survival; Micelles; Membrane Potential, Mitochondrial
PubMed: 38877119
DOI: 10.1038/s41598-024-64629-w -
RSC Advances Jun 2024Conventional surfactants encounter limitations for application in oil reservoirs; however, combining surface-active ionic liquids (SAILs) with conventional surfactants...
Conventional surfactants encounter limitations for application in oil reservoirs; however, combining surface-active ionic liquids (SAILs) with conventional surfactants presents an opportunity to enhance the interfacial properties of crude oil-water systems, giving also economic benefits. Accordingly, blends of a long-chain cationic imidazolium-based SAIL, namely, 1-dodecyl-3-methylimidazolium chloride, [Cmim][Cl], and the anionic conventional surfactant, sodium dodecyl sulfate were investigated here. Initial experiments with individual surfactants revealed efficient adsorption and consistent adsorption parameters. Subsequently, the use of mixtures showed synergistic effects for interfacial tension reduction of up to 86.0%, and critical micelle concentration reduction of 72.1% compared to the linear contribution of individual components. These improvements were observed at the optimal SAIL mole fraction of 0.3 and the mixture concentration of 0.003 mol dm, resulting in interfacial tension reduction from 29.1 to 1.6 mN m as well as achieving a low critical micelle concentration of 2.7 × 10 mol dm coinciding with 83.6% synergy. These findings underscore the favorable interactions between oppositely charged components in the mixtures, amplifying their activity beyond the linear contributions of the individual surfactants. Additionally, theoretical assessments using the Gibbs adsorption equation and the Rosen model provided insight into the adsorption behavior of both the individual surfactants and their mixtures, together with reasonable variations in the corresponding parameters.
PubMed: 38873546
DOI: 10.1039/d4ra02092h -
Journal of Molecular Biology Jun 2024TolC is the outer membrane protein responsible for antibiotic efflux in E. coli. Compared to other outer membrane proteins it has an unusual fold and has been shown to...
TolC is the outer membrane protein responsible for antibiotic efflux in E. coli. Compared to other outer membrane proteins it has an unusual fold and has been shown to fold independently of commonly used periplasmic chaperones, SurA and Skp. Here we find that the assembly of TolC involves the formation of two folded intermediates using circular dichroism, gel electrophoresis, site-specific disulfide bond formation and radioactive labeling. First the TolC monomer folds, and then TolC assembles into a trimer both in detergent-free buffer and in the presence of detergent micelles. We find that a TolC trimer also forms in the periplasm and is present in the periplasm before it inserts in the outer membrane. The monomeric and trimeric folding intermediates may be used in the future to develop a new approach to antibiotic efflux pump inhibition by targeting the assembly pathway of TolC.
PubMed: 38871177
DOI: 10.1016/j.jmb.2024.168652 -
Food Chemistry Jun 2024This study explored how co-oleogelator type, concentration, and water addition affect lipid digestion and β-carotene (βC) bioaccessibility in corn oil oleogels....
This study explored how co-oleogelator type, concentration, and water addition affect lipid digestion and β-carotene (βC) bioaccessibility in corn oil oleogels. Oleogels containing 0.1% βC, 20% glyceryl stearate (GS), with lecithin (L) or hydrogenated lecithin (HL) (at 0, 0.5, or 2.5%) and their water-filled counterparts (1% water) were examined. In vitro intestinal digestion revealed HL-oleogels experienced higher lipolysis due to their smaller crystal size enhancing surface area for lipase action, whereas L-oleogels presented lower digestibility, attributed to larger oil droplets and a minimized surface area. Water addition didn't significantly change lipid digestibility. βC bioaccessibility was inversely related to co-oleogelator concentration, with L-oleogels demonstrating the largest decrease, likely due to less free fatty acids released for micelle formation. However, water-filled oleogels enhanced βC bioaccessibility. These findings highlight that tailored microstructure in oleogels can control lipid digestion and βC bioaccessibility, paving the way for designing efficient delivery systems for targeted nutrient delivery.
PubMed: 38870810
DOI: 10.1016/j.foodchem.2024.139978 -
Nanomaterials (Basel, Switzerland) May 2024Small-size gold nanoparticles (AuNPs) are showing large potential in various fields, such as photothermal conversion, sensing, and medicine. However, current synthesis...
Small-size gold nanoparticles (AuNPs) are showing large potential in various fields, such as photothermal conversion, sensing, and medicine. However, current synthesis methods generally yield lower, resulting in a high cost. Here, we report a novel uni-micelle method for the controlled synthesis of monodisperse gold nanocrystals, in which there is only one kind micelle containing aqueous solution of reductant while the dual soluble Au (III) precursor is dissolved in oil phase. Our synthesis includes the reversible phase transfer of Au (III) and "uni-micelle" synthesis, employing a Au (III)-OA complex as an oil-soluble precursor. Size-controlled monodisperse AuNPs with a size of 4-11 nm are synthesized by tuning the size of the micelles, in which oleylamine (OA) is adsorbed on the shell of micelles and enhances the rigidity of the micelles, depressing micellar coalescence. Monodisperse AuNPs can be obtained through a one-time separation process with a higher yield of 61%. This method also offers a promising way for the controlled synthesis of small-size alloy nanoparticles and semiconductor heterojunction quantum dots.
PubMed: 38869525
DOI: 10.3390/nano14110900