RESUMEN
BACKGROUND: Melanoma is a highly invasive skin cancer with limited treatment strategies. Bupivacaine, a commonly used local anesthetic recognized for its safety, has shown promise in combating tumors. 3'-phosphoadenosine 5'-phosphosulfate synthase 2 (PAPSS2) is a key enzyme in the sulfation process and is associated with the development and metastasis of various tumors. This study aimed to explore the mechanism by which bupivacaine inhibits melanoma proliferation and metastasis by targeting PAPSS2. METHODS: The effects of bupivacaine on the proliferation of A375 and A2058 melanoma cells were evaluated using Cell Counting Kit-8 (CCK-8), 5-Ethynyl-2'-deoxyuridine (EdU) labeling, and clonogenic assays. Cell migration, invasion, and PAPSS2 expression were evaluated using Transwell experiments and Quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR) analysis. Additionally, an in vivo melanoma tumor model in nude mice was constructed to evaluate the impact of bupivacaine on melanoma growth and metastasis. Immunohistochemistry was used to assess tumor metastasis and PAPSS2 expression levels in the nude mouse model. RESULTS: Experimental results demonstrated that bupivacaine significantly inhibited melanoma proliferation and invasion compared to the control group. Notably, this inhibitory effect was partially reversed by PAPSS2 overexpression. In vivo experiments demonstrated that bupivacaine-treated nude mice exhibited reduced tumor volumes, weights, and fewer lung metastatic foci. Molecular analysis via qRT-PCR and immunohistochemistry analysis further indicated that bupivacaine significantly reduced PAPSS2 in tumor tissues. CONCLUSION: This study confirms that bupivacaine, a local anesthetic, can inhibit melanoma proliferation and metastasis by targeting the PAPSS2 signaling pathway. These findings suggest its potential as an anti-tumor medication and present new treatment strategies for melanoma.
Asunto(s)
Anestésicos Locales , Bupivacaína , Proliferación Celular , Melanoma , Ratones Desnudos , Animales , Humanos , Proliferación Celular/efectos de los fármacos , Melanoma/patología , Melanoma/tratamiento farmacológico , Bupivacaína/farmacología , Ratones , Línea Celular Tumoral , Anestésicos Locales/farmacología , Metástasis de la Neoplasia , Movimiento Celular/efectos de los fármacos , Ensayos Antitumor por Modelo de Xenoinjerto , Neoplasias Cutáneas/patología , Neoplasias Cutáneas/tratamiento farmacológicoRESUMEN
Plasma membranes not only maintain the intracellular microenvironment through their phospholipid bilayer but also eliminate exogenous compounds outside the cell membranes. Most drugs especially with high polarity are prevented from entering into cells to exert their effects. Therefore, it is of great significance to design effective drug carriers with a penetrating ability toward plasma membranes. In this study, a dual-templated MIP (dt-MIPs) carrier with controllable microstructure and high drug loading capacity was prepared using highly expressed sphingomyelin on the plasma membrane and tenofovir (TFV), a first-line drug for HIV and chronic hepatitis B, as template molecules. The drug release experiments performed in vitro under simulated physiological conditions demonstrated that sustained and stable adsorption of TFV on dt-MIPs was more than 80% over 50 h. By a combination of flow cytometry and confocal microscopy, dt-MIPs were found to have efficient cell permeability. Furthermore, mass-spectrometry-based intracellular pharmacokinetic studies demonstrated that TFV was delivered completely into cells within 30 min with the delivery of dt-MIPs. The study presented above suggested that dt-MIPs are expected to be alternative nanoscale drug carriers for enhanced drug permeability and controlled release.
Asunto(s)
Membrana Celular , Portadores de Fármacos , Esfingomielinas , Esfingomielinas/química , Portadores de Fármacos/química , Membrana Celular/metabolismo , Membrana Celular/química , Humanos , Tenofovir/química , Tenofovir/farmacocinética , Liberación de FármacosRESUMEN
The HIV-1 envelope is a heavily glycosylated class 1 trimeric fusion protein responsible for viral entry into CD4+ immune cells. Developing neutralizing antibodies against the specific envelope glycans is an alternative method for antiviral therapies. This work presents the first-ever development and characterization of artificial neutralizing antibodies using molecular imprinting technology to recognize and bind to the envelope protein of HIV-1. The prepared envelope glycan-imprinted nanoparticles (GINPs) can successfully prevent HIV-1 from infecting target cells by shielding the glycans on the envelope protein. In vitro experiments showed that GINPs have strong affinity toward HIV-1 (Kd = 36.7 ± 2.2 nM) and possess high anti-interference and specificity. GINPs demonstrate broad inhibition activity against both tier 1 and tier 2 HIV-1 strains with a pM-level IC50 and exhibit a significant inhibitory effect on long-term viral replication by more than 95%. The strategy provides a promising method for the inhibition and therapy of HIV-1 infection.
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Infecciones por VIH , VIH-1 , Humanos , Anticuerpos Neutralizantes , Anticuerpos Anti-VIH/metabolismo , Glicosilación , Infecciones por VIH/tratamiento farmacológico , Polisacáridos/metabolismoRESUMEN
Molecular imprinting polymers (MIPs) are synthetic receptors as biomimetic materials for various applications ranging from sensing to separation and catalysis. However, currently existing MIPs are stuck to some of the issues including the longer preparation steps and poor performance. In this report, a facile and one-pot strategy by integrating the in-situ growth of magnetic nanoparticles and reversed phase microemulsion oriented molecularly imprinting strategy to develop magnetic molecular imprinted nanocomposites was proposed. Through self-assembling of the template, it brought up highly ordered and uniform arrangement of the imprinting structure, which offered faster adsorption kinetic as adsorption equilibrium was achived within 15 min, higher adsorption capacity (Qmax = 48.78 ± 1.54 µmol/g) and high affinity (Kd = 127.63 ± 9.66 µM) toward paradigm molecule-adenosine monophosphate (AMP) compared to the conventional bulk imprinting. The developed MIPs offered better affinity and superior specificity which allowed the specific enrichment toward targeted phosphorylated peptides from complex samples containing 100-fold more abundant interfering peptides. Interestingly, different types of MIPs can be developed which could targetly enrich the specific phosphorylated peptides for mass spectrometry analysis by simply switching the templates, and this strategy also successfully achieved imprinting of macromolecular peptides. Collectively, the approach showed broad applicability to target specific enrichment from metabolites to phosphorylated peptides and providing an alternative choice for selective recognition and analysis from complex biological systems.
Asunto(s)
Impresión Molecular , Polímeros , Polímeros/química , Péptidos , Sustancias Macromoleculares , Adsorción , Impresión Molecular/métodosRESUMEN
Phospholipids, as fundamental building blocks of the cell membrane, play important roles for molecule transportation, cell recognition, etc. However, due to the structural diversity and amphipathic nature, there are few methods for the specific recognition of lipids as compared to other biomolecules such as proteins and glycans. Herein, we developed a molecular imprinting strategy for controllable imprinting toward the polar head of phospholipid exposed on the surface of cellular membranes for recognition. Phosphatidylserine, as unique lipid on the outer membrane leaflet of exosome and also hallmark for cell apoptosis, was imprinted with the developed method. The phosphatidylserine imprinted materials showed high efficiency and specific targeting capability not only for apoptotic cell imaging but also for the isolation of exosomes. Collectively, the synthesized molecularly imprinted materials have great potential for selective plasma membrane recognition for targeted drug delivery and biomarker discovery.
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Impresión Molecular , Fosfolípidos , Epítopos/química , Fosfatidilserinas , Membrana Celular , Impresión Molecular/métodosRESUMEN
Phosphorylation is one of the most important post-translational modifications of proteins, but due to the low abundance of phosphopeptides, enrichment is an essential step before mass spectrometric analysis. Although there are a number of enrichment methods developed targeting different forms of proteins phosphorylations, there are few reports on specific recognition and capture of single phosphopeptide. Herein, based on the advantages of dual affinity of TiO2 and urea to a phosphate group and molecular imprinting towards the peptide sequence, the precise recognition of intact phosphorylated peptides was successfully achieved. The same peptide sequence with different phosphorylation forms (c.a. Ser, Thr and Tyr) were used as templates for proof-of-principle study, and the imprinted particles were successfully synthesized, characterized, and have the capacity to specifically recognize the targeted unique phosphorylation excluding even its isoforms. In addition, the produced molecularly imprinted nanoparticles have numerous important advantages, including strong affinity, high specificity toward single phosphopeptides, tolerance to interferences, fast binding kinetics, substantial binding capacity, excellent stability and reusability, making them an ideal sorbent for specific enrichment of unique phosphopeptides. Finally, different phosphorylation forms were specifically enriched from both standard peptides' mixture and casein/milk digests.
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Impresión Molecular , Nanopartículas , Espectrometría de Masas , Nanopartículas/química , Fosfopéptidos/análisis , Fosforilación , Isoformas de Proteínas/metabolismo , Titanio/químicaRESUMEN
Surface coatings are extensively applied on biomedical devices to provide protection against biofouling and infections. However, most surface coatings prevent both bacteria and cells interactions with the biomaterials, limiting their uses as implants. Furthermore, damage to the surface such as scratches and abrasions can happen during transport and clinical usage, resulting in the loss of antibacterial property. In this work, we introduce an efficient method to fabricate stable anti-infective and self-healable multilayer coatings on stainless steel surface via a three-step procedue. Firstly, modified polyethyleneimine (PEI) and poly(acrylic acid) (PAA), both contain pendant furan groups, were deposited on the surface using Layer-by-Layer (LbL) self-assembly technique. Secondly, the polymer layers were cross-linked, via Diels-Alder cycloaddition, using a bismaleimide poly(ethylene glycol) linker, to enhance the stability of the coatings. Thirdly, the Diels-Alder adduct was utilised in the thiol-ene click reaction for post-modification of the coatings, which allowed for the grafting of antimicrobial poly(hexamethylene biguanide) (PHMB) and ε-poly(lysine) (EPL). The resultant multilayer coatings not only exhibited rapid self-healing property, with complete scratch closure within 30 min, but also demonstrated effective antibacterial activity against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). In addition, biofouling of bovine serum albumin was found to be inhibited on the coated surfaces. Furthermore, these coatings showed no toxicity effect towards seeded osteoblastic cells (MC3T3-E1) and evidence of anti-inflamatory activity when tested against macrophage cell line U-937. Our coating method thus represents an effective strategy for the anti-infective protection of biomedical-devices having direct contact with tissues.
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Antiinfecciosos , Incrustaciones Biológicas , Antibacterianos/farmacología , Antiinfecciosos/farmacología , Materiales Biocompatibles Revestidos/farmacología , Escherichia coli , Staphylococcus aureus , Propiedades de SuperficieRESUMEN
Dielectric metasurfaces, which are capable of manipulating incident light, have been a novel branch of flat optics. This modulation ability is realized by nanostructures with space-variant geometrical parameters such as height and diameter. Therefore, accurate profile measurement of metasurfaces is of great importance. White-light scanning interferometry is widely used for profile measurement. The step height is retrieved by locating the envelope's peak. However, spurious fringes attached to the desired fringes were observed at the measured area near the edge of nanostructures. Their amplitude distributions vary with the density of nanostructures as well as distance to the edge. Further, anomalous coherence signals with two fringe envelopes are produced, which result in inaccurate measurement results. We attributed this phenomenon to the complex light modulation by the nanostructures. When referring to the anomalous coherence signals for the top of the nanostructures, one envelope is produced by the top, and the other is produced by the bottom; however, it is difficult to distinguish these two, which is the same case for the bottom of the nanostructures. To automatically solve these obstacles, a signal processing method, which integrates the image segmentation technology to identify and divide the anomalous coherence signals, along with a Morlet wavelet transform to extract the fringe envelope, suitable for any measured area of the dielectric metasurface, is proposed. One metasurface belt consisting of seven kinds of nanopillars with varying arrayed densities that produce different coherence signals is measured. The diameter distribution ranges from 500 to 1250 nm with a constant height of 1850 nm. The local periods in the X and Y directions are 3020 and 1740 nm, respectively. Measurement results demonstrate the validity of the proposed method for spurious fringes processing.
RESUMEN
Tyrosine phosphorylation regulates the upstream signaling pathway but accounts for less than 0.1% of total phosphorylation in human cells. Herein, molecularly imprinted mesoporous materials were first synthesized to recognize the phosphorylated tyrosine residue from other phosphorylated residues.