RESUMEN
Cu2ZnSnS4 (CZTS) was synthesized following hot injection method and the process was optimized by varying temperature conditions. Four samples at different temperatures viz., 200, 250, 300 and 350 °C were prepared and analyzed using different characterization techniques. Based on the correlation between XRD, Raman and XPS, we conclude that the formation of ZnS and SnS2 occurs at 350 °C but at 200 °C there is no breakdown of the complex as per XRD. According to Raman and XPS analysis, as the temperature rises, the bonds between the metals become weaker, which is visibly seen in Raman and XPS due to the minor peaks of copper sulfide. Scanning electron microscopic analysis confirmed nanometric particles which increase in size with temperature. The photocatalytic evaluation showed that CZTS synthesized at 200 °C performed efficiently in the removal of the two colorants, methylene blue and Rhodamine 6G, achieving 92.80% and 90.65%, respectively. The photocatalytic degradation efficiencies decreased at higher temperatures due to bigger sized CZTS particles as confirmed by SEM results. Computational simulations confirm that CZTS has a highly negative energy -25,764 Ry, confirming its structural stability and higher covalent than ionic character.
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Cobre , Azul de Metileno , Rodaminas , Sulfuros , Rodaminas/química , Azul de Metileno/química , Sulfuros/química , Cobre/química , Catálisis , Compuestos de Estaño/química , Calor , Contaminantes Químicos del Agua/químicaRESUMEN
Polymersomes are synthetic vesicles with potential use in healthcare, chemical transformations in confined environment (nanofactories), and in the construction of artificial cells and organelles. In this framework, one of the most important features of such supramolecular structures is the permeability behavior allowing for selective control of mass exchange between the inner and outer compartments. The use of biological and synthetic nanopores in this regard is the most common strategy to impart permeability nevertheless, this typically requires fairly complex strategies to enable porosity. Yet, investigations concerning the permeability of polymer vesicles to different analytes still requires further exploration and, taking these considerations into account, we have detailed investigated the permeability behavior of a variety of polymersomes with regard to different analytes (water, protons, and rhodamine B) which were selected as models for solvents, ions, and small molecules. Polymersomes based on hydrophilic blocks of poly[N-(2-hydroxypropyl)methacrylamide] (PHPMA) or PEO (poly(ethylene oxide)) linked to the non-responsive blocks poly[N-(4-isopropylphenylacetamide)ethyl methacrylate] (PPPhA) or poly(methyl methacrylate) (PMMA), or to the stimuli pH-responsive block poly[2-(diisopropylamino)ethyl methacrylate] (PDPA) have been investigated. Interestingly, the produced PEO-based vesicles are notably larger than the ones produced using PHPMA-containing block copolymers. The experimental results reveal that all the vesicles are inherently permeable to some extent with permeability behavior following exponential profiles. Nevertheless, polymersomes based on PMMA as the hydrophobic component were demonstrated to be the least permeable to the small molecule rhodamine B as well as to water. The synthetic vesicles based on the pH-responsive PDPA block exhibited restrictive and notably slow proton permeability as attributed to partial chain protonation upon acidification of the medium. The dye permeability was evidenced to be much slower than ion or solvent diffusion, and in the case of pH-responsive assemblies, it was demonstrated to also depend on the ionic strength of the environment. These findings are understood to be highly relevant towards polymer selection for the production of synthetic vesicles with selective and time-dependent permeability, and it may thus contribute in advancing biomimicry and nanomedicine.
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Permeabilidad , Polímeros , Rodaminas , Rodaminas/química , Polímeros/química , Células Artificiales/química , Tamaño de la Partícula , Interacciones Hidrofóbicas e Hidrofílicas , Concentración de Iones de Hidrógeno , Propiedades de Superficie , Agua/químicaRESUMEN
Ubiquitin-specific protease 7 (USP7) is a deubiquitinase enzyme that plays a critical role in regulating various cellular processes by cleaving ubiquitin molecules from target proteins. The C-terminal loop (CTL) motif is a specific region at the C-terminal end of the USP7 enzyme. Recent experiments suggest that the CTL motif plays a role in USP7's catalytic activity by contributing to the enzyme's structural stability, substrate recognition, and catalytic efficiency. The objective of this work is to elucidate these roles through the utilization of computational methods for molecular simulations. For this, we conducted extensive molecular dynamics (MD) simulations to investigate the conformational dynamics and protein-protein interactions within the USP7 enzyme-substrate complex with the substrate consisting of the ubiquitin tagged with the fluorescent label rhodamine 110-gly (Ub-Rho). Our results demonstrate that the CTL motif plays a crucial role in stabilizing the Ubl domains' conformation and augmenting the stability of active conformations within the enzyme-substrate complex. Conversely, the absence of the CTL motif results in increased flexibility and variability in Ubl domains' motion, leading to a reduced percentage of active conformations. Furthermore, our analysis of protein-protein interactions highlights the significance of the CTL motif in anchoring the Ubl45 domains to the catalytic domain (CD), thereby facilitating stable interactions with the substrate. Overall, our findings provide valuable insights into the conformational dynamics and protein-protein interactions inherent in the USP7 enzyme-substrate complex. These insights shed light on some mechanistic details of USP7 concerning the substrate's recognition before its catalytic action.
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Simulación de Dinámica Molecular , Unión Proteica , Peptidasa Específica de Ubiquitina 7 , Ubiquitina , Peptidasa Específica de Ubiquitina 7/metabolismo , Peptidasa Específica de Ubiquitina 7/química , Especificidad por Sustrato , Ubiquitina/metabolismo , Ubiquitina/química , Dominios Proteicos , Humanos , Rodaminas/química , Rodaminas/metabolismo , Conformación ProteicaRESUMEN
This study investigated the single and multicomponent adsorption of three emerging pollutants, the basic dyes Rhodamine 6G (R6G), Auramine-O (AO), and Brilliant Green (BG) by using hydroxyapatite synthesized from Pirarucu scales as adsorbent (HAP). The adsorption process was studied using seven different systems: AO-single, R6G-single, BG-single, R6G + AO, BG + AO, BG + R6G, and R6G + AO + BG. For kinetics, the initial concentration of each adsorbate per system was 50 mg/L, the results showed that the singular adsorption of these dyes was best-represented by the pseudo-second-order model (qAO = 62.54 mg/g, qR6G = 7.91 mg/g, qBG = 62.40 mg/g), however, the multicomponent adsorption was well-fitted by a pseudo-first-order model (ternary system: qAO = 56.21 mg/g, qR6G = 14.95 mg/g, qBG = 60.62 mg/g). For equilibrium, the initial concentration of each adsorbate per system was 10-300 mg/L, and the single adsorption systems were best represented by the Langmuir model. Nonetheless, the results displayed in the multicomponent mixture showed the presence of inflection points of AO and R6G whenever BG was present in solution with C0 > 150 mg/L, thus indicating that BG has greater affinity with HAP. The presence of inflection points in the curves represented a limitation for applying traditional equilibrium models, thus, an artificial neural network (ANN) was applied to non-linear curve fit this process and satisfactorily predicted the kinetics and equilibrium data. Finally, the analysis of thermodynamics for the ternary mixture revealed that the adsorption process is spontaneous (ΔG < 0), endothermic (ΔH > 0), and increases to a disorganized state as the temperature rises (ΔS > 0).
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Colorantes , Durapatita , Compuestos de Amonio Cuaternario , Contaminantes Químicos del Agua , Adsorción , Cinética , Durapatita/química , Colorantes/química , Contaminantes Químicos del Agua/química , Rodaminas/químicaRESUMEN
Excited-state chemistry relies on the communication between molecules, making it a crucial aspect of the field. One important question that arises is whether intermolecular communication and its rate can be modified when a molecule is confined. To explore the interaction in such systems, we investigated the ground and excited states of 4'-N,N-diethylaminoflavonol (DEA3HF) in an octa acid-based (OA) confined medium and in ethanolic solution, both in the presence of Rhodamine 6G (R6G). Despite the observed spectral overlap between the flavonol emission and the R6G absorption, as well as the fluorescence quenching of the flavonol in the presence of R6G, the almost constant fluorescence lifetime at different amounts of R6G discards the presence of FRET in the studied systems. Steady-state and time-resolved fluorescence indicate the formation of an emissive complex between the proton transfer dye encapsulated within water-soluble supramolecular host octa acid (DEA3HF@(OA)2) and R6G. A similar result was observed between DEA3HF:R6G in ethanolic solution. The respective Stern-Volmer plots corroborate with these observations, suggesting a static quenching mechanism for both systems.
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Éteres Cíclicos , Rodaminas/química , Análisis Espectral/métodosRESUMEN
A nanosensor based on magnetic core-shell nanoparticles functionalized with rhodamine derivative, N-(3-carboxy)acryloyl rhodamine B hydrazide (RhBCARB), using (3-aminopropyl)triethoxysilane (APTES) as a linker, has been synthesized for detection of Cu(II) ions in water. The magnetic nanoparticle and the modified rhodamine were fully characterized, showing a strong orange emission sensitive to Cu(II) ions. The sensor shows a linear response from 10 to 90 µg L-1, detection limit of 3 µg L-1 and no interference of Ni(II), Co(II), Cd(II), Zn(II), Pb(II), Hg(II) and Fe(II) ions. The nanosensor performance is similar to those described in the literature, being a viable option for the determination of Cu(II) ions in natural waters. In addition, the magnetic sensor can be easily removed from the reaction medium with the aid of a magnet and its signal recovered in acidic solution, allowing its reuse in subsequent analysis.
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Cobre , Colorantes Fluorescentes , Cobre/análisis , Rodaminas , Iones , Fenómenos MagnéticosRESUMEN
The use of fluorescent dyes in microscopy studies is frequent. Therefore, it is important to investigate whether these compounds may alter the physicochemical properties of materials in which they are incorporated to avoid methodological biases. This study evaluated the physicochemical properties of two endodontic sealers containing dry or diluted rhodamine B. Six groups were evaluated: AH Plus and MTA Fillapex in their original composition and mixed with 0.1% dry or diluted rhodamine. Push-out test was applied to assess the bond strength to root dentin. ANSI-ADA No. 57 and ISO No. 6876:2012 specifications were followed to evaluate flowability, setting time, and solubility of the sealers. pH changes were assessed after 24 h, 7, and 30 days. Intergroup comparisons were analyzed by ANOVA complemented by Tukey's post-hoc test; comparisons among periods were analyzed by Wilcoxon and Friedman tests. The incorporation of dry rhodamine decreased the bond strength and prolonged the setting time of AH Plus. The incorporation of diluted rhodamine decreased the setting time and increased the flowability of MTA Fillapex. Diluted rhodamine promoted pH reduction of AH Plus after 24 h and dry rhodamine after 7 days. Rhodamine incorporation promoted pH reduction after 30 days for MTA Fillapex. Solubility was not affected. In conclusion, the incorporation of diluted rhodamine changed the properties of MTA Fillapex, and the incorporation of dry rhodamine changed the properties of AH Plus. Previous dilution of rhodamine did not severely compromise the physicochemical properties of AH Plus and may be suggested to assess its penetrability.
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Materiales de Obturación del Conducto Radicular , Materiales de Obturación del Conducto Radicular/química , Resinas Epoxi/química , Compuestos de Calcio/química , Óxidos/química , Rodaminas , Solubilidad , Combinación de Medicamentos , Silicatos/química , Ensayo de MaterialesRESUMEN
Heterogeneous chemical processes occupy a pivotal position in many fields of applied chemistry. Monitoring reaction kinetics in such heterogeneous systems together with challenges associated with ex-situ analytical methodologies can lead to inaccurate information about the nature of the catalyst surfaces as well as information about the steps involved. The present work explores the possibility of kinetic measurements of chemical reactions and adsorption processes of homogeneous and heterogeneous systems through the variation of RGB intensities of digital images using a smartphone combined with a program written in Python to accelerate and facilitate data acquisition. In order to validate the method proposed, the base promoted hydrolysis of 4-nitrophenyl acetate was initially investigated. The rate constants obtained through RGB analysis (0.01854 min-1) is almost identical to that using traditional UV-Vis spectroscopy (0.01848 min-1). The proposed method was then applied to monitor the kinetics of three heterogeneous processes: (1) reduction of 4-nitrophenolate in the presence of dispersed Pd/C; (2) decomposition of methyl orange with TiO2; and (3) adsorption of rhodamine on montmorillonite. In general, the method via digital images showed high reproducibility and analytical frequency, allowing the execution of simultaneous analyses, with an accuracy comparable to UV-Vis spectrophotometry. The method developed herein is a practical and valuable alternative for obtaining kinetic data of heterogeneous reactions and processes where a color change is involved, bypassing sampling collection and processing which decreases analytical frequency and may lead to data errors.
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Bentonita , Teléfono Inteligente , Cinética , Reproducibilidad de los Resultados , RodaminasRESUMEN
Molecular machines, as exemplified by the kinesin and microtubule system, are responsible for molecular transport in cells. The monitoring of the cellular machinery has attracted much attention in recent years, requiring sophisticated techniques such as optical tweezers, and dark field hyperspectral and fluorescence microscopies. It also demands suitable procedures for immobilization and labeling with functional agents such as dyes, plasmonic nanoparticles and quantum dots. In this work, microtubules were co-polymerized by incubating a tubulin mix consisting of 7 biotinylated tubulin to 3 rhodamine tubulin. Rhodamine provided the fluorescent tag, while biotin was the anchoring group for receiving streptavidin containing species. To control the microtubule alignment and consequently, the molecular gliding directions, functionalized iron oxide nanoparticles were employed in the presence of an external magnet field. Such iron oxide nanoparticles, (MagNPs) were previously coated with silica and (3-aminopro-pyl)triethoxysilane (APTS) and then modified with streptavidin (SA) for linking to the biotin-functionalized microtubules. In this way, the binding has been successfully performed, and the magnetic alignment probed by Inverted Fluorescence Microscopy. The proposed strategy has proved promising, as tested with one of the most important biological structures of the cellular machinery.
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Biotina , Tubulina (Proteína) , Biotina/análisis , Biotina/química , Biotina/metabolismo , Óxido Ferrosoférrico/análisis , Óxido Ferrosoférrico/metabolismo , Fenómenos Magnéticos , Microscopía Fluorescente , Microtúbulos/química , Microtúbulos/metabolismo , Rodaminas/análisis , Rodaminas/metabolismo , Estreptavidina/análisis , Estreptavidina/química , Estreptavidina/metabolismo , Tubulina (Proteína)/análisis , Tubulina (Proteína)/metabolismoRESUMEN
The colocalization of taurine and zinc transporters (TAUT, ZnTs) has not been explored in retina. Our objective is to evaluate the effect of the intracellular zinc chelator N,N,N,N-tetrakis-(2-pyridylmethyl) ethylenediamine (TPEN) on zinc localization and colocalization TAUT and ZnT-1 (of plasma membrane), 3 (vesicular), and 7 (vesicular and golgi apparatus) in layers of retina by immunohistochemistry. To mark zinc, it was used cell-permeable fluorescent Zinquin ethyl ester. Specific first and secondary antibodies, conjugated with rhodamine or fluorescein-isothiocyanate were used to mark TAUT and ZnTs. The fluorescence results were reported as integrated optical density (IOD). Zinc was detected in all layers of the retina. The treatment with TPEN produced changes in the distribution of zinc in layers of retina less in the outer nuclear layer compared with the control. TAUT was detected in all layers of retina and TPEN chelator produced decrease of IOD in all layers of retina except in the photoreceptor compared with the control. ZnT 1, 3, and 7 were distributed in all retina layers, with more intensity in ganglion cell layer (GCL) and in the layers where there is synaptic connection. For all transporters, the treatment with TPEN produced significant decrease of IOD in layers of retina least in the inner nuclear layer for ZnT1, in the photoreceptor for ZnT3 and in the GCL and outer plexiform layer for ZnT7. The distribution of zinc, TAUT, and ZnTs in the layers of retina is indicative of the interaction of taurine and zinc for the function of the retina and normal operation of said layers. HIGHLIGHTS: Taurine and zinc are two molecules highly concentrated in the retina and with relevant functions in this structure. Maintaining zinc homeostasis in this tissue is necessary for the normal function of the taurine system in the retina. The study of the taurine transporter and the different zinc transporters in the retina (responsible for maintaining adequate levels of taurine and zinc) is relevant and novel, since it is indicative of the interactions between both molecules in this structure.
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Etilenodiaminas , Zinc , Animales , Proteínas Portadoras , Quelantes/análisis , Ésteres/análisis , Ésteres/metabolismo , Ésteres/farmacología , Etilenodiaminas/química , Etilenodiaminas/metabolismo , Etilenodiaminas/farmacología , Fluoresceínas/metabolismo , Isotiocianatos/análisis , Isotiocianatos/metabolismo , Isotiocianatos/farmacología , Ratas , Retina , Rodaminas/análisis , Taurina/análisis , Taurina/metabolismo , Taurina/farmacología , Zinc/químicaRESUMEN
This investigation reports the photocatalytic performance of the tungsten doped titania (TiO2:W or TW) with and without coating of chlorophyll (Chl) for the removal of the RhB dye from the drinking water. These particles were also supported on recycled plastic bottle caps (Bcap) to form other photocatalytic composites (TW/Bcap and TW + Chl/Bcap). The SEM images demonstrated that the TW particles without Chl had irregular shapes and sizes of 0.8-12 µm. The TW particles coated by the Chl presented shapes of quasi-rounded grains and smaller particle sizes of 0.8-1.8 µm. The photocatalytyic experiments showed that the photocatalyst powders containing Chl removed completely the RhB dye from the water after 2h under UV-VIS light, while the photocatalyst without Chl removed a maximum of 95% of the RhB. Interestingly, the TW/Bcap and TW + Chl/Bcap composites removed 94-100% of the RhB after 2h. Those ones removed such dye by photocatalysis and by physical adsorption at the same time (as confirmed by the absorbance and FTIR measurements), therefore, the removal of RhB was still very high. Scavenger experiments were also achieved and found that the â¢OH radicals are the main oxidizing species generated by the photocatalysts with and without Chl. The â¢O2- radicals and holes (h+) were the secondary oxidizing species. The presence of the chlorophyll on the photocatalyst increased in general the light absorption and the photocurrent. Overall, our work demonstrated that making composites with recycled plastic bottle caps is a feasible alternative to remove dyes from contaminated drinking water with high efficiency and low cost.
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Agua Potable , Catálisis , Clorofila , Colorantes , Plásticos , Rodaminas , TitanioRESUMEN
The present work aims to evaluate the removal capacity of Rhodamine B dye (RhB) using nano-porous chitosan (NC) from shrimp shells. NC was characterized by XRD, SEM-EDS, N2 porosimetry, zeta potential (ZP), FTIR, DLS, and zero charge point (pHZCP). Compound central rotational design (CCRD) was used to determine the ideal condition and antimicrobial activity was evaluated against different strains. NC showed characteristic of semi-crystalline material with negative charge surface (around - 21.13 mV), and SBET = 1.12 m2 g-1, Vp = 0.0064 cm3 g-1, Dp = 32.09 nm and pHZCP ≈ 7.98. Kinetic adsorption showed the pseudo first-order model had the best fit, with adsorption capacity (q1) between 3.78 and 64.43 mg g-1 and pseudo first-order kinetic constant (k1) between 0.066 and 0.052 min-1. Sips model best described the equilibrium data, with a maximum adsorption capacity of 505.131 mg g-1. Antimicrobial activity was observed at 0.25 mg mL-1 for different strains. Therefore, NC has potential application in the removal of the dye, combining sustainable development associated with nanotechnology.
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Antiinfecciosos , Quitosano , Contaminantes Químicos del Agua , Adsorción , Concentración de Iones de Hidrógeno , Cinética , RodaminasRESUMEN
Reconstituted high-density lipoprotein (rHDL) nanoparticles are excellent transporters of molecules and very useful for targeted therapy as they specifically recognize the scavenger receptor, class B1 (SR-B1) that is present on the surface of a wide range of tumor cells. However, they have rarely been employed to transport photosensitizers (PS) for photodynamic therapy (PDT). Rhodamine (R) compounds have been dismissed as useful PSs for PDT due to their low 1O2 production, excitation wavelengths with little tissue penetration, and poor selectivity for tumor cells. It was recently demonstrated that when irradiating at 532 nm or with Cerenkov radiation (CR) from a ß-emitting radionuclide, R123, R6G, and RB undergo electron transfer reactions (type I reaction) with folic acid. R6G also produces type I reactions with O2. In this work, the photodynamic effects of the rHDL-R system were evaluated in vitro. rHDL nanoparticles loaded with R123, R6G, and RB were synthesized, and the PS was internalized into T47D tumor cells. When cells were irradiated with a 532-nm laser in the presence of an rHDL-R systems, a cytotoxic photodynamic effect was obtained in the order R6G > R123 > RB. In the presence of CR from a 177Lu source, cytotoxicity showed the order R6G > RB > R123. The higher cytotoxicity induced by R6G in both cases corresponds to higher cellular internalization and larger production of type I and II reactions. Thus, in this work, it is proposed that rHDL-R/177Lu system can be applied in theragnostics as a multimodal radiotherapy-PDT-imaging system (imaging by SPECT or Cerenkov) and in hypoxic solid tumors in which external radiation is not effective and 177Lu-CR acts as light source.
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Nanopartículas , Fotoquimioterapia , Línea Celular Tumoral , Humanos , Lipoproteínas HDL , Fotoquimioterapia/métodos , Fármacos Fotosensibilizantes/farmacología , Fármacos Fotosensibilizantes/uso terapéutico , RodaminasRESUMEN
BACKGROUND: Crotalicidin (Ctn), a snake venom cathelicidin-related antimicrobial peptide, is a 34-residue-long linear lysine-rich vipericidin obtained from the South American rattlesnake, Crotalus durissus terrificus. Ctn contains tandem repeats of nine amino acid residues (1KRFKKFFKK9 and 16KRLKKIFKK24; consensus: 1KRhKKhFKK9, h = hydrophobic amino acid) as an integral part of its structure. OBJECTIVES: The aim of this study was to evaluate the antimicrobial activity of the encrypted vipericidin nonapeptide KRFKKFFKK, designated as Ctn[1-9], and its structural analogue, rhodamine- Bâconjugated Ctn[1-9], designated as RhoB-Ctn[1-9]. METHODS: The susceptibility of representative pathogenic bacteria and yeasts to antimicrobial agents was determined using the broth microdilution minimum inhibitory concentration (MIC) method. Cytotoxicity was estimated using a hemolytic assay. The accumulation of RhoB-Ctn[1-9] in microbial cells was observed by fluorescence microscopy. The antimicrobial synergism of RhoB-Ctn[1-9] with antimicrobials was evaluated using a checkerboard analysis. RESULTS: RhoB-conjugated Ctn[1-9] displayed selective antimicrobial activity against infectious gram-negative bacteria such as Escherichia coli, Pseudomonas aeruginosa, and pathogenic species of Candida with low hemolytic effects on human erythrocytes which were not observed with unconjugated Ctn[1-9]. RhoB-Ctn[1-9] could permeate cell membranes and accumulate intracellularly in microbial cells. RhoB-Ctn[1-9] exhibits synergistic effects when used with antibiotics or antifungal agents and reduced the MICs of the peptide and antimicrobials. CONCLUSION: These findings indicate the potential of crotalicidin-related short peptides as structural motifs for the diversification of biological functionalities. Further, they set the stage to investigate the molecular mechanisms by which chemically modified vipericidin repeats modulate cell fate.
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Antiinfecciosos , Antibacterianos/farmacología , Péptidos Antimicrobianos , Bacterias Gramnegativas , Humanos , Pruebas de Sensibilidad Microbiana , Fragmentos de Péptidos , RodaminasRESUMEN
We demonstrate a novel structure based on smart carbon nanocomposites intended for fabricating laser-triggered drug delivery devices (DDDs). The performance of the devices relies on nanocomposites' photothermal effects that are based on polydimethylsiloxane (PDMS) with carbon nanoparticles (CNPs). Upon evaluating the main features of the nanocomposites through physicochemical and photomechanical characterizations, we identified the main photomechanical features to be considered for selecting a nanocomposite for the DDDs. The capabilities of the PDMS/CNPs prototypes for drug delivery were tested using rhodamine-B (Rh-B) as a marker solution, allowing for visualizing and quantifying the release of the marker contained within the device. Our results showed that the DDDs readily expel the Rh-B from the reservoir upon laser irradiation and the amount of released Rh-B depends on the exposure time. Additionally, we identified two main Rh-B release mechanisms, the first one is based on the device elastic deformation and the second one is based on bubble generation and its expansion into the device. Both mechanisms were further elucidated through numerical simulations and compared with the experimental results. These promising results demonstrate that an inexpensive nanocomposite such as PDMS/CNPs can serve as a foundation for novel DDDs with spatial and temporal release control through laser irradiation.
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Portadores de Fármacos/química , Nanocompuestos/química , Materiales Inteligentes/química , Carbono/química , Dimetilpolisiloxanos/química , Portadores de Fármacos/efectos de la radiación , Elasticidad , Rayos Láser , Luz , Fenómenos Mecánicos , Nanocompuestos/efectos de la radiación , Rodaminas/química , Materiales Inteligentes/efectos de la radiaciónRESUMEN
Photosensitizer (PS) molecules play a critical role in photodynamic therapy of cancer and the understanding of the molecular mechanism involved in the photophysics of these compounds, and their reactions in the excited state are, therefore, of great interest for the development of this technique. In this article, the photophysics of the cationic PS 4,5-dibromorodamine methyl ester (TH9402), its electron- and energy-transfer reactions in the excited triplet state, with molecular oxygen, nitric oxide, guanosine-5'-monophosphate (GMP), and guanine, and the interaction with DNA were evaluated. Time-dependent density functional theory calculations at the TPSSh/Def2-TZVP//B3LYP/Def2-TZVP level of theory in water solution reveals that the PS has a bright S1 state 2.33 eV above the ground state that produces a fluorescent rate constant of 5.40 × 107 s-1, calculated using Fermi's golden rule within a path integral formalism. Once excited to the bright state, the main intersystem crossing (ISC) channel involves the coupling with the T2 state just below S1 (S1 â T2 â T1) with an overall ISC rate constant of 10.1 × 107 s-1, in good agreement with the experimental data. Excited-state reaction thermodynamics, computed at the M06-2X/Def2-TZVP//B3LYP/Def2-TZVP level of theory in water, showed that from all the excited-state electron-transfer reactions studied, only the transfer from GMP to the PS is thermodynamically favorable, independent of the protonation state of guanosine, which indicates a possible DNA photo-oxidation mechanism for the PS. Triplet-triplet energy-transfer reactions from TH9402 to molecular oxygen, producing reactive singlet oxygen, and to the deprotonated guanosine, producing 3GMP2-, are also thermodynamically favorable, with ΔG = -2.0 and -24.0 kcal//mol, respectively. However, the energy transfer to the monoprotonated guanosine is not favorable, (ΔG = 36.1), suggesting that in the DNA double-strand environment, this energy-transfer process may not be observed. The results show that the PS can act through electron transfer and triplet-triplet energy-transfer reactions involved in mechanism types I and II in photodynamic therapy. Interactions of TH9402 with the d(AGACGTCT)2 octanucleotide revealed that the PS can intercalate between the d(GpC)-d(CpG) base pairs in three different orientations and, upon intercalation, the π â π* transition of the PS shows a bathochromic shift up to 90 nm and up to 60% decrease in intensity. Interactions through groove binding showed a smaller bathochromic shift of 52.2 nm and a 56% decrease in intensity of the main transition band.
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Fotoquimioterapia , Fármacos Fotosensibilizantes , ADN , Ésteres , RodaminasRESUMEN
We synthesized twelve hybrids based on curcumin and resveratrol, and their structures were elucidated by spectroscopic analysis. The chemopreventive potential of these compounds was evaluated against SW480 human colon adenocarcinoma cells, its metastatic derivative SW620, along with the non-malignant CHO-K1 cell line. Among the tested compounds, hybrids 3e and 3i (for SW480) and 3a, 3e and 3k (for SW620) displayed the best cytotoxic activity with IC50 values ranging from 11.52 ± 2.78 to 29.33 ± 4.73 µM for both cell lines, with selectivity indices (SI) higher than 1, after 48 h of treatment. Selectivity indices were even higher than those reported for the reference drug, 5-fluorouracil (SI = 0.96), the starting compound resveratrol (SI = 0.45) and the equimolar mixture of curcumin plus resveratrol (SI = 0.77). The previous hybrids showed good antiproliferative activity.
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Antineoplásicos/síntesis química , Neoplasias Colorrectales/patología , Curcumina/farmacología , Resveratrol/farmacología , Animales , Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Células CHO , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Neoplasias Colorrectales/tratamiento farmacológico , Cricetinae , Cricetulus , Curcumina/síntesis química , Diseño de Fármacos , Ensayos de Selección de Medicamentos Antitumorales , Fluorouracilo/farmacología , Humanos , Resveratrol/síntesis química , Rodaminas/farmacologíaRESUMEN
BACKGROUND: Molecular and genetic studies of blood-stage Plasmodium falciparum parasites require limiting dilution cloning and prolonged cultivation in microplates. The entire process is laborious and subject to errors due to inaccurate dilutions at the onset and failed detection of parasite growth in individual microplate wells. METHODS: To precisely control the number of parasites dispensed into each microplate well, parasitaemia and total cell counts were determined by flow cytometry using parasite cultures stained with ethidium bromide or SYBR Green I. Microplates were seeded with 0.2 or 0.3 infected cells/well and cultivated with fresh erythrocytes. The c-SNARF fluorescent pH indicator was then used to reliably detect parasite growth. RESULTS: Flow cytometry required less time than the traditional approach of estimating parasitaemia and cell numbers by microscopic examination. The resulting dilutions matched predictions from Poisson distribution calculations and yielded clonal lines. Addition of c-SNARF to media permitted rapid detection of parasite growth in microplate wells with high confidence. CONCLUSION: The combined use of flow cytometry for precise dilution and the c-SNARF method for detection of growth improves limiting dilution cloning of P. falciparum. This simple approach saves time, is scalable, and maximizes identification of desired parasite clones. It will facilitate DNA transfection studies and isolation of parasite clones from ex vivo blood samples.
Asunto(s)
Benzopiranos/química , Clonación Molecular/métodos , Citometría de Flujo , Naftoles/química , Plasmodium falciparum/aislamiento & purificación , Rodaminas/química , Malaria Falciparum/diagnósticoRESUMEN
Catalytic processes based on Fenton-like reactions on the degradation of organic pollutants have been improved by accelerating the redox cycling of metal ions. This work presents, at first, the results obtained for the heterogeneous degradation of rhodamine B (RhB) by copper ferrite (CuFe2O4) in presence of hydrogen peroxide (H2O2) and hydrazine (N2H4) as redox cycle accelerator. Atomic absorption spectroscopy showed small amounts of Cu2+ are leached from ferrite highlighting the influence of the homogeneous catalysis in the whole process. The data obtained for the homogeneous process using Cu2+ in solution containing both N2H4 and H2O2 indicated such system is highly efficient mineralizing 73% of RhB within only 10 min of reaction and having H2O and CO2 as major products. Using tert-butyl alcohol as radical scavenger, it was confirmed hydroxyl radical (HOâ¢) is the active oxidant species regarding the RhB degradation. The homogeneous catalyst was applied to a real sample of textile effluent spiked with RhB and showed reasonable efficiency, although lower than that obtained for the standard solutions of RhB. This result was assigned to the interference of salts in the medium that react with HO⢠thus acting as radical scavenger.
Asunto(s)
Cobre , Peróxido de Hidrógeno , Catálisis , Hidrazinas , Oxidación-Reducción , Estrés Oxidativo , RodaminasRESUMEN
Melanin-concentrating hormone (MCH) is a hypothalamic neuropeptide synthesized by posterior hypothalamic and incerto-hypothalamic neurons that project throughout the central nervous system. The MCHergic system modulates several important functions such as feeding behavior, mood and sleep. MCH exerts its biological functions through interaction with the MCHR-1 receptor, the only functional receptor present in rodents. The internalization process of MCHR-1 triggered by MCH binding was described in vitro in non-neuronal heterologous systems with over-expression of MCHR-1. Reports of in vivo MCHR-1 internalization dynamics are scarce, however, this is an important process to explore based on the critical functions of the MCHergic system. We had previously determined that 60â¯min after intracerebroventricular (i.c.v.) microinjections of MCH conjugated with fluorophore rhodamine (R-MCH), the dorsal and median raphe nucleus presented R-MCH positive labeled neurons. In the present work, we further studied the in vivo uptake process focusing on the distribution and time-dependent pattern of R-MCH positive cells 10, 20 and 60â¯min (T10, T20 and T60, respectively) after i.c.v. microinjection of R-MCH. We also explored this uptake process to see whether it was receptor- and clathrin-dependent and examined the phenotype of R-MCH positive cells and their proximity to MCHergic fibers. We found a great number of R-MCH positive cells with high fluorescence intensity in the lateral septum, nucleus accumbens and hippocampus at T20 and T60 (but not at T10), while a lower number with low intensity was observed in the dorsal raphe nucleus. At T20, in rats pre-treated with a MCHR-1 antagonist (ATC-0175) or with phenylarsine oxide (PAO), a clathrin endocytosis inhibitor, a robust decrease (> 50 %) of R-MCH uptake occurred in these structures. The R-MCH positive cells were identified as neurons (NeuN positive, GFAP negative) and some MCHergic fibers run in the vicinities of them. We concluded that neurons localized at structures that were close to the ventricular surfaces could uptake R-MCH in vivo through a receptor-dependent and clathrin-mediated process. Our results support volume transmission of MCH through the cerebrospinal fluid to reach distant targets. Finally, we propose that R-MCH would be an effective tool to study MCH-uptake in vivo.