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1.
J Struct Biol ; 216(2): 108084, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38479547

RESUMO

In humans, the growth pattern of the acellular extrinsic fibre cementum (AEFC) has been useful to estimate the age-at-death. However, the structural organization behind such a pattern remains poorly understood. In this study tooth cementum from seven individuals from a Mexican modern skeletal series were analyzed with the aim of unveiling the AEFC collagenous and mineral structure using multimodal imaging approaches. The organization of collagen fibres was first determined using: light microscopy, transmission electron microscopy (TEM), electron tomography, and plasma FIB scanning electron microscopy (PFIB-SEM) tomography. The mineral properties were then investigated using: synchrotron small-angle X-ray scattering (SAXS) for T-parameter (correlation length between mineral particles); synchrotron X-ray diffraction (XRD) for L-parameter (mineral crystalline domain size estimation), alignment parameter (crystals preferred orientation) and lattice parameters a and c; as well as synchrotron X-ray fluorescence for spatial distribution of calcium, phosphorus and zinc. Results show that Sharpey's fibres branched out fibres that cover and uncover other collagen bundles forming aligned arched structures that are joined by these same fibres but in a parallel fashion. The parallel fibres are not set as a continuum on the same plane and when they are superimposed project the AEFC incremental lines due to the collagen birefringence. The orientation of the apatite crystallites is subject to the arrangement of the collagen fibres, and the obtained parameter values along with the elemental distribution maps, revealed this mineral tissue as relatively homogeneous. Therefore, no intrinsic characteristics of the mineral phase could be associated with the alternating AEFC incremental pattern.


Assuntos
Cemento Dentário , Minerais , Difração de Raios X , Humanos , Cemento Dentário/ultraestrutura , Cemento Dentário/química , Cemento Dentário/metabolismo , Difração de Raios X/métodos , Minerais/metabolismo , Minerais/química , Colágeno/química , Colágeno/metabolismo , Microscopia Eletrônica de Transmissão/métodos , Espalhamento a Baixo Ângulo , Microscopia Eletrônica de Varredura/métodos , Tomografia com Microscopia Eletrônica/métodos , Feminino , Adulto , Masculino , Pessoa de Meia-Idade
2.
Part Fibre Toxicol ; 21(1): 26, 2024 05 22.
Artigo em Inglês | MEDLINE | ID: mdl-38778339

RESUMO

BACKGROUND: During inhalation, airborne particles such as particulate matter ≤ 2.5 µm (PM2.5), can deposit and accumulate on the alveolar epithelial tissue. In vivo studies have shown that fractions of PM2.5 can cross the alveolar epithelium to blood circulation, reaching secondary organs beyond the lungs. However, approaches to quantify the translocation of particles across the alveolar epithelium in vivo and in vitro are still not well established. In this study, methods to assess the translocation of standard diesel exhaust particles (DEPs) across permeable polyethylene terephthalate (PET) inserts at 0.4, 1, and 3 µm pore sizes were first optimized with transmission electron microscopy (TEM), ultraviolet-visible spectroscopy (UV-VIS), and lock-in thermography (LIT), which were then applied to study the translocation of DEPs across human alveolar epithelial type II (A549) cells. A549 cells that grew on the membrane (pore size: 3 µm) in inserts were exposed to DEPs at different concentrations from 0 to 80 µg.mL- 1 ( 0 to 44 µg.cm- 2) for 24 h. After exposure, the basal fraction was collected and then analyzed by combining qualitative (TEM) and quantitative (UV-VIS and LIT) techniques to assess the translocated fraction of the DEPs across the alveolar epithelium in vitro. RESULTS: We could detect the translocated fraction of DEPs across the PET membranes with 3 µm pore sizes and without cells by TEM analysis, and determine the percentage of translocation at approximatively 37% by UV-VIS (LOD: 1.92 µg.mL- 1) and 75% by LIT (LOD: 0.20 µg.cm- 2). In the presence of cells, the percentage of DEPs translocation across the alveolar tissue was determined around 1% at 20 and 40 µg.mL- 1 (11 and 22 µg.cm- 2), and no particles were detected at higher and lower concentrations. Interestingly, simultaneous exposure of A549 cells to DEPs and EDTA can increase the translocation of DEPs in the basal fraction. CONCLUSION: We propose a combination of analytical techniques to assess the translocation of DEPs across lung tissues. Our results reveal a low percentage of translocation of DEPs across alveolar epithelial tissue in vitro and they correspond to in vivo findings. The combination approach can be applied to any traffic-generated particles, thus enabling us to understand their involvement in public health.


Assuntos
Material Particulado , Alvéolos Pulmonares , Emissões de Veículos , Humanos , Emissões de Veículos/toxicidade , Emissões de Veículos/análise , Células A549 , Material Particulado/toxicidade , Material Particulado/análise , Alvéolos Pulmonares/efeitos dos fármacos , Alvéolos Pulmonares/metabolismo , Tamanho da Partícula , Microscopia Eletrônica de Transmissão , Polietilenotereftalatos/química , Polietilenotereftalatos/toxicidade , Células Epiteliais Alveolares/efeitos dos fármacos , Células Epiteliais Alveolares/metabolismo , Poluentes Atmosféricos/toxicidade , Poluentes Atmosféricos/análise
3.
Luminescence ; 39(1): e4634, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38286605

RESUMO

In this study, cellulose nanocrystals (CNCs) were synthesized from celery stalks to be used as the platform for quercetin delivery. Additionally, CNCs and CNCs-quercetin were characterized using the results of scanning electron microscope (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, and zeta potential, while their interactions with human holo-transferrin (HTF) were also investigated. We examined their interaction under physiological conditions through the exertion of fluorescence, resonance light scattering, synchronized fluorescence spectroscopy, circular dichroism, three-dimensional fluorescence spectroscopy, and fluorescence resonance energy transfer techniques. The data from SEM and TEM exhibited the spherical shape of CNCs and CNCs-quercetin and also, a decrease was detected in the size of quercetin-loaded CNCs from 676 to 473 nm that indicated the intensified water solubility of quercetin. The success of cellulose acid hydrolysis was confirmed based on the XRD results. Apparently, the crystalline index of CNCs-quercetin was reduced by the interaction of CNCs with quercetin, which also resulted in the appearance of functional groups, as shown by FTIR. The interaction of CNCs-quercetin with HTF was also demonstrated by the induced quenching in the intensity of HTF fluorescence emission and Stern-Volmer data represent the occurrence of static quenching. Overall, the effectiveness of CNCs as quercetin vehicles suggests its potential suitability for dietary supplements and pharmaceutical products.


Assuntos
Apium , Nanopartículas , Humanos , Celulose/química , Quercetina , Transferrina/química , Adsorção , Nanopartículas/química , Digestão
4.
J Liposome Res ; 34(1): 113-123, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-37493091

RESUMO

Spherical structures built from uni- and multilamellar lipid bilayers (LUV and MLV) are nowadays considered not just as nanocarriers of various kinds of therapeutics, but also as the vehicles that, when coupled with gold (Au) nanoparticles (NPs), can also serve as a tool for imaging and discriminating healthy and diseased tissues. Since the presence of Au NPs or their aggregates may affect the properties of the drug delivery vehicle, we investigated how the shape and position of Au NP aggregates adsorbed on the surface of MLV affect the arrangement and conformation of lipid molecules. By preparing MLVs constituted from 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) in the presence of uncoated Au NP aggregates found i) both within liposome core and on the surface of the outer lipid bilayer, or ii) adsorbed on the outer lipid bilayer surface only, we demonstrated the maintenance of lipid bilayer integrity by microscopic techniques (cryo-TEM, and AFM). The employment of SERS and FTIR-ATR techniques enabled us not only to elucidate the lipid interaction pattern and their orientation in regards to Au NP aggregates but also unequivocally confirmed the impact of Au NP aggregates on the persistence/breaking of van der Waals interactions between hydrocarbon chains of DPPC.


Assuntos
Nanopartículas Metálicas , Fosfatidilcolinas , Fosfatidilcolinas/química , Lipossomos/química , Bicamadas Lipídicas/química , Ouro/química
5.
Int J Mol Sci ; 25(2)2024 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-38256239

RESUMO

Ropinirole is a non-ergolinic dopamine agonist used to manage Parkinson's disease and it is characterized by poor oral bioavailability. This study aimed to design and develop advanced drug delivery systems composed of poloxamer 407, a non-ionic surfactant (Tween 80), and cyclodextrins (methyl-ß-CD or hydroxy-propyl-ß-CD) for possible brain targeting of ropinirole after nasal administration for the treatment of Parkinson's disease. The hybrid systems were formed by the thin-film hydration method, followed by an extensive physicochemical and morphological characterization. The in vitro cytotoxicity of the systems on HEK293 cell lines was also tested. In vitro release and ex vivo mucosal permeation of ropinirole were assessed using Franz cells at 34 °C and with phosphate buffer solution at pH 5.6 in the donor compartment, simulating the conditions of the nasal cavity. The results indicated that the diffusion-controlled drug release exhibited a progressive increase throughout the experiment, while a proof-of-concept experiment on ex vivo permeation through rabbit nasal mucosa revealed a better performance of the prepared hybrid systems in comparison to ropinirole solution. The encouraging results in drug release and mucosal permeation indicate that these hybrid systems can serve as attractive platforms for effective and targeted nose-to-brain delivery of ropinirole with a possible application in Parkinson's disease. Further ex vivo and in vivo studies to support the results of the present work are ongoing.


Assuntos
Indóis , Doença de Parkinson , Surfactantes Pulmonares , Humanos , Animais , Coelhos , Tensoativos , Polímeros , Células HEK293 , Doença de Parkinson/tratamento farmacológico , Encéfalo , Lipoproteínas , Mucosa Nasal
6.
Microsc Microanal ; 29(6): 2090-2098, 2023 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-37966971

RESUMO

The detailed characterization of fouling in membranes is essential to understand any observed improvement or reduction on filtration performance. Electron microscopy allows detailed structural characterization, and its combination with labeling techniques, using electron-dense probes, typically allows for the differentiation of biomolecules. Developing specific protocols that allow for differentiation of biomolecules in membrane fouling by electron microscopy is a major challenge due to both as follows: the necessity to preserve the native state of fouled membranes upon real filtration conditions as well as the inability of the electron-dense probes to penetrate the membranes once they have been fouled. In this study, we present the development of a heavy metal staining technique for identification and differentiation of biomolecules in membrane fouling, which is compatible with cryofixation methods. A general contrast enhancement of biomolecules and fouling is achieved. Our observations indicate a strong interaction between biomolecules: A tendency of proteins, both in solution as well as in the fouling, to surround the lipids is observed. Using transmission electron microscopy and scanning electron microscopy at cryogenic conditions, cryo-SEM, in combination with energy-dispersive X-ray spectroscopy, the spatial distribution of proteins and lipids within fouling is shown and the role of proteins in fouling discussed.


Assuntos
Membranas Artificiais , Metais Pesados , Temperatura , Microscopia Eletrônica de Varredura , Proteínas , Lipídeos
7.
Int J Mol Sci ; 24(2)2023 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-36674712

RESUMO

Liposomes have been successfully used as drug-delivery vehicles, but there are no clinical studies on improved fertility and the few reported experimental studies have been performed in animal models far from humans. The aim of this paper was to study the effects of treatment with cationic, anionic and zwitterionic liposomes on our superior mammalian model of porcine prepubertal Sertoli cells (SCs) to find a carrier of in vitro test drugs for SCs. Porcine pre-pubertal SCs cultures were incubated with different liposomes. Viability, apoptosis/necrosis status (Annexin-V/Propidium iodide assay), immunolocalisation of ß-actin, vimentin, the phosphorylated form of AMP-activated protein Kinase (AMPK)α and cell ultrastructure (Transmission Electron Microscopy, TEM) were analysed. Zwitterionic liposomes did not determine changes in the cell cytoplasm. The incubation with anionic and cationic liposomes modified the distribution of actin and vimentin filaments and increased the levels of the phosphorylated form of AMPKα. The Annexin/Propidium Iodide assay suggested an increase in apoptosis. TEM analysis highlighted a cytoplasmic vacuolisation. In conclusion, these preliminary data indicated that zwitterionic liposomes were the best carrier to use in an in vitro study of SCs to understand the effects of molecules or drugs that could have a clinical application in the treatment of certain forms of male infertility.


Assuntos
Lipossomos , Células de Sertoli , Humanos , Masculino , Animais , Suínos , Lipossomos/química , Vimentina , Células de Sertoli/metabolismo , Propídio , Apoptose , Mamíferos/metabolismo
8.
J Biol Chem ; 296: 100645, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33839156

RESUMO

Interactions of membrane-bound mammalian cytochromes P450 (CYPs) with NADPH-cytochrome P450 oxidoreductase (POR), which are required for metabolism of xenobiotics, are facilitated by membrane lipids. A variety of membrane mimetics, such as phospholipid liposomes and nanodiscs, have been used to simulate the membrane to form catalytically active CYP:POR complexes. However, the exact mechanism(s) of these interactions are unclear because of the absence of structural information of full-length mammalian CYP:POR complexes in membranes. Herein, we report the use of amphipols (APols) to form a fully functional, soluble, homogeneous preparation of full-length CYP:POR complexes amenable to biochemical and structural study. Incorporation of CYP2B4 and POR into APols resulted in a CYP2B4:POR complex with a stoichiometry of 1:1, which was fully functional in demethylating benzphetamine at a turnover rate of 37.7 ± 2.2 min-1, with a coupling efficiency of 40%. Interestingly, the stable complex had a molecular weight (Mw) of 338 ± 22 kDa determined by multiangle light scattering, suggestive of a tetrameric complex of 2CYP2B4:2POR embedded in one APol nanoparticle. Moreover, negative stain electron microscopy (EM) validated the homogeneity of the complex and allowed us to generate a three-dimensional EM map and model consistent with the tetramer observed in solution. This first report of the full-length mammalian CYP:POR complex by transmission EM not only reveals the architecture that facilitates electron transfer but also highlights a potential use of APols in biochemical and structural studies of functional CYP complexes with redox partners.


Assuntos
Hidrocarboneto de Aril Hidroxilases/metabolismo , NADPH-Ferri-Hemoproteína Redutase/metabolismo , Polímeros/metabolismo , Propilaminas/metabolismo , Animais , Hidrocarboneto de Aril Hidroxilases/química , Catálise , Família 2 do Citocromo P450/química , Família 2 do Citocromo P450/metabolismo , NADPH-Ferri-Hemoproteína Redutase/química , Ligação Proteica , Conformação Proteica , Multimerização Proteica , Coelhos
9.
Anal Biochem ; 647: 114692, 2022 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-35461801

RESUMO

Despite the important role of membrane proteins in biological function and physiology, studying them remains challenging because of limited biomimetic systems for the protein to remain in its native membrane environment. Cryo electron microscopy (Cryo-EM) is emerging as a powerful tool for analyzing the structure of membrane proteins. However, Cryo-EM and other membrane protein analyses are better studied in a native lipid bilayer. Although traditional, mimetic systems have disadvantages that limit their use in the study of membrane proteins. As an alternative, styrene-maleic acid copolymers are used to form nanoparticles with POPC:POPG lipids. Traditional characterization of these styrene maleic acid lipid nanoparticles (SMALPs) includes dynamic light scattering (DLS), electron paramagnetic resonance (EPR), nuclear magnetic resonance (NMR), and transmission electron microscopy (TEM). In this study a new method was developed that utilizes SMALPs using a styrene-maleic acid copolymer (SMA) thin film on a TEM grid, acting as a substrate. By directly adding POPC:POPG lipid vesicles to the SMA coated grid SMALPs can be formed, visualized, and characterized by TEM without the need to make them in solution prior to imaging. We envision these functionalized grids could aid in single particle specimen preparation, increasing the efficiency of structural biology and biophysical techniques such as Cryo-EM.


Assuntos
Maleatos , Nanopartículas , Lipossomos , Maleatos/química , Proteínas de Membrana/química , Nanopartículas/química
10.
Arch Microbiol ; 204(8): 507, 2022 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-35859215

RESUMO

Vancomycin (VAN) is unable to penetrate the outer membrane of Gram-negative bacteria and reach the target site. One approach to overcome this limitation is to associate it with compounds with permeabilizing or antimicrobial properties. Eudragit E100® (Eu) is a cationic polymer insufficiently characterized for its potential antimicrobial action. Eu-VAN combinations were characterized, the antimicrobial efficacy against Pseudomonas aeruginosa was evaluated and previous studies on the effects of Eu on bacterial envelopes were extended. Time-kill assays showed eradication of P. aeruginosa within 3-6 h exposure to Eu-VAN, whilst VAN was ineffective. Eu showed regrowth in 24 h and delayed colony pigmentation. Although permeabilization of bacterial envelopes or morphological alterations observed by TEM and flow cytometry after exposure to Eu were insufficient to cause bacterial death, they allowed access of VAN to the target site, since Eu-VAN/Van-FL-treated cultures showed fluorescent staining in all bacterial cells, indicating Van-FL internalization. Consequently, Eu potentiated the activity of an otherwise inactive antibiotic against P. aeruginosa. Moreover, Eu-VAN combinations exhibited improved physicochemical properties and could be used in the development of therapeutic alternatives in the treatment of bacterial keratitis.


Assuntos
Pseudomonas aeruginosa , Vancomicina , Antibacterianos/química , Antibacterianos/farmacologia , Testes de Sensibilidade Microbiana , Polímeros/farmacologia , Vancomicina/farmacologia
11.
Environ Sci Technol ; 56(5): 3011-3020, 2022 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-35133799

RESUMO

The radionuclide selenium-79 (Se-79) is predicted to be a key contributor to the long-term radiologic hazards associated with geological high-level waste (HLW) repositories; hence its release is of pertinent concern in the safety assessment of repositories. However, interactions of reduced Se species with aqueous Fe(II) species and solid phases arising from the corrosion of a steel overpack could play a role in mitigating its migration to the surrounding environment. In this study, we examined the immobilization mechanisms of Se(-II) during its interaction with aqueous Fe(II) and freshly precipitated Fe(OH)2 at circumneutral and alkaline conditions, respectively, its response to changes in pH, and its behavior during aging at 90 °C. Using microscopic and spectroscopic techniques, we observed ß-FeSe precipitation, regardless of whether Se(-II) reacts with aqueous species or solid phases, and that modifying the pH following initial immobilization did not remobilize Se(-II). These observations indicate that Se(-II) migration beyond the overpack can be effectively and rapidly retarded via interactions with Fe(II) species arising from overpack corrosion. Thermodynamic calculations, however, showed that iron selenides became metastable at alkaline conditions and will dissolve in the long term. Aging experiments at 90 °C showed that Se(-II) can be completely retained via the crystallization of ferroselite at circumneutral conditions, while it will be largely remobilized at alkaline conditions. Our results show that Se(-II) mobility can be significantly influenced by its interactions with the corrosion products of the steel overpack and that these behaviors will have to be considered in repository safety assessments.


Assuntos
Ferro , Radioisótopos , Corrosão , Compostos Ferrosos/química , Ferro/química , Aço
12.
Proc Natl Acad Sci U S A ; 116(45): 22491-22499, 2019 11 05.
Artigo em Inglês | MEDLINE | ID: mdl-31636174

RESUMO

Rational design of supramolecular nanomaterials fundamentally depends upon an atomic-level understanding of their structure and how it responds to chemical modifications. Here we studied a series of crystalline diblock copolypeptoids by a combination of sequence-controlled synthesis, cryogenic transmission electron microscopy, and molecular dynamics simulation. This family of amphiphilic polypeptoids formed free-floating 2-dimensional monolayer nanosheets, in which individual polymer chains and their relative orientations could be directly observed. Furthermore, bromine atom side-chain substituents in nanosheets were directly visualized by cryogenic transmission electron microscopy, revealing atomic details in position space inaccessible by conventional scattering techniques. While the polypeptoid backbone conformation was conserved across the set of molecules, the nanosheets exhibited different lattice packing geometries dependent on the aromatic side chain para substitutions. Peptoids are inherently achiral, yet we showed that sequences containing an asymmetric aromatic substitution pattern pack with alternating rows adopting opposite backbone chiralities. These atomic-level insights into peptoid nanosheet crystal structure provide guidance for the future design of bioinspired nanomaterials with more precisely controlled structures and properties.


Assuntos
Nanoestruturas/química , Peptoides/química , Microscopia Crioeletrônica , Simulação de Dinâmica Molecular , Nanoestruturas/ultraestrutura , Peptoides/síntese química , Polímeros/síntese química , Polímeros/química
13.
Microsc Microanal ; : 1-8, 2022 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-35659819

RESUMO

The tongue is a fundamental organ in feeding, vocalization, and grooming. It is characterized by evolutionary adaptations reflected by diet, habitat, and function. Rodents are a very diverse mammalian order and the tongue's morphology varies in size, form, and presence of papillae. This work aimed to describe the morphological and ultrastructural aspects of the tongue of Spix's yellow-toothed cavy (Galea spixii, Wagler, 1831). Tongues of Spix's yellow-toothed cavies were analyzed with light microscopy, scanning, and transmission electron microscopy. The results showed that the tongue was divided into apex, body, and root. There were different types of papillae, such as vallate, foliate, laterally placed fungiform, fungiform, filiform, and robust filiform. The epithelium was organized into layers, including keratinized, granulous, spinous, and basal, below were lamina propria, and musculature, which evolved mucous and serous gland clusters. The tongue of Spix's yellow-toothed cavy was structurally and ultrastructurally similar to other rodents and had papillae with similar morphologies to other Caviidae species. However, the presence of robust filiform papillary lines and laterally placed fungiform papillae showed the main differences from other species. This was the first description of the tongue of Spix's yellow-toothed cavy.

14.
Anaerobe ; 75: 102515, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35026418

RESUMO

OBJECTIVES: The dysbiosis of bacteria and horizontal transfer of antibiotic resistance genes (ARGs) could be highly problematic particularly in the oral environment. Here, we aimed to identify the anaerobic species from patients with periodontitis and to screen the isolates for the ß-lactamase resistance genes, blaTEM, cfxA, its variants, and mobA. METHODS: The 129 samples from periodontal pockets were subjected to anaerobic culture, followed by 16S rRNA gene sequencing, PCR assays for the cfxA, blaTEM, and mobA. The minimum inhibitory concentration (MIC) of amoxicillin, ampicillin, amoxicillin/clavulanate, ampicillin/sulbactam, and cefixime was determined against CfxA producing isolates using MIC Test Strips. RESULTS: The species with frequency higher than 10% were Lactobacillus spp. (26.3%), Streptococcus spp. (18.8%), Leptotrichia wadei (14%) and Veillonella spp. (11.4%). The blaTEM was not found in any of the isolates whereas cfxA was found in 12.5% of isolates including V. parvula, V. rogosae, Prevotella nigrescens and Campylobacter concisus. Of CfxA variants, CfxA2 (90%) was the most frequent one. Among the CfxA producing isolates, the resistance to ampicillin and amoxicillin was observed only in two isolates of P. nigrescens and V. rogosae. CONCLUSIONS: This study showed that various anaerobes species may be involved in the development of periodontitis. Of them, Prevotella and Veillonella species were found to commonly carry cfxA even though they are susceptible to beta-lactams and its combination.


Assuntos
Bactérias Anaeróbias , Periodontite , Amoxicilina/farmacologia , Combinação Amoxicilina e Clavulanato de Potássio/farmacologia , Antibacterianos/farmacologia , Humanos , Irã (Geográfico) , Testes de Sensibilidade Microbiana , Periodontite/microbiologia , RNA Ribossômico 16S/genética , Resistência beta-Lactâmica/genética , beta-Lactamases/genética
15.
Molecules ; 27(22)2022 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-36432127

RESUMO

The new method of antibacterial-drug-activity investigation in vitro is proposed as a powerful strategy for understanding how carriers affect drug action during long periods (7 days). In this paper, we observed fluoroquinolone moxifloxacin (MF) antibacterial-efficiency in non-covalent complexes, with the sulfobutyl ether derivative of ß-cyclodextrin (SCD) and its polymer (SCDpol). We conducted in vitro studies on two Escherichia coli strains that differed in surface morphology. It was found that MF loses its antibacterial action after 3-4 days in liquid media, whereas the inclusion of the drug in SCD led to the increase of MF antibacterial activity by up to 1.4 times within 1-5 days of the experiment. In the case of MF-SCDpol, we observed a 12-fold increase in the MF action, and a tendency to prolonged antibacterial activity. We visualized this phenomenon (the state of bacteria, cell membrane, and surface morphology) during MF and MF-carrier exposure by TEM. SCD and SCDpol did not change the drug's mechanism of action. Particle adsorption on cells was the crucial factor for determining the observed effects. The proteinaceous fimbriae on the bacteria surface gave a 2-fold increase of the drug carrier adsorption, hence the strains with fimbriae are more preferable for the proposed treatment. Furthermore, the approach to visualize the CD polymer adsorption on bacteria via TEM is suggested. We hope that the proposed comprehensive method will be useful for the studies of drug-delivery systems to uncover long-term antibacterial action.


Assuntos
Antibacterianos , Infecções por Escherichia coli , Humanos , Antibacterianos/farmacologia , Escherichia coli , Portadores de Fármacos/farmacologia , Bactérias , Polímeros/farmacologia
16.
Small ; 17(30): e2100307, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34146389

RESUMO

Norovirus and Rotavirus are among the pathogens causing a large number of disease outbreaks due to contaminated water. These viruses are nanoscale particles that are difficult to remove by common filtration approaches which are based on physical size exclusion, and require adsorption-based filtration methods. This study reports the pH-responsive interactions of viruses with cationic-modified nanocellulose and demonstrates a filter material that adsorbs nanoscale viruses and can be regenerated by changing the solution's pH. The bacteria viruses Qbeta and MS2, with diameters below 30 nm but different surface properties, are used to evaluate the pH-dependency of the interactions and the filtration process. Small angle X-ray scattering, cryogenic transmission electron microscopy, and ζ-potential measurements are used to study the interactions and analyze changes in their nanostructure and surface properties of the virus upon adsorption. The virus removal capacity of the cationic cellulose-based aerogel filter is 99.9% for MS2 and 93.6% for Qbeta, at pH = 7.0; and desorption of mostly intact viruses occurs at pH = 3.0. The results contribute to the fundamental understanding of pH-triggered virus-nanocellulose self-assembly and can guide the design of sustainable and environmentally friendly adsorption-based virus filter materials as well as phage and virus-based materials.


Assuntos
Celulose , Vírus , Filtração , Concentração de Íons de Hidrogênio , Água
17.
Nanotechnology ; 32(48)2021 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-34407513

RESUMO

Liquid cell electron microscopy is an imaging technique allowing for the investigation of the interaction of liquids and solids at nanoscopic length scales. Suchin situobservations are increasingly in-demand in an array of fields, from biological sciences to medicine to batteries. Graphene liquid cells (GLCs), in particular, have generated a great interest as a low-scattering window material with the potential for increasing the quality of both imaging and spectroscopy. However, preserving the stability of the liquid and of the sample in the GLC remains a considerable challenge. In the present work we encapsulate water and hydroxyapatite (HAP), a pH-sensitive biological material, in GLCs to observe the interactions between the graphene, HAP, and the electron beam. HAP was chosen for several reasons. One is its ubiquity in biological specimens such as bones and teeth, and the second is the presence of phosphate ions in common buffer solutions. Finally, there is its sensitivity to changes in pH, which result from beam-induced hydrolysis in liquid cells. A dynamic process of dissolution and recrystallization of HAP was observed, which correlated with the production of H+ions by the beam during imaging. In addition, a large increase in the stability of the GLC under irradiation was noted. Specifically, no stable hydrogen bubbles were detected under the electron fluxes routinely exceeding 170 e-Å-2s-1. With the measured threshold dose for the bubble formation in pure water equaling 9 e-Å-2s-1, it was concluded that the presence of HAP increases the resistance of water against radiolysis in the GLC by more than an order of magnitude. These results confirm the possibility of using biological materials, such as HAP, as stabilizers in liquid cell electron microscopy. They outline a potential route for stabilization of specimens in liquid cells through the addition of a scavenger of reactive species generated by the beam-induced hydrolysis of water. These improvements are essential for enhancing both the resolution of imaging and the available imaging time, as well as avoiding the beam-induced artifacts.

18.
Luminescence ; 36(2): 481-488, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33080101

RESUMO

In the present study, CaZrO3 nanophosphors were sensitized with lanthanum (La) at different concentrations (0.5, 1.0, 1.5, 2.0, and 2.5) prepared using polyvinyl alcohol as the chelating agent through the sol-gel method. To study their structural and optical properties, samples were characterized by X-ray diffractometry (XRD), field emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM) and photoluminescence (PL). The XRD results revealed that samples were well crystallized and average crystallite sizes were calculated. The average crystallite size value was in good agreement with the value obtained from TEM analysis. Energy dispersive spectroscopy and FE-SEM confirmed the existence of La in the prepared samples. In the PL spectra, La-sensitized samples exhibited three bands at 402 nm, 438 nm, and 463 nm in the visible range when excited at the 260 nm wavelength. As the proportion of La increased, the intensity of bands at 438 nm and 463 nm decreased, whereas the band at 402 nm remained stable. Time-resolved PL spectra illustrated the lifetime of the samples. Corresponding CIE co-ordinates for La-sensitized CaZrO3 were calculated.


Assuntos
Lantânio , Álcool de Polivinil , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Espectrometria por Raios X
19.
Nano Lett ; 20(6): 4292-4297, 2020 06 10.
Artigo em Inglês | MEDLINE | ID: mdl-32453587

RESUMO

Solvated soft matter, both biological and synthetic, can now be imaged in liquids using liquid-cell transmission electron microscopy (LCTEM). However, such systems are usually composed solely of organic molecules (low Z elements) producing low contrast in TEM, especially within thick liquid films. We aimed to visualize liposomes by LCTEM rather than requiring cryogenic TEM (cryoTEM). This is achieved here by imaging in the presence of aqueous metal salt solutions. The increase in scattering cross-section by the cation gives a staining effect that develops in situ, which could be captured by real space TEM and verified by in situ energy dispersive x-ray spectroscopy (EDS). We identified beam-induced staining as a time-dependent process that enhances contrast to otherwise low contrast materials. We describe the development of this imaging method and identify conditions leading to exceptionally low electron doses for morphology visualization of unilamellar vesicles before beam-induced damage propagates.


Assuntos
Corantes , Lipossomos , Microscopia Eletrônica de Transmissão , Coloração e Rotulagem , Água
20.
Int J Mol Sci ; 22(9)2021 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-34063086

RESUMO

Polyphenols are natural substances that have been shown to provide various health benefits. Antioxidant, anti-inflammatory, and anti-carcinogenic effects have been described. At the same time, they inhibit the actions of bacteria, viruses, and fungi. Thus, studies have also examined their effects within the oral cavity. This review provides an overview on the different polyphenols, and their structure and interactions with the tooth surface and the pellicle. In particular, the effects of various tea polyphenols on bioadhesion and erosion have been reviewed. The current research confirms that polyphenols can reduce the growth of cariogenic bacteria. Furthermore, they can decrease the adherence of bacteria to the tooth surface and improve the erosion-protective properties of the acquired enamel pellicle. Tea polyphenols, especially, have the potential to contribute to an oral health-related diet. However, in vitro studies have mainly been conducted. In situ studies and clinical studies need to be extended and supplemented in order to significantly contribute to additive prevention measures in caries prophylaxis.


Assuntos
Odontologia , Polifenóis/farmacologia , Animais , Película Dentária/efeitos dos fármacos , Dieta , Humanos , Redes e Vias Metabólicas/efeitos dos fármacos , Chá
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