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1.
Nat Commun ; 11(1): 5062, 2020 10 08.
Artigo em Inglês | MEDLINE | ID: mdl-33033254

RESUMO

Septins are GTP-binding proteins involved in diverse cellular processes including division and membrane remodeling. Septins form linear, palindromic heteromeric complexes that can assemble in filaments and higher-order structures. Structural studies revealed various septin architectures, but questions concerning assembly-dynamics and -pathways persist. Here we used high-speed atomic force microscopy (HS-AFM) and kinetic modeling which allowed us to determine that septin filament assembly was a diffusion-driven process, while formation of higher-order structures was complex and involved self-templating. Slightly acidic pH and increased monovalent ion concentrations favor filament-assembly, -alignment and -pairing. Filament-alignment and -pairing further favored diffusion-driven assembly. Pairing is mediated by the septin N-termini face, and may occur symmetrically or staggered, likely important for the formation of higher-order structures of different shapes. Multilayered structures are templated by the morphology of the underlying layers. The septin C-termini face, namely the C-terminal extension of Cdc12, may be involved in membrane binding.


Assuntos
Microscopia de Força Atômica , Septinas/metabolismo , Simulação por Computador , Difusão , Concentração de Íons de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Cinética , Lipídeos/química , Domínios Proteicos , Septinas/ultraestrutura , Eletricidade Estática
2.
Nat Commun ; 11(1): 5066, 2020 10 08.
Artigo em Inglês | MEDLINE | ID: mdl-33033255

RESUMO

The inducible co-stimulator (ICOS) is a member of the CD28/B7 superfamily, and delivers a positive co-stimulatory signal to activated T cells upon binding to its ligand (ICOS-L). Dysregulation of this pathway has been implicated in autoimmune diseases and cancer, and is currently under clinical investigation as an immune checkpoint blockade. Here, we describe the molecular interactions of the ICOS/ICOS-L immune complex at 3.3 Å resolution. A central FDPPPF motif and residues within the CC' loop of ICOS are responsible for the specificity of the interaction with ICOS-L, with a distinct receptor binding orientation in comparison to other family members. Furthermore, our structure and binding data reveal that the ICOS N110 N-linked glycan participates in ICOS-L binding. In addition, we report crystal structures of ICOS and ICOS-L in complex with monoclonal antibodies under clinical evaluation in immunotherapy. Strikingly, antibody paratopes closely mimic receptor-ligand binding core interactions, in addition to contacting peripheral residues to confer high binding affinities. Our results uncover key molecular interactions of an immune complex central to human adaptive immunity and have direct implications for the ongoing development of therapeutic interventions targeting immune checkpoint receptors.


Assuntos
Anticorpos/uso terapêutico , Complexo Antígeno-Anticorpo/química , Ligante Coestimulador de Linfócitos T Induzíveis/química , Proteína Coestimuladora de Linfócitos T Induzíveis/química , Mimetismo Molecular/imunologia , Sequência de Aminoácidos , Complexo Antígeno-Anticorpo/metabolismo , Antígenos CD28/metabolismo , Humanos , Interações Hidrofóbicas e Hidrofílicas , Ligante Coestimulador de Linfócitos T Induzíveis/metabolismo , Proteína Coestimuladora de Linfócitos T Induzíveis/metabolismo , Cinética , Ligantes , Modelos Moleculares , Ligação Proteica , Multimerização Proteica
3.
Water Sci Technol ; 82(7): 1296-1303, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-33079710

RESUMO

Melamine foam is an important material in production and life. A series of porous carbon foams were obtained through a simple carbonization process of melamine foam at different temperatures. The carbon foams obtained at the carbonization temperature of 400 and 600 °C reveal a hydrophobic and even super-hydrophobic property (water contact angle larger than 150°) with a hexane adsorption much larger than that of melamine foam. However, the carbon foam obtained at the carbonization temperature of 800 °C reveals a super-hydrophilic property (water contact angle smaller than 5°) due to its severest shrinkage during the carbonization process. Interestingly, this series of carbon foams have an excellent performance in oil adsorption. However, the carbon membranes derived from the 800 °C carbon foam reveals oleophobicity under water (the adsorbed water at the surface was extremely important), which allows the penetration of water and blocks the infiltration of hexane at the same time. These different carbon forms have reversed applications in hexane/water separation.


Assuntos
Hexanos , Água , Adsorção , Carbono , Interações Hidrofóbicas e Hidrofílicas
4.
Water Sci Technol ; 82(2): 207-226, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32941164

RESUMO

Biological creatures with unique surface wettability have long served as a source of inspiration for scientists and engineers. More specifically, certain beetle species in the Namib Desert have evolved to collect water from fog on their backs by way of wettability patterns, which attracted an ongoing interest in biomimetic studies. Bioinspired materials exhibiting extreme wetting properties, such as superhydrophilic and superhydrophobic surfaces, have attracted considerable attention because of their potential use in various applications. Combining these two extreme states of superhydrophilicity and superhydrophobicity on the same surface in precise two-dimensional micropatterns opens exciting new functionalities and possibilities for a wide variety of applications. In this review we briefly describe the water-harvesting mechanisms of a genus of Namib Desert beetle, Stenocarpa, consisting of the theory of wetting and transporting. Then we describe the methods for fabricating superhydrophilic-superhydrophobic patterns and highlight some of the newer and emerging applications of these patterned substrates that are currently being explored. Finally, we provide conclusions and outlook concerning the future development of bioinspired surfaces of patterned wettability.


Assuntos
Materiais Biomiméticos , Besouros , Animais , Interações Hidrofóbicas e Hidrofílicas , Propriedades de Superfície , Água
5.
Waste Manag ; 118: 471-480, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32979778

RESUMO

Waste polystyrene (PS) and polycarbonate (PC) are crucial components arising from mixtures of plastic products, whose recycling is significantly limited by separation efficiency. In this work, to assist the flotation separation of PC and PS, we proposed a novel modification technology of surface alkoxylation pretreatment (SAP) where PC surface reacted with glycerol and urea. The SAP could selectively transform the hydrophobic PC into hydrophilic plastic, while the PS remained its hydrophobic surface owing to the exclusion from SAP process. Benefiting from the hydrophilic PC, the separation efficiency of PS and PC could reach the maximum of 99.34% under optimum conditions (urea dosage of 5 g, pretreatment temperature of 130 °C, pretreatment time of 10 min, flotation time of 2.5 min, frother concentration of 16.5 mg/L, and airflow rate of 7.2 mL/min). The mechanism of SAP was systematically analyzed by wettability, surface morphology, molecular weight, and chemical reactions. Compared with PS plastic, the pretreated PC presented better wettability, rougher surface, and significantly reducing molecular weight. The improvement of PC hydrophilicity can be attributed to the cleavage of ester bonds on backbone chains and the introduction of hydrophilic hydroxyl groups. The effective SAP process proves that chemical recycling of waste plastic can provide a novel strategy for surface modification and flotation separation of PS and PC.


Assuntos
Poliestirenos , Eliminação de Resíduos , Interações Hidrofóbicas e Hidrofílicas , Plásticos , Cimento de Policarboxilato
6.
PLoS One ; 15(8): e0238150, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32866159

RESUMO

Immunogenicity is an important concern for therapeutic antibodies during drug development. By analyzing co-crystal structures of idiotypic antibodies and their antibodies, we found that anti-idiotypic antibodies usually bind the Complementarity Determining Regions (CDR) of idiotypic antibodies. Sequence and structural features were identified for distinguishing immunogenic antibodies from non-immunogenic antibodies. For example, non-immunogenic antibodies have a significantly larger cavity volume at the CDR region and a more hydrophobic CDR-H3 loop than immunogenic antibodies. Antibodies containing no Gly at the turn of CDR-H2 loop are often immunogenic. We integrated these features together with a machine learning platform to Predict Immunogenicity for humanized and full human THerapeutic Antibodies (PITHA). This method achieved an accuracy of 83% in leave-one-out experiment for 29 therapeutic antibodies with available crystal structures. The accuracy decreased to 65% for 23 test antibodies with modeled structures, because their crystal structures were not available, and the prediction was made with modeled structures. The server of this method is accessible at http://mabmedicine.com/PITHA.


Assuntos
Anticorpos Monoclonais Humanizados/química , Anticorpos Monoclonais Humanizados/imunologia , Formação de Anticorpos/imunologia , Cristalografia por Raios X/métodos , Desenvolvimento de Medicamentos/métodos , Humanos , Interações Hidrofóbicas e Hidrofílicas , Conformação Proteica
7.
PLoS One ; 15(9): e0239531, 2020.
Artigo em Inglês | MEDLINE | ID: covidwho-781673

RESUMO

The worldwide shortage of single-use N95 respirators and surgical masks due to the COVID-19 pandemic has forced many health care personnel to use their existing equipment for as long as possible. In many cases, workers cover respirators with available masks in an attempt to extend their effectiveness against the virus. Due to low mask supplies, many people instead are using face coverings improvised from common fabrics. Our goal was to determine what fabrics would be most effective in both practices. Under laboratory conditions, we examined the hydrophobicity of fabrics (cotton, polyester, silk), as measured by their resistance to the penetration of small and aerosolized water droplets, an important transmission avenue for the virus causing COVID-19. We also examined the breathability of these fabrics and their ability to maintain hydrophobicity despite undergoing repeated cleaning. Laboratory-based tests were conducted when fabrics were fashioned as an overlaying barrier for respirators and when constructed as face coverings. When used as material in these two situations, silk was more effective at impeding the penetration and absorption of droplets due to its greater hydrophobicity relative to other tested fabrics. We found that silk face coverings repelled droplets in spray tests as well as disposable single-use surgical masks, and silk face coverings have the added advantage over masks such that they can be sterilized for immediate reuse. We show that silk is a hydrophobic barrier to droplets, can be more breathable than other fabrics that trap humidity, and are re-useable via cleaning. We suggest that silk can serve as an effective material for making hydrophobic barriers that protect respirators, and silk can now be tested under clinical conditions to verify its efficacy for this function. Although respirators are still the most appropriate form of protection, silk face coverings possess properties that make them capable of repelling droplets.


Assuntos
Betacoronavirus , Infecções por Coronavirus/prevenção & controle , Máscaras/normas , Pandemias/prevenção & controle , Equipamento de Proteção Individual/normas , Pneumonia Viral/prevenção & controle , Seda/normas , Têxteis/normas , Filtração/métodos , Humanos , Interações Hidrofóbicas e Hidrofílicas , Equipamento de Proteção Individual/virologia , Dispositivos de Proteção Respiratória
8.
Biomolecules ; 10(9)2020 09 21.
Artigo em Inglês | MEDLINE | ID: mdl-32967116

RESUMO

We report the results of our in silico study of approved drugs as potential treatments for COVID-19. The study is based on the analysis of normal modes of proteins. The drugs studied include chloroquine, ivermectin, remdesivir, sofosbuvir, boceprevir, and α-difluoromethylornithine (DMFO). We applied the tools we developed and standard tools used in the structural biology community. Our results indicate that small molecules selectively bind to stable, kinetically active residues and residues adjoining them on the surface of proteins and inside protein pockets, and that some prefer hydrophobic sites over other active sites. Our approach is not restricted to viruses and can facilitate rational drug design, as well as improve our understanding of molecular interactions, in general.


Assuntos
Antivirais/farmacologia , Infecções por Coronavirus/tratamento farmacológico , Pandemias , Pneumonia Viral/tratamento farmacológico , Monofosfato de Adenosina/análogos & derivados , Monofosfato de Adenosina/química , Monofosfato de Adenosina/farmacologia , Alanina/análogos & derivados , Alanina/química , Alanina/farmacologia , Anticorpos Antivirais/imunologia , Reações Antígeno-Anticorpo , Antivirais/química , Antivirais/uso terapêutico , Betacoronavirus , Sítios de Ligação , Cloroquina/química , Cloroquina/farmacologia , Infecções por Coronavirus/prevenção & controle , Reposicionamento de Medicamentos , Eflornitina/química , Eflornitina/farmacologia , Humanos , Interações Hidrofóbicas e Hidrofílicas , Ivermectina/química , Ivermectina/farmacologia , L-Lactato Desidrogenase/química , L-Lactato Desidrogenase/efeitos dos fármacos , Modelos Moleculares , Simulação de Acoplamento Molecular , Pandemias/prevenção & controle , Peptidil Dipeptidase A/química , Peptidil Dipeptidase A/efeitos dos fármacos , Pneumonia Viral/prevenção & controle , Prolina/análogos & derivados , Prolina/química , Prolina/farmacologia , Ligação Proteica , Conformação Proteica , Mapeamento de Interação de Proteínas , Receptores da Glicina/química , Receptores da Glicina/efeitos dos fármacos , Saposinas/química , Saposinas/efeitos dos fármacos , Sofosbuvir/química , Sofosbuvir/farmacologia , Glicoproteína da Espícula de Coronavírus/química , Glicoproteína da Espícula de Coronavírus/efeitos dos fármacos , Relação Estrutura-Atividade
9.
PLoS One ; 15(9): e0239531, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32946526

RESUMO

The worldwide shortage of single-use N95 respirators and surgical masks due to the COVID-19 pandemic has forced many health care personnel to use their existing equipment for as long as possible. In many cases, workers cover respirators with available masks in an attempt to extend their effectiveness against the virus. Due to low mask supplies, many people instead are using face coverings improvised from common fabrics. Our goal was to determine what fabrics would be most effective in both practices. Under laboratory conditions, we examined the hydrophobicity of fabrics (cotton, polyester, silk), as measured by their resistance to the penetration of small and aerosolized water droplets, an important transmission avenue for the virus causing COVID-19. We also examined the breathability of these fabrics and their ability to maintain hydrophobicity despite undergoing repeated cleaning. Laboratory-based tests were conducted when fabrics were fashioned as an overlaying barrier for respirators and when constructed as face coverings. When used as material in these two situations, silk was more effective at impeding the penetration and absorption of droplets due to its greater hydrophobicity relative to other tested fabrics. We found that silk face coverings repelled droplets in spray tests as well as disposable single-use surgical masks, and silk face coverings have the added advantage over masks such that they can be sterilized for immediate reuse. We show that silk is a hydrophobic barrier to droplets, can be more breathable than other fabrics that trap humidity, and are re-useable via cleaning. We suggest that silk can serve as an effective material for making hydrophobic barriers that protect respirators, and silk can now be tested under clinical conditions to verify its efficacy for this function. Although respirators are still the most appropriate form of protection, silk face coverings possess properties that make them capable of repelling droplets.


Assuntos
Betacoronavirus , Infecções por Coronavirus/prevenção & controle , Máscaras/normas , Pandemias/prevenção & controle , Equipamento de Proteção Individual/normas , Pneumonia Viral/prevenção & controle , Seda/normas , Têxteis/normas , Filtração/métodos , Humanos , Interações Hidrofóbicas e Hidrofílicas , Equipamento de Proteção Individual/virologia , Dispositivos de Proteção Respiratória
10.
Chemosphere ; 254: 126873, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32957285

RESUMO

The removal of organic pollutants from water is highly desired because of the development of industrial and social economy. Superhydrophilic and underwater superoleophobic membranes are emerging materials for effective oil/water separation. In this paper, superhydrophilic and underwater superoleophobic polypropylene (PP) melt-blown membranes were prepared through melt-blown and in situ growth method, achieving highly efficient oil/water separation. After in situ growth, polydopamine (PDA) grows on the surface of PP fibers, and the addition of coupling agent (3-aminopropyltriethoxysilane, APTES) can improve the stability of the membrane in harsh environments (1 M HCl, 1 M NaOH, 1 M NaCl). The PDA/APTES@PP membrane could dramatically enhance the wetting (water contact angle ∼0, underwater oil contact angle∼154°) compare with the pristine PP melt-blown membrane (water contact angle ∼130°, underwater oil contact angle ∼0). Moreover, the filtration performance is at a high level (∼99%). The behaviors are comparable or even superior to the typical reported results in the references (such as the mussel-inspired superhydrophilic PVDF membrane and copper mesh). This method provides a facile route to prepared multi-functional membrane for highly efficiency oil/water separation and industrial oily wastewater remediation.


Assuntos
Indóis/análise , Polímeros/análise , Cobre , Interações Hidrofóbicas e Hidrofílicas , Membranas Artificiais , Óleos , Águas Residuárias , Água , Purificação da Água , Molhabilidade
11.
Yakugaku Zasshi ; 140(8): 1001-1006, 2020.
Artigo em Japonês | MEDLINE | ID: mdl-32741857

RESUMO

Ascertaining the absorption, distribution, metabolism, and excretion (ADME) profile of drugs is one of the most crucial factors in the process of drug discovery. Since it is important to combine water solubility and cell permeability within the compound to achieve the desired ADME properties, an appropriate balance between lipophilicity and hydrophilicity is required. It is often necessary to facilitate hydrophilicity of very hydrophobic candidates, because quite lipophobic molecules are rarely hit as positive in molecular-targeted or cell-based screenings. For that purpose, it has been popular to conjugate hydrophobic molecules with polyethylene glycol (PEG). However, PEG is a polymer, and PEG-conjugated molecules are not uniform. Besides, the dosage should be much increased compared with the original molecule due to the increase in molecular weight. Therefore we have been developing alternative ways to endow hydrophobic compounds with extra hydrophilicity by conjugating with symmetrically branched glycerol oligomers. This technology is versatile and easily applicable to various hydrophobic compounds. Water-solubility of fenofibrate, one of the most hydrophobic medicines in clinical use, was facilitated by a factor of more than 2000, and its lipid-lowering effect in vivo improved more than ten-fold, by simply conjugating with branched glycerol trimer, for instance. Here we will briefly introduce the basic concepts and our successful experiences of applying branched glycerol oligomers including antitumor agents in terms of water-solubility, pharmacological effects, and pharmacokinetics, and merits and current issues will be discussed in this review.


Assuntos
Antineoplásicos , Glicerol/química , Interações Hidrofóbicas e Hidrofílicas , Antineoplásicos/farmacocinética , Antineoplásicos/farmacologia , Ácidos Fíbricos , Polietilenoglicóis/química , Polímeros , Solubilidade , Água
12.
Nat Commun ; 11(1): 4137, 2020 08 18.
Artigo em Inglês | MEDLINE | ID: mdl-32811827

RESUMO

The class B secretin GPCR (SecR) has broad physiological effects, with target potential for treatment of metabolic and cardiovascular disease. Molecular understanding of SecR binding and activation is important for its therapeutic exploitation. We combined cryo-electron microscopy, molecular dynamics, and biochemical cross-linking to determine a 2.3 Å structure, and interrogate dynamics, of secretin bound to the SecR:Gs complex. SecR exhibited a unique organization of its extracellular domain (ECD) relative to its 7-transmembrane (TM) core, forming more extended interactions than other family members. Numerous polar interactions formed between secretin and the receptor extracellular loops (ECLs) and TM helices. Cysteine-cross-linking, cryo-electron microscopy multivariate analysis and molecular dynamics simulations revealed that interactions between peptide and receptor were dynamic, and suggested a model for initial peptide engagement where early interactions between the far N-terminus of the peptide and SecR ECL2 likely occur following initial binding of the peptide C-terminus to the ECD.


Assuntos
Subunidades alfa Gs de Proteínas de Ligação ao GTP/química , Simulação de Dinâmica Molecular , Receptores Acoplados a Proteínas-G/química , Receptores dos Hormônios Gastrointestinais/química , Secretina/química , Sequência de Aminoácidos , Animais , Sítios de Ligação/genética , Linhagem Celular , Cricetinae , Microscopia Crioeletrônica , Cristalografia por Raios X , Cisteína/química , Cisteína/metabolismo , Subunidades alfa Gs de Proteínas de Ligação ao GTP/ultraestrutura , Humanos , Ligação de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Insetos , Modelos Moleculares , Ligação Proteica , Domínios Proteicos/genética , Estrutura Secundária de Proteína , Receptores Acoplados a Proteínas-G/metabolismo , Receptores Acoplados a Proteínas-G/ultraestrutura , Receptores dos Hormônios Gastrointestinais/metabolismo , Receptores dos Hormônios Gastrointestinais/ultraestrutura , Secretina/metabolismo
13.
Nat Commun ; 11(1): 4141, 2020 08 18.
Artigo em Inglês | MEDLINE | ID: mdl-32811830

RESUMO

Members of the Herpesviridae, including the medically important alphaherpesvirus varicella-zoster virus (VZV), induce fusion of the virion envelope with cell membranes during entry, and between cells to form polykaryocytes in infected tissues. The conserved glycoproteins, gB, gH and gL, are the core functional proteins of the herpesvirus fusion complex. gB serves as the primary fusogen via its fusion loops, but functions for the remaining gB domains remain unexplained. As a pathway for biological discovery of domain function, our approach used structure-based analysis of the viral fusogen together with a neutralizing antibody. We report here a 2.8 Å cryogenic-electron microscopy structure of native gB recovered from VZV-infected cells, in complex with a human monoclonal antibody, 93k. This high-resolution structure guided targeted mutagenesis at the gB-93k interface, providing compelling evidence that a domain spatially distant from the gB fusion loops is critical for herpesvirus fusion, revealing a potential new target for antiviral therapies.


Assuntos
Anticorpos Neutralizantes/química , Herpesvirus Humano 3/química , Proteínas do Envelope Viral/química , Internalização do Vírus , Anticorpos Neutralizantes/imunologia , Anticorpos Neutralizantes/ultraestrutura , Microscopia Crioeletrônica , Humanos , Interações Hidrofóbicas e Hidrofílicas , Modelos Moleculares , Conformação Proteica em Folha beta/genética , Domínios Proteicos/genética , Proteínas do Envelope Viral/imunologia , Proteínas do Envelope Viral/ultraestrutura
14.
PLoS One ; 15(8): e0237789, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32810188

RESUMO

Aquaporins are water-permeable membrane-channel proteins found in biological cell membranes that selectively exclude ions and large molecules and have high water permeability, which makes them promising candidates for water desalination systems. To effectively apply the properties of aquaporins in the desalination process, many studies have been conducted on aquaporin-lipid membrane systems using phospholipids, which are the main component of cell membranes. Many parametric studies have evaluated the permeability of such systems with various aquaporin types and lipid compositions. In this study, we performed molecular dynamics simulations for four cases with different protein-lipid molar ratios (1:50, 1:75, 1:100, and 1:150) between aquaporin Z and the phospholipids, and we propose a possibility of the existence of optimal protein-lipid molar ratio to maximize water permeability. Elucidating these simulation results from a structural viewpoint suggests that there is a relationship between the permeability and changes in the hydrophobic thickness of the lipid membrane adjacent to the aquaporin as a structural parameter. The results of this study can help optimize the design of an aquaporin-lipid membrane by considering its molar ratio at an early stage of development.


Assuntos
Aquaporinas/metabolismo , Proteínas de Escherichia coli/metabolismo , Bicamadas Lipídicas/metabolismo , Fosfolipídeos/metabolismo , Purificação da Água/métodos , Água/metabolismo , Aquaporinas/química , Proteínas de Escherichia coli/química , Interações Hidrofóbicas e Hidrofílicas , Bicamadas Lipídicas/química , Modelos Químicos , Simulação de Dinâmica Molecular , Pressão Osmótica , Fosfolipídeos/química , Salinidade , Água/química
15.
Int J Nanomedicine ; 15: 5629-5643, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32801706

RESUMO

Purpose: Lecithin/chitosan nanoparticles have shown great promise in the transdermal delivery of therapeutic agents. Baicalein, a natural bioactive flavonoid, possesses multiple biological activities against dermatosis. However, its topical application is limited due to its inherently poor hydrophilicity and lipophilicity. In this study, the baicalein-phospholipid complex was prepared to enhance the lipophilicity of baicalein and then lecithin/chitosan nanoparticles loaded with the baicalein-phospholipid complex were developed to improve the transdermal retention and permeability of baicalein. Methods: Lecithin/chitosan nanoparticles were prepared by the solvent-injection method and characterized in terms of particle size distribution, zeta potential, and morphology. The in vitro release, the ex vivo and in vivo permeation studies, and safety evaluation of lecithin/chitosan nanoparticles were performed to evaluate the effectiveness in enhancing transdermal retention and permeability of baicalein. Results: The lecithin/chitosan nanoparticles obtained by the self-assembled interaction of chitosan and lecithin not only efficiently encapsulated the drug with high entrapment efficiency (84.5%) but also provided sustained release of baicalein without initial burst release. Importantly, analysis of the permeation profile ex vivo and in vivo demonstrated that lecithin/chitosan nanoparticles prolonged the retention of baicalein in the skin and efficiently penetrated the barrier of stratum corneum without displaying skin irritation. Conclusion: These results indicate the potential of drug-phospholipid complexes in enhancing the entrapment efficiency and self-assembled lecithin/chitosan nanoparticles based on phospholipid complexes in the design of a rational transdermal delivery platform to improve the efficiency of transdermal therapy by enhancing its percutaneous retention and penetration in the skin.


Assuntos
Flavanonas/administração & dosagem , Nanopartículas/administração & dosagem , Fosfolipídeos/química , Administração Cutânea , Animais , Quitosana/química , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos/métodos , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Flavanonas/farmacocinética , Interações Hidrofóbicas e Hidrofílicas , Lecitinas/química , Masculino , Nanopartículas/efeitos adversos , Nanopartículas/química , Permeabilidade , Ratos Sprague-Dawley , Pele/efeitos dos fármacos , Pele/patologia , Absorção Cutânea/efeitos dos fármacos , Testes de Irritação da Pele
16.
J Chromatogr A ; 1627: 461402, 2020 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-32823107

RESUMO

Surfactants are used in various applications: cosmetics, pharmaceuticals, petrochemicals, environmental, etc. Many of these compounds are polydisperse, and because of this intrinsic polydispersity, it is essential to have a universal detector with a uniform response to quantify them in a simple way. Indeed, Charged Aerosol Detector (CAD) was presented as a universal detector with a uniform response. Thus, in the present study, the CAD response, in a High-Performance Liquid Chromatography - CAD configuration (HPLCCAD), was evaluated using purified alcohol ethoxylated surfactants. A semi-preparative liquid chromatography step using a Hydrophilic interaction chromatography (HILIC) bare silica column (150 mm, 4.6 mm, 2.6 µm) was implemented to prepare eleven homologues of BrijC10, a nonionic surfactant. These homologues differed only by the number of ethylene oxide units. BrijC10 homologues were analyzed by HPLCCAD, using a HILIC bare silica column (150 mm, 2.1 mm, 2.6 µm) to determine the HPLCCAD response factors of purified homologues. From the calibration curves (from 100 to 500 mg.kg-1), their response factors were estimated: differences in response factors were observed and a maximum difference in response factors of 3.6 was obtained. Thus, it could be concluded that CAD hyphenated to HILIC separation did not present a uniform response for this homologue's distribution.


Assuntos
Aerossóis/química , Cromatografia Líquida de Alta Pressão/métodos , Calibragem , Cromatografia Líquida de Alta Pressão/normas , Etil-Éteres/química , Interações Hidrofóbicas e Hidrofílicas , Dióxido de Silício/química , Tensoativos/química
17.
J Chromatogr A ; 1628: 461444, 2020 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-32822983

RESUMO

The effect of bead and ligand structure on protein adsorption was investigated for multimodal anion exchangers combining a quaternary ammonium ion group with hydrophobic moieties: Nuvia aPrime 1 and aPrime 2, based on a 54 µm diameter polymeric bead, and Capto Adhere ImpRes and Capto Adhere, based on agarose beads 51 and 78 µm diameter, respectively. Bovine serum albumin (BSA) monomer, BSA dimer, and thyroglobulin (Tg) were used as model proteins. Based on TEM imaging and iSEC, the Nuvia resins have a microgranular structure and large pores (110 nm radius), while the Capto resins have a fibrous structure and smaller pores (32-36 nm radius). Comparable binding capacities (80-110 mg/mL), decreasing as salt is added, are observed for all three proteins on the Nuvia resins. Higher capacities (110-130 mg/mL), also decreasing as salt is added, are observed for BSA monomer and dimer on the Capto resins. However, the Tg binding capacity is very low in this case and increases as salt is added. Confocal laser scanning microscopy show that the kinetics are controlled by pore diffusion for all four resins, but with diffusivities that decrease as the protein size increases especially for the Capto resins. For Tg at low salt, binding is restricted to a thin shell close to the bead surface for both Capto resins. The ratio of effective and free diffusivity is about 0.30, 0.18, and 0.08 for BSA monomer, BSA dimer, and Tg, respectively, on the Nuvia resin. These values decrease to about 0.11, 0.04, and 0.01, respectively, for the Capto resins as a result of diffusional hindrance. Dynamic binding capacities are consistent with the equilibrium and rate behaviors.


Assuntos
Resinas de Troca de Ânions/química , Cromatografia por Troca Iônica , Proteínas/metabolismo , Adsorção , Ânions/química , Difusão , Interações Hidrofóbicas e Hidrofílicas , Cinética , Ligantes , Polímeros/química , Sefarose/química , Soroalbumina Bovina/química
18.
Nat Protoc ; 15(9): 2980-3008, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32839575

RESUMO

High-surface-area mesoporous materials expose abundant functional sites for improved performance in applications such as gas storage/separation, catalysis, and sensing. Recently, soft templates composed of amphiphilic surfactants and block copolymers have been used to introduce mesoporosity in various materials, including metals, metal oxides and carbonaceous compounds. In particular, mesoporous metals are attractive in electrocatalysis because their porous networks expose numerous unsaturated atoms on high-index facets that are highly active in catalysis. In this protocol, we describe how to create mesoporous metal films composed of gold, palladium, or platinum using block copolymer micelle templates. The amphiphilic block copolymer micelles are the sacrificial templates and generate uniform structures with tunable pore sizes in electrodeposited metal films. The procedure describes the electrodeposition in detail, including parameters such as micelle diameters, deposition potentials, and deposition times to ensure reproducibility. The micelle diameters can be controlled by swelling the micelles with different solvent mixtures or by using block copolymer micelles with different molecular weights. The deposition potentials and deposition times allow further control of the mesoporous structure and its thickness, respectively. Procedures for example applications are included: glucose oxidation, ethanol oxidation and methanol oxidation reactions. The synthetic methods for preparation of mesoporous metal films will take ~4 h; the subsequent electrochemical tests will take ~5 h for glucose sensing and ~3 h for alcohol oxidation reaction.


Assuntos
Ouro/química , Paládio/química , Platina/química , Álcoois/química , Catálise , Técnicas de Química Sintética , Eletroquímica , Interações Hidrofóbicas e Hidrofílicas , Micelas , Modelos Moleculares , Conformação Molecular , Oxirredução , Polímeros/química
19.
PLoS One ; 15(8): e0237888, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32813716

RESUMO

Norovirus, the leading cause of non-bacterial food poisoning, is responsible for several outbreaks associated with bivalves and ready-to-eat food products worldwide. As norovirus is resistant to alcohol, which is commonly used in food manufacturing processes, sodium hypochlorite is used for its inactivation. However, sodium hypochlorite has two disadvantages: it cannot be added to foods, and its effect is significantly reduced in the presence of organic compounds. Thus, a novel disinfectant against norovirus is urgently required for food hygiene. Thermally denatured egg white lysozyme inactivates norovirus; however, the optimal inactivating conditions and the underlying mechanism are unclear. In the present study, the inactivating mechanism of heat-denatured lysozyme against norovirus was analyzed using murine norovirus strain 1 (MNV-1). We found that the inactivating effect was enhanced by adjusting the pH of the lysozyme solution before thermal denaturation to 6.5 or higher. The reaction of heat-denatured lysozyme and MNV-1 was irreversible, and norovirus was completely inactivated after exposure to heat-denatured lysozyme. Furthermore, it was found that lysozyme residues 5-39 contributed to the norovirus-inactivating effect. Notably, the hydrophobicity and positive charges in this region contributed to the norovirus-inactivating effect, as evidenced by the norovirus inactivation test using mutated residues 5-39. These findings are novel and highlight the possible application of heat-denatured lysozyme as a disinfectant against norovirus in a wide range of food processes.


Assuntos
Temperatura Alta , Interações Hidrofóbicas e Hidrofílicas , Muramidase/metabolismo , Norovirus/fisiologia , Desnaturação Proteica , Inativação de Vírus , Sequência de Aminoácidos , Animais , Regulação da Expressão Gênica , Concentração de Íons de Hidrogênio , Macrófagos/virologia , Camundongos , Muramidase/química , Peptídeos/química , Domínios Proteicos , Células RAW 264.7
20.
PLoS One ; 15(8): e0237884, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32841243

RESUMO

The Solanum tuberosum plant specific insert (StPSI) has a defensive role in potato plants, with the requirements of acidic pH and anionic lipids. The StPSI contains a set of three highly conserved disulfide bonds that bridge the protein's helical domains. Removal of these bonds leads to enhanced membrane interactions. This work examined the effects of their sequential removal, both individually and in combination, using all-atom molecular dynamics to elucidate the role of disulfide linkages in maintaining overall protein tertiary structure. The tertiary structure was found to remain stable at both acidic (active) and neutral (inactive) pH despite the removal of disulfide linkages. The findings include how the dimer structure is stabilized and the impact on secondary structure on a residue-basis as a function of disulfide bond removal. The StPSI possesses an extensive network of inter-monomer hydrophobic interactions and intra-monomer hydrogen bonds, which is likely the key to the stability of the StPSI by stabilizing local secondary structure and the tertiary saposin-fold, leading to a robust association between monomers, regardless of the disulfide bond state. Removal of disulfide bonds did not significantly impact secondary structure, nor lead to quaternary structural changes. Instead, disulfide bond removal induces regions of amino acids with relatively higher or lower variation in secondary structure, relative to when all the disulfide bonds are intact. Although disulfide bonds are not required to preserve overall secondary structure, they may have an important role in maintaining a less plastic structure within plant cells in order to regulate membrane affinity or targeting.


Assuntos
Dissulfetos/metabolismo , Simulação de Dinâmica Molecular , Proteínas de Plantas/metabolismo , Saposinas/metabolismo , Solanum tuberosum/metabolismo , Cisteína/metabolismo , Ligação de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Proteínas de Plantas/química , Multimerização Proteica , Estabilidade Proteica , Estrutura Secundária de Proteína , Sais/química , Enxofre/metabolismo
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