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1.
Biotechnol Bioeng ; 2023 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-37163237

RESUMO

Membrane fouling by monoclonal antibodies (mAbs) is one of the main challenges in virus-filtration processes. Previous publications attributed membrane fouling to the presence of mAb aggregates in the solution, which block the membrane pores. This fouling mechanism can be solved by a prefilter; however, it was shown that there are mAbs that severely foul the membranes (reduce permeability by 90% and more) even after prefiltering the aggregates, while other mAbs foul the membrane weakly (reduce permeability by ~10% and less). Unfortunately, the differences between the fouling- and the nonfouling mAbs have never been convincingly explained. To get a deeper insight on these differences, we measured the fouling of chemically modified Isoprene-Styrene-4-vinylpyridine (ISV) membranes (TeraPore Technologies) by 8 mAbs exhibiting different hydrophobicity and charge. The results show that mAb solutions with low concentration of aggregates foul ISV membranes via an adsorptive mechanism, and the adsorption is driven mainly by hydrophobic forces between the mAb and the membrane. The charge of the mAbs plays a secondary role in fouling. We want to emphasize that the conclusions pertain to ISV membranes; the insights presented in this paper can potentially be used to engineer new surface chemistries to mitigate fouling of other virus-filtration and/or ultrafiltration membranes.

2.
Proc Natl Acad Sci U S A ; 115(32): 8070-8075, 2018 08 07.
Artigo em Inglês | MEDLINE | ID: mdl-30026197

RESUMO

Understanding the fundamental wetting behavior of liquids on surfaces with pores or cavities provides insights into the wetting phenomena associated with rough or patterned surfaces, such as skin and fabrics, as well as the development of everyday products such as ointments and paints, and industrial applications such as enhanced oil recovery and pitting during chemical mechanical polishing. We have studied, both experimentally and theoretically, the dynamics of the transitions from the unfilled/partially filled (Cassie-Baxter) wetting state to the fully filled (Wenzel) wetting state on intrinsically hydrophilic surfaces (intrinsic water contact angle <90°, where the Wenzel state is always the thermodynamically favorable state, while a temporary metastable Cassie-Baxter state can also exist) to determine the variables that control the rates of such transitions. We prepared silicon wafers with cylindrical cavities of different geometries and immersed them in bulk water. With bright-field and confocal fluorescence microscopy, we observed the details of, and the rates associated with, water penetration into the cavities from the bulk. We find that unconnected, reentrant cavities (i.e., cavities that open up below the surface) have the slowest cavity-filling rates, while connected or non-reentrant cavities undergo very rapid transitions. Using these unconnected, reentrant cavities, we identified the variables that affect cavity-filling rates: (i) the intrinsic contact angle, (ii) the concentration of dissolved air in the bulk water phase (i.e., aeration), (iii) the liquid volatility that determines the rate of capillary condensation inside the cavities, and (iv) the presence of surfactants.


Assuntos
Interações Hidrofóbicas e Hidrofílicas , Termodinâmica , Molhabilidade , Ar , Fluoresceína/química , Menisco/química , Transição de Fase , Pressão , Silício/química , Solubilidade , Propriedades de Superfície , Tensoativos/química , Volatilização , Água/química
3.
Langmuir ; 35(26): 8709-8715, 2019 07 02.
Artigo em Inglês | MEDLINE | ID: mdl-31244251

RESUMO

Supported lipid bilayers with incorporated membrane proteins have promising potential for diverse applications, such as filtration processes, drug delivery, and biosensors. For these applications, the continuity (lack of defects), electrical resistivity, and charge capacitance of the lipid bilayers are crucial. Here, we highlight the effects of temperature changes and the rate of temperature changes on the vertical and lateral expansion and contraction of lipid bilayers, which in turn affect the lipid bilayer resistivity and capacitance. We focused on lipid bilayers that consist of 50 mol % dimyristoyl- sn-glycero-3-phosphocholine (zwitterionic lipid) and 50 mol % dimyristoyl-3-trimethylammonium-propane (positively charged lipid) lipids. This lipid mixture is known to self-assemble into a continuous lipid bilayer on silicon wafers. It is shown experimentally and explained theoretically that slow cooling (e.g., -0.4 °C min-1) increases the resistivity significantly and reduces the capacitance of lipid bilayers, and these trends are reversed by heating. However, fast cooling (∼ -10 °C min-1 or faster) damages the membrane and reduces the resistivity and capacitance of lipid bilayers to practically zero. Importantly, the addition of 50 mol % cholesterol to lipid bilayers prevents the resistivity and capacitance reduction after fast cooling. It is argued that the ratio of lipid diffusion coefficient to thermal expansion/contraction rate (proportional to the heating/cooling rate) is the crucial parameter that determines the effects of temperature changes on lipids bilayers. A high ratio (fast lipid diffusion) increases the lipid bilayer resistivity and decreases the capacitance upon cooling and vice versa. Similar trends are expected for lipid membranes that consist of other lipids or lipidlike mixtures.


Assuntos
Bicamadas Lipídicas/química , Temperatura , Capacitância Elétrica , Impedância Elétrica , Imagem Óptica , Tamanho da Partícula , Semicondutores , Silício/química , Propriedades de Superfície
4.
Langmuir ; 35(48): 15526-15534, 2019 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-31469282

RESUMO

Dust particles can adhere to surfaces, thereby decreasing the efficiency of diverse processes, such as light absorption by solar panels. It is well known that superhydrophobicity reduces the friction between water droplets and the surface, thus allowing water drops to slide/roll and detach (clean) particles from surfaces. However, the forces that attach and detach particles from surfaces during the self-cleaning mechanism and the effect of nanotextures on these forces are not fully understood. To shed light on these forces and the effect of nanotexture on them, we prepared four Si-based samples (relevant to solar panels): (1) smooth or (2) nanotextured hydrophilic surfaces and (3) smooth or (4) nanotextured hydrophobic surfaces. In agreement with previous publications, it is shown that the efficiency of particle removal increases with hydrophobicity. Furthermore, nanotexture enhances the hydrophobicity, whereby particle removal is further increased. Specifically, hydrophilic particle removal increased from ∼41%, from hydrophilic smooth Si wafers to 98% from superhydrophobic Si-based nanotextured surfaces. However, the reason for the increased particle removal is not low friction between the droplets and the superhydrophobic surfaces; it is the reduction of the adhesion force between the particle and the surface and the altered geometry of the water-particle-air line tension acting on the particles on superhydrophobic surfaces, which increases the force that can detach particles from the surfaces. The experimental methods we used and the criterion for particle removal we derived can be implemented to engineer self-cleaning surfaces using other surfaces and dust particles, exhibiting different chemistries and/or textures.

5.
Langmuir ; 35(5): 1699-1713, 2019 02 05.
Artigo em Inglês | MEDLINE | ID: mdl-29641904

RESUMO

There is a need for the development of antifouling materials to resist adsorption of biomacromolecules. Here we describe the preparation of a novel zwitterionic block copolymer with the potential to prevent or delay the formation of microbial biofilms. The block copolymer comprised a zwitterionic (hydrophilic) section of alternating glutamic acid (negatively charged) and lysine (positively charged) units and a hydrophobic polystyrene section. Cryo-TEM and dynamic-light-scattering (DLS) results showed that, on average, the block copolymer self-assembled into 7-nm-diameter micelles in aqueous solutions (0 to 100 mM NaCl, pH 6). Quartz crystal microbalance with dissipation monitoring (QCM-D), atomic force microscopy (AFM), and contact angle measurements demonstrated that the block copolymer self-assembled into a brush-like monolayer on polystyrene surfaces. The brush-like monolayer produced from a 100 mg/L block copolymer solution exhibited an average distance, d, of approximately 4-8 nm between each block copolymer molecule (center to center). Once the brush-like monolayer self-assembled, it reduced EPS adsorption onto the polystyrene surface by ∼70% (mass), reduced the rate of bacterial attachment by >80%, and inhibited the development of thick biofilms. QCM-D results revealed that the EPS molecules penetrate between the chains of the brush and adsorb onto the polystyrene surface. Additionally, AFM analyses showed that the brush-like monolayer prevents the adhesion of large (> d) hydrophilic colloids onto the surface via hydration repulsion; however, molecules or colloids small enough to fit between the brush polymers (< d) were able to be adsorbed onto the surface via van der Waals interactions. Overall, we found that the penetration of extracellular organelles, as well as biopolymers through the brush, is critical for the failure of the antifouling coating, and likely could be prevented through tuning of the brush density. Stability and biofilm development testing on multiple surfaces (polypropylene, glass, and stainless steel) support practical applications of this novel block copolymer.


Assuntos
Incrustação Biológica/prevenção & controle , Materiais Revestidos Biocompatíveis/química , Ácido Poliglutâmico/análogos & derivados , Polilisina/análogos & derivados , Adsorção , Biofilmes/efeitos dos fármacos , Micelas , Ácido Poliglutâmico/química , Polilisina/química , Poliestirenos/química , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/fisiologia
6.
Langmuir ; 35(1): 41-50, 2019 01 08.
Artigo em Inglês | MEDLINE | ID: mdl-30509072

RESUMO

Over the past few decades, field- and laboratory-scale studies have shown enhancements in oil recovery when reservoirs, which contain high-salinity formation water (FW), are waterflooded with modified-salinity salt water (widely referred to as the low-salinity, dilution, or SmartWater effect for improved oil recovery). In this study, we investigated the time dependence of the physicochemical processes that occur during diluted seawater (i.e., SmartWater) waterflooding processes of specific relevance to carbonate oil reservoirs. We measured the changes to oil/water/rock wettability, surface roughness, and surface chemical composition during SmartWater flooding using 10-fold-diluted seawater under mimicked oil reservoir conditions with calcite and carbonate reservoir rocks. Distinct effects due to SmartWater flooding were observed and found to occur on two different timescales: (1) a rapid (<15 min) increase in the colloidal electrostatic double-layer repulsion between the rock and oil across the SmartWater, leading to a decreased oil/water/rock adhesion energy and thus increased water wetness and (2) slower (>12 h to complete) physicochemical changes of the calcite and carbonate reservoir rock surfaces, including surface roughening via the dissolution of rock and the reprecipitation of dissolved carbonate species after exchanging key ions (Ca2+, Mg2+, CO32-, and SO42- in carbonates) with those in the flooding SmartWater. Our experiments using crude oil from a carbonate reservoir reveal that these reservoir rock surfaces are covered with organic-ionic preadsorbed films (ad-layers), which the SmartWater removes (detaches) as flakes. Removal of the organic-ionic ad-layers by SmartWater flooding enhances oil release from the surfaces, which was found to be critical to increasing the water wetness and significantly improving oil removal from carbonates. Additionally, the increase in water wetness is further enhanced by roughening of the rock surfaces, which decreases the effective contact (interaction) area between the oil and rock interfaces. Furthermore, we found that the rate of these slower physicochemical changes to the carbonate rock surfaces increases with increasing temperature (at least up to an experimental temperature of 75 °C). Our results suggest that the effectiveness of improved oil recovery from SmartWater flooding depends strongly on the formation of the organic-ionic ad-layers. In oil reservoirs where the ad-layer is fully developed and robust, injecting SmartWater would lead to significant removal of the ad-layer and improved oil recovery.

7.
Environ Sci Technol ; 53(9): 5202-5211, 2019 05 07.
Artigo em Inglês | MEDLINE | ID: mdl-30955329

RESUMO

Silica scaling of membranes used in reverse osmosis desalination processes is a severe problem, especially during the desalination of brackish groundwater due to high silica concentrations. This problem limits the water supply in inland arid and semiarid regions. Here, we investigated the influence of surface-exposed organic functional groups on silica precipitation and scaling. A test solution simulating the mineral content of brackish groundwater desalination brine at 75% recovery was used. The mass and chemical composition of the precipitated silica was monitored using a quartz crystal microbalance, X-ray photoelectron spectroscopy, and infrared spectroscopy, showing that surfaces with positively charged groups induced rapid silica precipitation, and the rate of silica precipitation followed the order -NH2 ∼ -N+(CH3)3 > -NH2/-COOH > -H2PO3 ∼ -OH > -COOH > -CH3. Force vs distance AFM measurements showed that the adhesion energy between a silica colloid glued to AFM cantilever and the studied surfaces increased as the surface charge changed from negative to positive. Thus, for the first time direct measurements of molecular forces and specific chemical groups that govern silica scaling during brackish water desalination is reported here. The influence of the different functional groups and the effect of the surface charge on silica precipitation that were found here can be used to design membranes that resist silica scaling in membrane-based desalination processes.


Assuntos
Dióxido de Silício , Purificação da Água , Filtração , Osmose , Águas Salinas
8.
Nat Mater ; 15(4): 407-412, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26779881

RESUMO

Polyelectrolyte complexation is critical to the formation and properties of many biological and polymeric materials, and is typically initiated by aqueous mixing followed by fluid-fluid phase separation, such as coacervation. Yet little to nothing is known about how coacervates evolve into intricate solid microarchitectures. Inspired by the chemical features of the cement proteins of the sandcastle worm, here we report a versatile and strong wet-contact microporous adhesive resulting from polyelectrolyte complexation triggered by solvent exchange. After premixing a catechol-functionalized weak polyanion with a polycation in dimethyl sulphoxide (DMSO), the solution was applied underwater to various substrates whereupon electrostatic complexation, phase inversion, and rapid setting were simultaneously actuated by water-DMSO solvent exchange. Spatial and temporal coordination of complexation, inversion and setting fostered rapid (∼25 s) and robust underwater contact adhesion (Wad ≥ 2 J m(-2)) of complexed catecholic polyelectrolytes to all tested surfaces including plastics, glasses, metals and biological materials.


Assuntos
Adesivos/química , Poliquetos/química , Água/química , Animais , Dimetil Sulfóxido/química , Poliaminas/química , Poliaminas/metabolismo , Poliquetos/metabolismo , Polieletrólitos , Polímeros/química , Polímeros/metabolismo
9.
Langmuir ; 33(38): 10041-10050, 2017 09 26.
Artigo em Inglês | MEDLINE | ID: mdl-28745509

RESUMO

Measuring truly equilibrium adhesion energies or contact angles to obtain the thermodynamic values is experimentally difficult because it requires loading/unloading or advancing/receding boundaries to be measured at rates that can be slower than 1 nm/s. We have measured advancing-receding contact angles and loading-unloading adhesion energies for various systems and geometries involving molecularly smooth and chemically homogeneous surfaces moving at different but steady velocities in both directions, ±V, focusing on the thermodynamic limit of ±V → 0. We have used the Bell Theory (1978) to derive expressions for the dynamic (velocity-dependent) adhesion energies and contact angles suitable for both (i) dynamic adhesion measurements using the classic Johnson-Kendall-Roberts (JKR, 1971) theory of "contact mechanics" and (ii) dynamic contact angle hysteresis measurements of both rolling droplets and syringe-controlled (sessile) droplets on various surfaces. We present our results for systems that exhibited both steady and varying velocities from V ≈ 10 mm/s to 1 nm/s, where in all cases but one, the advancing (V > 0) and receding (V < 0) adhesion energies and/or contact angles converged toward the same theoretical (thermodynamic) values as V → 0. Our equations for the dynamic contact angles are similar to the classic equations of Blake & Haynes (1969) and fitted the experimental adhesion data equally well over the range of velocities studied, although with somewhat different fitting parameters for the characteristic molecular length/dimension or area and characteristic bond formation/rupture lifetime or velocity. Our theoretical and experimental methods and results unify previous kinetic theories of adhesion and contact angle hysteresis and offer new experimental methods for testing kinetic models in the thermodynamic, quasi-static, limit. Our analyses are limited to kinetic effects only, and we conclude that hydrodynamic, i.e., viscous, and inertial effects do not play a role at the interfacial velocities of our experiments, i.e., V < (1-10) mm/s (for water and hexadecane, but for viscous polymers it may be different), consistent with previously reported studies.

10.
Proc Natl Acad Sci U S A ; 111(8): E768-75, 2014 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-24516125

RESUMO

The surface forces apparatus and atomic force microscope were used to study the effects of lipid composition and concentrations of myelin basic protein (MBP) on the structure of model lipid bilayers, as well as the interaction forces and adhesion between them. The lipid bilayers had a lipid composition characteristic of the cytoplasmic leaflets of myelin from "normal" (healthy) and "disease-like" [experimental allergic encephalomyelitis (EAE)] animals. They showed significant differences in the adsorption mechanism of MBP. MBP adsorbs on normal bilayers to form a compact film (3-4 nm) with strong intermembrane adhesion (∼0.36 mJ/m(2)), in contrast to its formation of thicker (7-8 nm) swelled films with weaker intermembrane adhesion (∼0.13 mJ/m(2)) on EAE bilayers. MBP preferentially adsorbs to liquid-disordered submicron domains within the lipid membranes, attributed to hydrophobic attractions. These results show a direct connection between the lipid composition of membranes and membrane-protein adsorption mechanisms that affects intermembrane spacing and adhesion and has direct implications for demyelinating diseases.


Assuntos
Encefalomielite Autoimune Experimental/metabolismo , Bicamadas Lipídicas/metabolismo , Modelos Moleculares , Proteína Básica da Mielina/metabolismo , Bainha de Mielina/metabolismo , Neurônios/citologia , Adsorção , Animais , Callithrix , Microscopia de Força Atômica , Estrutura Terciária de Proteína , Sus scrofa
11.
Nano Lett ; 16(10): 6709-6715, 2016 10 12.
Artigo em Inglês | MEDLINE | ID: mdl-27673480

RESUMO

Despite the need for molecularly smooth self-assembled monolayers (SAMs) on silicon dioxide surfaces (the most common dielectric surface), current techniques are limited to nonideal silane grafting. Here, we show unique bioinspired zwitterionic molecules forming a molecularly smooth and uniformly thin SAM in "water" in <1 min on various dielectric surfaces, which enables a dip-coating process that is essential for organic electronics to become reality. This monomolecular layer leads to high mobility of organic field-effect transistors (OFETs) based on various organic semiconductors and source/drain electrodes. A combination of experimental and computational techniques confirms strong adsorption (Wad > 20 mJ m-2), uniform thickness (∼0.5 or ∼1 nm) and orientation (all catechol head groups facing the oxide surface) of the "monomolecular" layers. This robust (strong adsorption), rapid, and green SAM represents a promising advancement toward the next generation of nanofabrication compared to the current nonuniform and inconsistent polysiloxane-based SAM involving toxic chemicals, long processing time (>10 h), or heat (>80 °C).

12.
Small ; 11(17): 2058-68, 2015 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-25504803

RESUMO

Surfactant self-assembly on surfaces is an effective way to tailor the complex forces at and between hydrophobic-water interfaces. Here, the range of structures and forces that are possible at surfactant-adsorbed hydrophobic surfaces are demonstrated: certain long-chain bolaform surfactants-containing a polydimethylsiloxane (PDMS) mid-block domain and two cationic α, ω-quarternary ammonium end-groups-readily adsorb onto thin PDMS films and form dynamically fluctuating nanostructures. Through measurements with the surface forces apparatus (SFA), it is found that these soft protruding nanostructures display polymer-like exploration behavior at the PDMS surface and give rise to a long-ranged, temperature- and rate-dependent attractive bridging force (not due to viscous forces) on approach to a hydrophilic bare mica surface. Coulombic interactions between the cationic surfactant end-groups and negatively-charged mica result in a rate-dependent polymer bridging force during separation as the hydrophobic surfactant mid-blocks are pulled out from the PDMS interface, yielding strong adhesion energies. Thus, (i) the versatile array of surfactant structures that may form at hydrophobic surfaces is highlighted, (ii) the need to consider the interaction dynamics of such self-assembled polymer layers is emphasized, and (iii) it is shown that long-chain surfactants can promote robust adhesion in aqueous solutions.

13.
Biomacromolecules ; 16(3): 1002-8, 2015 Mar 09.
Artigo em Inglês | MEDLINE | ID: mdl-25692318

RESUMO

Mussel foot protein-1 (mfp-1) is an essential constituent of the protective cuticle covering all exposed portions of the byssus (plaque and the thread) that marine mussels use to attach to intertidal rocks. The reversible complexation of Fe(3+) by the 3,4-dihydroxyphenylalanine (Dopa) side chains in mfp-1 in Mytilus californianus cuticle is responsible for its high extensibility (120%) as well as its stiffness (2 GPa) due to the formation of sacrificial bonds that help to dissipate energy and avoid accumulation of stresses in the material. We have investigated the interactions between Fe(3+) and mfp-1 from two mussel species, M. californianus (Mc) and M. edulis (Me), using both surface sensitive and solution phase techniques. Our results show that although mfp-1 homologues from both species bind Fe(3+), mfp-1 (Mc) contains Dopa with two distinct Fe(3+)-binding tendencies and prefers to form intramolecular complexes with Fe(3+). In contrast, mfp-1 (Me) is better adapted to intermolecular Fe(3+) binding by Dopa. Addition of Fe(3+) did not significantly increase the cohesion energy between the mfp-1 (Mc) films at pH 5.5. However, iron appears to stabilize the cohesive bridging of mfp-1 (Mc) films at the physiologically relevant pH of 7.5, where most other mfps lose their ability to adhere reversibly. Understanding the molecular mechanisms underpinning the capacity of M. californianus cuticle to withstand twice the strain of M. edulis cuticle is important for engineering of tunable strain tolerant composite coatings for biomedical applications.


Assuntos
Proteínas/química , Sequência de Aminoácidos , Animais , Concentração de Íons de Hidrogênio , Ferro/química , Mytilus/química , Ligação Proteica
14.
Biofouling ; 31(2): 221-7, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25875963

RESUMO

Mussel (Mytilus californianus) adhesion to marine surfaces involves an intricate and adaptive synergy of molecules and spatio-temporal processes. Although the molecules, such as mussel foot proteins (mfps), are well characterized, deposition details remain vague and speculative. Developing methods for the precise surveillance of conditions that apply during mfp deposition would aid both in understanding mussel adhesion and translating this adhesion into useful technologies. To probe the interfacial pH at which mussels buffer the local environment during mfp deposition, a lipid bilayer with tethered pH-sensitive fluorochromes was assembled on mica. The interfacial pH during foot contact with modified mica ranged from 2.2 to 3.3, which is well below the seawater pH of ~ 8. The acidic pH serves multiple functions: it limits mfp-Dopa oxidation, thereby enabling the catecholic functionalities to adsorb to surface oxides by H-bonding and metal ion coordination, and provides a solubility switch for mfps, most of which aggregate at pH ≥ 7-8.


Assuntos
Bicamadas Lipídicas/química , Mytilus/fisiologia , Proteínas/química , Adesividade , Silicatos de Alumínio , Animais , Concentração de Íons de Hidrogênio
15.
Biomacromolecules ; 15(6): 2128-37, 2014 Jun 09.
Artigo em Inglês | MEDLINE | ID: mdl-24835578

RESUMO

Bacterial outer membrane components play a critical role in bacteria-surface interactions (adhesion and repulsion). Sphingomonas species (spp.) differ from other Gram-negative bacteria in that they lack lipopolysaccharides (LPSs) in their outer membrane. Instead, Sphingomonas spp. outer membrane consists of glycosphingolipids (GSLs). To delineate the properties of the outer membrane of Sphingomonas spp. and to explain the adhesion of these cells to surfaces, we employed a single-component-based approach of comparing GSL vesicles to LPS vesicles. This is the first study to report the formation of vesicles containing 100% GSL. Significant physicochemical differences between GSL and LPS vesicles are reported. Composition-dependent vesicle adherence to different surfaces using quartz crystal microbalance with dissipation monitoring (QCM-D) technology was observed, where higher GSL content resulted in higher mass accumulation on the sensor. Additionally, the presence of 10% GSL and above was found to promote the relative rigidity of the vesicle obtaining viscoelastic ratio of 30-70% higher than that of pure LPS vesicles.


Assuntos
Glicoesfingolipídeos/metabolismo , Lipopolissacarídeos/metabolismo , Nylons/metabolismo , Dióxido de Silício/metabolismo , Substâncias Viscoelásticas/metabolismo , Adsorção , Glicoesfingolipídeos/química , Lipopolissacarídeos/química , Nylons/química , Dióxido de Silício/química , Sphingomonas/metabolismo , Substâncias Viscoelásticas/química
16.
Nat Commun ; 13(1): 7831, 2022 12 20.
Artigo em Inglês | MEDLINE | ID: mdl-36539423

RESUMO

Protein micropatterning enables robust control of cell positioning on electron-microscopy substrates for cryogenic electron tomography (cryo-ET). However, the combination of regulated cell boundaries and the underlying electron-microscopy substrate (EM-grids) provides a poorly understood microenvironment for cell biology. Because substrate stiffness and morphology affect cellular behavior, we devised protocols to characterize the nanometer-scale details of the protein micropatterns on EM-grids by combining cryo-ET, atomic force microscopy, and scanning electron microscopy. Measuring force displacement characteristics of holey carbon EM-grids, we found that their effective spring constant is similar to physiological values expected from skin tissues. Despite their apparent smoothness at light-microscopy resolution, spatial boundaries of the protein micropatterns are irregular at nanometer scale. Our protein micropatterning workflow provides the means to steer both positioning and morphology of cell doublets to determine nanometer details of punctate adherens junctions. Our workflow serves as the foundation for studying the fundamental structural changes governing cell-cell signaling.


Assuntos
Processamento de Imagem Assistida por Computador , Proteínas , Processamento de Imagem Assistida por Computador/métodos , Microscopia Crioeletrônica/métodos , Carbono/química , Transdução de Sinais
17.
Biomacromolecules ; 12(4): 1169-77, 2011 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-21361342

RESUMO

Microbial biofilms and their components present a major obstacle for ensuring the long-term effectiveness of membrane processes. Graft polymerization on membrane surfaces, in general, and grafting with oppositely charged monomers, have been shown to reduce biofouling significantly. In this study, surface forces and macromolecular properties of graft copolymers that possess oppositely charged groups were related to their potent antibiofouling behavior. Graft polymerization was performed using the negatively charged 3-sulphopropyl methacrylate (SPM) and positively charged [2-(methacryloyloxy)ethyl]-trimethylammonium (MOETMA) monomers to yield a copolymer layer on polyvinylidene fluoride (PVDF) surface. Quartz crystal microbalance with dissipation monitoring (QCM-D) technology was used to monitor the reduced adsorption of extracellular polymeric substances (EPS) extracted from a membrane bioreactor (MBR) wastewater treatment facility. Complemented measurements of attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy provided evaluation of the antifouling properties of the surface. Increase in water content in grafted layer exposed to 100 mM aqueous NaCl solution was observed by QCM-D. Therefore, the grafted copolymer layer is swelled in the presence of 100 mM NaCl because of reversing of polymer self-association by counterions. Force measurements by atomic force microscopy (AFM) showed an increased repulsion between a carboxylate-modified latex (CML) particle probe and a modified PVDF surface, especially in the presence of 100 mM NaCl. The hydration and swelling of the grafted polymer layer are shown to repel EPS and reduce their adsorption. Delineating the surface properties of antifouling grafted layers may lead to the design of novel antifouling surfaces.


Assuntos
Polivinil/química , Microscopia de Força Atômica , Compostos de Amônio Quaternário/química , Espectroscopia de Infravermelho com Transformada de Fourier , Ácidos Sulfônicos/química , Propriedades de Superfície
18.
NPJ Biofilms Microbiomes ; 7(1): 26, 2021 03 17.
Artigo em Inglês | MEDLINE | ID: mdl-33731698

RESUMO

The bacteriophage infection cycle has been extensively studied, yet little is known about the nanostructure and mechanical changes that lead to bacterial lysis. Here, atomic force microscopy was used to study in real time and in situ the impact of the canonical phage T4 on the nanotopography and biomechanics of irreversibly attached, biofilm-forming E. coli cells. The results show that in contrast to the lytic cycle in planktonic cells, which ends explosively, anchored cells that are in the process of forming a biofilm undergo a more gradual lysis, developing distinct nanoscale lesions (~300 nm in diameter) within the cell envelope. Furthermore, it is shown that the envelope rigidity and cell elasticity decrease (>50% and >40%, respectively) following T4 infection, a process likely linked to changes in the nanostructure of infected cells. These insights show that the well-established lytic pathway of planktonic cells may be significantly different from that of biofilm-forming cells. Elucidating the lysis paradigm of these cells may advance biofilm removal and phage therapeutics.


Assuntos
Bacteriófago T4/patogenicidade , Biofilmes/crescimento & desenvolvimento , Escherichia coli/fisiologia , Aderência Bacteriana , Bacteriólise , Fenômenos Biomecânicos , Escherichia coli/ultraestrutura , Escherichia coli/virologia , Microscopia de Força Atômica
19.
Langmuir ; 26(10): 7388-95, 2010 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-20099798

RESUMO

Some biological plasma membranes pass water with a permeability and selectivity largely exceeding those of commercial membranes for water desalination using specialized trans-membrane proteins aquaporins. However, highly selective transport of water through aquaporins is usually driven by an osmotic rather mechanical pressure, which is not as attractive from the engineering point of view. The feasibility of adopting biomimetic membranes for water purification driven by a mechanical pressure, i.e., filtration is explored in this paper. Toward this goal, it is proposed to use a commercial nanofiltration (NF) membrane as a support for biomimetic lipid bilayer membranes to render them robust enough to withstand the required pressures. It is shown in this paper for the first time that by properly tuning molecular interactions supported phospholipid bilayers (SPB) can be prepared on a commercial NF membrane. The presence of SPB on the surface was verified and quantified by several spectroscopic and microscopic techniques, which showed morphology close to the desired one with very few defects. As an ultimate test it is shown that hydraulic permeability of the SPB supported on the NF membrane (NTR-7450) approaches the values deduced from the typical osmotic permeabilities of intact continuous bilayers. This permeability was unaffected by the trans-membrane flow of water and by repeatedly releasing and reapplying a 10 bar pressure. Along with a parallel demonstration that aquaporins could be incorporated in a similar bilayer on mica, this demonstrates the feasibility of the proposed approach. The prepared SPB structure may be used as a platform for preparing biomimetic filtration membranes with superior performance based on aquaporins. The concept of SPBs on permeable substrates of the present type may also be useful in the future for studying transport of various molecules through trans-membrane proteins.


Assuntos
Materiais Biomiméticos/química , Bicamadas Lipídicas/química , Membranas Artificiais , Nanocompostos/química , Osmose , Água/química , Potenciais da Membrana , Estrutura Molecular , Tamanho da Partícula , Propriedades de Superfície
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