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The standard model of pore formation was introduced more than fifty years ago, and it has been since, despite some refinements, the cornerstone for interpreting experiments related to pores in membranes. A central prediction of the model concerning pore opening under an electric field is that the activation barrier for pore formation is lowered proportionally to the square of the electric potential. However, this has only been scarcely and inconclusively confronted to experiments. In this paper, we study the electropermeability of model lipid membranes composed of 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine (POPC) containing different fractions of POPC-OOH, the hydroperoxidized form of POPC, in the range 0 to 100 mol %. By measuring ion currents across a 50-µm-diameter black lipid membrane (BLM) with picoampere and millisecond resolution, we detect hydroperoxidation-induced changes to the intrinsic bilayer electropermeability and to the probability of opening angstrom-size or larger pores. Our results over the full range of lipid compositions show that the energy barrier to pore formation is lowered linearly by the absolute value of the electric field, in contradiction with the predictions of the standard model.
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Eletricidade , Fosforilcolina , Transporte de Íons , Membranas , LipídeosRESUMO
A cell-penetrating peptide (CPP) is a short amino-acid sequence capable of efficiently translocating across the cellular membrane of mammalian cells. However, the potential of CPPs as a delivery vector is hampered by the strong reduction of its translocation efficiency when it bears an attached molecular cargo. To overcome this problem, we used previously developed diblock copolymers of elastin-like polypeptides (ELPBCs), which we end functionalized with TAT (transactivator of transcription), an archetypal CPP built from a positively charged amino acid sequence of the HIV-1 virus. These ELPBCs self-assemble into micelles at a specific temperature and present the TAT peptide on their corona. These micelles can recover the lost membrane affinity of TAT and can trigger interactions with the membrane despite the presence of a molecular cargo. Herein, we study the influence of membrane surface charge on the adsorption of TAT-functionalized ELP micelles onto giant unilamellar vesicles (GUVs). We show that the TAT-ELPBC micelles show an increased binding constant toward negatively charged membranes compared to neutral membranes, but no translocation is observed. The affinity of the TAT-ELPBC micelles for the GUVs displays a stepwise dependence on the lipid charge of the GUV, which, to our knowledge, has not been reported previously for interactions between peptides and lipid membranes. By unveiling the key steps controlling the interaction of an archetypal CPP with lipid membranes, through regulation of the charge of the lipid bilayer, our results pave the way for a better design of delivery vectors based on CPPs.
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Peptídeos Penetradores de Células , Micelas , Animais , Polipeptídeos Semelhantes à Elastina , Adsorção , Bicamadas Lipídicas/química , Peptídeos/química , Lipossomas Unilamelares/química , Peptídeos Penetradores de Células/química , Mamíferos/metabolismoRESUMO
The propagation of frost in an assembly of supercooled dew droplets takes place by the formation of ice protrusions that bridge ice particles and still-liquid droplets. In this work, we develop a Kinetic Monte Carlo (KMC) model to study the formation kinetics of the ice protrusions. The KMC simulations reproduce well the experimental results reported in the literature. The elongation speed of the ice protrusions does not depend on the droplet size but increases when the interdroplet distance decreases, the temperature increases, or the substrate wettability increases. While 2D diffusion of the water molecules on the substrate surface is sufficient to explain the process kinetics, high 3D (vapor) water-molecule concentration can lead to the development of 3D lateral branches on the ice protrusions. A 1D analytical model based on the water-molecule concentration gradient between a droplet and a nearby ice particle reproduces well the simulation results and highlights the relation between the protrusion elongation kinetics and parameters like the interdroplet distance, the water diffusivity, and the concentration gradient. The bridge-formation time has a quadratic dependence on the droplet-ice distance. Comparing the simulations, the analytical model, and the experimental results of the literature, we conclude that the propagation of frost on a flat substrate in an assembly of supercooled dew droplets with interdroplet spacing larger than about 1 µm is limited by water-molecule diffusivity.
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We report on a new mechanism of nanowire formation: during Au deposition on Si(110) substrates, Au-Si droplets grow, move spontaneously, and fabricate a Si nanowire behind them in the absence of Si external flux. Nanowires are formed by Si dissolved from the substrate at the advancing front of the droplets and transported backward to the crystallization front. The droplet shape is determined by the Si etching anisotropy. The nanowire formation can be tuned by changing experimental parameters like substrate temperature and Au deposition rate.
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We have adsorbed plasmid pUc19 DNA on a supported bilayer. By varying the fraction of cationic lipids in the membrane, we have tuned the surface charge. Plasmid conformations were imaged by Atomic Force Microscopy (AFM). We performed two sets of experiments: deposition from salt free solution on charged bilayers and deposition from salty solutions on neutral bilayers. Both sets show similar trends: at low surface charge density or low bulk salt concentration, the internal electrostatic repulsion forces plasmids to adopt completely opened structures, while at high surface charge density or higher bulk salt concentration, usual supercoiled plectonemes are observed. We experimentally demonstrate the equivalence of surface screening by mobile interfacial charges and bulk screening from salt ions. At low to medium screening, the electrostatic repulsion at plasmid crossings is predominant, leading to a number of crossovers decreasing linearly with the characteristic screening length. We compare our data with an analytical 2D-equilibrated model developed recently for the system and extract the DNA effective charge density when strands are adsorbed at the surface.
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DNA Super-Helicoidal/química , DNA Super-Helicoidal/metabolismo , Íons/química , Microscopia de Força Atômica , Conformação de Ácido Nucleico , Plasmídeos/química , Plasmídeos/metabolismo , Sais/química , Eletricidade Estática , Propriedades de Superfície , Água/químicaRESUMO
As the primary products of lipid oxidation, lipid hydroperoxides constitute an important class of lipids generated by aerobic metabolism. However, despite several years of effort, the structure of the hydroperoxidized bilayer has not yet been observed under electron microscopy. Here we use a 200 kV Cryo-TEM to image small unilamellar vesicles (SUVs) made (i) of pure POPC or SOPC, (ii) of their pure hydroperoxidized form, and (iii) of their equimolar mixtures. We show that the challenges posed by the determination of the thickness of the hydroperoxidized bilayers under these observation conditions can be addressed by an image analysis method that we developed and describe here.
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Microscopia Crioeletrônica , Bicamadas Lipídicas , Fosfatidilcolinas , Lipossomas Unilamelares , Bicamadas Lipídicas/química , Bicamadas Lipídicas/metabolismo , Microscopia Crioeletrônica/métodos , Lipossomas Unilamelares/química , Lipossomas Unilamelares/metabolismo , Fosfatidilcolinas/química , Oxirredução , Processamento de Imagem Assistida por Computador/métodos , Peróxidos Lipídicos/química , Peróxidos Lipídicos/análiseRESUMO
Epitaxial growth of WTe2 offers significant advantages, including the production of high-quality films, possible long-range in-plane ordering, and precise control over layer thicknesses. However, the mean island size of WTe2 grown by molecular beam epitaxy (MBE) in the literature is only a few tens of nanometers, which is not suitable for the implementation of devices at large lateral scales. Here we report the growth of Td -WTe2 ultrathin films by MBE on monolayer (ML) graphene, reaching a mean flake size of ≃110 nm, which is, on overage, more than three times larger than previous results. WTe2 films thicker than 5 nm have been successfully synthesized and exhibit the expected Td phase atomic structure. We rationalize the epitaxial growth of Td-WTe2 and propose a simple model to estimate the mean flake size as a function of growth parameters that can be applied to other transition metal dichalcogenides (TMDCs). Based on nucleation theory and the Kolmogorov-Johnson-Meh-Avrami (KJMA) equation, our analytical model supports experimental data showing a critical coverage of 0.13 ML above which WTe2 nucleation becomes negligible. The quality of monolayer WTe2 films is demonstrated by electronic band structure analysis using angle-resolved photoemission spectroscopy (ARPES), which is in agreement with first-principles calculations performed on free-standing WTe2 and previous reports. We found electron pockets at the Fermi level, indicating a n-type doping of WTe2 with an electron density of n = 2.0 ± 0.5 × 1012 cm-2 for each electron pocket.
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Entry of micro- or nanosized objects into cells or vesicles made of lipid membranes occurs in many processes such as entry of viruses into host cells, microplastics pollution, drug delivery, or biomedical imaging. Here we investigate the microparticle crossing of lipid membranes in giant unilamellar vesicles in the absence of strong binding interactions (e.g., streptavidin-biotin binding). In these conditions, we observe that organic and inorganic particles can always penetrate inside the vesicles provided an external piconewton force is applied and for relatively low membrane tensions. In the limit of vanishing adhesion, we identify the role of the membrane area reservoir and show that a force minimum exists when the particle size is comparable to the bendocapillary length.
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Pinças Ópticas , Plásticos , Lipossomas Unilamelares , Membranas , LipídeosRESUMO
Lipid peroxidation is a process which is key in cell signaling and disease, it is exploited in cancer therapy in the form of photodynamic therapy. The appearance of hydrophilic moieties within the bilayer's hydrocarbon core will dramatically alter the structure and mechanical behavior of membranes. Here, we combine viscosity sensitive fluorophores, advanced microscopy, and X-ray diffraction and molecular simulations to directly and quantitatively measure the bilayer's structural and viscoelastic properties, and correlate these with atomistic molecular modelling. Our results indicate an increase in microviscosity and a decrease in the bending rigidity upon peroxidation of the membranes, contrary to the trend observed with non-oxidized lipids. Fluorescence lifetime imaging microscopy and MD simulations give evidence for the presence of membrane regions of different local order in the oxidized membranes. We hypothesize that oxidation promotes stronger lipid-lipid interactions, which lead to an increase in the lateral heterogeneity within the bilayer and the creation of lipid clusters of higher order.
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Minimal surface problems arise naturally in many soft matter systems whose free energies are dominated by surface or interface energies. Of particular interest are the shapes, stability, and mechanical stresses of minimal surfaces spanning specific geometric boundaries. The "catenoid" is the best-known example where an analytical solution is known which describes the form and stability of a minimal surface held between two parallel, concentric circular frames. Here we extend this problem to nonaxisymmetric, parallel frame shapes of different orientations by developing a perturbation approach around the known catenoid solution. We show that the predictions of the perturbation theory are in good agreement with experiments on soap films and finite element simulations (Surface Evolver). Combining theory, experiment, and simulation, we analyze in depth how the shapes, stability, and mechanical properties of the minimal surfaces depend on the type and orientation of elliptic and three-leaf clover shaped frames. In the limit of perfectly aligned nonaxisymmetric frames, our predictions show excellent agreement with a recent theory established by Alimov et al. [Phys. Fluids 33, 052104 (2021)1070-663110.1063/5.0047461]. Moreover, we put in evidence the intriguing capacity of minimal surfaces between nonaxisymmetric frames to transmit a mechanical torque despite being completely liquid. These forces could be interesting to exploit for mechanical self-assembly of soft matter systems or as highly sensitive force captors.
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Investigations of free-standing liquid films enjoy an increasing popularity due to their relevance for many fundamental and applied scientific problems. They constitute soap bubbles and foams, serve as membranes for gas transport or as model membranes in biophysics. More generally, they provide a convenient tool for the investigation of numerous fundamental questions related to interface- and confinement-driven effects in soft matter science. Several approaches and devices have been developed in the past to characterise reliably the thinning and stability of such films, which were commonly created from low-viscosity, aqueous solutions/dispersions. With an increasing interest in the investigation of films made from strongly viscoelastic and complex fluids that may also solidify, the development of a new generation of devices is required to manage reliably the constraints imposed by these formulations. We therefore propose here a microfluidic chip design which allows for the reliable creation, control and characterisation of free-standing films of complex fluids. We provide all technical details and we demonstrate the device functioning for a larger range of systems via a selection of illustrative examples, including films of polymer melts and gelling hydrogels.
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PURPOSE: Manual feedback from senior surgeons observing less experienced trainees is a laborious task that is very expensive, time-consuming and prone to subjectivity. With the number of surgical procedures increasing annually, there is an unprecedented need to provide an accurate, objective and automatic evaluation of trainees' surgical skills in order to improve surgical practice. METHODS: In this paper, we designed a convolutional neural network (CNN) to classify surgical skills by extracting latent patterns in the trainees' motions performed during robotic surgery. The method is validated on the JIGSAWS dataset for two surgical skills evaluation tasks: classification and regression. RESULTS: Our results show that deep neural networks constitute robust machine learning models that are able to reach new competitive state-of-the-art performance on the JIGSAWS dataset. While we leveraged from CNNs' efficiency, we were able to minimize its black-box effect using the class activation map technique. CONCLUSIONS: This characteristic allowed our method to automatically pinpoint which parts of the surgery influenced the skill evaluation the most, thus allowing us to explain a surgical skill classification and provide surgeons with a novel personalized feedback technique. We believe this type of interpretable machine learning model could integrate within "Operation Room 2.0" and support novice surgeons in improving their skills to eventually become experts.
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Competência Clínica , Retroalimentação , Cirurgia Geral/educação , Cirurgia Geral/instrumentação , Aprendizado de Máquina , Redes Neurais de Computação , Fenômenos Biomecânicos , Análise por Conglomerados , Humanos , Cadeias de Markov , Modelos Estatísticos , Movimento (Física) , Análise de Regressão , Procedimentos Cirúrgicos Robóticos , CirurgiõesRESUMO
OBJECTIVE: The analysis of surgical motion has received a growing interest with the development of devices allowing their automatic capture. In this context, the use of advanced surgical training systems makes an automated assessment of surgical trainee possible. Automatic and quantitative evaluation of surgical skills is a very important step in improving surgical patient care. MATERIAL AND METHOD: In this paper, we present an approach for the discovery and ranking of discriminative and interpretable patterns of surgical practice from recordings of surgical motions. A pattern is defined as a series of actions or events in the kinematic data that together are distinctive of a specific gesture or skill level. Our approach is based on the decomposition of continuous kinematic data into a set of overlapping gestures represented by strings (bag of words) for which we compute comparative numerical statistic (tf-idf) enabling the discriminative gesture discovery via its relative occurrence frequency. RESULTS: We carried out experiments on three surgical motion datasets. The results show that the patterns identified by the proposed method can be used to accurately classify individual gestures, skill levels and surgical interfaces. We also present how the patterns provide a detailed feedback on the trainee skill assessment. CONCLUSIONS: The proposed approach is an interesting addition to existing learning tools for surgery as it provides a way to obtain a feedback on which parts of an exercise have been used to classify the attempt as correct or incorrect.
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Gestos , Reconhecimento Automatizado de Padrão/métodos , Procedimentos Cirúrgicos Operatórios/educação , Algoritmos , Fenômenos Biomecânicos , Competência Clínica , Feedback Formativo , Humanos , Análise e Desempenho de Tarefas , Estudos de Tempo e MovimentoRESUMO
Nano-droplets on a foreign substrate have received increasing attention because of their technological possible applications, for instance to catalyse the growth of nanowires. In some cases the droplets can move as a result of a reaction with the substrate. In this work we show that the substrate orientation, the surface morphology and the shape of the pits etched in the substrate by the droplets affect the droplet motion, so that a single mechanism (droplet-induced substrate dissolution) may lead to several unexpected droplet dynamics. The experiments are carried out by low energy electron microscopy on Au-Si and Au-Ge, which are model systems for studying liquid droplet alloys. Studying in-situ the behaviour of Au droplets on various Si and Ge surfaces, we describe a subtle interplay between the substrate orientation, the surface defects, and the droplet motion. Our observations allow a deep understanding of the interfacial mechanisms at the origin of the alloy formation and the associated droplet motion. These mechanisms are based on events of substrate dissolution/recrystallization. The outcomes of this work highlight the importance of the etching anisotropy on the droplet-substrate behaviours, and are essential in the perspective of positioning liquid alloy droplets used for instance as nanowire catalysts.
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Although cationic cell-penetrating peptides (CPPs) are able to bind to cell membranes, thus promoting cell internalization by active pathways, attachment of cargo molecules to CPPs invariably reduces their cellular uptake. We show here that CPP binding to lipid bilayers, a simple model of the cell membrane, can be recovered by designing cargo molecules that self-assemble into spherical micelles and increase the local interfacial density of CPP on the surface of the cargo. Experiments performed on model giant unilamellar vesicles under a confocal laser scanning microscope show that a family of thermally responsive elastin-like polypeptides that exhibit temperature-triggered micellization can promote temperature triggered attachment of the micelles to membranes, thus rescuing by self-assembly the cargo-induced loss of the CPP affinity to bio-membranes.
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Peptídeos Penetradores de Células/metabolismo , Lipídeos de Membrana/metabolismo , Bicamadas Lipídicas/metabolismo , Microscopia Confocal , Ligação ProteicaRESUMO
BACKGROUND: Approximately 30% of lung cancer cases are diagnosed in patients > 70 years of age. Standard chemotherapy regimens are generally considered too toxic for elderly patients. We conducted a multicenter phase II trial to determine the efficacy and safety of carboplatin combined with vinorelbine every 4 weeks as first-line treatment for advanced non-small-cell lung cancer (NSCLC) in elderly patients. PATIENTS AND METHODS: Patients were eligible if they were aged >OR= 70 years, had stage IIIB (with pleural effusion) or stage IV NSCLC, had a performance status of 0/1, had not previously received chemotherapy, and had normal organ function. Forty patients (31 men and 9 women) were enrolled and received 3-5 courses of treatment. Median age was 72 years (range, 70-82 years). Eighty percent of patients had stage IV NSCLC, with squamous cell (n=21), adenocarcinoma (n=12), and undifferentiated (n=7) histologies. RESULTS: Forty patients were assessable for toxicity and 32 for treatment response. Among these 32 patients, 8 had a partial response (intent-to-treat response rate, 20%), and 10 (25%) had stable disease. The median survival was 7.8 months (range, 4-11.6 months). The 1- and 2-year survival rates were 25% and 7%, respectively; median time to progression was 4.3 months (range, 0.2-13.8 months). Grade 3/4 neutropenia was seen in 27 patients (68%), and grade 3/4 anemia was seen in 5 patients (13%). One patient died of febrile neutropenia during treatment. The main nonhematologic adverse effect was fatigue (grade 3/4 in 18% of patients). CONCLUSION: Carboplatin/vinorelbine is well tolerated by elderly patients with extensive-stage NSCLC. Efficacy is low but similar to that of other treatments used in this setting.
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Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Neoplasias Pulmonares/tratamento farmacológico , Qualidade de Vida , Idoso , Idoso de 80 Anos ou mais , Carboplatina/administração & dosagem , Carboplatina/efeitos adversos , Carcinoma Pulmonar de Células não Pequenas/mortalidade , Carcinoma Pulmonar de Células não Pequenas/patologia , Feminino , França/epidemiologia , Humanos , Neoplasias Pulmonares/mortalidade , Neoplasias Pulmonares/patologia , Masculino , Estadiamento de Neoplasias , Perfil de Impacto da Doença , Estatística como Assunto , Análise de Sobrevida , Resultado do Tratamento , Vimblastina/administração & dosagem , Vimblastina/efeitos adversos , Vimblastina/análogos & derivados , VinorelbinaRESUMO
Elastically mediated interactions between surface domains are classically described in terms of point forces. Such point forces lead to local strain divergences that are usually avoided by introducing a poorly defined cut-off length. In this work, we develop a self-consistent approach in which the strain field induced by the surface domains is expressed as the solution of an integral equation that contains surface elastic constants, S ij . For surfaces with positive S ij the new approach avoids the introduction of a cut-off length. The classical and the new approaches are compared in case of 1-D periodic ribbons.
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The regional distribution of inhaled gas within the lung is affected in part by normal variations in airway geometry or by obstructions resulting from disease. In the present work, the effects of heterogeneous airway obstructions on the distribution of air and helium-oxygen were examined using an in vitro model, the two compartments of a dual adult test lung. Breathing helium-oxygen resulted in a consistently more uniform distribution, with the gas volume delivered to a severely obstructed compartment increased by almost 80%. An engineering approach to pipe flow was used to analyze the test lung and was extrapolated to a human lung model to show that the in vitro experimental parameters are relevant to the observed in vivo conditions. The engineering analysis also showed that helium-oxygen can decrease the relative weight of the flow resistance due to obstructions if they are inertial in nature (i.e., density dependent) due to either turbulence or laminar convective losses.
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Obstrução das Vias Respiratórias/fisiopatologia , Hélio , Pulmão/fisiopatologia , Modelos Biológicos , Oxigênio , Ventilação Pulmonar , Adulto , HumanosRESUMO
The morphological evolution of an initially straight stripe assimilated to a straight line of infinite length lying on a semi-infinite substrate has been investigated in the linear regime when the mass transport mechanism is the diffusion of adatoms along stripe edges and when the heteroepitaxy between the line and the substrate is taken to be anisotropic. It is found that contrary to the isotropic case where serpentine-like morphology is favored, antiphase fluctuations grows faster than in-phase ones for selected values of epitaxial stress components such that a pinched shape preferentially emerges.