RESUMO
Lignin is the second most abundant polymer after cellulose in lignocellulosic biomass. Its aromatic composition and recalcitrant nature make its valorization a major challenge for obtaining low molecular weight aromatics compounds with high value-added from the enzymatic depolymerization of industrial lignins. The oxidation reaction of lignin polymer using laccases alone remains inefficient. Therefore, researches are focused on the use of a laccase-mediator system (LMS) to facilitate enzymatic depolymerization. Until today, the LMS system was studied using water-soluble lignin only (commercial lignins, modified lignins, or lignin model compounds). This work reports a study of three LMS systems to depolymerize the three major industrial lignins (organosolv lignin, Kraft lignin, and sodium lignosulfonate). We show that an enzymatic depolymerization of these lignins can be achieved by LMS using laccase from Trametes versicolor, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt as mediator and a cosolvent (25% of 1,4-dioxane) to enhance the solubilization of lignins.
Assuntos
Lacase/metabolismo , Lignina/metabolismo , Polyporaceae/enzimologia , Biocatálise , Dioxanos/metabolismo , Oxirredução , Polyporaceae/metabolismo , Solubilidade , Ácidos Sulfônicos/metabolismoRESUMO
We present an experimental study of stick-jump (SJ) evaporation of strongly pinned nanoliter volume sessile water droplets drying on micropatterned surfaces. The evaporation is studied on surfaces composed of photolithographically micropatterned negative photoresist (SU-8). The micropatterning of the SU-8 enables circular, smooth, trough-like features to be formed which causes a very strong pinning of the three phase (liquid-vapor-solid) contact line of an evaporating droplet. This is ideal for studying SJ evaporation as it contains sequential constant contact radius (CCR) evaporation phases during droplet evaporation. The evaporation was studied in nonconfined conditions, and forced convection was not used. Micropatterned concentric circles were defined having an initial radius of 1000 µm decreasing by a spacing ranging from 500 to 50 µm. The droplet evaporates, successively pinning and depinning from circle to circle. For each pinning radius, the droplet contact angle and volume are observed to decrease quasi-linearly with time. The experimental average evaporation rates were found to decrease with decreasing pining radii. In contrast, the experimental average evaporation flux is found to increase with decreasing droplet radii. The data also demonstrate the influence of the initial contact angle on evaporation rate and flux. The data indicate that the total evaporation time of a droplet depends on the specific micropattern spacing and that the total evaporation time on micropatterned surfaces is always less than on flat, homogeneous surfaces. Although the surface patterning is observed to have little effect on the average droplet flux-indicating that the underlying evaporation physics is not significantly changed by the patterning-the total evaporation time is considerably modified by patterning, up to a factor or almost 2 compared to evaporation on a flat, homogeneous surface. The closely spaced concentric circle pinning maintains a large droplet radius and small contact angle from jump to jump; the result is a large evaporation rate leading to faster evaporation.
RESUMO
Nanostructure wetting is a key problem when developing superhydrophobic surfaces. Conventional methods do not allow us to draw conclusions about the partial or complete wetting of structures on the nanoscale. Moreover, advanced techniques are not always compatible with an in situ, real time, multiscale (from macro to nanoscale) characterization. A high-frequency (1 GHz) acoustic method is used for the first time to characterize locally partial wetting and the wetting transition between nanostructures according to the surface tension of liquids (the variation is obtained by ethanol concentration modification). We can see that this method is extremely sensitive both to the level of liquid imbibition and to the impalement dynamic. We thus demonstrate the possibility to evaluate the critical surface tension of a liquid for which total wetting occurs according to the aspect ratio of the nanostructures. We also manage to identify intermediate states according to the height of the nanotexturation. Finally, our measurements revealed that the drop impalement depending on the surface tension of the liquid also depends on the aspect ratio of the nanostructures. We do believe that our method may lead to new insights into nanoscale wetting characterization by accessing the dynamic mapping of the liquid imbibition under the droplet.
Assuntos
Acústica , Nanoestruturas/química , MolhabilidadeRESUMO
Many applications involving superhydrophobic materials require accurate control and monitoring of wetting states and wetting transitions. Such monitoring is usually done by optical methods, which are neither versatile nor integrable. This letter presents an alternative approach based on acoustic measurements. An acoustic transducer is integrated on the back side of a superhydrophobic silicon surface on which water droplets are deposited. By analyzing the reflection of longitudinal acoustic waves at the composite liquid-solid-vapor interface, we show that it is possible to track the local evolution of the Cassie-to-Wenzel wetting transition efficiently, as induced by evaporation or the electrowetting actuation of droplets.
RESUMO
When a drop sits on a highly liquid-repellent surface (super-hydrophobic or super-omniphobic) made of periodic micrometer-sized posts, its contact-line can recede with very weak mechanical retention providing that the liquid stays on top of the microsized posts. Occurring in both sliding and evaporation processes, the achievement of low-contact-angle hysteresis (low retention) is required for discrete microfluidic applications involving liquid motion or self-cleaning; however, careful examination shows that during receding, a minute amount of liquid is left on top of the posts lying at the receding edge of the drop. For the first time, the heterogeneities of these deposits along the drop-receding contact-line are underlined. Both nonvolatile liquid and particle-laden water are used to quantitatively characterize what rules the volume distribution of deposited liquid. The experiments suggest that the dynamics of the liquid de-pinning cascade is likely to select the volume left on a specific post, involving the pinch-off and detachment of a liquid bridge. In an applied prospective, this phenomenon dismisses such surfaces for self-cleaning purposes, but offers an original way to deposit controlled amounts of liquid and (bio)-particles at well-targeted locations.
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This article aims to show the influence of surface characteristics (microtopography, chemistry, mechanical properties) and seawater parameters on the settlement of marine micro- and macroorganisms. Polymers with nine microtopographies, three distinct mechanical properties, and wetting characteristics are immersed for one month into two contrasting coastal sites (Toulon and Kristineberg Center) and seasons (Winter and Summer). Influence of microtopography and chemistry on wetting is assessed through static contact angle and captive air bubble measurements over 3-weeks immersion in artificial seawater. Microscopic analysis, quantitative flow cytometry, metabarcoding based on the ribulose biphosphate carboxylase (rbcL) gene amplification, and sequencing are performed to characterize the settled microorganisms. Quantification of macrofoulers is done by evaluating the surface coverage and the type of organism. It is found that for long static in situ immersion, mechanical properties and non-evolutive wettability have no major influence on both abundance and diversity of biofouling assemblages, regardless of the type of organisms. The apparent contradiction with previous results, based on model organisms, may be due to the huge diversity of marine environments, both in terms of taxa and their size. Evolutive wetting properties with wetting switching back and forth over time have shown to strongly reduce the colonization by macrofoulers.
Assuntos
Incrustação Biológica , Incrustação Biológica/prevenção & controle , Polímeros/química , Molhabilidade , Propriedades de SuperfícieRESUMO
The discovery of novel antihelmintic molecules to combat the development and spread of schistosomiasis, a disease caused by several Schistosoma flatworm species, mobilizes significant research efforts worldwide. With a limited number of biochemical assays for measuring the viability of adult worms, the antischistosomicidal activity of molecules is usually evaluated by a microscopic observation of worm mobility and/or integrity upon drug exposure. Even if these phenotypical assays enable multiple parameters analysis, they are often conducted during several days and need to be associated with image-based analysis to minimized subjectivity. We describe here a self-purifying microfluidic system enabling the selection of healthy adult worms and the identification of molecules acting instantly on the parasite. The worms are assayed in a dynamic environment that eliminates unhealthy worms that cannot attach firmly to the chip walls prior to being exposed to the drug. The detachment of the worms is also used as second step readout for identifying active compounds. We have validated this new fluidic screening approach using the two major antihelmintic drugs, praziquantel and artemisinin. The reported dynamic system is simple to produce and to parallelize. Importantly, it enables a quick and sensitive detection of antischistosomal compounds in no more than one hour.
RESUMO
Following the achievement of superhydrophobicity which prevents water adhesion on a surface, superomniphobicity extends this high repellency property to a wide range of liquids, including oils, solvents, and other low surface energy liquids. Recent theoretical approaches have yield to specific microstructures design criterion to achieve such surfaces, leading to superomniphobic structured silicon substrate. To transfer this technology on a flexible substrate, we use a polydimethylsiloxane (PDMS) molding process followed by surface chemical modification. It results in so-called sticky superomniphobic surfaces, exhibiting large apparent contact angles (>150°) along with large contact angle hysteresis (>10°). We then focus on the modified Cassie equation, considering the 1D aspect of wetting, to explain the behavior of droplets on these surfaces and compare experimental data to previous works to confirm the validity of this model.
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Ex vivo explant culture models offer unique properties to study complex mechanisms underlying tissue growth, renewal, and disease. A major weakness is the short viability depending on the biopsy origin and preparation protocol. We describe an interphase microfluidic culture system to cultivate full thickness murine colon explants which keeps morphological structures of the tissue up to 192 h. The system was composed of a central well on top of a porous membrane supported by a microchannel structure. The microfluidic perfusion allowed bathing the serosal side while preventing immersion of the villi. After eight days, up to 33% of the samples displayed no histological abnormalities. Numerical simulation of the transport of oxygen and glucose provided technical solutions to improve the functionality of the microdevice.
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Nanoscale probes have been developed for the online characterization of the electrical properties of biological cells by dielectric spectroscopy. Two types of sensors have been designed and fabricated. The first one is devoted to low (<10 MHz) frequency range analysis and consists of gold nanoelectrodes. The second one works for high (>40 Hz) frequency range analysis and consists of a gold nanowire. The patterning of the sensors is performed by electron beam lithography. These devices are integrated in a microfluidic channel network for the manipulation of the cells and for the improvement of the performances of the sensors. These devices are used for the analysis of a well-characterized biological model in the area of the ligand-receptor interaction. The purpose is to monitor the interaction between the lactoferrin (the ligand) and the nucleolin and sulfated proteoglycans (the receptors) present or not on a set of mutant Chinese hamster ovary cell lines and their following internalization into the cytoplasm. Initial measurements have been performed with this microsystem and they demonstrate its capability for label-free, real-time, analysis of a dynamic mechanism involving biological cells.
Assuntos
Nanoestruturas/química , Nanotecnologia/instrumentação , Sistemas On-Line , Análise Espectral/métodos , Animais , Células CHO , Simulação por Computador , Cricetinae , Cricetulus , Impedância Elétrica , Ouro/metabolismo , Humanos , Lactoferrina/metabolismo , Microfluídica , Microscopia Eletrônica de Varredura , Microscopia de FluorescênciaRESUMO
Concentrating diluted samples is a key step to improve detection capabilities. The wise use of scaling laws shows the advantages of working with sub-microliter-sized samples. Rapid progress in MEMS technologies has driven the design of integrated platforms performing many biochemical operations. Here we report a new concentrator device based on electro-hydrodynamic forces which can be easily integrated into electrowetting-on-dielectric (EWOD) platforms. This approach is label-free and applicable to a wide range of micro-objects. The detection and analysis of two common waterborne parasites, Cryptosporidium and Giardia, is a perfect test case due to their global health relevance. By fully controlling the interplay of the various forces acting on the micron-sized Cryptosporidium parvum and Cryptosporidium muris oocysts, we show that it is possible to concentrate them on the side of a 10 µL initial drop and then extract them efficiently from a droplet of a few hundred nanoliters. We performed a finite element modeling of the forces acting on the parasites' oocysts to optimize the electrodes' shapes. We obtained state-of-the-art concentration factors of 12 ± 0.4 times and 2 to 4 times in the sub-region of the drop and the extracted droplet, respectively, with an efficiency of 70 ± 6%. Furthermore, this device had the ability to selectively concentrate parasites of different species out of a mix. We demonstrated this by segregating C. parvum oocysts from either Giardia lamblia cysts or its related species, C. muris oocysts.
Assuntos
Cryptosporidium parvum/isolamento & purificação , Hidrodinâmica , Dispositivos Lab-On-A-Chip , Água/parasitologia , Animais , Cryptosporidium parvum/citologia , Impedância Elétrica , Eletrodos , Eletroforese , Monitoramento Ambiental , Oocistos/citologiaRESUMO
Conducting polymers demonstrate an interesting ability to change their wettability at ultralow voltage (<1 V). While the conducting hydrogel poly(3,4-ethylenedioxythiophene) polystyrenesulfonate (PEDOT:PSS) is increasingly used as an interface with biology partly thanks to its mechanical properties, little is known about the electrical control of its wettability. We rely on the captive bubble technique to study this hydrogel property under relevant conditions (fully immerged). We here report that the wettability variations of PEDOT:PSS are driven by an electrowetting phenomenon in contrast to other conducting polymers which are thought to undergo wettability changes due to oxido-reduction reactions. In addition, we propose a modified electrowetting model to describe the wettability variations of PEDOT:PSS in aqueous solution under ultralow voltage and we show how these variations can be tuned in different ranges of contact angles (above or under 90°) by coating the PEDOT:PSS surface.
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In vitro models of the liver microvasculature, especially with respect to cancer cell extravasation, should include not only endothelial and cancer cells but also surrounding cells to mimic the physiological situation. To this end, in the present study, we established a physiologically-relevant hierarchical co-culture model by stacking layers of primary rat hepatocytes (Hep), hepatic stellate cells embedded in collagen gel (LX-2) and endothelial cells (HUVECs) on a specially designed oxygen-permeable polydimethylsiloxane PDMS bottom plate. The model was used to investigate the role and contribution of each of the three cell types in pancreatic cancer and promyeloblast cell adhesion. In particular, we showed an increase in albumin production by the primary hepatocytes and in the consumption of the produced vascular endothelial growth factors (VEGFs). Furthermore, in co-culture, the HUVECs exhibited a mature vascular endothelial and non-inflamed phenotype, as evidenced by Stabilin-1, lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), intercellular adhesion molecule (ICAM-1), and vascular adhesion protein-1 (VAP-1) expression. The HUVECs were also successfully activated with an inflammatory cytokine and their ICAM-1 response was found to be higher in monoculture compared to co-culture. Additionally, the adhesion of MiaPaCa-2 pancreatic cancer cells and HL60 promyeloblasts was tested in both cases (i.e.: activation or not by an inflammatory cytokine). It has been found that their adhesion was always reduced in the co-culture model. These results highlight the importance of integrating hepatic stellate cells in the design of biomimetic models of the hepatic endothelial barrier.
Assuntos
Carcinoma/metabolismo , Neoplasias Hepáticas/metabolismo , Neoplasias Pancreáticas/metabolismo , Animais , Biomimética , Carcinoma/patologia , Adesão Celular , Técnicas de Cocultura , Citocinas/metabolismo , Citoplasma/metabolismo , Dimetilpolisiloxanos/química , Células HL-60 , Células Estreladas do Fígado/citologia , Células Estreladas do Fígado/metabolismo , Hepatócitos/citologia , Hepatócitos/metabolismo , Células Endoteliais da Veia Umbilical Humana/citologia , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Inflamação , Neoplasias Hepáticas/patologia , Masculino , Membranas Artificiais , Modelos Biológicos , Neoplasias Pancreáticas/patologia , Fenótipo , Ratos , Ratos Wistar , Fator de Necrose Tumoral alfa/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Cryptosporidium parvum is a major cause of diarrheal illness and was recently potentially associated with digestive carcinogenesis. Despite its impact on human health, Cryptosporidium pathogenesis remains poorly known, mainly due to the lack of a long-term culture method for this parasite. Thus, the aim of the present study was to develop a three-dimensional (3D) culture model from adult murine colon allowing biological investigations of the host-parasite interactions in an in vivo-like environment and, in particular, the development of parasite-induced neoplasia. Colonic explants were cultured and preserved ex vivo for 35 days and co-culturing was performed with C. parvum. Strikingly, the resulting system allowed the reproduction of neoplastic lesions in vitro at 27 days post-infection (PI), providing new evidence of the role of the parasite in the induction of carcinogenesis. This promising model could facilitate the study of host-pathogen interactions and the investigation of the process involved in Cryptosporidium-induced cell transformation.
Assuntos
Técnicas de Cultura de Células/métodos , Colo/parasitologia , Neoplasias do Colo/parasitologia , Criptosporidiose/complicações , Criptosporidiose/parasitologia , Cryptosporidium parvum/patogenicidade , Modelos Animais de Doenças , Animais , Proliferação de Células , Interações Hospedeiro-Parasita , Humanos , Técnicas In Vitro , Camundongos , Camundongos SCID , Transdução de SinaisRESUMO
Cryptosporidium is the main origin of worldwide waterborne epidemic outbreaks caused by protozoan parasites. Its resilience to water chemical treatments and the absence of therapy led to consider it as a reference pathogen to assess water quality and as a possible bioterrorism agent. We here show that an electrical impedance-based device is able to get insights on Cryptosporidium development on a cell culture and to quantify sample infectivity. HCT-8 cells were grown to confluency on Interdigitated Microelectrode Arrays (IMA's) during 76h and then infected by Cryptosporidium parvum during 60h. The impedimetric response was measured at frequencies ranging from 100Hz to 1MHz and a 7min sampling period. As the infection progresses the impedance signal shows a reproducible distinct succession of peaks at 12h post infection (PI), 23h PI and 31h PI and local minima at 9h PI, 19h PI and 28h PI. An equivalent circuit modeling-based approach indicates that these features are mostly originated from paracellular pathway modifications due to host-parasite interactions. Furthermore, our data present for the first time a real-time monitoring of early parasitic stage development with alternating zoite and meront predominances, observed respectively at peaks and local minima in the impedimetric signal. Finally, by quantifying the magnitude of the impedimetric response, we demonstrate this device can also be used as an infectivity sensor as early as 12h PI thus being at least 6 times faster than other state of the art techniques.