Perfis proteicos e desempenho fisiológico de sementes de café submetidas a diferentes métodos de processamento e secagem.

O objetivo deste trabalho foi avaliar os perfis proteicos e o desempenho fisiológico de sementes de café submetidas a diferentes métodos de processamento e secagem. Foram avaliados os processamentos por via seca e úmida, e as secagens natural, em terreiro, e artificial a 60ºC, ou a 60ºC até 30% de umidade e 40ºC até teor final de 11% (base úmida). Após serem processadas e secadas, as sementes foram avaliadas quanto ao desempenho fisiológico e submetidas a análises bioquímicas, por meio da eletroforese de proteínas resistentes ao calor LEA (?late embryogenesis abundant?) e das enzimas superóxido dismutase, catalase, peroxidase, esterase, polifenoloxidase, isocitrato desidrogenase, álcool desidrogenase e malato desidrogenase. O perfil proteico de sementes de café é afetado pelo método de processamento e de secagem. Os cafés processados por via úmida apresentam maior tolerância à secagem ? revelada pela maior atividade de enzimas antioxidativas e pelo melhor desempenho fisiológico ? do que os processados por via seca. A atividade de proteínas resistentes ao calor e de enzimas antioxidantes é variável promissora para diferenciar a qualidade dos cafés submetidos a diferentes manejos pós‑colheita

Y Engineering a 3D in vitro model of human skeletal muscle at the single fiber scale

The reproduction of reliable in vitro models of human skeletal muscle is made harder by the intrinsic 3D structural complexity of this tissue. Here we coupled engineered hydrogel with 3D structural cues and specific mechanical properties to derive human 3D muscle constructs (“myobundles”) at the scale of single fibers, by using primary myoblasts or myoblasts derived from embryonic stem cells. To this aim, cell culture was performed in confined, laminin-coated micrometric channels obtained inside a 3D hydrogel characterized by the optimal stiffness for skeletal muscle myogenesis. Primary myoblasts cultured in our 3D culture system were able to undergo myotube differentiation and maturation, as demonstrated by the proper expression and localization of key components of the sarcomere and sarcolemma. Such approach allowed the generation of human myobundles of ~10 mm in length and ~120 μm in diameter, showing spontaneous contraction 7 days after cell seeding. Transcriptome analyses showed higher similarity between 3D myobundles and skeletal signature, compared to that found between 2D myotubes and skeletal muscle, mainly resulting from expression in 3D myobundles of categories of genes involved in skeletal muscle maturation, including extracellular matrix organization. Moreover, imaging analyses confirmed that structured 3D culture system was conducive to differentiation/maturation also when using myoblasts derived from embryonic stem cells. In conclusion, our structured 3D model is a promising tool for modelling human skeletal muscle in healthy and diseases conditions

Intravital three-dimensional bioprinting

Fabrication of three-dimensional (3D) structures and functional tissues directly in live animals would enable minimally invasive surgical techniques for organ repair or reconstruction. Here, we show that 3D cell-laden photosensitive polymer hydrogels can be bioprinted across and within tissues of live mice, using bio-orthogonal two-photon cycloaddition and crosslinking of the polymers at wavelengths longer than 850 nm. Such intravital 3D bioprinting\u2014which does not create by-products and takes advantage of commonly available multiphoton microscopes for the accurate positioning and orientation of the bioprinted structures into specific anatomical sites\u2014enables the fabrication of complex structures inside tissues of live mice, including the dermis, skeletal muscle and brain. We also show that intravital 3D bioprinting of donor-muscle-derived stem cells under the epimysium of hindlimb muscle in mice leads to the de novo formation of myofibres in the mice. Intravital 3D bioprinting could serve as an in vivo alternative to conventional bioprinting

Intravital three-dimensional bioprinting

Fabrication of three-dimensional (3D) structures and functional tissues directly in live animals would enable minimally invasive surgical techniques for organ repair or reconstruction. Here, we show that 3D cell-laden photosensitive polymer hydrogels can be bioprinted across and within tissues of live mice, using bio-orthogonal two-photon cycloaddition and crosslinking of the polymers at wavelengths longer than 850 nm. Such intravital 3D bioprinting—which does not create by-products and takes advantage of commonly available multiphoton microscopes for the accurate positioning and orientation of the bioprinted structures into specific anatomical sites—enables the fabrication of complex structures inside tissues of live mice, including the dermis, skeletal muscle and brain. We also show that intravital 3D bioprinting of donor-muscle-derived stem cells under the epimysium of hindlimb muscle in mice leads to the de novo formation of myofibres in the mice. Intravital 3D bioprinting could serve as an in vivo alternative to conventional bioprinting

Post-harvest effects on beverage quality and physiological performance of coffee beans.

During coffee drying, different temperatures applied to the beans with varied humidity content levels can interfere in the membranes integrity, germination, organic acid and carbohydrate content resulting in coffees with distinct flavors. The quality control of the beans will be much more effective the earlier the alterations provoked in the postharvest are detected. This work has an objective to study alternative methods for the dehydration o f the coffee beans using ultra-drying followed by slow drying and its impact on the sensorial quality, chemical composition and physiology. For that purpose, coffee lots were processed by the methods, dry (natural coffee) and wet (fully washed coffee); and sun-dried and machine-dried at a constant 60°C temperature and alternating 60/40°C. The sensory quality of the samples was assessed by the Specialty Coffee Association of America (SCAA) analysis protocol. The sugar, total titratable acidity and the phenolic compound content was also analyzed. The physiological alterations of the coffee beans were analyzed by germination tests, emergence speed index, electrical conductivity and potassium leaching. The temperature of the drying air significantly altered the sensorial quality of the coffee beans. The processing way associated to drying methods causes many physiological alterations with the highest damage observed in the natural coffees. For the first time, we are showing that drying with heated air at 60/40ºC is promising for the fully washed coffee beans, which are more tolerant to dehydration than the natural coffee beans. Conversely, the natural coffee beans were much more sensitive to drying regardless the temperature, with very low performance in the physiological analyses. The drying at the constant 60ºC temperature is inappropriate for the natural coffee as well as for the fully washed coffee beans. In addition, the physiological tests used were shown effective for the early evaluation of coffee beans quality

A Combined Electrochemical-Microfluidic Strategy for the Microscale-Sized Selective Modification of Transparent Conductive Oxides

Surface chemical functionalization of transparent conductive oxides (TCOs) is helpful for a wide range of technological applications, ranging from solar cells to biomedical devices, as it allows to tune the electrical, optical, and morphological properties of TCOs toward the desired goal. The electrochemical grafting technique is a surface modification methodology affording robust coatings with tuneable properties and has the potential to be exploited for modifying TCO surfaces. However, due to technical limitations, like the use of a 3-electrode cell and the need for low pH-solutions, this approach has not been recurrently applied. Here a novel electrochemical-microfluidic combined methodology is used where the use of a microchannel drives the spatially controlled covalent grafting of reagents on a TCO surface. To corroborate the validity of this approach in producing more complex chemical structures localized on selected microscale-sized areas, where a first electrochemical grafting step takes place, an electrochemical glucose biosensor is realized through a layer-by-layer approach that shows a remarkable limit of detection in the micromolar concentration range. The sensing mechanism is based on an efficient electron transfer from glucose to the functionalized TCO surface. Biosensor performance is conveniently tuned by acting on the number of enzymatic units loaded onto the biosensor-tree

Surface Functionalization of Fluorine-Doped Tin Oxide Samples through Electrochemical Grafting

Transparent conductive oxides are emerging materials in several fields, such as photovoltaics, photoelectrochemistry, and optical biosensing. Their high chemical inertia, which ensured long-term stability on one side, makes challenging the surface modification of transparent conductive oxides; long-term robust modification, high yields, and selective surface modifications are essential prerequisite for any further developments. In this work, we aim at inducing chemical functionality on fluorine-doped tin oxide surfaces (one of the most inexpensive transparent conductive oxide) by means of electrochemical grafting of aryl diazonium cations. The grafted layers are fully characterized by photoemission spectroscopy, cyclic voltammetry, and atomic force microscopy showing linear correlation between surface coverage and degree of modification. The electrochemical barrier effect of modified surfaces was studied at different pH to characterize the chemical nature of the coating. We showed immuno recognition of biotin complex built onto grafted fluorine-doped tin oxides, which opens the perspective of integrating FTO samples with biological-based devices