Conservation Of Minimally Processed Apples Using Edible Coatings Made Of Turnip Extract And Xanthan Gum

The objective of this study was to evaluate the potential of turnip extract and xanthan gum in the conservation of minimally processed apples. The apples were washed, sanitized with sodium hypochlorite (200 ppm) for 15 minutes, peeled, and cut into eight pieces prior to being subjected to one of the following treatments in aqueous solution: A - water (control); B - turnip extract; C - turnip extract and CaCl2; D - xanthan gum, CaCl2 and glycerol; E - turnip extract, xanthan gum, CaCl2, and glycerol. Subsequently, the freshly cut apples were dried under ventilation on nylon screens to ensure drying of the coatings, and then packed in polystyrene trays, covered with polyvinylchloride films and stored at 4 ± 1 ° C for 13 days. The following parameters were evaluated: mass loss, firmness, colouration, pH value, soluble solids, and peroxidase/polyphenoloxidase activities. The edible coatings were found to be ineffective with respect to controlling mass loss, but the minimally processed apples coated with turnip extract maintained their initial levels of colouration, firmness and pH value. A considerable increase in peroxidase activity was registered for apples treated with turnip extract, suggesting that this effect may also be responsible for the reduction in browning. No advantage could be observed for the simultaneous presence of turnip extract and xanthan gum or calcium chloride. The turnip extract may represent an interesting alternative for applications to minimally processed apples, especially as it is a natural product, easily obtained, cost effective and contributes to the nutritional quality (e.g. as a source of calcium ions).1

Human pluripotent stem cell expansion in vertical-wheel bioreactors

Human induced pluripotent stem cells (hiPSC) have been regarded as an enormous breakthrough for medicine, since they can be derived from patients and be used to generate virtually all types of cells in the human body. One of the great bottlenecks in the usage of these cells for regenerative medicine or drug discovery applications is their expansion to relevant quantities. The Vertical-Wheel Bioreactors (PBS Biotech) present a novel scalable bioreactor configuration, whose agitation mechanism allows for homogeneous mixing conditions inside the single-use vessel, while conveying less shear stress to the cells when compared to traditional alternatives. These characteristics are advantageous for hiPSC expansion and thus, in this work, hiPSC were expanded in the Vertical-Wheel Bioreactor using different strategies, namely culturing the cells 1) on microcarriers and 2) as floating aggregates. In the first approach, cells were cultured under xeno-free conditions, using the Essential 8 medium together with microcarriers and coatings devoid of any animal-derived products [1]. The culture conditions were optimized in terms of initial cell/microcarrier ratio, inoculation method and agitation rate, in the PBS 0.1 vessel (working volume: 80 mL). The cells were successfully expanded, maintaining a normal karyotype, up to a 6.7-fold increase in cell number, after 6 days. These optimized culture conditions were successfully repeated in a larger vessel, the PBS 0.5 (300 mL working volume) demonstrating the scalability of the Vertical-Wheel system. In the second approach, hiPSC were expanded as floating aggregates, a methodology which does not require a separation step at the end of culture, to remove microcarriers, facilitating the downstream processing and Good Manufacturing Practice-compliance of the process. Cells were cultured in the PBS 0.1 (working volume: 60 mL), using mTeSR1, a serum-free medium and were monitored throughout culture regarding growth kinetics, aggregate size distribution and expression of pluripotency markers. The Vertical-Wheel Bioreactors were shown to efficiently keep the cell aggregates in suspension, under lower linear agitation speeds than an equivalent volume spinner flask (7 cm/s vs. 13 cm/s). Following 7 days of culture, cells were expanded up to a 5.2 ± 0.6-fold increase in cell number. The hiPSC aggregates increased in size over time, from an average diameter of 135 ± 61 µm to 397 ± 119 µm after 7 days. Pluripotency was maintained throughout time, as assessed by sustained high (\u3e 80%) expression of pluripotency markers OCT4, SOX2 and TRA-1-60, and low (\u3c 10%) expression of early differentiation marker SSEA-1. The results were validated using a second hiPSC line. This study revealed that the Vertical-Wheel Bioreactor allows hiPSC growth either on microcarriers and as aggregates and suggested it to have advantages versus other configurations. These results make the Vertical-Wheel Bioreactor a promising platform for hiPSC expansion and, prospectively, differentiation approaches, contributing for the generation of bona fide cells for various biomedical applications, namely drug screening, disease modelling, and, ultimately, for Regenerative Medicine. [1] Rodrigues CAV, Silva TP, Nogueira DES, Fernandes TG, Hashimura Y, Wesselschmidt R, Diogo MM, Lee B, Cabral JMS (2018), “Scalable Culture Of Human Induced Pluripotent Cells On Microcarriers Under Xeno‐Free Conditions Using Single‐Use Vertical‐Wheel™ Bioreactors”, Journal of Chemical Technology and Biotechnology, DOI: 10.1002/jctb.573

Alterações na ultra-estrutura de genótipos de soja em resposta ao fornecimento de manganês em solução nutritiva

The deleterious effects of Mn stress on many species have been studied, mainly concerning biochemical, physiological and growth parameters of plants. However, there are few studies relating the anatomical and ultrastructural changes in response to manganese (Mn) nutritional disorders, This study examined the leaf ultrastructure of Mn-inefficient (IAC-15, Santa Rosa) and Mn-efficient (IAC-Foscarin 31) soybean (Glycine max L.) genotypes in response to three rates of Mn (0.5, 2 and 200 µmol L-1) in the nutrient solution. Symptoms of Mn deficiency developed 12 days after transplanting in IAC-15 and Santa Rosa, followed by IAC-Foscarin 31 on the 15th day. Only IAC-15 and Santa Rosa leaves showed symptoms of Mn toxicity. The Mn concentration in leaves ranged from 8.6 (deficiency) to 886.3 mg kg-1 d.w. (toxicity). There were no changes either in stomata length or stomata number per unit of leaf surface. Cytoplasm disorganization was observed in IAC-15 under Mn-excess. In this case, the cytoplasm was amorphous, densely stained and extensively disorganized, with increased vacuolation. Mn effects were not found in mitochondria and nucleus in any of the genotypes tested. Under all Mn concentrations, many lipid globules were observed in the IAC15 chloroplasts. There was an increase in the number of plastids as well as in the size of starch grains within IAC-Foscarin 31 chloroplasts as Mn concentration in the nutrient solution increased. Genotypes had marked differences in the ultrastructure organization, mainly in leaf chloroplasts grown under conditions of both Mn deficiency and toxicity (the most sensitive genotype was IAC-15).Os efeitos negativos provocados tanto pela deficiência quanto pela toxidez de manganês (Mn) no desenvolvimento das plantas têm sido avaliados, considerando-se os aspectos bioquímicos e produtivos da parte aérea, particularmente, onde os sintomas visuais são manifestados. Entretanto, há poucas informações na literatura abordando as alterações anatômicas e de ultra-estrutura, em relação ao suprimento de Mn. Os objetivos do presente estudo foram avaliar os efeitos do fornecimento de três doses de Mn (0,5; 2 e 200 µmol L-1), em solução nutritiva, nas ultra-estruturas de folhas de cultivares de soja Glycine max (L.): Santa Rosa, IAC-15 e IAC-Foscarin 31, contrastantes quanto à aquisição e ao uso do Mn. Os sintomas visuais de deficiência foram observados primeiramente em Santa Rosa e IAC-15 (ineficientes), os únicos a exibirem sintomas de toxidez. As concentrações de Mn nas folhas com sintomas variaram de 8,6 (deficiência) a 886,3 mg kg-1 (toxidez). Não houve alterações no comprimento e no número de estômatos nos limbos foliares. Em condição de toxidez, constatou-se no IAC-15, citoplasma desorganizado, vacuolado em excesso e denso evidenciando alterações nas membranas dos tilacóides. Não ocorreram alterações ultra-estruturais nas mitocôndrias e no núcleo das células dos três genótipos. Constatou-se presença de glóbulos de lipídios nos cloroplastos do cultivar IAC-15, em todas as condições de fornecimento de Mn. Houve aumento no número de plastídeos e grãos de amido, bem como no tamanho destes no IAC-Foscarin 31 com o suprimento de Mn. Os genótipos, tanto na condição de deficiência quanto de excesso, exibiram distintos graus de organização das ultraestruturas, notadamente, os cloroplastos. O IAC-15 exibiu maiores alterações das ultra-estruturas em função das desordens nutricionais em manganês