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14 pages, 8 figures, 4 tables.-- This is an open access article distributed under the Creative Commons Attribution License, High molecular weight (Mw) collagen hydrolysates have been demonstrated to produce a higher synthesis of collagen type I mRNA. Mw determination is a key factor maximizing the effect of collagen hydrolysates on collagen type I synthesis by fibroblasts. This work aimed to achieve a high average Mw in Blue Shark Collagen Hydrolysate, studying different hydrolysis parameters by GPC-LS analysis and testing its effect on mRNA Type I collagen expression. Analysis revealed differences in blue shark collagen hydrolysates Mw depending on hydrolysis conditions. Papain leads to obtaining a significantly higher Mw hydrolysate than Alcalase at different times of hydrolysis and at different enzyme/substrate ratios. Besides, the time of the hydrolysis factor is more determinant than the enzyme/substrate ratio factor for obtaining a higher or lower hydrolysate Mw when using Papain as the enzyme. Contrary, Alcalase hydrolysates resulted in similar Mw with no significant differences between different conditions of hydrolysis assayed. Blue shark collagen hydrolysate showing the highest Mw showed neither cytotoxic nor proliferation effect on fibroblast cell culture. Besides, it exhibited an increasing effect on both mRNA expression and pro-collagen I production, This research was funded by the project BlueHuman, EU_EAPA_151/2016 and Xunta de Galicia (Grupos de Potencial Crecimiento, IN607B2021/11)

16 pages, 6 figures, 2 tables, The smallest size fractions of plankton, nano- and pico-plankton, have been highlighted due to they accomplish key functions in marine ecosystems. However, the knowledge about some of them is scarce because they are difficult or impossible to be detected and identified with non-DNA-based methodologies. Here we have evaluated five DNA extraction protocols (MT1–MT5) and seven bioinformatic pipelines (P1–P7) to find the best protocol for detecting most of the eukaryotic species of nano- and pico-plankton present in an environmental sample using Ion Torrent technology. The protocol MT3 was the most reproducible methodology, showing less variation among samples, good DNA quality and sufficient quantity to amplify and sequence the eukaryote species, offering the best results after sequencing. For bioinformatic analyses, P1 and P7 resulted in the highest percentage of detection for the difficult-to-detect species in mock communities. However, only P1 avoided the confusion with other closed species during the taxonomic assignment. The final protocols, MT3-P1 (free) and MT3-P7 (private), showed good and consistent results when they were applied to an environmental sample, being a valuable tool to study the eukaryotes present in environmental samples of nano -and pico-plankton, even for the genera that are difficult to be detected by other techniques., This study was funded by the Spanish Ministry of Science and Innovation, Grant CTM2014-56119-R (i-SMALL project). MM-C holds a “Juan de la Cierva Formación” postdoctoral fellowship from the Spanish Ministry of Education with reference code FJCI-2017-32722.

Culturing axillary shoot explants of willow in liquid medium in plantform™ and RITA ® vessels increased the multiplication coefficient relative to that obtained by culturing the explants in semisolid medium in jars. A protocol for micropropagating willow by multiplying axillary shoots in liquid medium was developed, and the effects of the type of explant, medium composition, bioreactor type and frequency of immersion were investigated. The type of explant influenced shoot quality and the multiplication coefficient, with basal segments showing a higher capacity for proliferation than apical and mid-shoot segments. Hyperhydricity was not observed in shoots cultured with Murashige and Skoog medium with half-strength nitrates and supplemented with 0.22 µM BA and 3% sucrose. The willow shoots cultured in plantform™ and RITA® vessels yielded higher multiplication coefficients than shoots grown in semisolid medium. High proliferation rates were obtained by immersion of shoots for 1 min three or six times a day, with additional aeration of 1 min per hour in the plantform™ bioreactors. Transfer to medium with 1% sucrose after 4 weeks of culture in medium with 3% sucrose increased the multiplication coefficient of basal segments cultured in RITA® vessels, suggesting that the species could be micropropagated under photoautotrophic conditions. The willow shoots rooted spontaneously and were successfully acclimatized to greenhouse conditions. This is the first report of the use of a temporary immersion system to culture Salix viminalis. The study findings indicate the feasibility of the system as a method for the large-scale propagation of this species.

A protocol for culturing chestnut axillary shoots with the basal sections continuously immersed in liquid medium was developed and the influence of various parameters was assessed: explant type and size, support material, bioreactor size and aeration regime. Shoots excised from eight selected chestnut genotypes were successfully cultured in 10 L bioreactors in liquid Murashige and Skoog medium with half strength nitrates and supplemented with 0.05 mg/L N6-benzyladenine and 30 g/L sucrose. Forced ventilation and the use of a support material were essential for producing healthy, non-hyperhydric shoots. High proliferation rates were achieved and good quality shoots ready for multiplication, rooting and acclimatization were obtained. Larger leaves showing higher levels of photosynthetic pigments were observed in shoots cultured in bioreactors, suggesting a certain degree of photoautotrophy in shoots cultured under forced ventilation. This is the first demonstration of the production of chestnut shoots in stationary liquid medium, indicating the feasibility of the method for large-scale propagation of the species.

Fish discards and subproducts may represent an important source of raw material, not only for the food industry, but for other different kind of industries, such as the nutraceutical and cosmetic industries. Collagen, which is mainly obtained from animal skins, is an important structural protein in the animal kingdom having many different applications. It is well known that fish skins constitute a significant subproduct in the fishery industry, especially in the case of some species, where fish skins may represent up to 20% of the total body weight of fish. Peptides from collagen hydrolysates have been described to be useful for preventing skin aging and osteoarthritis, however, the mechanism for these biological activities is not well known. Fibroblasts are the main cell types involved in the collagen synthesis, and in the present work, human dermal fibroblasts have been exposed to the treatment of collagen peptides of two different molecular weight ranges. Results show that higher molecular weight collagen peptides produce higher synthesis of collagen type I mRNA and, therefore, it may suggest that prior molecular weight selection may be an important step to maximize the effect of collagen hydrolysates on collagen type I synthesis by dermal fibroblasts., The work was funded by MARMED project, supported by the EU Atlantic Area Program Project Number 2011-1/164. We thank Lumar S.L. for the skin samples used in this work.