Alternative protein sources such as seaweed can help relieve the pressure on land-based protein supply. This proof-of-concept study developed an extraction method to recover soluble and lipophilic proteins from the seaweed Ulva fenestrata. The method consisted of processing U. fenestrata with 0.1–0.5 % aqueous Triton X-114 solution and reprocessing the pellet with an alkaline aqueous solution. Then, the solubilized proteins were precipitated via acidification. The new method extracted 3.4-times more protein, measured as total amino acids, compared to the control with two alkaline aqueous extraction cycles. Triton disrupted the chloroplasts and likely solubilized lipophilic membrane proteins as supported by microstructure and polypeptide pattern analysis. Triton-derived protein extracts contained lipids inside the precipitates/aggregates and were richer in fatty acids typical of photosynthetic membranes. The higher extraction yields are proposed to result from membrane charge neutralization upon acidification, triggering interactions between the membrane lipids and their subsequent precipitation with the lipophilic membrane protein. 

Protein extraction from wheat bran is challenging due to its multi-layer and fiber-rich structure. Here, opening aleurone cells, via dry and wet milling, their combination and a novel ultrafine milling, and its effect on wheat bran’s protein recovery using the alkaline solubilization/isoelectric precipitation and protein structure, functionality, and phytate content were investigated. Wet milling and ultrafine milling improved protein recovery and purity but only ultrafine milling reduced bran particle size to the aleurone cells and exposed their structure. Despite this, ultrafine milling did not significantly increase protein yield compared to wet milling, which partially opened the aleurone cells, meaning that opening the cells per se is not enough for extracting their protein. Proteins extracted with the aid of ultrafine milling had smaller particle sizes with significantly better water solubility (>2-fold) and rheological properties. Both wet milling and ultrafine milling significantly improved the removal of phytate during the wet fractionation process. Altogether, optimizing milling techniques offers a promising path to enhance accessibility to wheat bran proteins and their quality if carefully fine-tuned but other assistant technologies are necessary for boosting the recovery of the released protein from aleurone cells. 

Dietary fibre intake is essential for all human beings and has been correlated to beneficial health effects. Pea hull fibres (PF) are generally seen as a side stream during extraction of protein and starch from yellow pea but could be used in various food products to boost fibre content. In this study, the thermal treatment of pea hull fibres was investigated in terms of physicochemical properties and in vitro colonic fermentation. The PF that was subjected to heating showed an increase of fibres solubilised in the liquid and particle size. Results also showed that viscosity and storage modulus increased with thermal treatment, possibly due to the swelling of the PF. The pea fibre was readily fermentable based on total gas production and pH. However, the susceptibility to fermentation of PF did not increase with thermal treatment. Total gas production and short chain fatty acid produced were similar independent of thermal treatment. Conclusively, heating of the PF resulted in increased ability to structure water suspension, owing to increased fibre particle size, but is not sufficient to increase short chain fatty acid production during colonic fermentation. To explain this, we propose that the changes in cell wall structure were not major enough to induce higher fermentability. 

The incorporation of fibre into pea protein matrices influences their microstructure, yet our understanding of their gut fermentability remains unexplored. In this study, dietary fibres and protein from yellow pea were investigated for their physico-chemical properties and impact on in vitro colonic fermentation using human inoculum. Pea fibre and pea protein blends were studied at different pH and after thermal treatment at 95 °C for 30 min with oscillatory rheology, static light scattering and confocal laser scanning microscopy. The effect on in vitro colonic fermentation was evaluated measuring gas production, ammonia, and short chain fatty acid (SCFA) production. Rheology indicated that during thermal treatment a firmer gel is formed close to the protein isoelectric point with a structure characterised by aggregation, but less particle swelling compared to other pH. Addition of fibre led to higher storage modulus (G′), with the fibre dominating the rheological properties. Fermentation of samples containing protein led to higher levels of ammonia and SCFA compared to only fibres. Blends produced higher amounts of valerate, i-valerate and caproate, and lower amounts of ammonia. Reduced fermentation of proteins in the presence of fibres was also reflected in a more intact microstructure of the protein particles in the digesta. Although thermal treatment of blends caused particle swelling and induced gelation, only small differences could be discerned in the in vitro colonic fermentation outcomes. Our results highlight that potentially harmful fermentation products from protein, such as ammonia, were reduced in the presence of pea hull fibre. 

The relationship between hemolysis and lipid oxidation was explored in red blood cell (RBCs)-spiked washed cod mince (WCM). At pH 6.8 and 3 ± 1 °C, intact RBCs (71 µM Hb) delayed lipid oxidation by 1 day compared to WCM with partly or fully lysed RBCs which oxidized immediately. Intact RBCs also lowered peak peroxide value (PV) and thiobarbituric acid reactive substances (TBARS) with up to 59.5% and 48.1%, respectively. Adding 3% (v/w) blood plasma to RBC-spiked WCM delayed the lipid oxidation onset from 1 to 3–4 days without delaying hemolysis. At pH 6.4 the oxidation onset in RBC-WCM was the same as for pH 6.8 while at pH 7.2–7.6 lipid oxidation was suppressed for 7 days. Micrographs revealed RBC-lysis from day 2 at pH 6.4 but at pH 7.6, RBC stayed intact for ≥ 7 days. Thus, assuring presence of plasma-derived antioxidants and/or elevating muscle pH to avoid hemolysis can aid valorization of blood rich underutilized fish raw materials. 

Tissue engineering is a promising methodology to produce advanced therapy medicinal products (ATMPs). We have developed personalized tissue engineered veins (P-TEV) as an alternative to autologous or synthetic vascular grafts utilized in reconstructive vein surgery. Our hypothesis is that individualization through reconditioning of a decellularized allogenic graft with autologous blood will prime the tissue for efficient recellularization, protect the graft from thrombosis, and decrease the risk of rejection. In this study, P-TEVs were transplanted to vena cava in pig, and the analysis of three veins after six months, six veins after 12 months and one vein after 14 months showed that all P-TEVs were fully patent, and the tissue was well recellularized and revascularized. To confirm that the ATMP product had the expected characteristics one year after transplantation, gene expression profiling of cells from P-TEV and native vena cava were analyzed and compared by qPCR and sequencing. The qPCR and bioinformatics analysis confirmed that the cells from the P-TEV were highly similar to the native cells, and we therefore conclude that P-TEV is functional and safe in large animals and have high potential for use as a clinical transplant graft.

Patients with cardiovascular disease often need replacement or bypass of a diseased blood vessel. With disadvantages of both autologous blood vessels and synthetic grafts, tissue engineering is emerging as a promising alternative of advanced therapy medicinal products for individualized blood vessels. By reconditioning of a decellularized blood vessel with the recipient's own peripheral blood, we have been able to prevent rejection without using immunosuppressants and prime grafts for efficient recellularization in vivo. Recently, decellularized veins reconditioned with autologous peripheral blood were shown to be safe and functional in a porcine in vivo study as a potential alternative for vein grafting. In this study, personalized tissue engineered arteries (P-TEA) were developed using the same methodology and evaluated for safety in a sheep in vivo model of carotid artery transplantation. Five personalized arteries were transplanted to carotid arteries and analyzed for safety and patency as well as with histology after four months in vivo. All grafts were fully patent without any occlusion or stenosis. The tissue was well cellularized with a continuous endothelial cell layer covering the luminal surface, revascularized adventitia with capillaries and no sign of rejection or infection. In summary, the results indicate that P-TEA is safe to use and has potential as clinical grafts. 

The purpose of this study was to develop and evaluate finger food components as part of a complete meal for older adults with major motoric eating difficulties. Overall, the evaluation of sensory characteristics as well as texture analysis forms a valuable basis for further development of a meal that can be eaten without cutlery, comprising flatbread, beef rolls and brown sauce. The nutritionally enriched flatbreads were generally perceived as neutral in odour and flavour, while higher concentrations of protein and fat influenced the texture negatively. Although bread was not commonly eaten with a meal, the consumer evaluation stressed the importance of texture of flatbreads intended for wrapping. Differences between meat cuts were not pronounced; however, beef rolls made from inner thigh were perceived as more tender and crumblier than beef rolls made from outer thigh. Moreover, the odour and flavour intensity were thought to be higher in beef rolls braised in rolls due to the caramelised surface. However, tenderness was considered the most important parameter for beef rolls whereas crumbliness and dryness in tender meat can be compensated for by serving the meat with sauce. In fact, sauce was found to play an important role in a well-accepted meal. The addition of sweet, sour, or bitter tastes to brown sauces, to investigate the effect of basic tastes, reduced the perceived intensity of the original flavour profile of the brown sauces. Finally, combinations of the developed meal components could be investigated further to create attractive finger food meals for those unable to eat with knife and fork. © 2022 The Authors

Brewers’ spent grain (BSG), the by‐product of brewing, was subjected to a xylanase treatment followed by fermentation with Lactiplantibacillus plantarum PU1. Bioprocessed BSG has been used as ingredient to obtain a fortified semolina pasta which can be labeled as “high fiber” and “source of protein” according to the European Community Regulation No. 1924/2006. Compared to native BSG, the use of bioprocessed BSG led to higher protein digestibility and quality indices (es-sential amino acid index, biological value, protein efficiency ratio, nutritional index), as well as lower predicted glycemic index. Bioprocessing also improved the technological properties of fortified pasta. Indeed, brightfield and confocal laser scanning microscopy revealed the formation of a more homogeneous protein network, resulting from the degradation of the arabinoxylan structure of BSG, and the release of the components entrapped into the cellular compartments. The extensive cell wall disruption contributed to the release of phenols, and conferred enhanced antioxidant activity to the fortified pasta. The persistence of the activity was demonstrated after in vitro‐mimicked digestion, evaluating the protective effects of the digested pasta towards induced oxidative stress in Caco‐2 cells cultures. The fortified pasta showed a peculiar sensory profile, markedly improved by the pre‐treatment, thus confirming the great potential of bioprocessed BSG as health‐promoting food ingredient. © 2021 by the authors.

Conventional analysis of fluorescence recovery after photobleaching (FRAP) data for diffusion coefficient estimation typically involves fitting an analytical or numerical FRAP model to the recovery curve data using non-linear least squares. Depending on the model, this can be time consuming, especially for batch analysis of large numbers of data sets and if multiple initial guesses for the parameter vector are used to ensure convergence. In this work, we develop a completely new approach, DeepFRAP, utilizing machine learning for parameter estimation in FRAP. From a numerical FRAP model developed in previous work, we generate a very large set of simulated recovery curve data with realistic noise levels. The data are used for training different deep neural network regression models for prediction of several parameters, most importantly the diffusion coefficient. The neural networks are extremely fast and can estimate the parameters orders of magnitude faster than least squares. The performance of the neural network estimation framework is compared to conventional least squares estimation on simulated data, and found to be strikingly similar. Also, a simple experimental validation is performed, demonstrating excellent agreement between the two methods. We make the data and code used publicly available to facilitate further development of machine learning-based estimation in FRAP. Lay description: Fluorescence recovery after photobleaching (FRAP) is one of the most frequently used methods for microscopy-based diffusion measurements and broadly used in materials science, pharmaceutics, food science and cell biology. In a FRAP experiment, a laser is used to photobleach fluorescent particles in a region. By analysing the recovery of the fluorescence intensity due to the diffusion of still fluorescent particles, the diffusion coefficient and other parameters can be estimated. Typically, a confocal laser scanning microscope (CLSM) is used to image the time evolution of the recovery, and a model is fit using least squares to obtain parameter estimates. In this work, we introduce a new, fast and accurate method for analysis of data from FRAP. The new method is based on using artificial neural networks to predict parameter values, such as the diffusion coefficient, effectively circumventing classical least squares fitting. This leads to a dramatic speed-up, especially noticeable when analysing large numbers of FRAP data sets, while still producing results in excellent agreement with least squares. Further, the neural network estimates can be used as very good initial guesses for least squares estimation in order to make the least squares optimization convergence much faster than it otherwise would. This provides for obtaining, for example, diffusion coefficients as soon as possible, spending minimal time on data analysis. In this fashion, the proposed method facilitates efficient use of the experimentalist's time which is the main motivation to our approach. The concept is demonstrated on pure diffusion. However, the concept can easily be extended to the diffusion and binding case. The concept is likely to be useful in all application areas of FRAP, including diffusion in cells, gels and solutions. © 2020 The Authors.