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Clinical determination of MP counts using flow cytometry has not been fully accepted yet due to the lack of standardization protocols. In the past 5 years, we have proposed two versions of a method with reproducible PMP counts in plasma samples. Both methods use forward scatter (FSC)-based threshold set with reference beads of appropriate sizes; first using 0.5 µm beads and later with 0.3 µm beads. Both systems provide reproducible PMP counts. However, this technique works only with some of currently used commercial flow cytometers. Instruments with limited resolution or generating heterogeneous FSC signals are excluded. Such performances are incompatible with the required interinstrument standardization. Here we show that (i) flow cytometers with sub-optimal FSC capabilities generally have higher SSC resolution and background rejection capacity, and (ii) that the same biological entities, 'dim and bright PMP,' both can be counted using alternative strategies, either as previously described, based on FSC measurements, or as presented here, based on SSC detection. The critical element in the standardization protocol is the use of different sizes of reference beads. This study was designed to permit simultaneous access to both FSC- and SSC-optimized platforms. A new range of about 0.17-0.6 µm eq. (µm-equivalents) is proposed for an alternative SSC-based MP gate generating the same PMP counts as those obtained in the previously proposed 0.3-1 µm eq. FSC-based MP gate. The two equivalent standardization options reconcile intrinsically different scattering behaviors between SSC- and FCS - triggered instruments and open the opportunity for multicenter studies in the future. © 2015 International Society for Advancement of Cytometry [ABSTRACT FROM AUTHOR]

Importance: Only 1 randomized clinical trial has shown the superiority of immune checkpoint inhibitors in patients with deficient mismatch repair and/or microsatellite instability (dMMR/MSI) metastatic colorectal cancer (mCRC) in the first-line setting., Objectives: To determine whether avelumab (an anti-programmed cell death ligand 1 antibody) improves progression-free survival (PFS) compared with standard second-line chemotherapy in patients with dMMR/MSI mCRC., Design, Setting, and Participants: The SAMCO-PRODIGE 54 trial is a national open-label phase 2 randomized clinical trial that was conducted from April 24, 2018, to April 29, 2021, at 49 French sites. Patients with dMMR/MSI mCRC who experienced progression while receiving standard first-line therapy were included in the analysis., Interventions: Patients were randomized to receive standard second-line therapy or avelumab every 2 weeks until progression, unacceptable toxic effects, or patient refusal., Main Outcome and Measures: The primary end point was PFS according to RECIST (Response Evaluation Criteria in Solid Tumours), version 1.1, evaluated by investigators in patients with mCRC and confirmed dMMR and MSI status who received at least 1 dose of treatment (modified intention-to-treat [mITT] population)., Results: A total of 122 patients were enrolled in the mITT population. Median age was 66 (IQR, 56-76) years, 65 patients (53.3%) were women, 100 (82.0%) had a right-sided tumor, and 52 (42.6%) had BRAF V600E-mutated tumors. There was no difference in patients and tumor characteristics between treatment groups. No new safety concerns in either group were detected, with fewer treatment-related adverse events of at least grade 3 in the avelumab group than in the chemotherapy group (20 [31.7%] vs 34 [53.1%]; P = .02). After a median follow-up of 33.3 (95% CI, 28.3-34.8) months, avelumab was superior to chemotherapy with or without targeted agents with respect to PFS (15 [24.6%] vs 5 [8.2%] among patients without progression; P = .03). Rates of PFS rates at 12 months were 31.2% (95% CI, 20.1%-42.9%) and 19.4% (95% CI, 10.6%-30.2%) in the avelumab and control groups, respectively, and 27.4% (95% CI, 16.8%-39.0%) and 9.1% (95% CI, 3.2%-18.8%) at 18 months. Objective response rates were similar in both groups (18 [29.5%] vs 16 [26.2%]; P = .45). Among patients with disease control, 18 (75.7%) in the avelumab group compared with 9 (19.1%) in the control group had ongoing disease control at 18 months., Conclusions: The SAMCO-PRODIGE 54 phase 2 randomized clinical trial showed, in patients with dMMR/MSI mCRC, better PFS and disease control duration with avelumab over standard second-line treatment, with a favorable safety profile., Trial Registration: ClinicalTrials.gov Identifier: NCT03186326.

Background: The fortification of wheat-based staple foods, such as pasta, with pseudocereal and legume flours has received growing research interest in recent years. While it is associated with many challenges regarding technological and sensory quality of the products, it promises a substantial improvement of the nutritional value of pasta. However, investigations of the nutritional quality of fortified pasta often focus on the carbohydrate/starch fraction, and information on changes in protein quality is relatively scarce. This study evaluates the nutritional profile of a high-protein hybrid pasta (HPHP) formulation in which a combination of three high-protein ingredients (HPIs) from buckwheat, faba bean and lupin is used to partially replace wheat semolina. The formulation's macronutrient composition, protein quality and the content of antinutritional compounds are assessed in comparison to regular wheat pasta., Results: The HPHP formulation represents a more favourable macronutrient profile compared to regular wheat pasta, particularly in relation to the isocaloric replacement of wheat starch by non-wheat protein. Furthermore, a more balanced amino acid profile, improved N utilisation and increased protein efficiency ratio (in vivo) were determined for HPHP, which conclusively suggests a substantially enhanced protein quality. The cooking process was shown to significantly reduce levels of vicine/convicine and trypsin inhibitor activity originating from HPIs. The small remaining levels seem not to adversely affect HPHP's nutritional quality., Conclusion: This significant upgrade of pasta's nutritional value identifies HPHP, and similar hybrid formulations, as a healthy food choice and valuable alternative to regular wheat pasta, specifically for a protein supply of adequate quality in mostly plant-based diets. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Background: The amino acid composition, and rheological, thermal and colloidal stability of plant protein-based oil-in-water emulsion systems containing 1.90, 3.50 and 7.70 g 100 mL -1 protein, fat and carbohydrate, respectively, using quinoa and lentil protein ratios of 100:0 and 60:40 were investigated. The emulsion containing lentil protein showed lower initial, peak and final viscosity values (22.7, 61.7 and 61.6 mPa s, respectively) than the emulsion formulated with quinoa protein alone (34.3, 102 and 80.0 mPa s, respectively) on heat treatment., Results: Particle size analysis showed that both samples had small particle sizes (~1.36 μm) after homogenization; however, the sample with 60:40 quinoa:lentil protein ratio showed greater physical stability, likely related to the superior emulsifying properties of lentil protein. However, upon heat treatment, large aggregates (~100 μm) were formed in both samples, reducing the physical stability of the samples. This physical stability was increased with the addition of 0.20% sodium dodecyl sulfate (SDS), whereas it was negatively affected by the addition of α-amylase. Addition of α-amylase led to lower viscosity for both emulsion samples, with measured values of 41.8 and 46.0 mPa s for the 100:0 and 60:40 samples, respectively. This suggests that the heat-induced increases in particle size were partially due to hydrophobic interactions between the proteins as SDS disrupts hydrophobic bonds between proteins., Conclusion: These results demonstrated that using a mixture of lentil and quinoa proteins positively affected the physical stability of plant protein-based emulsions, in addition to contributing to a more nutritionally complete amino acid profile - both important considerations in the development of plant-based beverages. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Background: In the European Union proteins for food are largely animal based, consisting of meat and dairy products. Almost all soy but also a larger part of pulses and cereals consumed in the European Union are used for animal nutrition. While livestock is an important source of proteins, it also creates substantial environmental impacts. The food and feed system is closely linked to the planetary and health boundaries and a transformation to healthy diets will require substantial dietary shifts towards healthy foods, such as nuts, fruits, vegetables and legumes., Results: Extrudated vegetable meat alternatives consisting of protein combined with amaranth or buckwheat flour and a vegetable milk alternative made from lentil proteins were shown to have the potential to generate significantly less environmental impact than their animal-based counterparts in most of the environmental indicators examined, taking into account both functional units (mass and protein content). The underlying field-to-fork life cycle assessment models include several variants for both plant and animal foods. The optimized plant-based foods show a clear potential for improvement in the environmental footprints., Conclusions: Development of higher processed and therefore higher performing products is crucial for appealing to potential user groups beyond dedicated vegetarians and vegans and ultimately achieving market expansion. The Protein2Food project showed that prototypes made from European-grown legumes and pseudocereals are a valuable source for high-quality protein foods, and despite being substantially processed they could help reduce the environmental impact of food consumption. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

Background: The protein-rich fractions of pulses and pseudocereals exhibit a well-balanced amino acid profile, particularly when combined in different portions, and are therefore high-value ingredients for the production of extruded snacks. However, the impact of a combination of pulses and pseudocereals on the physical and sensory qualities of extruded snacks has not been investigated up to now. Native or preconditioned protein isolates and concentrates from pulses - as single ingredients or in combination with protein-rich flours of pseudocereals - were analyzed regarding their thermal and functional properties in relation to extrusion characteristics. Low moisture extrusion cooking was used to investigate the impact of protein source (lentil, lupin, faba bean), pseudocereal source (quinoa, amaranth, buckwheat) and protein content (30%, 50%, 70%) on sectional expansion, specific hardness, density and sensory properties of the snacks., Results: With increasing protein content from 30% to 50%, the sectional expansion decreased and the density and specific hardness of the extrudates increased, which could be counteracted by preconditioning of the protein-rich ingredients. Lupin protein-based extrudates exhibited satisfactory texture and sensory properties. Extruded mixtures of pulses and pseudocereals (70% protein) exhibited a smaller sectional expansion compared to pulses as single ingredients (30%, 50%), regardless of pseudocereal type. However, the texture and sensory properties of the extruded blends were satisfactory., Conclusions: We show for the first time that protein-rich fractions of pulses and pseudocereals can be processed into expanded snacks with favorable texture and nutritional properties such as increased protein contents (70%) and balanced amino acid profiles. © 2020 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2020 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Background: The fortification of cereal foods, like pasta, with pseudocereal and legume ingredients promises a substantial improvement of their nutritional quality. However, partial replacement of wheat by pseudocereals and legumes in pasta formulations bears challenges regarding the products' technological and sensory quality. This study investigates the partial replacement of wheat semolina by a combination of high-protein ingredients (HPIs) from buckwheat, faba bean and lupin to reach a protein level of 20% of calories provided by protein. This high-protein hybrid pasta (HPHP) formulation was subjected to a thorough evaluation of technological quality characteristics and compared to regular wheat pasta and pasta formulations containing the single HPIs. Additionally, descriptive sensory profiling was performed to compare organoleptic properties of HPHP with regular wheat pasta., Results: The quality of pasta formulations containing single HPIs was significantly reduced with regard to at least one of the determined quality characteristics. For the HPHP formulation containing all three HPIs, the technological quality was found to be equal to regular wheat pasta. No significant differences were detected for the most indicative quality characteristics cooking loss, firmness and stickiness. This was attributed primarily to compensating effects of the HPIs with respect to different quality characteristics. Sensory analysis revealed only slightly inferior overall quality of HPHP in comparison to regular wheat pasta, especially promoted by similar textural properties., Conclusion: The combination of selected HPIs offers the opportunity to produce high-protein hybrid pasta with technological and sensory quality similar to regular wheat pasta at a level of wheat semolina replacement of 25%. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Background: Infant formula is a human milk substitute for consumption during the first months of life. The protein component of such products is generally of dairy origin. Alternative sources of protein, such as those of plant origin, are of interest due to dairy allergies, intolerances, and ethical and environmental considerations. Lentils have high levels of protein (20-30%) with a good amino acid profile and functional properties. In this study, a model lentil protein-based formula (LF), in powder format, was produced and compared to two commercial plant-based infant formulae (i.e., soy; SF and rice; RF) in terms of physicochemical properties and digestibility., Results: The macronutrient composition was similar between all the samples; however, RF and SF had larger volume-weighted mean particle diameters (D[4,3] of 121-134 μm) than LF (31.9 μm), which was confirmed using scanning electron and confocal laser microscopy. The larger particle sizes of the commercial powders were attributed to their agglomeration during the drying process. Regarding functional properties, the LF showed higher D[4,3] values (17.8 μm) after 18 h reconstitution in water, compared with the SF and RF (5.82 and 4.55 μm, respectively), which could be partially attributed to hydrophobic protein-protein interactions. Regarding viscosity at 95 °C and physical stability, LF was more stable than RF. The digestibility analysis showed LF to have similar values (P < 0.05) to the standard SF., Conclusion: These results demonstrated that, from the nutritional and physicochemical perspectives, lentil proteins represent a good alternative to other sources of plant proteins (e.g., soy and rice) in infant nutritional products. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Pembrolizumab, a PD1 immune checkpoint inhibitor (ICI), was recently reported to be very effective in patients with microsatellite instable/deficient mismatch repair metastatic colorectal cancer (MSI/dMMR mCRC), unlike patients with microsatellite stable/proficient MMR (MSS/pMMR) mCRC, in whom ICIs are generally ineffective. However, about 15% of MSS/pMMR CRCs are highly infiltrated by tumour infiltrating lymphocytes. In addition, both oxaliplatin and bevacizumab have been shown to have immunomodulatory properties that may increase the efficacy of an ICI. We formulated the hypothesis that patients with MSS/pMMR mCRC with a high immune infiltrate can be sensitive to ICI plus oxalipatin and bevacizumab-based chemotherapy. POCHI is a multicenter, open-label, single-arm phase II trial to evaluate efficacy of Pembrolizumab with Capox Bevacizumab as first-line treatment of MSS/pMMR mCRC with a high immune infiltrate for which we plan to enrol 55 patients. Primary endpoint is progression-free survival (PFS) at 10 months, which is expected greater than 50%, but a 70% rate is hoped for. Main secondary objectives are overall survival, secondary resection rate and depth of response. Patients must have been resected of their primary tumour so as to evaluate two different immune scores (Immunoscore® and TuLIS) and are eligible if one score is "high". The first patient was included on April 20, 2021., (Copyright © 2021. Published by Elsevier Ltd.)

The substitution of animal protein with proteins of plant origin is a viable way to decrease the negative impact caused by animal husbandry on the environment. Pulse consumption has been widely promoted as a nutritious contribution to protein supplementation. In this study, an emulsion of lentil ( Lens culinaris ) protein isolate is fermented with lactic acid bacteria (LAB) to manufacture a yoghurt alternative and the techno-functional properties compared to a dairy- and a soy-based product with similar protein contents. The yoghurt-like products are subjected to large and small deformation analysis, quantification of fermentable oligosaccharides, disaccharides, monosaccharides and polyols (FODMAP), water holding capacity tests, protein profile analysis and the gel structure is visualised by confocal laser scanning microscopy (CLSM). The lentil yoghurt alternative shows good water holding capacity, high firmness and consistency values in large deformation analysis, with cohesiveness and viscosity not significantly different from that of dairy yoghurt. The high gel strength and rigidity of the lentil yoghurt gels measured by small deformation analysis is well-reflected in the dense protein matrix in the CLSM graphs. FODMAP content of the lentil yoghurt is very low, making it suitable for consumption by irritable bowel syndrome (IBS) patients. Our results show that lentil protein isolate is an excellent base material for producing a plant-based yoghurt alternative.

This study addresses an innovative approach to generate aerated foods with appealing texture through the utilization of lupin protein isolate (LPI) in combination with edible fats. We show the impact of transglutaminases (TGs; SB6 and commercial), glycerol (Gly), soy lecithin (Lec) and linoleic acid (LA) on the micro- and nanostructure of health promoting solid foods created from LPI and fats blends. 3-D tomographic images of LPI with TG revealed that SB6 contributed to an exceptional bubble spatial organization. The inclusion of Gly and Lec decreased protein polymerization and also induced the formation of a porous layered material. LA promoted protein polymerization and formation of homogeneous thick layers in the LPI matrix. Thus, the LPI is a promising protein resource which when in blend with additives is able to create diverse food structures. Much focus has been placed on the great foamability of LPI and here we show the resulting microstructure of LPI foams, and how these were improved with addition of TGs. New food applications for LPI can arise with the addition of food grade dispersant Lec and essential fatty-acid LA, by improved puffiness, and their contributing as replacer of chemical leavening additives in gluten-free products.

Immune checkpoint inhibitors have failed in treating metastatic colorectal cancer (mCRC) patients except those with dMMR/MSI tumors. However, until very recently we had only non-comparative promising data in this population with anti-programmed cell death 1/ programmed cell death ligand 1 (PD1/PD-L1) antibodies alone or combined with anti- cytotoxic T-lymphocyte-associated protein 4 (CTLA4) antibodies. This comparative phase II trial (NCT03186326), conducted in more than 100 centers in France, will include dMMR/MSI mCRC patients with progression after a first-line treatment with chemotherapy ± targeted therapies, to evaluate efficacy and safety of the anti-PDL1 Avelumab versus a standard second-line treatment. Main inclusion criteria were patients aged 18 to 75 years, ECOG performance status ≤2, dMMR/MSI mCRC and failure of a standard first-line regimen. Patient will be randomised to receive Avelumab 10 mg/kg versus standard second-line doublet chemotherapy plus a targeted agent according to tumor RAS status. Patients will be followed for 4 years. A gain of 5 months in median PFS is expected in favour of the Avelumab arm (12 vs 7 months; HR=0.58). Secondary endpoints include objective response rate, overall survival, quality of life and toxicity. In addition, circulating tumour DNA and microbiota will be explored to test their potential prognostic and predictive values. The study was opened in March 2018., Competing Interests: Declaration of Competing Interest JT has received honoraria for speaker or advisory role from Sanofi, Roche, Merck Serono, Amgen, Servier, Pierre Fabre, Lilly, Astra Zeneca and MSD. LE has received honoraria for speaker or advisory role Servier, Merck Serono, Amgen. PLP is a consultant/advisory board member for Merck Serono, Astrazeneca Amgen, Boehringer Ingelheim, Biocartis, Roche, Bristol-Myers Squibb, Pierre Fabre, Servierand MSD. CL has served in a consulting/advisory role and or received honoraria for, Amgen, Pierre Fabre, Novartis, AAA. FE has received honoraria for speaker and/or advisory role from Ipsen, Merck, Bayer, Sanofi and Servier. OB has received honoraria for speaker and/or advisory role from Merck KGaA, Roche Genentech, Bayer, Astra-Zeneca, Grunenthal, MSD, Amgen, Servier, and Pierre Fabre, TA has served in a consulting/advisory role and/or received honoraria from Amgen, Bristol-Myers Squibb, Chugai, Clovis, Halliodx, MSD, Pierre Fabre, Roche/Ventana, Sanofi, Servier and has received travel, accommodations, and expenses from Roche/Ventana, MSD Oncology, and Bristol-Myers Squibb. DT has served in a consulting/advisory role and/or received honoraria from Amgen, Bristol-Myers Squibb, MSD Oncology, Roche/Ventana, Sanofi, Servier, Novartis, Merck Serono and Astra Zeneca. HP has served in a consulting/advisory role and/or received honoraria from Amgen, Sanofi, Servier, Merck Serono and Roche. SK has received honoraria for speaker and/or advisory role from Amgen, Ipsen, MSD, Pfizer, Sanofi, and Servier. EB, JFE, JB, MC and FD have nothing to disclose. CT has received honoraria for speaker or advisory role from Merck, Sanofi, MSD, Ipsen, Amgen, Servier, and Bristol-Myers Squibb. This research was financially supported by Merck Santé S.A.S., an affiliate of Merck KGaA, Darmstadt, Germany, as part of an alliance between Merck KGaA and Pfizer., (Copyright © 2020 Editrice Gastroenterologica Italiana S.r.l. Published by Elsevier Ltd. All rights reserved.)

Lentil ( Lens culinaris ) is a high-protein crop with a promising potential as a plant-based protein source for human nutrition. This study investigated nutritional and anti-nutritional properties of whole seed lentil flour (LF) compared to lentil protein isolates (LPIs) prepared in pilot-scale by isoelectric precipitation (LPI-IEP) and ultrafiltration (LPI-UF). Fermentable oligosaccharides, disaccharides, monosaccharides, and polyols (FODMAPs) profiles showed significant reductions in total galacto-oligosaccharides (GOS) contents by 58% and 91% in LPI-IEP and LPI-UF, respectively, compared to LF. Trypsin inhibitor activity (TIA) levels based on dry protein mass were lowered by 81% in LPI-IEP and 87% in LPI-UF relative to LF. Depending on the stage of digestion, the in vitro protein digestibility (IVPD) of LPIs was improved by 35-53% compared to LF, with both products showing a similar long-term protein digestibility to that of bovine serum albumin (BSA). This work supports the use of purified LPI products as a novel source of high quality protein for food applications., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 The Authors.)

Grain legumes, such as faba beans, have been investigated as promising ingredients to enhance the nutritional value of wheat bread. However, a detrimental effect on technological bread quality was often reported. Furthermore, considerable amounts of antinutritional compounds present in faba beans are a subject of concern. Sourdough-like fermentation can positively affect baking performance and nutritional attributes of faba bean flours. The multifunctional lactic acid bacteria strain Leuconostoc citreum TR116 was employed to ferment two faba bean flours with different protein contents (dehulled flour (DF); high-protein flour (PR)). The strain's fermentation profile (growth, acidification, carbohydrate metabolism and antifungal phenolic acids) was monitored in both substrates. The fermentates were applied in regular wheat bread by replacing 15% of wheat flour. Water absorption, gluten aggregation behaviour, bread quality characteristics and in vitro starch digestibility were compared to formulations containing unfermented DF and PR and to a control wheat bread. Similar microbial growth, carbohydrate consumption as well as production of lactic and acetic acid were observed in both faba bean ingredients. A less pronounced pH drop as well as a slightly higher amount of antifungal phenolic acids were measured in the PR fermentate. Fermentation caused a striking improvement of the ingredients' baking performance. GlutoPeak measurements allowed for an association of this observation with an improved gluten aggregation. Given its higher potential to improve protein quality in cereal products, the PR fermentate seemed generally more promising as functional ingredient due to its positive impact on bread quality and only moderately increased starch digestibility in bread.

In recent years, dietary products with quinoa and buckwheat have attracted attention mostly due to the high nutritive value of their protein fraction. However, their dietary effect on intestinal microbiota activity and related systemic responses are still poorly understood. Therefore, a 2 week study of twenty-eight growing male Wistar rats was conducted to investigate the effects of quinoa (QU) and buckwheat (BK) protein-rich flours on the growth parameters, intestinal microbial activity, plasma lipid profile, and inflammatory markers. The biological value of protein and body weight gain were considerably increased in the QU and BK groups compared with those in the soy protein isolate group. Moreover, both flours increased the microbial activity of α-glucosidase, β-glucosidase, and α-galactosidase and the concentration of short-chain fatty acids in the caecum. The studied flours favourably reduced the plasma total cholesterol and LDL cholesterol. In rats fed a diet with QU, elevated levels of plasma interleukin 6 and alanine transaminase were observed. The effect of QU on inflammatory markers may be related to the increased expression of aryl hydrocarbon receptor in the liver and to the decreased level of plasma albumin. In conclusion, quinoa and buckwheat protein-rich flours are valuable sources of proteins that favourably affect growth parameters, gut metabolism, and blood lipid profile in rats; however, only the buckwheat flour has no effect on inflammatory processes.

Plant proteins, and legume proteins in particular, have become the centre of attention moving towards a more sustainable and, therefore, more plant-based human diet. Especially hybrid products, containing wheat and legume proteins, promise a balanced amino acid composition and an upgraded nutritional value of both protein sources. This study investigates a high-protein hybrid bread (HPHB) formulation, where wheat flour was partially replaced by high-protein ingredients from faba bean, carob and gluten. In addition to a detailed characterisation of technological quality and sensory profile, also the formulation's nutritional value was examined in comparison to regular wheat bread. Therefore, macronutrient composition, antioxidant potential, amino acid profile and contents of antinutritional compounds were analysed. Furthermore, protein digestibility was determined in an in vitro model and in vivo. Dough analysis revealed significant differences of the HPHB formulation compared to regular wheat dough. However, results obtained for bread quality characteristics prove HPHB to be equal to regular wheat bread and sensory results and the determined sensory attributes suggest high consumer acceptance. Nutritional analyses of HPHB showed a more favourable macronutrient composition in comparison to regular wheat bread; as well as low contents of antinutritional compounds and high antioxidant potential linked to high levels of phenolics. Also an improved amino acid profile, increased nitrogen utilisation rate (by 69%) and higher protein efficiency ratio were determined, which are associated with enhanced protein quality. This suggests HPHB, and similar formulations of its kind, as a valuable and healthy food choice, which can contribute to adequate protein supply in predominantly plant-based diets.

Oil-in-water emulsion systems formulated with plant proteins are of increasing interest to food researchers and industry due to benefits associated with cost-effectiveness, sustainability and animal well-being. The aim of this study was to understand how the stability of complex model emulsions formulated using lentil proteins are influenced by calcium fortification (0 to 10 mM CaCl 2 ) and thermal processing (95 or 140 °C). A valve homogeniser, operating at first and second stage pressures of 15 and 3 MPa, was used to prepare emulsions. On heating at 140 °C, the heat coagulation time (pH 6.8) for the emulsions was successively reduced from 4.80 to 0.40 min with increasing CaCl 2 concentration from 0 to 10 mM, respectively. Correspondingly, the sample with the highest CaCl 2 addition level developed the highest viscosity during heating (95 °C × 30 s), reaching a final value of 163 mPa·s. This was attributed to calcium-mediated interactions of lentil proteins, as confirmed by the increase in the mean particle diameter (D[4,3]) to 36.5 µm for the sample with 6 mM CaCl 2 , compared to the unheated and heated control with D[4,3] values of 0.75 and 0.68 µm, respectively. This study demonstrated that the combination of calcium and heat promoted the aggregation of lentil proteins in concentrated emulsions.

Dry fractionated faba bean protein-rich flour (FPR) produced by milling/air classification, and faba bean protein isolate (FPI) produced by acid extraction/isoelectric precipitation were compared in terms of composition, techno-functional properties, nutritional properties and environmental impacts. FPR had a lower protein content (64.1%, dry matter (DM)) compared to FPI (90.1%, DM), due to the inherent limitations of air classification. Of the two ingredients, FPR demonstrated superior functionality, including higher protein solubility (85%), compared to FPI (32%) at pH 7. Foaming capacity was higher for FPR, although foam stability was similar for both ingredients. FPR had greater gelling ability compared to FPI. The higher carbohydrate content of FPR may have contributed to this difference. An amino acid (AA) analysis revealed that both ingredients were low in sulfur-containing AAs, with FPR having a slightly higher level than FPI. The potential nutritional benefits of the aqueous process compared to the dry process used in this study were apparent in the higher in vitro protein digestibility (IVPD) and lower trypsin inhibitor activity (TIA) in FPI compared to FPR. Additionally, vicine/convicine were detected in FPR, but not in FPI. Furthermore, much lower levels of fermentable oligo-, di- and monosaccharides, and polyols (FODMAPs) were found in FPI compared to FPR. The life cycle assessment (LCA) revealed a lower environmental impact for FPR, partly due to the extra water and energy required for aqueous processing. However, in a comparison with cow's milk protein, both FPR and FPI were shown to have considerably lower environmental impacts.