Multi-criteria assessment of pea protein quality in rats: a comparison between casein, gluten and pea protein alone or supplemented with methionine.

The objective of this study was to assess the nutritional quality of pea protein isolate in rats and to evaluate the impact of methionine (Met) supplementation. Several protein diets were studied: pea protein, casein, gluten, pea protein-gluten combination and pea protein supplemented with Met. Study 1: Young male Wistar rats (n 8/group) were fed the test diets ad libitum for 28 d. The protein efficiency ratio (PER) was measured. Study 2: Adult male Wistar rats (n 9/group) were fed the test diets for 10 d. A protein-free diet group was used to determine endogenous losses of N. The rats were placed in metabolism cages for 3 d to assess N balance, true faecal N digestibility and to calculate the Protein Digestible-Corrected Amino Acid Score (PDCAAS). They were then given a calibrated meal and euthanised 6 h later for collection of digestive contents. The true caecal amino acid (AA) digestibility was determined, and the Digestible Indispensable Amino Acid Score (DIAAS) was calculated. Met supplementation increased the PER of pea protein (2·52 v. 1·14, P < 0·001) up to the PER of casein (2·55). Mean true caecal AA digestibility was 94 % for pea protein. The DIAAS was 0·88 for pea protein and 1·10 with Met supplementation, 1·29 for casein and 0·25 for gluten. Pea protein was highly digestible in rats under our experimental conditions, and Met supplementation enabled generation of a mixture that had a protein quality that was not different from that of casein.

Registered dietitians' perceptions and use of the plant-based recommendations in the 2019 Canada's Food Guide.

BACKGROUND: In 2019, Canada's Food Guide underwent its first transformation in over a decade. The change received considerable attention given the increased focus on plant-based proteins and the decision to eliminate the milk and alternatives food group. Yet little is known about health professionals' views of these updates. In response, the present study examined Canadian dietitians' attitudes and behaviours towards the guide's expanded plant-based recommendations. METHODS: A pan-Canadian cross-sectional online survey was administered from January to March 2020 to currently practicing registered dietitians. The study was pre-registered, piloted and received ethical approval. Descriptive and inferential statistics were performed and open-text responses underwent thematic analysis. RESULTS: In total, 411 dietitians from 10 provinces and various work settings participated. Most dietitians (82.8%) consider the food guide's recommendation to choose protein foods that come from plants more often is evidence-based. Compared to its predecessor, dietitians encourage their patients/clients to choose protein foods that come from plants (p < 0.001) and non-dairy sources of calcium (p < 0.001) significantly more frequently under the new guide. A slight majority of dietitians (57.7%) agree with the decision to omit the standalone milk and alternatives group in favour of bringing dairy into the protein category. CONCLUSIONS: Canadian dietitians generally look favourably upon the new plant-based recommendations and have adjusted their nutrition counselling in response. The findings are considered to be the first to characterise how dietitians view changes to the plant-based content of one of the most recognisable diet-related educational tools in Canada.

A Systematic Review of the Effects of Plant Compared with Animal Protein Sources on Features of Metabolic Syndrome.

Dietary protein may play an important role in the prevention of metabolic dysfunctions. However, the way in which the protein source affects these dysfunctions has not been clearly established. The aim of the current systematic review was to compare the impact of plant- and animal-sourced dietary proteins on several features of metabolic syndrome in humans. The PubMed database was searched for both chronic and acute interventional studies, as well as observational studies, in healthy humans or those with metabolic dysfunctions, in which the impact of animal and plant protein intake was compared while using the following variables: cholesterolemia and triglyceridemia, blood pressure, glucose homeostasis, and body composition. Based on data extraction, we observed that soy protein consumption (with isoflavones), but not soy protein alone (without isoflavones) or other plant proteins (pea and lupine proteins, wheat gluten), leads to a 3% greater decrease in both total and LDL cholesterol compared with animal-sourced protein ingestion, especially in individuals with high fasting cholesterol concentrations. This observation was made when animal proteins were provided as a whole diet rather than given supplementally. Some observational studies reported an inverse association between plant protein intake and systolic and diastolic blood pressure, but this was not confirmed by intervention studies. Moreover, plant protein (wheat gluten, soy protein) intake as part of a mixed meal resulted in a lower postprandial insulin response than did whey. This systematic review provides some evidence that the intake of soy protein associated with isoflavones may prevent the onset of risk factors associated with cardiovascular disease, i.e., hypercholesterolemia and hypertension, in humans. However, we were not able to draw any further conclusions from the present work on the positive effects of plant proteins relating to glucose homeostasis and body composition.

Nutritive quality and protein production from grain legumes in a boreal climate.

BACKGROUND: Boreal cropping systems are heavily focused on the production of small-grain cereals; to improve their resilience to climate change and to achieve food and feed security, diversification is needed. This study investigated the potential of faba bean, narrow-leafed lupin and lentil as protein crops in southern Finland, where faba bean is traditional but the other two are novel. RESULTS: Early cultivars of narrow-leafed lupin and lentil matured adequately. Protein concentration in faba bean was, at 32%, higher than the world average of 29%, while those of narrow-leafed lupin and lentil were close to their world averages. Protein yields decreased in the order faba bean > narrow-leafed lupin > lentil. Lipid content of faba bean and lentil was about 1.2% and that of narrow-leafed lupin about 5.5%, and fatty acid composition was largely oleic and linoleic in all three species. CONCLUSION: Both lentil and narrow-leafed lupin can be added to the range of feed and food crops produced at high latitudes in Europe. While faba bean produces the greatest protein yield and lysine concentration, the higher sulfur amino acid concentration in lupin, its oil content and its adaptation to acid, sandy soils not suitable for faba bean make it an attractive alternative.

Comparative proteome analysis of glutenin synthesis and accumulation in developing grains between superior and poor quality bread wheat cultivars.

BACKGROUND: Wheat glutenins are the major determinants of wheat quality. In this study, grains at the development stage from three wheat cultivars (Jimai 20, Jin 411 and Zhoumai 16) with different bread-making quality were harvested based on thermal times from 150 °C(d) to 750 °C(d) , and were used to investigate glutenin accumulation patterns and their relationships with wheat quality. RESULTS: High and low molecular weight glutenin subunits (HMW-GSs and LMW-GSs) were synthesised concurrently. No obvious correlations between HMW/LMW glutenin ratios and dough property were observed. Accumulation levels of HMW-GSs and LMW-GSs as well as 1Bx13 + 1By16 and 1Dx4 + 1Dy12 subunits were higher in superior gluten quality cultivar Jimain 20 than in poor quality cultivar Jing 411 and Zhoumai 16. According to the results of two-dimensional gel electrophoresis, six types of accumulation patterns in LMW-GSs were identified and classified. The possible relationships between individual LMW-GSs and gluten quality were established. CONCLUSION: The high accumulation level of HMW-GSs and LMW-GSs as well as 1Bx13 + 1By16 and 1Dx4 + 1Dy12 subunits contributed to the superior gluten quality of Jimai 20. Two highly expressed and 16 specifically expressed LMW glutenin subunits in Jimain 20 had positive effects on dough quality, while 17 specifically expressed subunits in Zhoumai 16 and Jing 411 appeared to have negative effects on gluten quality.

High antioxidant activity mixture of extruded whole quality protein maize and common bean flours for production of a nutraceutical beverage elaborated with a traditional Mexican formulation.

The objective of this study was to determine the best combination of extrusion process variables for the production of whole quality protein maize (EQPMF) and common bean (ECBF) flours to prepare a high antioxidant activity mixture (EQPMF + ECBF) suitable to produce a nutraceutical beverage with high acceptability elaborated with a traditional Mexican formulation. Processing conditions were obtained from a factorial combination of barrel temperature (BT = 120-170 °C) and screw speed (SS = 120-200 rpm). Response surface methodology was applied to obtain maximum values for antioxidant activity (A ( ox ) A) of the flour mixture (EQPMF + ECBF) and acceptability (A) of the nutraceutical beverage. The best combinations of extrusion process variables for EQPMF and ECBF to prepare an optimized mixture (60%EQPMF + 40%ECBF) were BT = 98 °C/SS = 218 rpm and BT = 105 °C/SS = 83 rpm, respectively. The optimized mixture had A ( ox ) A = 14,320 µmol Trolox equivalent (TE)/100 g sample dry weight (dw) and a calculated protein efficiency ratio (C-PER) of 2.17. A 200 ml portion of a beverage prepared with 25 g of the optimized flour mixture had A ( ox ) A = 3,222 µmol TE, and A = 89 (level of satisfaction "I like it extremely"). This nutraceutical beverage could be used as an alternative to beverages with low nutritional/nutraceutical value, such as those prepared with water, simple sugars, artificial flavoring and colorants, which are widely offered in the market.

Selection and validation of reference genes for normalization of qRT-PCR data to study the cannabinoid pathway genes in industrial hemp.

There has been significant interest in researching the pharmaceutical applications of Industrial hemp since its legalization three years ago. The crop is mostly dioecious and known for its production of phytocannabinoids, flavonoids, and terpenes. Although many scientific reports have showed gene expression analysis of hemp through OMICs approaches, unreliable reference genes for normalization of qRT-PCR data make it difficult to validate the OMICs data. Four software packages: geNorm, NormFinder, BestKeeper, and RefFinder were used to evaluate the differential gene expression patterns of 13 candidate reference genes under osmotic, heavy metal, hormonal, and UV stresses. EF-1α ranked as the most stable reference gene across all stresses, TUB was the most stable under osmotic stress, and TATA was the most stable under both heavy metal stress and hormonal stimuli. The expression patterns of two cannabinoid pathway genes, AAE1 and CBDAS, were used to validate the reliability of the selected reference genes. This work provides useful information for gene expression characterization in hemp and future research in the synthesis, transport, and accumulation of secondary metabolites.

A survey of quantitative real-time polymerase chain reaction internal reference genes for expression studies in Brassica napus.

Eight reference genes of Brassica napus were evaluated using quantitative real-time polymerase chain reaction (qRT-PCR) data, focusing on vegetative tissues and developing embryos. Analyses of expression stability indicated that UP1, UBC9, UBC21, and TIP41 were the top four choices as stably expressed reference genes for vegetative tissues, whereas ACT7, UBC21, TIP41, and PP2A were the top four choices for maturing embryos. In addition, radiolabeling of overall messenger RNA (mRNA) of maturing embryos indicated that the expression patterns of the top four ranked reference genes reflected the overall mRNA content changes in maturing embryos.

Candidate reference genes for gene expression studies in water lily.

The selection of an appropriate reference gene(s) is a prerequisite for the proper interpretation of quantitative Real-Time polymerase chain reaction data. We report the evaluation of eight candidate reference genes across various tissues and treatments in the water lily by the two software packages geNorm and NormFinder. Across all samples, clathrin adaptor complexes medium subunit (AP47) and actin 11 (ACT11) emerged as the most suitable reference genes. Across different tissues, ACT11 and elongation factor 1-alpha (EF1alpha) exhibited a stable expression pattern. ACT11 and AP47 also stably expressed in roots subjected to various treatments, but in the leaves of the same plants the most stably expressed genes were ubiquitin-conjugating enzyme 16 (UBC16) and ACT11.

Protein and RNA Quality Control by Autophagy in Plant Cells.

Eukaryotic cells use conserved quality control mechanisms to repair or degrade defective proteins, which are synthesized at a high rate during proteotoxic stress. Quality control mechanisms include molecular chaperones, the ubiquitin-proteasome system, and autophagic machinery. Recent research reveals that during autophagy, membrane-bound organelles are selectively sequestered and degraded. Selective autophagy is also critical for the clearance of excess or damaged protein complexes (e.g., proteasomes and ribosomes) and membrane-less compartments (e.g., protein aggregates and ribonucleoprotein granules). As sessile organisms, plants rely on quality control mechanisms for their adaptation to fluctuating environments. In this mini-review, we highlight recent work elucidating the roles of selective autophagy in the quality control of proteins and RNA in plant cells. Emphasis will be placed on selective degradation of membrane-less compartments and protein complexes in the cytoplasm. We also propose possible mechanisms by which defective proteins are selectively recognized by autophagic machinery.

Validation of ELISA methods for quantification of the major birch allergen Bet v 1 (BSP090).

To date, the potency of allergen products in Europe is expressed in manufacturer-specific units relative to a product-specific in-house reference. Consequently, cross-product comparability of allergen products from different manufacturers with respect to strength and efficacy is impossible. The Biological Standardisation Programme (BSP) project BSP090 addresses this issue via the establishment of reference standards in conjunction with ELISA methods for the quantification of major allergens in allergen products. Since the initiation of BSP090, the recombinant major allergen Bet v 1 has been adopted by the European Pharmacopoeia Commission as a Chemical Reference Substance (CRS). In parallel, two sandwich ELISA systems for quantification of Bet v 1 were found suitable in preliminary phases of BSP090 to be validated in a large collaborative study. In this study, the candidate ELISA systems were compared with respect to accuracy, precision and variability. Thirteen participating laboratories tested model samples containing the CRS as well as spiked and unspiked birch pollen extracts. Both in pre-testing and in the collaborative study, the 2 candidate ELISA systems confirmed their suitability to quantify recombinant and native Bet v 1. As no clear-cut decision for one of the ELISA systems could be made based on the results of the collaborative study, a post-study testing was performed. Bet v 1 content of 30 birch pollen allergen products was determined in parallel in both ELISA systems. Consequently, 1 candidate ELISA system was selected to be proposed as the future European Pharmacopoeia standard method for Bet v 1 quantification.

[Study on temperature correctional models of quantitative analysis with near infrared spectroscopy].

Effect of enviroment temperature on near infrared spectroscopic quantitative analysis was studied. The temperature correction model was calibrated with 45 wheat samples at different environment temperaturs and with the temperature as an external variable. The constant temperature model was calibated with 45 wheat samples at the same temperature. The predicted results of two models for the protein contents of wheat samples at different temperatures were compared. The results showed that the mean standard error of prediction (SEP) of the temperature correction model was 0.333, but the SEP of constant temperature (22 degrees C) model increased as the temperature difference enlarged, and the SEP is up to 0.602 when using this model at 4 degrees C. It was suggested that the temperature correctional model improves the analysis precision.

[Applied study on support vector machine (SVM) regression method in quantitative analysis with near-infrared spectroscopy].

This paper introduced the application of support vector machines(SVM) regression method based on statistics studytheory to the quantitative analysis with near-infrared (NIR) spectroscopy. Sixty-six wheat samples were used as experimental materials, and thirty-three of them were used as calibration samples. The protein contents and NIR spectra of the calibration samples were used to build SVM regression models by four different kernel functions. The protein content of the predicting samples are estimated by four different SVM regression models. All of the correlation coefficients between the estimated values by different SVM regression models and the standard chemical values of protein content by Kjeldahl's method are more than 0.97. The average absolute error is less than 0.32. To investigate the predicting effect, it is compared with PLS regression models. The result suggested that the SVM regression, which was built to estimate the protein content of wheat samples, can also be used in the quantitative analysis of real samples by NIR.

Vitamin B-6 requirement and status assessment: young women fed a depletion diet followed by a plant- or animal-protein diet with graded amounts of vitamin B-6.

The vitamin B-6 requirement of young women consuming a high-protein diet (1.55 g/kg body wt) and the effect of protein quality on this requirement was studied. In addition, the response of clinical, functional, and biochemical measures of vitamin B-6 nutriture to short-term depletion and step-wise repletion of vitamin B-6 were evaluated. Eight healthy young women resided in a metabolic unit and were fed a formula depletion diet (< 0.05 mg vitamin B-6/d) for 11-28 d followed by either an animal-protein (AP) or plant-protein (PP) diet with successively increasing vitamin B-6 intakes (0.5, 1.0, 1.5, and 2.0 mg/d) for periods of 14-21 d. Animal proteins were primarily from dairy and poultry sources and plant proteins were primarily from legumes. Vitamin B-6 status measures were assessed at weekly intervals. Results showed that a PP diet does not elevate the vitamin B-6 requirement over that required for an AP diet given the high amount of dietary protein used in this study. It was also found that 0.015 mg vitamin B-6/g protein intake normalized most biochemical indexes of vitamin B-6 status (including those indicative of functional status), and that 0.020 mg/g protein normalized all biochemical measures except total urinary vitamin B-6. Adding a margin of safety to either the 0.015 or 0.020 mg/g protein intake would raise the vitamin B-6 requirement for young women above the currently recommended dietary allowance of 0.016 mg/g protein.

Determination of Ole e 1 by enzyme immunoassay and scanning densitometry. Validation by skin-prick testing.

Ole e 1 is an important allergen in Olea europaea pollen extracts. This study describes the development of two new methods that can be used to estimate the Ole e 1 content in olive tree pollen extracts. They are based on (1) an enzyme immunoassay that uses rabbit polyclonal, monospecific antibodies and purified Ole e 1, and (2) scanning densitometry of SDS-PAGE gels. Twelve extracts were evaluated by in vivo and in vitro methods. The in vivo biological potency was estimated by prick skin testing 17 allergic individuals; the in vitro allergenic potency by direct IgE and IgE inhibition assays. The enzyme immunoassay showed an operative range of 0.03-100 microg/ml and demonstrated to be specific for Ole e 1. The Ole e 1 content ranged from 1% to 5% of the total protein in the 12 extracts. The amount of Ole e 1, assessed by gel scanning densitometry significantly correlated with the Ole e 1 content obtained by the immunoassay (r = 0.92; p < 0.001). The Ole e 1 content showed a significant correlation with the total allergenic potency of the extracts, evaluated by direct IgE, specific IgE inhibition and skin-prick testing. These two methods can be used to determine the Ole e 1 content in olive pollen extracts. The content of Ole e 1 can vary from 1% to 5% of the total protein in the extracts.

Nutritive evaluation of the seed, germinated seed, and string bean of Erythrina americana and the detoxification of the material by boiling.

The nutritive and toxicological values of the dry seeds, germinated seeds, and string beans of Erythrina americana were studied using raw and boiled samples. Raw germinated seeds had a higher protein content and lower fiber content than dry seeds. The whole string bean had lower protein content and higher fiber content. However, the seeds of the green pod showed the same protein concentration as the dry seeds (dry basis). Boiling and elimination of broth was beneficial in diminishing the alkaloid concentration in all the samples. The trypsin inhibitors, lectins, and tannins were also diminished as was expected. The raw string bean showed the lowest LD(50). Although the total essential amino acids content of the boiled germinated seeds was increased, the quality of protein, protein efficiency ratio (PER), was lower than in boiled dry seeds, and in these, the PER was similar to the control (casein). The present results suggest that for the protein quality and low alkaloid content, the boiled dry seeds and string beans could be used for animal feeding. It could be interesting to test the raw string beans in ruminants since in this stage E. americana showed the lowest toxicity.

Opportunities for nutritionally enhanced maize and wheat varieties to combat protein and micronutrient malnutrition.

Naturally occurring variation detected in the germplasm of maize and wheat, two of the top three cereal crops in the world, provides options for incorporating higher levels of iron, zinc, and beta-carotene into these grains. In addition, quality protein maize (QPM) has been developed from naturally occurring variation; its seed contains enhanced levels of lysine and tryptophan, two essential amino acids lacking in cereals. The International Maize and Wheat Improvement Center, along with its many partners, has identified several maize and wheat varieties with 25% to 30% higher grain iron and zinc concentrations. Wild relatives of wheat have been found to contain some of the highest iron and zinc concentrations in the grains. Although these accessions are often low yielding and have poor grain quality, backcrossing to bread wheat could result in highly nutritious cultivars. Options are now available for conventional and biotechnology-assisted improvement of the nutritional content of maize and wheat germplasm.

Establishment of recombinant major allergens Bet v 1 and Phl p 5a as Ph. Eur. reference standards and validation of ELISA methods for their measurement. Results from feasibility studies.

The potency of allergen extracts is determined as total allergenic activity without consideration of their composition and the units differ from one manufacturer to another, making it very difficult to compare the different products. Recently, purified major allergens have been obtained by recombinant DNA technology and produced under Good Manufacturing Practice (GMP) conditions. In principle, such recombinant allergens could be established as reference standards and could help for the standardisation of the major allergen content of allergen extracts. Two recombinant major allergens, one from birch pollen, rBet v 1, and one from Timothy grass pollen, Phl p 5a, have been selected at the end of the CREATE programme as a potential starting point for the establishment as European Pharmacopoeia (Ph. Eur.) Reference Standards through a project run by the Biological Standardisation Programme (BSP) of the European Directorate for the Quality of Medicines & HealthCare (EDQM). To this end, bulk candidate recombinant materials, produced under GMP conditions, were procured from two European manufacturers and subsequently formulated and lyophilised. Four ELISA systems from three different manufacturers were included in the project, two for Bet v 1 and two for Phl p 5a with the aim of establishing reference methods for determination of the respective major antigens both in natural allergen extracts as well as in recombinant allergen products. The project was run in 3 phases: a preparatory and preliminary testing phase (feasibility phase or Phase 1), an extended feasibility phase carried out in 3 laboratories (Phase 2) to confirm the transferability of the methods and an international collaborative study with a large number of participating laboratories (Phase 3). This article describes the work done in Phase 1 and Phase 2, i.e. the physico-chemical and biological characterisation of the recombinant candidate reference standards, the assessment of their suitability for the intended purpose as well as the evaluation of the candidate ELISA systems. The results show that both candidate reference standards are suitable for the intended purpose. In addition, three out of the four ELISA systems that were included in the preliminary phase were found to be appropriate for further evaluation in the collaborative study which was organised in 2011. The results of the collaborative study will be published separately.