Novel insights into the mechanism(s) of silicon-induced drought stress tolerance in lentil plants revealed by RNA sequencing analysis.

BACKGROUND: Lentil is an essential cool-season food legume that offers several benefits in human nutrition and cropping systems. Drought stress is the major environmental constraint affecting lentil plants' growth and productivity by altering various morphological, physiological, and biochemical traits. Our previous research provided physiological and biochemical evidence showing the role of silicon (Si) in alleviating drought stress in lentil plants, while the molecular mechanisms are still unidentified. Understanding the molecular mechanisms of Si-mediated drought stress tolerance can provide fundamental information to enhance our knowledge of essential gene functions and pathways modulated by Si during drought stress in plants. Thus, the present study compared the transcriptomic characteristics of two lentil genotypes (drought tolerant-ILL6002; drought sensitive-ILL7537) under drought stress and investigated the gene expression in response to Si supplementation using high-throughput RNA sequencing. RESULTS: This study identified 7164 and 5576 differentially expressed genes (DEGs) from drought-stressed lentil genotypes (ILL 6002 and ILL 7537, respectively), with Si treatment. RNA sequencing results showed that Si supplementation could alter the expression of genes related to photosynthesis, osmoprotection, antioxidant systems and signal transduction in both genotypes under drought stress. Furthermore, these DEGs from both genotypes were found to be associated with the metabolism of carbohydrates, lipids and proteins. The identified DEGs were also linked to cell wall biosynthesis and vasculature development. Results suggested that Si modulated the dynamics of biosynthesis of alkaloids and flavonoids and their metabolism in drought-stressed lentil genotypes. Drought-recovery-related DEGs identified from both genotypes validated the role of Si as a drought stress alleviator. This study identified different possible defense-related responses mediated by Si in response to drought stress in lentil plants including cellular redox homeostasis by reactive oxygen species (ROS), cell wall reinforcement by the deposition of cellulose, lignin, xyloglucan, chitin and xylan, secondary metabolites production, osmotic adjustment and stomatal closure. CONCLUSION: Overall, the results suggested that a coordinated interplay between various metabolic pathways is required for Si to induce drought tolerance. This study identified potential genes and different defence mechanisms involved in Si-induced drought stress tolerance in lentil plants. Si supplementation altered various metabolic functions like photosynthesis, antioxidant defence system, osmotic balance, hormonal biosynthesis, signalling, amino acid biosynthesis and metabolism of carbohydrates and lipids under drought stress. These novel findings validated the role of Si in drought stress mitigation and have also provided an opportunity to enhance our understanding at the genomic level of Si's role in alleviating drought stress in plants.

Silicon modulates nitro-oxidative homeostasis along with the antioxidant metabolism to promote drought stress tolerance in lentil plants.

Lentil is the fifth most important grain legume growing in arid/semi-arid regions of the world. Drought is one of the major constraints leading up to 50% of production losses just in lentil. Application of silicon (Si) has been shown to be a promising solution to improve drought tolerance; however, the biochemical mechanisms and interactions involved are not fully understood, especially in legumes. This study was designed to evaluate the effects of Si on drought stress tolerance of lentil genotypes. Seven lentil genotypes with different drought tolerance levels (tolerant, moderately tolerant and sensitive) were subjected to moderate and severe drought stress at the onset of the reproductive stage. Results showed that different drought stress treatments significantly decreased the above ground biomass, water status and the concentration of chlorophyll pigments, whereas Si supplementation of drought stressed lentil genotypes significantly improved the same traits, irrespective of their drought tolerant levels. On the other hand, Si effect on osmoregulation leads to a decline in the membrane damage and osmolytes (proline and glycine betaine) concentration in drought-stressed lentil. Application of Si to drought-stressed lentil plants significantly maintained the nitro-oxidative homeostasis by balancing the concentrations of reactive oxygen/nitrogen species, superoxide anion, hydrogen peroxide and nitrous oxide, thereby reducing the oxidative damage caused due to drought stress. Furthermore, Si supplementation also stimulated the efficiency of the glutathione (GSH)-ascorbate (ASC) cycle by increasing the concentrations of GSH and ASC as well as the activities of antioxidant enzymes like ascorbate peroxidase, guaiacol peroxidase, catalase, superoxide dismutase, glutathione reductase, dehydro-ascorbate reductase and nitrate reductase for better protection of cell membranes from reactive oxygen species. Although Si showed the same regulatory mechanisms in all the studied genotypes to protect lentil plants from moderate and severe drought stress, the defensive role of Si against drought stress was more conspicuous in drought sensitive genotypes than in the tolerant ones. Thus, this study suggests the protective role of Si on drought-stressed lentil genotypes through the modulation of nitro-oxidative homeostasis and antioxidant defence responses.

Fundamental study on changes in the FODMAP profile of cereals, pseudo-cereals, and pulses during the malting process.

Whole grains and pulses are rich in nutrients but often avoided by individuals with gastrointestinal disorders, due to high levels of fermentable oligo-, di-, monosaccharides and polyols (FODMAPs). This study investigated the impact of malting as delivery-system for endogenous enzymes. Malts from barley and wheat (naturally high in fructans), lentils and chickpeas (high in galactooligosaccharides), oat and buckwheat (low in FODMAPs) were produced. While barley and wheat malts had slightly elevated fructan-levels, in oat malt 0.8 g/100 g DM fructans were de novo synthesized. In lentils and chickpeas galactooligosaccharides diminished by 80-90%. Buckwheat did not contain any FODMAPs commonly investigated, but fagopyritols which may have a similar physiological effect. Also fagopyritols were degraded. While malted pulses and buckwheat are directly suitable for low FODMAP applications, using the combined approach of malting and fermentation, malted cereals could contribute to high nutritional values of such products.

Genotoxicity of the food additive E171, titanium dioxide, in the plants Lens culinaris L. and Allium cepa L.

E171 (titanium dioxide, TiO2), an authorized foods and beverage additive, is also used in food packaging and in pharmaceutical and cosmetic preparations. E171 is considered to be an inert and non-digestible material, not storable in animal tissues, but the possible presence of TiO2 nanoparticles (NP) may present a risk to human health and the environment. We determined the presence of 15% TiO2 NP in a commercial E171 food additive product, by electron microscopy. The biological effects of E171 were assessed in Lens culinaris and Allium cepa for the following endpoints: percentage of germination, root elongation, mitotic index, presence of chromosomal abnormalities, and micronuclei. The results indicated low phytotoxicity but dose-dependent genotoxicity. We also observed internalization of TiO2 NP and ultrastructural alterations in the root systems.

In Vitro Colonic Fermentation of Saponin-Rich Extracts from Quinoa, Lentil, and Fenugreek. Effect on Sapogenins Yield and Human Gut Microbiota.

In vitro colonic fermentation of saponin-rich extracts from quinoa, lentil, and fenugreek was performed. Production of sapogenins by human fecal microbiota and the impact of extracts on representative intestinal bacterial groups were evaluated. The main sapogenins were found after fermentation (soyasapogenol B for lentil; oleanolic acid, hederagenin, phytolaccagenic acid, and serjanic acid for quinoa; and sarsasapogenin, diosgenin, and neotigogenin acetate for fenugreek). Interindividual differences were observed, but the highest production of sapogenins corresponded to quinoa (90 µg/mL) and fenugreek (70 µg/mL) extracts, being minor for lentil (4 µg/mL). Lentil and quinoa extracts showed a general antimicrobial effect, mainly on lactic acid bacteria and Lactobacillus spp. Significant increases of Bifidobacterium spp. and Lactobacillus spp. were observed for fenugreek in one volunteer. Thus, the transformation of saponin-rich extracts of quinoa, lentil, and fenugreek to sapogenins by human gut microbiota is demonstrated, exhibiting a modulatory effect on the growth of selected intestinal bacteria.

Insights into acetate-mediated copper homeostasis and antioxidant defense in lentil under excessive copper stress.

Gradual contamination of agricultural land with copper (Cu), due to the indiscriminate uses of fungicides and pesticides, and the discharge of industrial waste to the environment, poses a high threat for soil degradation and consequently food crop production. In this study, we combined morphological, physiological and biochemical assays to investigate the mechanisms underlying acetate-mediated Cu toxicity tolerance in lentil. Results demonstrated that high dose of Cu (3.0 mM CuSO4. 5H2O) reduced seedling growth and chlorophyll content, while augmenting Cu contents in both roots and shoots, and increasing oxidative damage in lentil plants through disruption of the antioxidant defense. Principle component analysis clearly indicated that Cu accumulation and increased oxidative damage were the key factors for Cu toxicity in lentil seedlings. However, acetate pretreatment reduced Cu accumulation in roots and shoots, increased proline content and improved the responses of antioxidant defense (e.g. increased catalase and glutathione-S-transferase activities, and improved action of glutathione-ascorbate metabolic pathway). As a result, excess Cu-induced oxidative damage was reduced, and seedling growth was improved under Cu stress conditions, indicating the role of acetate in alleviating Cu toxicity in lentil seedlings. Taken together, exogenous acetate application reduced Cu accumulation in lentil roots and shoots and mitigated oxidative damage by activating the antioxidant defense, which were the major determinants for alleviating Cu toxicity in lentil seedlings. Our findings provide mechanistic insights into the protective roles of acetate in mitigating Cu toxicity in lentil, and suggest that application of acetate could be a novel and economical strategy for the management of heavy metal toxicity and accumulation in crops.

Cooked Red Lentils Dose-Dependently Modulate the Colonic Microenvironment in Healthy C57Bl/6 Male Mice.

Dietary pulses, including lentils, are protein-rich plant foods that are enriched in intestinal health-promoting bioactives, such as non-digestible carbohydrates and phenolic compounds. The aim of this study was to investigate the effect of diets supplemented with cooked red lentils on the colonic microenvironment (microbiota composition and activity and epithelial barrier integrity and function). C57Bl/6 male mice were fed one of five diets: a control basal diet (BD), a BD-supplemented diet with 5, 10 or 20% cooked red lentils (by weight), or a BD-supplemented diet with 0.7% pectin (equivalent soluble fiber level as found in the 20% lentil diet). Red lentil supplementation resulted in increased: (1) fecal microbiota α-diversity; (2) abundance of short-chain fatty acid (SCFA)-producing bacteria (e.g., Prevotella, Roseburia and Dorea spp.); (3) concentrations of fecal SCFAs; (4) mRNA expression of SCFA receptors (G-protein-coupled receptors (GPR 41 and 43) and tight/adherens junction proteins (Zona Occulden-1 (ZO-1), Claudin-2, E-cadherin). Overall, 20% lentil had the greatest impact on colon health outcomes, which were in part explained by a change in the soluble and insoluble fiber profile of the diet. These results support recent public health recommendations to increase consumption of plant-based protein foods for improved health, in particular intestinal health.

Arbuscular mycorrhizal fungi reduce arsenic uptake and improve plant growth in Lens culinaris.

Arsenic (As) is a carcinogenic and hazardous substance that poses a serious risk to human health due to its transport into the food chain. The present research is focused on the As transport in different lentil genotypes and the role of Arbuscular Mycorrhizal Fungi (AMF) in mitigation of As phyto-toxicity. Arsenic transport from soil to root, shoot and grains in different lentil genotypes was analyzed by flow injection hydride generation atomic absorption spectrophotometry. AMF were applied for the reduction of As uptake as well as the improvement of plant growth in lentil genotypes. Arsenic phyto-toxicity was dose-dependent as evidenced by relatively higher shoot length, fresh and dry weight of root and shoot in 5 and 15 mgkg-1 As-treated lentil plants than that in 100 mgkg-1 As-treated lentil. Arsenic accumulation occurred in roots and shoots of all BARI-released lentil genotypes. Arsenic accumulation in grains was found higher in BARI Mashur 1 than other lentil genotypes. AMF treatment significantly increased growth and biomass accumulation in lentil compared to that in non-AMF plants. Furthermore, AMF effectively reduced the As concentrations in roots and shoots of lentil plants grown at 8 and 45 mgkg-1 As-contaminated soils. This study revealed remarkable divergence in As accumulation among different BARI-released lentil genotypes; however, AMF could reduce As uptake and mitigate As-induced phyto-toxicity in lentil. Taken together, our results suggest a great potential of AMF in mitigating As transfer in root and shoot mass and reallocation to grains, which would expand lentil cultivation in As-affected areas throughout the world.

Iron-fortified lentils to improve iron (Fe) status among adolescent girls in Bangladesh - study protocol for a double-blind community-based randomized controlled trial.

BACKGROUND: Lentils are generally considered to be a nutrient-dense food, and a good source of iron (Fe). This study aims to establish novel evidence of the effectiveness of the consumption of Fe-fortified lentils in improving the body Fe status and thus cognitive performance in non-pregnant adolescent girls in rural Bangladesh, compared to consumption of ordinary lentils. METHODS: We have designed a double-blind (both trial participants and outcome assessors), community-based, cluster-randomized controlled trial among 1260 Bangladeshi adolescent girls between the ages of 10-17 years who are non-smoking, not married, not pregnant, not breastfeeding, and generally healthy at the time of enrollment. The intervention will include three arms who will receive: (1) Fe-fortified lentils; (2) unfortified lentils; or (3) usual intake. Participants will be served a thick preparation of cooked Fe-fortified lentils (37.5 g raw lentils, approximately 200 g cooked lentils) 5 days per week for 85 feeding days (around 4 months) using a locally acceptable recipe. Lentils were fortified with Fe in the laboratory at the Department of Plant Sciences at the University of Saskatchewan in Canada. A subsample of participants (n = 360) will be randomly invited to be included in cognitive testing. DISCUSSION: Data on socio-demographic characteristics, household food security status, adolescent food habits and cognitive testing will be collected at baseline and endline (4 months). Venous blood samples will be collected at baseline, midline (2 months) and endline to measure adolescents' Fe status. Computerized cognitive testing will include five common measures of attentional (three of attention) and mnemonic functioning (two of memory) carried out using DMDX software. The results of this study will be used to garner support for and to substantiate large-scale production and market expansion of Fe-fortified lentils, and will contribute to knowledge about how to enhance Fe status in adolescents worldwide in resource-poor settings, using staple food crops. TRIAL REGISTRATION: ClinicalTrials.gov NCT03516734 . Registered on 24 May 2018.

Influence of extruded lentil containing high chromium nutritional yeast on the main physiological factors associated with metabolic syndrome in rodent models.

Insulin resistance, obesity and dyslipidemia are the main physiological factors associated with metabolic syndrome. The objectives of this study were to understand the effects of diets containing extruded lentil fortified with high chromium nutritional yeast (YCr) or chromium picolinate on glucose tolerance, clearance and fasting blood glucose concentrations in Normal and Obese (Ob/Ob) mice and to determine the effects of the diets on the mice plasma lipid profiles. Diets A, B and C contained YCr in different doses and concentrations, as follows: Diet A = 15.7 g and 16 ppm, B = 157.1 g and 16 ppm, and C = 299.3 g and 27 ppm, respectively. Diet D contained chromium picolinate at a dose and concentration of 15.7 g and 16 ppm, respectively. Intraperitoneal glucose tolerance tests and intraperitoneal insulin tolerance tests were conducted at 4-weeks and 8-weeks post diet initiation, in addition to, plasma lipoprotein profiles and organ weights. Normal mice showed only slight variability with respect to the studied biological parameters compared to the Ob/Ob mice group. Results indicated that following 4-weeks of diet supplementation, Ob/Ob mice fed diets A, C and D had significantly (p < 0.05) lower fasting blood glucose (FBG) than Ob/Ob mice fed Diet B. However, after 8-weeks Ob/Ob mice fed Diet C, containing YCr, had a significantly (p < 0.05) lower FBG than mice supplemented with Diet D, containing chromium picolinate. Therefore, based on these findings, it was concluded that YCr at the highest concentration and dose was more effective than chromium picolinate. These results indicate that ready-to-eat snacks and breakfast cereal type products supplemented with chromium in the form of YCr could be used as vehicles for the amelioration of main physiological factors associated with metabolic syndrome.

Relative Bioavailability of Iron in Bangladeshi Traditional Meals Prepared with Iron-Fortified Lentil Dal.

Due to low Fe bioavailability and low consumption per meal, lentil must be fortified to contribute significant bioavailable Fe in the Bangladeshi diet. Moreover, since red lentil is dehulled prior to consumption, an opportunity exists at this point to fortify lentil with Fe. Thus, in the present study, lentil was Fe-fortified (using a fortificant Fe concentration of 2800 µg g-1) and used in 30 traditional Bangladeshi meals with broad differences in concentrations of iron, phytic acid (PA), and relative Fe bioavailability (RFeB%). Fortification with NaFeEDTA increased the iron concentration in lentil from 60 to 439 µg g-1 and resulted in a 79% increase in the amount of available Fe as estimated by Caco-2 cell ferritin formation. Phytic acid levels were reduced from 6.2 to 4.6 mg g-1 when fortified lentil was added, thereby reducing the PA:Fe molar ratio from 8.8 to 0.9. This effect was presumably due to dephytinization of fortified lentil during the fortification process. A significant (p ≤ 0.01) Pearson correlation was observed between Fe concentration and RFeB% and between RFeB% and PA:Fe molar ratio in meals with fortified lentil, but not for the meal with unfortified lentil. In conclusion, fortified lentil can contribute significant bioavailable Fe to populations at risk of Fe deficiency.

Content of Minerals and Fatty Acids and Their Correlation with Phytochemical Compounds and Antioxidant Activity of Leguminous Seeds.

The aim of the study was to determine the mineral composition and fatty acid profile in the seeds of selected Fabaceae species and cultivars and to assess their correlations with phytochemicals and antioxidant activity. The Andean lupine was characterised by a particularly high level of Mg and K as well as Cu, Zn, and Fe (P < 0.05). There were various correlations (P < 0.05) between the total phenols and tannins and these elements. The highest contribution of α-linolenic acid (ALA, 18:3, n-3) in total fatty acids was noted in the lentil (13.8 in 100 g-1 fat), common bean (11.9 in 100 g-1 fat), and pea seeds (10.4 in 100 g-1 fat) (P = 0.028). In turn, the white lupine contained the highest content of ALA-0.67 g 100 g-1 seeds; its lowest level was determined in the broad bean-0.03 g 100 g-1 seeds. The seeds exhibited a high proportion of hypocholesterolemic fatty acids (on average 86%). The 2,2-diphenyl-1-picrylhydrazyl antiradical activity was positively correlated with UFA and PUFA (P < 0.05). This indicates great protective potential of legume seeds for prevention and treatment of diet-dependent diseases.

Critical evaluation of changes in the ratio of insoluble bound to soluble phenolics on antioxidant activity of lentils during germination.

A new indicator, the ratio of insoluble bound phenolics (IBPs) to soluble phenolics (SPs), is suggested as an effective means to monitor changes in the antioxidant activity of lentils during germination. This indicator may be used to monitor other process-induced changes in antioxidant potential of food phenolics in other foods. The antioxidant activity of SPs, IBPs, and total value, the sum of both free and esterified phenolics, of germinated CDC Richlea lentil variety was evaluated for 4 days. Total phenolic content (TPC), total flavonoid content (TFC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical cation scavenging ability were employed to record antioxidant activities. An incremental increase in IBPs was found in TPC, TFC, DPPH, and ABTS radical cation scavenging ability, whereas SPs showed a declining trend in TFC, DPPH, and ABTS, except TPC during 4 days of germination. The ratio of IBPs to SPs increased using most methods, and this may be possibly due to the changes of phenolic compound formation from soluble into insoluble bound form during germination process. The ratio can be used as a novel method for monitoring process-induced changes in the antioxidant activity of foods.

Coordinated response of sulfate transport, cysteine biosynthesis, and glutathione-mediated antioxidant defense in lentil (Lens culinaris Medik.) genotypes exposed to arsenic.

Response of sulfate transporters, thiol metabolism, and antioxidant defense system was studied in roots of two lentil (Lens culinaris Medik.) genotypes grown in arsenic (10, 25, and 40 µM As(V))-supplemented nutrient solution, and significant changes compared to control (0 µM As(V)) were observed mainly at 25 and 40 µM. In L 414, high glutathione (GSH) redox (0.8-0.9) was maintained with elevated thiol synthesis, powered by transcriptional up-regulation of LcSultr1;1 and LcSultr1;2 sulfate transporters and significant induction of LcSAT1;1 and LcSAT1;2 (serine acetyltransferase), OAS-TL (O-acetylserine(thiol)-lyase), γ-ECS (γ-glutamylcysteine synthetase), and PCS (phytochelatin synthase) genes predominantly within 12-24 h of As exposure at 25 µM and within 6-12 h at 40 µM. This thiolic potency in L 414 roots was effectively complemented by up-regulation of gene expressions and consequent enhanced activities of superoxide dismutase, ascorbate peroxidase (APX), dehydroascorbate reductase, glutathione reductase (GR), and glutathione-S-transferase (GST) isoforms at 25 and 40 µMAs, efficiently scavenging excess reactive oxygen species to prevent onset of As-induced oxidative stress and consequent inhibition of root growth in L 414. In contrast, down-regulation of vital sulfate-uptake transporters as well as entire thiol-metabolizing system and considerably low APX, GST, and GR expressions in DPL 59 not only resulted in reduced GSH redox but also led to over-accumulation of H2O2. This triggered membrane lipid peroxidations as the marks of As-induced oxidative damage. Results indicated coordinated response of thiol-metabolism and antioxidant defense in conferring As-tolerance in lentil, and GSH is the key point in this cascade.

Study of selenocompounds from selenium-enriched culture of edible sprouts.

Selenium is recognised as an essential micronutrient for humans and animals. One of the main sources of selenocompounds in the human diet is vegetables. Therefore, this study deals with the Se species present in different edible sprouts grown in Se-enriched media. We grew alfalfa, lentil and soy in a hydroponic system amended with soluble salts, containing the same proportion of Se, in the form of Se(VI) and Se(IV). Total Se in the sprouts was determined by acidic digestion in a microwave system and by ICP/MS. Se speciation was carried out by enzymatic extraction (Protease XIV) and measured by LC-ICP/MS. The study shows that the Se content of plants depends on the content in the growth culture, and that part of the inorganic Se was biotransformed mainly into SeMet. These results contribute to our understanding of the uptake of inorganic Se and its biotransformation by edible plants.

Beneficial effect of protracted sterilization of lentils on phytase production by Aspergillus ficuum in solid state fermentation.

Water addition to the solid substrate preceding autoclaving increased substrate porosity and phytase production in solid state fermentation. In comparison with dry sterilization, the phytase activity increased 6-, 8.5-, and 10-fold when the autoclaving time was 20, 40, and 60 min, respectively. Autoclaving increased the void space of sterilized lentils, and the increase was 16% higher when water was supplemented to the lentils before sterilization. Image analysis of SEM pictures of the solid substrate showed that water supplementation presterilization portended greater micro-fissure surface area, which also increased with increasing the sterilization time. SEM pictures of the fermentation product showed that fungal growth into the center of the solid substrate was ubiquitous when water was supplemented before sterilization but was absent when water was supplemented post sterilization. Similarly, spore formation on the substrate surface for the presterilization water supplementation samples far exceeded spore formation for samples that received supplementation poststerilization. This evidence suggests that improved mass transfer into the solid substrate resulting from additional pore volume and the formation of micro-fissures on the substrate surface is responsible for the observed gains in phytase productivity in solid state fermentation.

Effect of low- and high-glycemic-index meals on metabolism and performance during high-intensity, intermittent exercise.

Consuming carbohydrate-rich meals before continuous endurance exercise improves performance, yet few studies have evaluated the ideal preexercise meal for high-intensity intermittent exercise, which is characteristic of many team sports. The authors' purpose was to investigate the effects of low- and high-glycemic-index (GI) meals on metabolism and performance during high-intensity, intermittent exercise. Sixteen male participants completed three 90-min high-intensity intermittent running trials in a single-blinded random order, separated by ~7 d, while fasted (control) and 2 hr after ingesting an isoenergetic low-GI (lentil), or high-GI (potato and egg white) preexercise meal. Serum free fatty acids were higher and insulin lower throughout exercise in the fasted condition (p < .05), but there were no differences in blood glucose during exercise between conditions. Distance covered on a repeated-sprint test at the end of exercise was significantly greater in the low-GI and high-GI conditions than in the control (p < .05). Rating of perceived exertion was lower in the low-GI condition than in the control (p = .01). In a subsample of 5 participants, muscle glycogen availability was greater in the low-and high-GI conditions versus fasted control before the repeated-sprint test (p < .05), with no differences between low and high GI. When exogenous carbohydrates are not provided during exercise both low- and high-GI preexercise meals improve high-intensity, intermittent exercise performance, probably by increasing the availability of muscle glycogen. However, the GI does not influence markers of substrate oxidation during high-intensity, intermittent exercise.

IgE-mediated cross-reactivity among leguminous seed proteins in peanut allergic children.

The immunological cross-reactivity among major protein- and oil-crops, including lupin, lentil, pea, peanut, kidney bean and soybean, has been studied by a combination of in vitro and in vivo experimental approaches: SDS-PAGE separations of legume protein extracts and immuno-blot revelations with 12 peanut-sensitive subjects' sera, Immuno-CAP and Skin Prick tests on the same subjects. The immuno-blotting data showed a wide range of IgE-binding responses both displayed by one subject towards different plant extracts and among subjects. Differences were both quantitative and qualitative. The prevalent responses of most subjects' sera were seen with peanut polypeptides, as expected, as well as with various polypeptides of the other legumes, the most recurrent of which were the basic subunits of the 11S globulins. The distribution of in vivo responses generally paralleled those obtained by in vitro approaches with strong responses elicited by peanut, lentil and pea protein extracts, especially by most sensitive subjects, thus providing a consistent overall set of results. In this work, the comparison of various approaches has allowed us to get an overall broad picture of the immunological cross-reactivities among proteins of widely used different seed species and to hypothesize the role of most conserved specific polypeptides.

Label-free detection of lectins on carbohydrate-modified boron-doped diamond surfaces.

This paper describes the label-free detection of carbohydrate-lectin interactions. The sensor consists of a boron-doped diamond (BDD) electrode terminated with alkynyl surface groups, which have been functionalized via the CuACC (copper(I)-catalyzed azide-alkyne cycloaddition) "click" reaction with carbohydrate analogues bearing an azido-terminating arm. In this work, electrochemical impedance spectroscopy (EIS) was used as an effective technique to probe the specific interactions of the surface-bound carbohydrates with their complementary lectin partners, and the response was found to be dependent on the relative density of sugar units immobilized on the BDD surface. A BDD interface with 60% surface-bound mannose showed a detection limit of ∼5 ± 0.5 nM for Lens culinaris lectin, with an affinity constant of K(A) = (2.63 ± 0.5) × 10(6) M(-1).

Effects of enzymatic hydrolysis on lentil allergenicity.

Enzymatic hydrolysis and further processing are commonly used to produce hypoallergenic dietary products derived from different protein sources, such as cow's milk. Lentils and chickpeas seem to be an important cause of IgE-mediated hypersensitivity in the Mediterranean area and India. Some studies have investigated the effects of enzymatic treatments on the in vitro immunological reactivity of members of the Leguminosae family, such as soybean, chickpea, lentil, and lupine. Nevertheless, there are only a few studies carried out to evaluate the effect on IgE reactivity of these food-hydrolysis products with sera from patients with well-documented allergy to these foods. In this study, lentil protein extract was hydrolyzed by sequential action of an endoprotease (Alcalase) and an exoprotease (Flavourzyme). Immunoreactivity to raw and hydrolyzed lentil extract was evaluated by means of IgE immunoblotting and ELISA using sera from five patients with clinical allergy to lentil. The results indicated that sequential hydrolysis of lentil results in an important proteolytic destruction of IgE-binding epitopes shown by in vitro experiments. However, some allergenic proteins were still detected by sera from four out of five patients in the last step of sequential hydrolyzation.