Alcalase-Based Chickpea (Cicer arietinum L.) Protein Hydrolysates Efficiently Reduce Systolic Blood Pressure in Spontaneously Hypertensive Rats.

Studies on antihypertensive chickpea protein hydrolysates have rarely performed in vivo evaluations, limiting the entry of such hydrolysates into functional food development and clinical trials. Thus, our aim was to optimize the hydrolysis conditions to produce an alcalase-based chickpea hydrolysate with a hypotensive effect in vivo at convenient oral doses. The hydrolysis reaction time, temperature, and alcalase/substrate concentration were optimized using a response surface analysis (RSA). ACE-I inhibition was the response variable. The optimized hydrolysis conditions were time = 0.5 h, temperature = 40 °C, and E/S concentration = 0.254 (U/g). The IC50 of the optimized hydrolysate (OCPH) was 0.358 mg/mL. Five hydrolysates from the RSA worksheet (one of them obtained after 5 min of hydrolysis (CPH15)) had an ACE-I inhibitory potential similar to that of OCPH (p > 0.05). At 50 mg/kg doses, OCPH and CPH15 promoted a clinically relevant hypotensive effect in spontaneously hypertensive rats, up to -47.35 mmHg and -28.95 mmHg, respectively (p < 0.05 vs. negative control). Furthermore, the hypotensive effect was sustained for at least 7 h post-supplementation. Overall, OCPH and CPH15 are promising ingredients for functional food development and as test materials for clinical trials.

A Food Matrix Triggers a Similar Allergic Immune Response in BALB/c Mice Sensitized with Native, Denatured, and Digested Ovalbumin.

The search for an animal model to evaluate the allergenic potential of processed food products is still ongoing. Both the sensitization to ovalbumin (OVA) in different structural states and the allergic response triggered after intragastric or food challenges were assessed. BALB/c mice were sensitized intraperitoneally to OVA (50 µg) in different structural states (native OVA, N-OVA; denatured OVA, D-OVA; formaldehyde- and lysine-treated OVA, FK-OVA; denatured OVA-FK, OVA-DFK; peptides from pepsin digestion, Pep-OVA). Anti-OVA-specific IgE responses were evaluated using ELISA. Anaphylactic signs and mMCP-1 serum levels were evaluated after intragastric (2.0 mg/OVA) and food (0.41 mg/OVA) challenges. IgE reactivities to N-OVA and D-OVA were similar among groups (p > 0.05). After the challenges, all OVA-sensitized mice developed mild to severe anaphylactic signs (p < 0.05 vs. control). Mice sensitized to N-OVA and D-OVA had the highest mMCP-1 serum levels after challenges (p < 0.05 vs. control). Allergic responses were similar despite the different OVA doses used for the challenges. The N-OVA-sensitized murine model of egg allergy proposed in the present study holds the potential for evaluating the impact of food matrix composition and processing on the threshold of egg-allergic responses.

Dipeptidyl Peptidase IV Inhibitory Peptides from Chickpea Proteins (Cicer arietinum L.): Pharmacokinetics, Molecular Interactions, and Multi-Bioactivities.

Chickpea (Cicer arietinum L.) peptides can inhibit dipeptidyl peptidase IV (DPP-IV), an important type 2 diabetes mellitus therapeutic target. The molecular interactions between the inhibitory peptides and the active site of DPP-IV have not been thoroughly examined, nor have their pharmacokinetic properties. Therefore, the predictions of legumin- and provicilin-derived DPP-IV inhibitory peptides, their molecular interactions with the active site of DPP-IV, and their pharmacokinetic properties were carried out. Ninety-two unique DPP-IV inhibitory peptides were identified. Papain and trypsin were the enzymes with the highest AE (0.0927) and lowest BE (6.8625 × 10-7) values, respectively. Peptide binding energy values ranged from -5.2 to -7.9 kcal/mol. HIS-PHE was the most potent DPP-IV inhibitory peptide and interacts with residues of the active sites S1 (TYR662) and S2 (GLU205/ARG125 (hydrogen bonds: <3.0 Å)), S2 (GLU205/GLU206 (electrostatic interactions: <3.0 Å)), and S2' pocket (PHE357 (hydrophobic interaction: 4.36 Å)). Most peptides showed optimal absorption (76.09%), bioavailability (89.13%), and were non-toxic (97.8%) stable for gastrointestinal digestion (73.9%). Some peptides (60.86%) could also inhibit ACE-I. Chickpea is a source of non-toxic and bioavailable DPP-IV-inhibitory peptides with dual bioactivity. Studies addressing the potential of chickpea peptides as therapeutic or adjunct agents for treating type 2 diabetes are warranted.

Prevalence of Parent-Reported Food Allergy in a Mexican Pre-School Population.

The magnitude and relevance of food allergies in the preschool population remain unknown in most regions of Mexico and Latin America. Thus, our aim was to estimate the parent-reported prevalence of food allergies in a Mexican preschool population and to characterize their clinical diagnosis and presentation. A cross-sectional survey was conducted in Culiacán City. A validated questionnaire was utilized. A total of 810 parents responded to the questionnaire (valid response rate, 40.7%). The estimated prevalence rates (95% CI) were: "physician-diagnosed Food Allergy (FA), ever" 5.30% (3.86-7.08); "immediate-type FA, ever" 2.96% (1.91-4.38); "immediate-type FA, current" 1.60% (0.86-2.73); and food-dependent anaphylaxis 1.11% (0.51-2.01). The main food allergens were milk (0.49%), strawberries (0.37%), egg, and soy (0.25% each). Atopic diseases and a family history of allergies were significantly associated with immediate-type FA. Among "immediate-type FA, current" cases, 76.9% required emergency room visits, but the prescription of epinephrine autoinjectors was reported in one case only. The food reactions occurred at home (92.35%), in a relative's house (38.5), and at restaurants (23%). Immediate-type FA reactions requiring emergency room visits are not uncommon among the studied population. Actions like proper anaphylaxis management and the prevention of cross-contamination of foods should be encouraged.

Extrusion Improves the Antihypertensive Potential of a Kabuli Chickpea (Cicer arietinum L.) Protein Hydrolysate.

Chickpea hydrolysates could have antihypertensive potential, but there are no evaluations in vivo. Thus, the antihypertensive potential of a chickpea protein hydrolysate obtained before and after extrusion (a process that modifies protein digestibility) was evaluated. Protein precipitates were obtained from extruded and unextruded chickpea flours by isoelectric precipitation and hydrolyzed (α-amylase/pepsin/pancreatin). Chemical composition was determined (standard methods). ACE-I inhibition assays were carried out using a colorimetric test. For antihypertensive effect evaluations, spontaneously hypertensive rats (n = 8) received the treatments intragastrically (extruded or unextruded hydrolysate (1.2 g/kg), captopril (25 mg/kg), or water only). Fat, ash, and carbohydrate contents were lower in extruded chickpea flour (p < 0.05 versus unextruded). The protein content varied between protein precipitates (91.03%/78.66% unextruded/extruded (dry basis)) (p < 0.05). The hydrolysates' IC50 values (mg/mL) were 0.2834 (unextruded)/0.3218 (extruded) (p > 0.05). All treatments lowered the blood pressure (p < 0.05 vs. water). The extruded hydrolysate showed a more potent antihypertensive effect than the unextruded one (p < 0.05), an effect similar to captopril (p > 0.05). The results suggest that protein extrusion can be used to generate protein hydrolysates with improved health benefits. The findings have implications for the design and production of functional foods that could help to prevent hypertension or serve as an adjunct in its treatment.

Prediction of ACE-I Inhibitory Peptides Derived from Chickpea (Cicer arietinum L.): In Silico Assessments Using Simulated Enzymatic Hydrolysis, Molecular Docking and ADMET Evaluation.

Chickpea (Cicer arietinum L.) peptides have shown in vitro potential to inhibit the angiotensin I-converting enzyme (ACE-I). However, the potential molecular interactions between chickpea peptides (CP) and ACE-I as well as their ADMET (absorption/distribution/metabolism/excretion/toxicity) characteristics remain unknown. Thus, our aim was to study the in silico interactions of CP with ACE-I and the CP ADMET characteristics. Legumin and provicilin sequences were submitted to in silico analysis to search for ACE-I inhibitory peptides. Simulated enzymatic hydrolysis was performed using the BIOPEP-UWM database, and the ACE-I inhibitory peptides generated (EC50 ≤ 200 µM) were selected to perform molecular docking and ADMET analysis. After hydrolysis, 59 out of 381 peptides with ACE-I inhibitory potential were released. Based on A and B parameters, the legumin peptides showed better ACE-I inhibitory potential than the provicilin ones. CP mainly interact with residues from pocket S1 (Ala354/Glu384) and S2 (His353/His513) through hydrogen bonds (distances < 3.0 Å) and hydrophobic interactions (binding energy from −5.7 to −9.2 kcal/mol). Through ADMET analysis, CP showed optimal values for inhibiting ACE-I in vivo. ACE-I inhibitory peptides from legumin and provicilin can bind strongly and tightly to the active site of ACE-I. Further studies to evaluate in vivo the antihypertensive effects of CP are warranted.

Assessment of the Route of Exposure to Ovalbumin and Cow's Milk Proteins on the Induction of IgE Responses in BALB/c Mice.

BALB/c mice can be orally sensitized to food proteins under acid suppressive medication, mimicking human exposure and triggering a human-like allergic immune response. However, the reproducibility of such an oral food allergy model remains questionable. Our aim was to evaluate the IgE responses triggered against ovalbumin (OVA) and cow's milk proteins (CMP) after intragastric (IG), either under gastric-acid suppression or not, or intraperitoneal (IP) sensitization in BALB/c mice. OVA (0.2 mg) and different concentrations of CMP were administered with/without the antacid sucralfate by the IG route. For IP sensitization, OVA or CMP (0.5 mg) were administered. ELISA was used to evaluate IgE responses. The IP sensitization protocols triggered more robust and consistent anti-OVA or anti-CMP IgE responses than the intragastric ones (with/without sucralfate) (p < 0.05). 2.7% (1/36), and 5.5% (3/54) of the mice that underwent the sucralfate-assisted IG protocol triggered IgE responses against OVA or CMP, respectively. All the mice were administered OVA or CMP via IP triggered detectable IgE responses. The IP sensitization model is more reliable than the IG one for evaluating the intrinsic sensitizing and/or allergenic potential of food proteins, even if IG immunizations are carried out under gastric-acid suppression.

Non-Celiac Gluten Sensitivity: An Update.

Non-celiac gluten sensitivity (NCGS) is a clinical entity characterized by the absence of celiac disease and wheat allergy in patients that trigger reproducible symptomatic responses to gluten-containing foods consumption. Due to the lack of sensitive and reproducible biomarkers for NCGS diagnosis, placebo-controlled gluten challenges must be carried out for its diagnosis. The gluten challenges can be either double- or single-blind, for research or clinical practice purposes, respectively. For improving our understanding about the magnitude and relevance of NCGS in different populations, epidemiological studies based on self-report have been carried out. However, the gluten challenge-based prevalence of NCGS remains to be estimated. Since NCGS was recently recognized as a clinical entity, more studies are needed to delve into NCGS pathogenesis, for instance, the molecular interactions between the suspected cereal grain components that trigger NCGS, such as fermentable oligo-, di-, monosaccharides, and polyols (FODMAPs) and amylase and trypsin inhibitors, and the immune system remains to be elucidated. Although still under debate, NCGS patients can be susceptible to only one or more than one of the NCGS triggers. The treatment of NCGS involves the dietary restriction of the suspected triggers of the disease, but there is controversial data about the effectiveness of different dietary interventions such as the gluten-free diet and low-FODMAP diet. Certainly, our understanding of NCGS is improving quickly due to the constant availability of new scientific information on this topic. Thus, the aim of the present narrative review is to present an up-to-date overview on NCGS from epidemiology to current therapy.

Prevalence of Wheat/Gluten-Related Disorders and Gluten-Free Diet in Paraguay: An Online Survey-Based Study.

Gluten-related disorders (GRDs) are increasing around the world, but their magnitude and relevance remain unknown in most Latin American countries. Thus, an online survey was conducted to estimate the prevalence of GRDs as well as adherence to a gluten-free diet (GFD) in Paraguayan adult population. There were 1058 individuals surveyed using a validated questionnaire (response rate of 93.9%). The self-reported prevalence rates were as follows (95% CI): gluten sensitivity (GS), 10.30% (8.53-12.29); non-celiac GS (NCGS), 5.19% (3.94-6.71); physician-diagnosed celiac disease (PD-CD), 3.11% (2.15-4.35); wheat allergy (WA), 2.07% (1.30-3.13); and adherence to GFD, 15.69% (13.55-18.02). Excluding CD, more women than men met the criteria for GRDs, adverse food reactions, and GFD (p < 0.05). Eight respondents reported the coexistence of NCGS with PD-CD and/or WA. Most cases on a GFD indicated medical/dietitian advice for following the diet (68.07%). Non-self-reported GS individuals indicated weight control (46.4%) and the notion that the GFD is healthier (20.2%) as the main motivations for following the diet. GRDs are not uncommon in Paraguayan adult population. It seems that there is awareness about GRDs and the GFD, but training about the diagnosis of GRDs is desirable because of the informed overlapping diagnoses of CD or WA with NCGS. Future studies involving face-to-face interviews are necessary.

Design, Assessment, and Validation of a Questionnaire to Estimate Food-Dependent Exercise-Induced Anaphylaxis Prevalence in Latin American Population.

There are no epidemiological data about food-dependent exercise-induced anaphylaxis (FDEIA) in Latin America. Our aim was to design, assess, and validate a questionnaire to identify potential FDEIA cases and/or estimate its prevalence by self-report. Questions were included in the instrument to address the main symptoms of FDEIA, type/intensity of physical activity, and anaphylaxis. The instrument's clarity, comprehension and repeatability were evaluated. These evaluations were carried out by Hispanic people (Argentinians/Colombians/Mexicans/Peruvians), including nine individuals with medical diagnosis of FDEIA, and Brazilians. The Flesch-Kincaid score was calculated using the INFLESZ software. The instrument was translated from Spanish to Brazilian Portuguese following the translation back-translation procedure. The participants rated the two versions of the questionnaire as clear and comprehensible (three-point ordinal scale) and very easy to understand [0.33; average (scale 0-10)]. For these evaluations, the Kendall's W coefficient showed strong agreement among raters (W = 0.80; average). The Flesch-Kincaid score was 63.5 in average (documents considered as readable). The Cohen's Kappa coefficient showed almost perfect agreement in repeatability (0.88; average). The validation process of two versions of an instrument, used to identify potential FDEIA cases, was successfully carried out and it was found applicable to Latin American countries for generating epidemiological data.

Characteristics of Allergen Labelling and Precautionary Allergen Labelling in Packaged Food Products Available in Latin America.

The characteristics of food allergen labelling are relevant for avoiding accidental exposure to the allergens of interest but no Latin American country has evaluated these characteristics. Our aim was to evaluate the characteristics of food allergen labelling and precautionary allergen labelling (PAL) in six Latin American countries. All data were collected directly from the supermarkets surveyed. A total of 10,254 packaged food products were analyzed, of which 63.3% (n = 6494) and 33.2% (n = 3405) featured allergen labelling and/or PAL, respectively. Most products complied with local regulations (≥87.4% for both locally produced and imported). Thirty-three types of PAL statements were detected; the most frequent was "may contain traces of…" (35.1%). Countries without regulations on the characteristics of allergen labelling had two-fold more products that contained allergens in their ingredients lists but no food allergen labelling. The use of PAL in countries that regulate it (38.2%) was as high as that in countries without PAL regulations (19.2%-44.7%). The findings suggest that the lack of regulations for the characteristics of allergen labeling increases the risk of accidental exposure to allergens of interest. Our findings also suggest that beyond regulations, a scientific approach is required for minimizing and standardizing the use of PAL.

Prevalence of Adverse Reactions to Gluten and People Going on a Gluten-Free Diet: A Survey Study Conducted in Brazil.

BACKGROUND: The prevalence of gluten-related disorders (GRD) and adherence to a gluten-free diet (GFD) remains unknown in Brazilian population and there is no published information on the scientific literature about the proportion of Brazilians that were diagnosed with a gluten-related disorder. Thus, the aim of this work was to estimate the prevalence of GRDs and adherence to a GFD by self-report in adult Brazilian population. MATERIALS AND METHODS: A questionnaire-based cross-sectional study was conducted in two Brazilian cities. RESULTS: The response rate was 93.2% (1630/1749). The self-reported prevalence rates were (95% CI): adverse reactions to gluten 10.06% (8.64-11.62); gluten sensitivity 2.33% (1.65-3.18); physician-diagnosed celiac disease 0.3% (0.09-0.71); non-celiac gluten sensitivity 1.71% (1.14-2.47); wheat allergy 0.79% (0.42-1.36); adherence to gluten-free diet 7.48% (6.25-8.87); gluten avoiders 15.21% (13.5-17.05). Among those who were following a GFD (n = 122), 65.6% (n = 80) of them reported that they did not develop symptoms after wheat/gluten ingestion and 50% (n = 61) were following the diet without medical/dietitian advice. The main motivation for following a GFD in the self-reported and non-self-reported gluten sensitivity groups were the symptoms triggered after wheat/gluten ingestion (86.8%) and weight control (57.1%), respectively. CONCLUSIONS: Implementation of programs to increase awareness about GRDs among healthcare professionals and giving scientifically sound information to the general population about the risks and benefits for following a GFD are desirable actions in Brazil. The results also add to the growing body of evidence for highlighting the under-diagnosis of GRD and the trend for following a GFD in Latin America.

Translation, Cultural Adaptation, and Evaluation of a Brazilian Portuguese Questionnaire to Estimate the Self-Reported Prevalence of Gluten-Related Disorders and Adherence to Gluten-Free Diet.

BACKGROUND: A Spanish version of a questionnaire intended to estimate, at the population level, the prevalence rates of self-reported gluten-related disorders and adherence to gluten-free diets has been applied in four Latin American countries. However, idiom issues have hampered the questionnaire application in the Brazilian population. Thus, the aim of the present study was to carry out a translation, cultural adaptation, and evaluation of a Brazilian Portuguese questionnaire to estimate the self-reported prevalence of gluten-related disorders and adherence to gluten-free diets in a Brazilian population. MATERIALS AND METHODS: Two bilingual Portuguese-Spanish health professionals carried out the translation of the original Spanish version of the questionnaire to Brazilian-Portuguese. Matching between the two translations was evaluated using the WCopyFind.4.1.5 software. Words in conflict were conciliated, and the conciliated version of the Brazilian Portuguese instrument was evaluated to determine its clarity, comprehension, and consistency. A pilot study was carried out using an online platform. RESULTS: The two questionnaires translated into Brazilian Portuguese were highly matched (81.8%-84.1%). The questions of the conciliated questionnaire were clear and comprehensible with a high agreement among the evaluators (n = 64) (average Kendall's W score was 0.875). The participants did not suggest re-wording of questions. The answers to the questions were consistent after two applications of the questionnaire (Cohen's k = 0.869). The pilot online survey yielded low response rates (9.0%) highlighting the need for face-to-face interviews. CONCLUSIONS: The translation and evaluation of a Brazilian Portuguese questionnaire to estimate the self-reported prevalence rates of gluten-related disorders and adherence to gluten-free diets was carried out. The instrument is clear, comprehensible, and generates reproducible results in the target population. Further survey studies involving face-to-face interviews are warranted.

Pasta Enrichment with an Amaranth Hydrolysate Affects the Overall Acceptability while Maintaining Antihypertensive Properties.

BACKGROUND: Alcalase-treated amaranth proteins generate angiotensin-1-converting enzyme (ACE-1) inhibitory peptides, which could be useful for functional foods development. Our aim was to evaluate the technological, sensory, and antihypertensive properties of pasta enriched with an amaranth hydrolysate. METHODS: Pasta with 11% (A; control), 15% (B), and 20% (C) of protein content were formulated. Pastas B and C were supplemented with an alcalase-treated amaranth protein concentrate. Cooking time, cooking lost, color, and texture were assessed. An untrained panel (n = 30) evaluated sensory attributes. The antihypertensive effect was evaluated in hypertensive rats. RESULTS: The hydrolysate IC50 was 0.014 mg/mL. Optimum cooking time and cooking loss decreased in products B and C vs. A (p < 0.05). The L* values decreased in pasta C. Firmness increased in pasta C vs. A (p < 0.05). Adhesiveness was different among groups (p < 0.05). Pasta A had the highest acceptability (p < 0.05). The products B and C, and captopril (positive control) showed antihypertensive properties after 3 h of supplementation (p < 0.05). This effect remained after 7 h, 8 h, or 9 h. CONCLUSIONS: The addition of amaranth hydrolysates to pasta negatively impacts on the overall acceptability and, to a lesser extent, on pasta taste. However, it is possible to maintain the antihypertensive properties of the supplemented pasta under physiological conditions.

Gluten Vehicle and Placebo for Non-Celiac Gluten Sensitivity Assessment.

Non-celiac gluten sensitivity (NCGS) is a syndrome characterized by gastrointestinal and extraintestinal manifestations triggered after gluten ingestion in the absence of celiac disease and wheat allergy. Because of the lack of biomarkers for NCGS diagnosis, the cornerstone for its assessment is a single- or double-blind placebo-controlled (DBPC) gluten challenge. However, there are some non-standardized points in the diagnostic approach proposed by the experts. This complicate comparisons among the results published by different research groups. The gluten vehicle and placebo must be indistinguishable from each other, which entails sensory and technological evaluations of the designed gluten vehicle and placebo products. At the moment, there is no standardized method for the preparation of the gluten vehicle and placebo for carrying out DBPC gluten challenges for NCGS assessment. This review focuses on the challenges that researchers have to face, either for the development of an accepted gluten vehicle and placebo or for identifying NCGS cases on the basis of DBPC gluten challenges.

Food Allergy Prevalence in Salvadoran Schoolchildren Estimated by Parent-Report.

The prevalence of food allergy (FA) has not been estimated at a population level in Central American countries and, consequently, the magnitude and relevance of the problem in the Central American region remains unknown. Thus, our aim was to evaluate the parent-reported prevalence of FA in a population of schoolchildren from the Central American country El Salvador. A Spanish version of a structured questionnaire was utilized. Five hundred and eight (508) parents returned the questionnaire with valid responses (response rate, 32%). The estimated prevalence rates (95% CI) were: adverse food reactions 15.9 (13.0⁻19.3), "perceived FA, ever" 11.6 (9.1⁻14.6), "physician-diagnosed FA, ever" 5.7% (4.0⁻8.0), "immediate-type FA, ever" 8.8% (6.6⁻11.6), "immediate-type FA, current" 5.3% (3.6⁻7.6), and anaphylaxis 2.5% (1.5⁻4.3). The most common food allergens were milk (1.7%), shrimp (1.3), chili (0.7%), chocolate (0.7%), and nuts (0.3%). Most of the "food-dependent anaphylaxis" cases (60.5%) sought medical attention, but only one case reported the prescription of an epinephrine autoinjector. Mild and severe FA cases are not uncommon among Salvadoran schoolchildren and both the prescription of epinephrine autoinjectors by healthcare personnel and the use of the autoinjectors by anaphylactic individuals should be encouraged.