Phase 3 Trial of Epicutaneous Immunotherapy in Toddlers with Peanut Allergy.

BACKGROUND: No approved treatment for peanut allergy exists for children younger than 4 years of age, and the efficacy and safety of epicutaneous immunotherapy with a peanut patch in toddlers with peanut allergy are unknown. METHODS: We conducted this phase 3, multicenter, double-blind, randomized, placebo-controlled trial involving children 1 to 3 years of age with peanut allergy confirmed by a double-blind, placebo-controlled food challenge. Patients who had an eliciting dose (the dose necessary to elicit an allergic reaction) of 300 mg or less of peanut protein were assigned in a 2:1 ratio to receive epicutaneous immunotherapy delivered by means of a peanut patch (intervention group) or to receive placebo administered daily for 12 months. The primary end point was a treatment response as measured by the eliciting dose of peanut protein at 12 months. Safety was assessed according to the occurrence of adverse events during the use of the peanut patch or placebo. RESULTS: Of the 362 patients who underwent randomization, 84.8% completed the trial. The primary efficacy end point result was observed in 67.0% of children in the intervention group as compared with 33.5% of those in the placebo group (risk difference, 33.4 percentage points; 95% confidence interval, 22.4 to 44.5; P<0.001). Adverse events that occurred during the use of the intervention or placebo, irrespective of relatedness, were observed in 100% of the patients in the intervention group and 99.2% in the placebo group. Serious adverse events occurred in 8.6% of the patients in the intervention group and 2.5% of those in the placebo group; anaphylaxis occurred in 7.8% and 3.4%, respectively. Serious treatment-related adverse events occurred in 0.4% of patients in the intervention group and none in the placebo group. Treatment-related anaphylaxis occurred in 1.6% in the intervention group and none in the placebo group. CONCLUSIONS: In this trial involving children 1 to 3 years of age with peanut allergy, epicutaneous immunotherapy for 12 months was superior to placebo in desensitizing children to peanuts and increasing the peanut dose that triggered allergic symptoms. (Funded by DBV Technologies; EPITOPE ClinicalTrials.gov number, NCT03211247.).

Nanobody-Based Electrochemical Immunoassay for Sensitive Detection of Peanut Allergen Ara h 1.

Peanut is widely used for food supplementation with potential allergic reactions in infants and adults, which prompted the development of reliable and accurate detection of peanut allergens with emphasis on Ara h 1. In this study, a nanobody (Nb)-based micro-total electrochemical immunoassay (Nb-µTEI) was proposed to be generated. Generally, an alpaca was immunized with Ara h 1 to yield a Nb reservoir for selection of four specific Nbs. Nb-mediated immunocapturing allowed the identification of the target as Ara h 1. The Nb-based electrochemical immunoassay was developed by constructing a capturing electrode with cycles of signal enhancement. After construction of the capturing electrode, Nb152 with HA-tag was directly applied to connect immobilized anti-HA IgG for the capture of different concentrations of Ara h 1, which was labeled by biotinylated Nb152 to facilitate signal development with alkaline phosphatase conjugated streptavidin (SA-ALP). A linear range from 4.5 to 55 ng/mL was acquired with LOD and LOQ of 0.86 and 2.10 ng/mL, respectively, with an 11-fold increase of the sensitivity compared with the established sandwich ELISA. The dedicated immunoassay was verified by detecting the antigen spiked in food samples and demonstrated the successful conjugation of Nb with advanced detecting techniques.

Synthesis of co-pyrolyzed biochar using red mud and peanut shell for removing phosphate from pickling wastewater: Performance and mechanism.

Iron (Fe)/iron oxide-modified biochar has practicable adsorption capability for phosphorus (P), but it is expensive. In this study, we synthesized novel low-cost and eco-friendly adsorbents co-pyrolyzed biochars using Fe-rich red mud (RM) and peanut shell (PS) wastes via a one-step pyrolysis process for removing P from pickling wastewater. The preparation conditions (heating rate, pyrolysis temperature, and feedstock ratio) and P adsorption behaviors were systematically investigated. In addition, a series of characterization and approximate site energy distribution (ASED) analyses were conducted to understand the P adsorption mechanisms. The magnetic biochar (BR7P3) with m (RM):m (PS) of 7:3 prepared at 900°C and 10 °C/min had a high surface area (164.43 m2/g) and different abundant ions (including Fe3+, and Al3+). In addition, BR7P3 exhibited the best P removal capability (142.6 mg/g). The Fe2O3 from RM was successfully reduced to Fe0, which was easily oxidized as Fe3+ to precipitate with H2PO4-. The electrostatic effect, Fe-O-P bonding, and surface precipitation were the main mechanisms of P removal. ASED analyses revealed that high distribution frequency and solution temperature led to a high P adsorption rate of the adsorbent. Therefore, this study provides new insight into the waste-to-wealth strategy by transforming PS and RM into mineral-biomass biochar with excellent P adsorption capability and environmental adaptability.

The ethanolic extract of peanut shell attenuates the depressive-like behaviors of mice through modulation of inflammation and gut microbiota.

Peanut shell is an agricultural byproduct being wasted on a large scale, which is in urgent need to be recycled. To fully utilize its pharmacological ingredients, e.g. luteolin, eriodyctiol, and 5,7-dihydroxychromone, we evaluated the curative effect of ethanol extract deriving from peanut shell (PSE) in treating chronic unpredictable mild stress (CUMS)-induced depressive mice. The chronic stress lasted for 10 weeks, and PSE at 100-900 mg/kg/day was gavaged to mice in the last 2 weeks of modeling. The depressive behaviors were assessed by analyses of sucrose preference, tail suspension, and forced swimming. The brain injury was demonstrated by Hematoxylin and Eosin (H&E), Nissl body, and TdT-mediated dUTP nick end labeling (TUNEL) stainings in the mouse hippocampus. Biochemical indicators were analyzed, including levels of neurotrophic factors, neurotransmitters, stress hormones, and inflammatory mediators. The feces were collected for the 16S rDNA sequencing of gut microbiome. Administration of PSE improved the sucrose water consumption of depressive mice, while it decreased the immobile time in tail suspension and forced swimming tests. Meanwhile, the anti-depressive effect of PSE was supported by ameliorated histochemical staining, increased levels of neurotrophic factors and neurotransmitters, as well as down-regulated stress hormones. Furthermore, the treatment of PSE was able to mitigate the levels of inflammatory cytokines in brain, serum, and small intestine. Besides, the tight junction proteins, e.g., occludin and ZO-1, of gut showed elevated expressions, which coincided with the elevated abundance and diversity of gut microbiota upon PSE treatment. This study validated the therapeutic efficacy of PSE in fighting against depression, as well as its modulatory action on inflammation and gut microbiota, which promoted the recycling of this agricultural waste to be health supplements of added value.

Pediatric peanut aspirations before and after 2015 recommendation for early peanut exposure.

OBJECTIVES: To investigate if there has been an increase in peanut foreign body aspirations (FBA) in children since the publication of the Learning Early About Peanut Allergy (LEAP) trial, which revealed that early exposure to peanut-containing foods prevented peanut allergies in children at risk of atopic disease. METHODS: Retrospective chart reviews were conducted separately at two pediatric institutions. Institutions One and Two reviewed children less than 7 years old who underwent bronchoscopy for FBA over ten-year periods between January 2007 and September 2017 and November 2008 and May 2018, respectively. The proportion of FBAs attributed to peanuts was compared before and after the publication LEAP. RESULTS: Out of 515 reviewed cases, there was no change in pediatric peanut aspirations prior to and following the LEAP trial and AAP guideline change (33.5% vs 31.4%, p = 0.70). At Institution One, 317 patients met inclusion criteria. When comparing FBAs before and after LEAP, there were no significant changes in the rate of peanut aspiration (53.5% vs. 45.1%, p = 0.17). Institution Two also found no significant increase in the rate of peanut aspirations before and after the Addendum Guidelines (41.4% vs. 28.6%, p = 0.65) upon review of 198 cases. CONCLUSIONS: Multiple institutions demonstrated a non-significant change in the rate of peanut FBAs following the AAP recommendation. Given that peanuts comprise a large proportion of FBAs, it is important to continue to track peanut aspirations. Longer term data tracking is needed from more institutions to further understand how recommendations from other specialties and the media impacts pediatric aspiration outcomes.

Oral Immunotherapy: An Overview.

Oral immunotherapy (OIT) is an alternative treatment of IgE-mediated food allergy that has been shown to increase tolerance threshold to many of the top food allergens, although this effect may be dependent on age, dose, frequency, and duration. OIT has been shown to be effective and safe in infants, and early initiation can improve rates of desensitization even for those foods whose natural history favors loss of allergy. Studies looking at protocol modification to improve OIT success are ongoing as is the evaluation of clinical tools to help monitor OIT effects.

The Effects of Peanut Oligopeptides on Exercise-Induced Fatigue in Mice and Its Underlying Mechanism.

The aim of this study was to clarify the anti-fatigue effect of peanut oligopeptides (POPs) in mice and to investigate its possible underlying mechanism. A total of 150 male ICR mice were randomly assigned into five groups: control, whey protein (0.50 g/kg·bw), and three peanut peptide groups (0.25, 0.50, and 1.00 g/kg·bw). All the mice were treated with intra-gastric administration for 30 days. Following the intervention, a weight-loaded swimming test, blood lactate concentration, glycogen content, the activities of antioxidant factors and energy metabolism enzymes, and the function of mitochondria in the skeletal muscle were examined. The results show that POP intervention significantly prolonged the exhaustive swimming time, decreased blood lactate concentration levels, regulated the process of energy metabolism, and increased the level of antioxidant enzymes, muscle glycogen, and expressions of mtTFA and NRF-1 in the mitochondria of the gastrocnemius muscle. The results suggest that POPs produce an anti-fatigue effect in the animals, and they may exert this effect through the mechanism of improving the animals' antioxidant capacity to reduce oxidative damage levels and regulating the process of energy metabolism.

"Green" Extraction and On-Site Rapid Detection of Aflatoxin B1, Zearalenone and Deoxynivalenol in Corn, Rice and Peanut.

The common mycotoxins in polluted grains are aflatoxin B1(AFB1), zearalenone (ZEN) and deoxynivalenol (DON). Because of the potential threat to humans and animals, it is necessary to detect mycotoxin contaminants rapidly. At present, later flow immunoassay (LFIA) is one of the most frequently used methods for rapid analysis. However, multistep sample pretreatment processes and organic solvents are also required to extract mycotoxins from grains. In this study, we developed a one-step and "green" sample pretreatment method without using organic solvents. By combining with LFIA test strips and a handheld detection device, an on-site method for the rapid detection of AFB1, ZEN and DON was developed. The LODs for AFB1, ZEN and DON in corn are 0.90 µg/kg, 7.11 µg/kg and 10.6 µg/kg, respectively, and the working ranges are from 1.25 µg/kg to 40 µg/kg, 20 µg/kg to 2000 µg/kg and 35 µg/kg to 1500 µg/kg, respectively. This method has been successfully applied to the detection of AFB1, ZEN and DON in corn, rice and peanut, with recoveries of 89 ± 3%-106 ± 3%, 86 ± 2%-108 ± 7% and 90 ± 2%-106 ± 10%, respectively. The detection results for the AFB1, ZEN and DON residues in certified reference materials by this method were in good agreement with their certificate values.

Influences of biochar with selenite on bacterial community in soil and Cd in peanut.

Cadmium (Cd) pollution in crops seriously affects the ecosystem and human health. Effective measures should be employed to reduce the absorption and accumulation of cadmium in crops. Currently, there are many pieces of research on the application of biochar (BC) and selenium (Se) alone to the remediation of soil Cd pollution; however, few investigations have been devoted to the application of BC and Se together to the remediation of soil Cd pollution. The peanut was taken as the target crop to explore the effects of exogenous selenium and biochar on the remediation of soil Cd pollution. The response of the soil bacterial community to two levels of Cd concentration and its relationship with soil properties and Cd availability are methodically investigated. This study sets two cadmium pollution concentrations of low Cd (5 mg/ kg) and high Cd (20 mg/kg), as well as six treatments: blank, BC, soil Se, soil Se-BC, leaf Se, and leaf Se-BC. The achieved results revealed that both Se and BC could noticeably enhance the yield of peanut seeds and reduce the Cd content in peanut seeds. Among them, Se-BC treatment on soil exhibits the most influence, which reduces the Cd content by 47.86%. Se and BC also affect the physical and chemical properties of soil and remarkably magnify the content of soil available phosphorus, organic matter, soil pH, and soil conductivity. For instance, then effect is detected in the case of applying selenium biochar to soil, leading to an increase of about 64.38%, 72.62%, 2.64%, and 61.15%, respectively, and reducing the content of soil available cadmium by 21.02%. Redundancy analysis confirms that these properties enhance the abundance of dominant bacteria Actinobacteria, Proteobacteria, and Chloroflexi. The correlation analysis also indicates that Saccharimonadales, Bacillus, Arthrobacter, and other bacteria with the function of reducing the bioavailability of cadmium in soil reveal a considerable positive correlation with the variations of physical and chemical properties. In general, exogenous Se and BC incorporate to drop the content of available Cd in the soil through direct passivation, passivation caused by soil environmental change, and passivation caused by altering the soil microbial community structure; as a result, the migration and enrichment of Cd in peanut seeds are blocked and reduced. Moreover, the mixed application of BC and soil Se exhibits the best effect.

Mapping of QTLs Associated with Biological Nitrogen Fixation Traits in Peanuts (Arachis hypogaea L.) Using an Interspecific Population Derived from the Cross between the Cultivated Species and Its Wild Ancestors.

Peanuts (Arachis hypogaea L.) are an allotetraploid grain legume mainly cultivated by poor farmers in Africa, in degraded soil and with low input systems. Further understanding nodulation genetic mechanisms could be a relevant option to facilitate the improvement of yield and lift up soil without synthetic fertilizers. We used a subset of 83 chromosome segment substitution lines (CSSLs) derived from the cross between a wild synthetic tetraploid AiAd (Arachis ipaensis × Arachis duranensis)4× and the cultivated variety Fleur11, and evaluated them for traits related to BNF under shade-house conditions. Three treatments were tested: without nitrogen; with nitrogen; and without nitrogen, but with added0 Bradyrhizobium vignae strain ISRA400. The leaf chlorophyll content and total biomass were used as surrogate traits for BNF. We found significant variations for both traits specially linked to BNF, and four QTLs (quantitative trait loci) were consistently mapped. At all QTLs, the wild alleles decreased the value of the trait, indicating a negative effect on BNF. A detailed characterization of the lines carrying those QTLs in controlled conditions showed that the QTLs affected the nitrogen fixation efficiency, nodule colonization, and development. Our results provide new insights into peanut nodulation mechanisms and could be used to target BNF traits in peanut breeding programs.

Genetic analysis and exploration of major effect QTLs underlying oil content in peanut.

KEY MESSAGE: AhyHOF1, likely encoding a WRI1 transcription factor, plays critical roles in peanut oil synthesis. Although increasing the oil content of peanut to meet growing demand has long been a primary aim of breeding programs worldwide, the mining of genetic resources to achieve this objective has obviously lagged behind that of other oil crops. In the present study, we developed an advanced recombinant inbred line population containing 192 F9:11 families derived from parents JH5 and KX01-6. We then constructed a high-resolution genetic map covering 3,706.382 cM, with an average length of 185.32 cM per linkage group, using 2840 polymorphic SNPs. Two stable QTLs, qCOA08_1 and qCOA08_2 having the highest contributions to genetic variation (16.1% and 20.7%, respectively), were simultaneously detected in multiple environments and closely mapped within physical intervals of approximately 2.9 Mb and 1.7 Mb, respectively, on chromosome A08. In addition, combined analysis of whole-genome and transcriptome resequencing data uncovered a strong candidate gene encoding a WRI1 transcription factor and differentially expressed between the two parents. This gene, designated as High Oil Favorable gene 1 in Arachis hypogaea (AhyHOF1), was hypothesized to play roles in oil accumulation. Examination of near-inbred lines of #AhyHOF1/#Ahyhof1 provided further evidence that AhyHOF1 increases oil content, mainly by affecting the contents of several fatty acids. Taken together, our results provide valuable information for cloning the favorable allele for oil content in peanut. In addition, the closely linked polymorphic SNP markers within qCOA08_1 and qCOA08_2 loci may be useful for accelerating marker-assisted selection breeding of peanut.

Food anaphylaxis diagnostic marker compilation in machine learning design and validation.

BACKGROUND: Traditional food allergy assessment of anaphylaxis remains limited in accuracy and accessibility. Current methods of anaphylaxis risk assessment are costly with low predictive accuracy. The Tolerance Induction Program (TIP) for anaphylactic patients undergoing TIP immunotherapy produced large-scale diagnostic data across biosimilar proteins, which was used to develop a machine learning model for patient-specific and allergen-specific anaphylaxis assessment. In explanation of construct, this work describes the algorithm design for assignment of peanut allergen score as a quantitative measure of anaphylaxis risk. Secondarily, it confirms the accuracy of the machine learning model for a specific cohort of food anaphylactic children. METHODS AND RESULTS: Machine learning model design for allergen score prediction utilized 241 individual allergy assays per patient. Accumulation of data across total IgE subdivision served as the basis of data organization. Two regression based Generalized Linear Models (GLM) were utilized to position allergy assessment on a linear scale. The initial model was further tested with sequential patient data over time. A Bayesian method was then used to improve outcomes by calculating the adaptive weights for the results of the two GLMs of peanut allergy score prediction. A linear combination of both provided the final hybrid machine learning prediction algorithm. Specific analysis of peanut anaphylaxis within one endotype model is estimated to predict the severity of possible anaphylactic reaction to peanut with a recall of 95.2% on a dataset of 530 juvenile patients with various food allergies, including but not limited to peanut allergy. Receiver Operating Characteristic analysis yielded over 99% AUC (area under curve) results within peanut allergy prediction. CONCLUSIONS: Machine learning algorithm design established from comprehensive molecular allergy data produces high accuracy and recall in anaphylaxis risk assessment. Subsequent design of additional food protein anaphylaxis algorithms is needed to improve the precision and efficiency of clinical food allergy assessment and immunotherapy treatment.

Identification of quantitative trait loci and development of diagnostic markers for growth habit traits in peanut (Arachis hypogaea L.).

KEY MESSAGE: QTLs for growth habit are identified on Arahy.15 and Arahy.06 in peanut, and diagnostic markers are developed and validated for further use in marker-assisted breeding. Peanut is a unique legume crop because its pods develop and mature underground. The pegs derive from flowers following pollination, then reach the ground and develop into pods in the soil. Pod number per plant is influenced by peanut growth habit (GH) that has been categorized into four types, including erect, bunch, spreading and prostrate. Restricting pod development at the plant base, as would be the case for peanut plants with upright lateral branches, would decrease pod yield. On the other hand, GH characterized by spreading lateral branches on the ground would facilitate pod formation on the nodes, thereby increasing yield potential. We describe herein an investigation into the GH traits of 521 peanut recombinant inbred lines grown in three distinct environments. Quantitative trait loci (QTLs) for GH were identified on linkage group (LG) 15 between 203.1 and 204.2 cM and on LG 16 from 139.1 to 139.3 cM. Analysis of resequencing data in the identified QTL regions revealed that single nucleotide polymorphism (SNP) or insertion and/or deletion (INDEL) at Arahy15.156854742, Arahy15.156931574, Arahy15.156976352 and Arahy06.111973258 may affect the functions of their respective candidate genes, Arahy.QV02Z8, Arahy.509QUQ, Arahy.ATH5WE and Arahy.SC7TJM. These SNPs and INDELs in relation to peanut GH were further developed for KASP genotyping and tested on a panel of 77 peanut accessions with distinct GH features. This study validates four diagnostic markers that may be used to distinguish erect/bunch peanuts from spreading/prostrate peanuts, thereby facilitating marker-assisted selection for GH traits in peanut breeding.

Performance of some early-maturing groundnut (Arachis hypogaea L.) genotypes and selection of high-yielding genotypes in the potato-fallow system.

India imports the most edible oils because domestic demand exceeds production. Horizontally expanding groundnut production in non-traditional areas especially in the potato-paddy rice-fallow system is possible for increasing production and it requires trait-specific cultivars. Only 1% of oilseeds are grown in non-traditional regions. Nine interspecific groundnut derivatives were tested in potato-fallow system at Deesa, Gujarat, and Mohanpura, West Bengal, and non-potato fallow areas in Junagadh during Kharif 2020 to examine their performance and adaptability. Genotype-by-environment (G×E) interaction significantly affected pod yield and its components in the combined ANOVA. "Mean vs. stability" showed that the interspecific derivative NRCGCS 446 and variety TAG 24 were the most stable and valuable genotypes. GG 7 yielded more pods in Junagadh, whereas NRCGCS 254 yielded more in Mohanpur. Low heritability estimates and strong G×E interaction for flowering days showed complicated inheritance and environmental effects. The shelling percentage was significantly correlated with days to 50% blooming, days to maturity, SCMR, HPW, and KLWR, demonstrating negative connections between maturity, component characteristics, and seed size realisation.

Effect of oil exposure stages on the heat resistance of Salmonella enterica serovar Enteritidis phage type 30 in peanut flour.

The oil in low-moisture foods (LMFs) shows protective effects on bacteria during thermal processing. However, the circumstances under which this protective effect strengthens remain unclear. This study aimed to understand which step of the oil exposure to bacterial cells (inoculation, isothermal inactivation, or recovery and enumeration step) in LMFs can enhance their heat resistance. Peanut flour (PF) and defatted PF (DPF) were selected as the oil-rich and oil-free LMF models. Salmonella enterica Enteritidis Phage Type 30 (S. Enteritidis) was inoculated into four designated PF groups representing different oil exposure stages. It was isothermally treated to obtain heat resistance parameters. At a constant moisture content (aw,25°C = 0.32 ± 0.02) and controlled aw,85°C (0.32 ± 0.02), S. Enteritidis exhibited significantly high (p < 0.05) D values in oil-rich sample groups. For instance, the heat resistance values of S. Enteritidis in the PF-DPF and DPF-PF groups were D80°C of 138.22 ± 7.45 min and 101.89 ± 7.82 min; however, the D80°C in the DPF-DPF group was 34.54 ± 2.07 min. The oil addition after the thermal treatment also helped injured bacterial recovery in the enumeration. For instance, the D80°C, D85°C, and D90°C values in the DFF-DPF oil groups were 36.86 ± 2.30, 20.65 ± 1.23, and 7.91 ± 0.52 min, respectively, which were higher than those in the DPF-DPF group at 34.54 ± 2.07, 17.87 ± 0.78, and 7.10 ± 0.52 min. We confirmed that the oil protected S. Enteritidis in PF in all three stages: desiccation process, heat treatment, and recovery of bacterial cells in plates.

Genomic relationships of the polyploid rhizoma peanut (Arachis glabrata Benth.) inferred by genomic in situ hybridization (GISH).

The rhizoma peanut (Arachis glabrata Benth., section Rhizomatosae) is a tetraploid perennial legume. Although several A. glabrata cultivars have been developed as forage and ornamental turf, the origin and genomic constitution of this species are still unknown. In this study, we evaluated the affinity between the genomes of A. glabrata and the probable diploid donors of the sections Rhizomatosae, Arachis, Erectoides and Procumbentes by genomic in situ hybridization (GISH). Single GISH analyses detected that species of the sections Erectoides (E2 subgenome) and Procumbentes (E3 subgenome) were the diploid species with the highest degree of genomic affinity with A. glabrata. Based on single GISH experiments and DNA sequence similarity, three species -A. duranensis, A. paraguariensis subsp. capibarensis, and A. rigonii-, which showed the most uniform and brightest hybridization patterns and lowest genetic distance, were selected as probes for double GISH experiments. Double GISH experiments showed that A. glabrata is constituted by four identical or very similar chromosome complements. In these assays, A. paraguariensis subsp. capibarensis showed the highest brightness onto A. glabrata chromosomes. Thus, our results support the autopolyploid origin of A. glabrata and show that the species with E2 subgenome are the most probable ancestors of this polyploid legume forage.