Angiotensin-Converting Enzyme and Renin-Inhibitory Activities of Protein Hydrolysates Produced by Alcalase Hydrolysis of Peanut Protein.

Hypertension is a major controllable risk factor associated with cardiovascular disease (CVD) and overall mortality worldwide. Most people with hypertension must take medications that are effective in blood pressure management but cause many side effects. Thus, it is important to explore safer antihypertensive alternatives to regulate blood pressure. In this study, peanut protein concentrate (PPC) was hydrolyzed with 3-5% Alcalase for 3-10 h. The in vitro angiotensin-converting enzyme (ACE) and renin-inhibitory activities of the resulting peanut protein hydrolysate (PPH) samples and their fractions of different molecular weight ranges were determined as two measures of their antihypertensive potentials. The results show that the crude PPH produced at 4% Alcalase for 6 h of hydrolysis had the highest ACE-inhibitory activity with IC50 being 5.45 mg/mL. The PPH samples produced with 3-5% Alcalase hydrolysis for 6-8 h also displayed substantial renin-inhibitory activities, which is a great advantage over the animal protein-derived bioactive peptides or hydrolysate. Remarkably higher ACE- and renin-inhibitory activities were observed in fractions smaller than 5 kDa with IC50 being 0.85 and 1.78 mg/mL. Hence, the PPH and its small molecular fraction produced under proper Alcalase hydrolysis conditions have great potential to serve as a cost-effective anti-hypertensive ingredient for blood pressure management.

Epidemiological characteristics and spatio-temporal analysis of brucellosis in Shandong province, 2015-2021.

BACKGROUND: Brucellosis is one of the major public health problems in China, it not only causes huge economic losses to the society, but also threatens the human's physical and mental health. The reported cases of brucellosis in Shandong province were at a high level, therefore, it is necessary for us to understand the epidemic characteristics and distribution trend of Brucellosis in Shandong province. This study aims to describe the epidemiological characteristics and spatial clustering characteristics of brucellosis in Shandong Province, provide a reference for the scientific prevention and control. METHODS: Human brucellosis data in Shandong province from 2015 to 2021 were obtained from the China Information System for Disease Control and Prevention, the data were analyzed by descriptive epidemiological methods, spatial autocorrelation analysis and spatial-temporal cluster analysis methods use ArcGIS and SaTScan software, the results were presented in ArcMap. RESULTS: A total of 22,251 human cases of brucellosis were reported, the annual incidence ranged between 2.41/100,000 and 4.07/100,000 from 2015 to 2021 in Shandong province, incidence has been decreasing year by year, while there was a significant increase in 2021. The distribution of brucellosis was of a seasonal trend, mainly concentrating during March to August. The age of the cases was mainly concentrated in the 30-74 age ranges, the average annual incidence rate was significantly higher in males than in females. The spatial analysis showed that the epidemics were mainly concentrated in the north and southwest. For the spatial autocorrelation analysis, a high global autocorrelation was observed at the county level, and the high-high clusters mainly distributed in the north and southwest region. For the spatio-temporal scanning, the most likely cluster areas mainly distributed in the north area, and then gradually moved southward, and the radius of clustered narrowed. CONCLUSIONS: Human brucellosis remains a common challenge, particularly in northern region in spring and summer. More disease prevention and control measures should be taken in high-risk populations, and such higher-risk susceptible areas to reduce the incidence of brucellosis and ensure the health of the people.

Function of Cytochrome P450s and Gut Microbiome in Biopesticide Adaptation of Grapholita molesta on Different Host Diets.

Insects that feed on various host plants possess diverse xenobiotic adaptations; however, the underlying mechanisms are poorly understood. In the present study, we used Grapholita molesta, which shifts feeding sites from peach shoots to apple fruits, as a model to explore the effects of shifts in host plant diet on the profiles of cytochrome P450s and the gut bacteria microbiome, as well as their effects on biopesticide adaptation. We found that the sensitivity of the fruit-feeding G. molesta to emamectin benzoate biopesticide was significantly lower than that of the shoot-feeding larvae. We also found that the P450 enzyme activity and the expression of nine cytochrome P450s were enhanced in G. molesta fed on Fuji apples compared to those fed on peach shoots. The survival rates of G. molesta exposed to emamectin benzoate significantly decreased as each of three of four emamectin benzoate-inducted cytochrome P450 genes were silenced. Furthermore, we discovered the gut bacteria dynamics of G. molesta changed with the host shift and the structure of the gut bacteria microbiome was determined by the final diet ingested; additionally, the dysbiosis of the gut microbiota induced by antibiotics could significantly increase the sensitivity to emamectin benzoate. Taken together, our results suggest that the expression of P450s and the composition of the gut bacteria microbiome promote adaptation to emamectin benzoate in G. molesta, providing new insights into the molecular mechanisms underlying xenobiotic adaptation in this notorious pest.

The effect of layers of high tunnel covering and soil mulching on tomato fruit quality.

BACKGROUND: High tunnels (HT) and plastic mulch can increase yield, extend growing seasons, protect plants from harsh weather conditions, and produce more marketable and attractive tomatoes. However, plastic covering reduces solar radiation, which may affect the quality of tomatoes. This study investigated the effects of single-layer versus double-layer HT and plastic mulch versus bare soil on the quality of fully ripe tomato fruits. The study was conducted over 2 years (2021 and 2022). The fruit color (L*, a*, and b*), vitamin C, pH, total titratable acid (TTA), total soluble solids (TSS), and lycopene content of the two tomato cultivars (Brandywine and Rebelski) were quantified. RESULTS: The results showed that quality parameters varied with the plastic layers of HT, plastic mulch, and planting year. The double-layer HT only showed a slight but significant influence on the color of tomatoes of both cultivars in the same year (P < 0.05). Brandywine tomatoes grown in the double-layer HT had significantly lower pH and lycopene content than those grown in single-layer HT, regardless of mulching. The effect of plastic layers on TTA, TSS, and lycopene depended on whether the soil was mulched. Tomatoes grown on bare soil had higher TTA and TSS values than those grown on mulched soil in double-layer HT. Tomatoes grown in single-layer HTs had significantly higher vitamin C content than those in the double-layer HT for both cultivars regardless of mulching. CONCLUSION: This study demonstrates that double-layer HT is unnecessary for the improvement of the overall quality of tomatoes. © 2023 Society of Chemical Industry.

Comparison of uterine, endometrial and subendometrial blood flows in predicting pregnancy outcomes between fresh and frozen-thawed embryo transfer after GnRH antagonist protocol: a retrospective cohort study.

This retrospective cohort study aimed to compare endometrial receptivity and pregnancy rate between fresh embryo transfer (ET) and frozen-thawed ET after gonadotrophin-releasing hormone (GnRH) antagonist protocol in normal ovarian responders. The patients were divided into two groups: the fresh ET group and the frozen-thawed ET group. Uterine artery resistance index (RI) and endometrial thickness were lower in the frozen-thawed ET group. The proportion of detectable endometrial-subendometrial flow was significantly higher in the frozen-thawed ET group. There was no significant difference in miscarriage rate between the two groups. Frozen-thawed ET group had a significantly higher CPR (56.0% vs. 48.1%), implantation rate (32.2% vs. 26.4%), and LBR (45.4% vs. 36.5%) than the fresh ET group. In GnRH antagonist protocol, elective frozen-thawed ET should be ideally taken, as this could improve embryo implantation rate, clinical pregnancy rate, and live birth rate, thus presenting an effective strategy to enhance the embryo utilization rate.IMPACT STATEMENTWhat is already known on this subject? The clinical pregnancy rate following fresh embryo transfer (ET) was lower than frozen-thawed ET after GnRH antagonist protocol. IVF success depends on embryo quality, embryo-endometrium interaction and endometrial receptivity. A good blood supply toward the endometrium is generally considered a requirement for implantation.What do the results of this study add? Uterine artery RI and endometrial thickness were significantly lower in the frozen-thawed ET group. The proportion of detectable endometrial-subendometrial flow was significantly higher in the frozen-thawed ET group. Frozen-thawed ET group had a significantly higher clinical pregnancy rate, implantation rate and live birth rate than the fresh ET group after GnRH antagonist protocol.What are the implications of these findings for clinical practice and/or further research? In GnRH antagonist protocol, elective frozen-thawed ET should be ideally taken, as this could improve embryo implantation rate, clinical pregnancy rate and live birth rate, thus presenting an effective strategy to enhance the embryo utilization rate.

Endometrial preparation protocol of the frozen-thawed embryo transfer in patients with polycystic ovary syndrome.

PURPOSE: To assess which is the optimal protocol in terms of endometrial preparation prior to frozen-thawed embryo transfer (FET) in women with polycystic ovarian syndrome (PCOS) and to explore the effect in stimulated cycle with the addition of vaginal 17-ß oestradiol. METHODS: Five hundred and seventy-six patients with PCOS were prepared for FET using artificial cycle induced with oestradiol and progesterone supplementation (n = 291) and stimulated cycle induced by human menopausal gonadotrophin (HMG) within or without the addition of vaginal 17-ß oestradiol (n = 285). Then the FET was performed in a receptive endometrium. RESULTS: Endometrial thickness was similar (9.03 ± 1.65 vs. 9.12 ± 1.58, P > 0.05) in artificial and stimulated cycle. The two protocols resulted in clinical pregnancy rate (41.0 % vs. 41.6 %, P > 0.05), ongoing pregnancy rate (36.6 % vs. 34.7 %, P > 0.05), live birth rate (30.0 % vs. 31.7 %, P > 0.05), which were not statistically different. Nevertheless, the cancelled cycle rate made a significant difference (2.2 % vs. 5.4 %, P < 0.05). There is no significant difference in the clinical pregnancy rate in HMG, HMG added with vaginal oestradiol and HMG switch to vaginal oestradiol group (42.6 %, 41.1 %, and 33.3 %, respectively). CONCLUSIONS: The mean endometrial thickness, clinical pregnancy rate, ongoing pregnancy rate, live birth rate and implantation rate were similar in artificial and stimulated cycle for endometrial preparation prior to FET in PCOS. It was fine to add vaginal 17-ß oestradiol to stimulated cycle when necessary. However, stimulated cycles had a significantly higher cancelled cycle rate. We should follow the principles of individualization, securitization and optimization in endometrial preparation of the FET in patients with PCOS.

Recent Advances in Physical Processing Techniques to Enhance the Resistant Starch Content in Foods: A Review.

The physical modification of starch to produce resistant starch (RS) is a viable strategy for the glycemic index (GI) lowering of foods and functionality improvement in starchy food products. RS cannot be digested in the small intestine but can be fermented in the colon to produce short-chain fatty acids rather than being broken down by human digestive enzymes into glucose. This provides major health advantages, like better blood sugar regulation, weight control, and a lower chance of chronic illnesses. This article provides a concise review of the recent developments in physical starch modification techniques, including annealing, extrusion, high-pressure processing, radiation, and heat-moisture treatment. Specifically, the focus of this paper is on the alteration of the crystalline structure of starch caused by the heat-moisture treatment and annealing and its impact on the resistance of starch to enzymatic hydrolysis, as well as the granular structure and molecular arrangement of starch caused by extrusion and high-pressure processing, and the depolymerization and crosslinking that results from radiation. The impacts of these alterations on starch's textural qualities, stability, and shelf life are also examined. This review demonstrates how physically modified resistant starch can be used as a flexible food ingredient with both functional and health benefits. These methods are economically and ecologically sustainable since they successfully raise the RS content and improve its functional characteristics without the need for chemical reagents. The thorough analysis of these methods and how they affect the structural characteristics and health advantages of RS emphasizes the material's potential as an essential component in the creation of functional foods that satisfy contemporary dietary and health requirements.

Selection of pecan shell-based activated carbons for removal of organic and inorganic impurities from water.

Activated carbons are a byproduct from pyrolysis and have value as a purifying agent. The effectiveness of activated carbons is dependent on feedstock selection and pyrolysis conditions that modify their surface properties. Therefore, pecan shell-based activated carbons (PSACs) were prepared by soaking shells in 50% (v/v) HPO or 25 to 50% of KOH-NaHCO followed by pyrolysis at 400 to 700°C under a N atmosphere. Physically activated PSACs were produced by pyrolysis at 700°C under N followed by activation with steam or CO at 700 to 900°C. Physicochemical, surface, and adsorption properties of the PSACs were compared with two commercially available activated carbons. The average mass yield of PSACs with respect to the initial mass of the biomass was about 20 and 34% for physically activated and chemically activated carbons, respectively. Acid-activated carbons exhibited higher surface area, higher bulk density, and lower ash content compared with steam- or CO-activated carbons and the two commercial products. Base activation led to the development of biochar with moderate to high surface area with surface charges suitable for adsorption of anionic species. Regardless of the activation method, PSACs had high total surface area ranging from 400 to 1000 m g, better pore size distribution, and more surface charges than commercial samples. Our results also showed that PSACs were effective in removing inorganic contaminants such as Cu and NO as well as organic contaminants such as atrazine and metolachlor. This study showed that pyrolysis conditions and activation had a large influence on the PSAC's surface characteristics, which can limit its effectiveness as a custom sorbent for targeted water contaminants.

Mycotoxins in Cereal-Based Products and Their Impacts on the Health of Humans, Livestock Animals and Pets.

Cereal grains are the most important food staples for human beings and livestock animals. They can be processed into various types of food and feed products such as bread, pasta, breakfast cereals, cake, snacks, beer, complete feed, and pet foods. However, cereal grains are vulnerable to the contamination of soil microorganisms, particularly molds. The toxigenic fungi/molds not only cause quality deterioration and grain loss, but also produce toxic secondary metabolites, mycotoxins, which can cause acute toxicity, death, and chronic diseases such as cancer, immunity suppression, growth impairment, and neural tube defects in humans, livestock animals and pets. To protect human beings and animals from these health risks, many countries have established/adopted regulations to limit exposure to mycotoxins. The purpose of this review is to update the evidence regarding the occurrence and co-occurrence of mycotoxins in cereal grains and cereal-derived food and feed products and their health impacts on human beings, livestock animals and pets. The effort for safe food and feed supplies including prevention technologies, detoxification technologies/methods and up-to-date regulation limits of frequently detected mycotoxins in cereal grains for food and feed in major cereal-producing countries are also provided. Some important areas worthy of further investigation are proposed.

Intercropping Okra and Castor Bean Reduces Recruitment of Oriental Fruit Moth, Grapholita molesta (Lepidoptera: Tortricidae) in a Pear Orchard.

Intercrops can lower pest densities by increasing plant diversity, altering chemical communication in the arthropod community, and integrating well with other IPM tactics. We used two years of field observations and Y-tube olfactometer assays to explore the effects of intercropping a pear orchard with okra and castor bean on the cosmopolitan fruit-boring pest Grapholita molesta (Lepidoptera: Tortricidae). Intercropping okra reduced G. molesta trap catches in the pear orchard in both years, and intercropping with castor bean reduced them in the second year. Hydrocarbons, phenols, and ketones predominated in the GC-MS assay of okra volatiles, whereas castor bean volatiles were rich in aldehydes, ketones, and esters. Five of the commercially available volatiles released by these plants exhibited repellency to G. molesta in olfactometer trials, especially cinnamaldehyde, dibutyl phthalate, and thymol; the former compound also exhibited attraction to the egg parasitoid Trichogamma dendrolimi (Hymenoptera: Trichogrammatidae). In addition to their repellent properties, okra and castor bean may enhance integrated control of G. molesta in orchards by hosting prey that support populations of generalist predators that either provide biological pest control services within the orchard ecosystem or generate non-consumptive effects that contribute to pest deterence. Among the plant volatiles evaluated, cinnamaldehyde has the best potential for deployment in orchards to repel G. molesta without disrupting augmentative releases of T. dendrolimi.

Preliminary Assessment of Repellency and Toxicity of Essential Oils against Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae) on Stored Organic Corn Grains.

Insect infestation of cereal grains during post-harvest storage not only causes significant grain loss, but also reduces grain quality and makes grains more susceptible to mold infection. Synthetic pesticides are banned from being used in organic grain storage setting due to their high toxicity. The main insect damaging stored corn grains is maize weevil, Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae). The purpose of this study was to evaluate insect repellency and insecticidal potentials of some generally recognized as safe (GRAS) essential oils (EOs) (including cinnamon, clove, thyme, oregano, and orange terpene oils) at concentrations of 1-20% against the maize weevil using an olfactometer and a simulated fumigation method, respectively. The olfactory tests show that cinnamon oil had the highest repellency (90%) to the weevils among the EOs tested. The insecticidal activity study indicates that maize weevil mortality increased with EO concentration and storage time with cinnamon, clove, and thyme oils being more effective. No weevil death was observed at 1% EOs; weevil mortality was 3.3-36% at 5%, which varied with the type of EO and storage time. At 10% or higher concentrations, all tested EO showed comparable or higher insecticidal activity than pirimiphos methyl-positive control at its recommended concentration (5 mg/kg corn). No significant increase in weevil mortality was observed with further increase in EO concentration, with exceptions of oregano oil and thyme oil. The highest weevil mortality levels were observed at week 7 for 15% cinnamon oil (100%) and eugenol (100%), followed by 20% thyme oil (93%). The study indicates that some EOs have great potential to serve as synthetic insecticide alternatives to protect organic corn grains from maize weevil damage during storage. This is important to food security, safety and environmental health.

Increased quality of bowel preparation via smartphone WeChat application: a multicenter randomized controlled trial.

Introduction: High-quality bowel preparation is an essential precondition for colonoscopy. Few studies have evaluated the smartphone WeChat application as a means of improving the quality of bowel preparation. Aim: To assess the effect of patient education by using smartphone WeChat application aids on the quality of bowel preparation. Material and methods: A multicenter prospective, endoscopist-blinded, randomized, controlled study was conducted. Patients were randomly assigned to three groups. A total of 478 patents in groups A were accepted for smartphone WeChat application, 477 in groups B were accepted for conventional education plus smartphone WeChat application while group C (473 patients) was a control group. The primary outcome was the quality of the bowel preparation according to the Boston Bowel Preparation Scale (BBPS). The secondary outcomes included polyp detection rate (PDR), cecal intubation rate, insertion and withdrawal time, anxiety score, self-rated sleep quality, and willingness to undergo another colonoscopy. Results: Total BBPS score was significantly higher in groups B and C than in the control group (7.5 ±1.2, 7.5 ±1.3 vs. 6.5 ±1.2, p < 0.001). PDR in group A (40.2%, 192/478) and group B (41.7%, 199/477) was higher than that in the control group (p = 0.003) and mean number of polyps per patient was higher too (p = 0.015). Moreover, a shorter cecal insertion time was recorded in group A and group B than in the control group (8.8 ±3.9, 8.9 ±3.8 vs. 10.5 ±4.2 min, p < 0.001). Intervention groups showed lower anxiety scores and better quality sleep and were more likely to be willing to repeat colonoscopy. Conclusions: Patient instruction via smartphone WeChat application efficiently improved bowel preparation for colonoscopy.

Enzymatic treatment of peanut kernels to reduce allergen levels.

This study investigated the use of enzymatic treatment to reduce peanut allergens in peanut kernels as affected by processing conditions. Two major peanut allergens, Ara h 1 and Ara h 2, were used as indicators of process effectiveness. Enzymatic treatment effectively reduced Ara h 1 and Ara h 2 in roasted peanut kernels by up to 100% under optimal conditions. For instance, treatment of roasted peanut kernels with α-chymotrypsin and trypsin for 1-3h significantly increased the solubility of peanut protein while reducing Ara h 1 and Ara h 2 in peanut kernel extracts by 100% and 98%, respectively, based on ELISA readings. Ara h 1 and Ara h 2 levels in peanut protein extracts were inversely correlated with protein solubility in roasted peanut. Blanching of kernels enhanced the effectiveness of enzyme treatment in roasted peanuts but not in raw peanuts. The optimal concentration of enzyme was determined by response surface to be in the range of 0.1-0.2%. No consistent results were obtained for raw peanut kernels since Ara h 1 and Ara h 2 increased in peanut protein extracts under some treatment conditions and decreased in others.

Allergenicity, antioxidant activity and ACE-inhibitory activity of protease hydrolyzed peanut flour.

Regularly consuming peanuts has been reported to have many health benefits. Peanut flour, a by-product of peanut oil processing, has higher protein and dietary fiber contents than peanut kernels, but its application as protein source in foods and dietary supplement is limited due to the fear of peanut allergy. This study indicates that hydrolysis of peanut flour (12% lipid) up to 4 h with Alcalase lowered the immunoreactivity of both soluble and insoluble portions of peanut flour, generated peanut flour hydrolysate (PFH) with good in vitro antioxidant and ACE-inhibitory activities. Importantly, the fraction smaller than 5 kDa did not bind IgE of peanut allergic patients, but exhibited higher ACE-inhibitory activity than the crude PFH. Thus, peanut flour could be an inexpensive protein source of antioxidant and anti-hypertensive ingredient. These findings are important for the value added application of peanut flour. However, studies with food and animal/human models are needed to confirm the benefits.

Effectiveness of different proteases in reducing allergen content and IgE-binding of raw peanuts.

The effectiveness of some non-specific proteases in reducing raw peanut allergenicity was investigated. Peanut kernels were treated by Alcalase, papain, Neutrase and bromelain, respectively. The residues of major peanut allergens Ara h 1, Ara h 2 and Ara h 6 were determined by sandwich ELISA and SDS-PAGE, and the allergenicities of treated peanuts were compared to that of untreated peanuts by western blot. All tested proteases were effective in reducing Ara h 1, but their effectiveness in hydrolyzing Ara h 2 and Ara h 6 varied greatly. The maximal reductions of extractable Ara h 1, Ara h 2 and Ara h 6 were 100%, 100% and 99.8%, respectively, achieved by Alcalase hydrolysis. Alcalase was more effective in overall allergenicity reduction; bromelain and Neutrase were the least effective in reducing Ara h 2 and Ara h 6, respectively. The hydrolysis of original allergens also produced some smaller peptides with strong IgE-binding.

Reducing Ochratoxin A Content in Grape Pomace by Different Methods.

Grape pomace (GP) is the residue of grapes after wine making and is a valuable source of dietary polyphenol and fiber for health promotion. However, studies found the presence of ochratoxin A (OTA) in GP at very high concentrations, which raises a safety issue in the value-added utilization of GP. This study evaluated the effects of thermal pressure, baking, acid and enzymatic treatments on OTA content in GP. Thermal pressure treatment was conducted with wet GP at 121 °C for 10-30 min in an autoclave; acid treatments were conducted with hydrochloric acid, acetic acid, citric acid, and lactic acid, respectively, at 50 °C for 24 h. Baking was conducted using a cookie model. For enzymatic treatment, purified OTA solution was treated with carboxypeptidase A, alcalase, flavourzyme, pepsin, and lipase, respectively, and the effective enzymes were selected to treat GP. Results show that autoclaving for 10-30 min reduced 19-80% of OTA, varying with treatment time and GP variety. The effectiveness of acid treatment was similar to that of autoclaving and varied with acid type and GP variety. Baking increased the detectable OTA. Among all tested enzymes, carboxypeptidase A was the most effective in reducing OTA, followed by lipase and flavourzyme, but their effects were significantly lower in GP samples.

Comparison of gut bacterial communities and their associations with host diets in four fruit borers.

BACKGROUND: Microbiota that live in the gut of insects have a wide range of effects on host nutrition, physiology, and behavior. They may shape the adaptation of their hosts to different habitats and lifestyles. To characterize the gut microbiota of fruit borers comprehensively, we compared bacterial communities among Grapholita molesta, Conogethes punctiferalis, Carposina sasakii, and Cydia pomonella, which are serious lepidopteran pests. We selected G. molesta as a representative pest to more explicitly test the influence of host dietary niche on the insect gut microbiome, and compared the bacterial microbial communities of G. molesta fed different diets (peach shoots and apple) using Illumina high-throughput sequencing technology. RESULTS: The results show that Proteobacteria and Firmicutes are dominant in their gut microbiota. The C. sasakii had the highest richness values and G. molesta (shoot-feeding) had the highest diversity, whereas C. pomonella and G. molesta (fruit-feeding) had the lowest bacterial richness and diversity, respectively. The ANOSIM analysis revealed significant differences in the structure of gut microbiota among different insects. In addition, G. molesta with a different feeding diet had significant differences in gut microbiota composition. PICRUSt analysis indicated that most functional prediction categories were related to metabolism. CONCLUSION: Our results show that gut microbiota composition is affected significantly not only by host species but also host diets. An enhanced understanding of these herbivore-associated microbial symbionts is essential for understanding the biology and ecology of the host insect, and may offer new possibilities to improve integrated pest-management strategies for efficient control of fruit borers. © 2019 Society of Chemical Industry.

Oxidative Stability of Protease Treated Peanut with Reduced Allergenicity.

Oxidative stability and allergenicity are two major concerns of peanuts. This study evaluated the impact of protease treatment of peanuts on its oxidative stability during storage. The raw and dry-roasted peanut kernels were hydrolyzed with Alcalase solution at pH 7.5 for 3 h. The contents of Ara h 1, Ara h 2, and Ara h 6 in peanuts were determined before and after enzyme treatment by a sandwich ELISA. After drying, the samples were packed in eight amber glass jars and stored at 37 °C for 1-8 weeks. Controls are untreated raw and dry-roasted peanuts packed and stored in the same way as their treated counterparts. Samples were taken biweekly to determine peroxide value (PV) and thiobarbituric acid reactive substances (TBARS) as indicators of oxidation (n = 3), and to determine antioxidant activity. Alcalase treatment reduced intact major allergens Ara h 1, Ara h 2, and Ara h 6 by 100%, 99.8%, and 85%, respectively. The PVs of Alcalase-treated raw and roasted peanuts was lower than those of untreated (p < 0.05) over the 8-week storage. The TBARS of Alcalase-treated raw peanuts were slightly higher than that of untreated (p < 0.05), but the TBARS of Alcalase-treated dry-roasted peanuts were slightly but significantly lower than that of untreated (p < 0.05). The protease treatment increased the antioxidant activities including reducing power, DPPH free radical scavenging capacity, and metal chelating capacity of peanuts.

Immunostimulant adjuvant patch enhances humoral and cellular immune responses to DNA immunization.

The focus of this report is on the development of an improved DNA immunization protocol, which takes advantage of the strengths of DNA immunization, as well as those associated with adjuvant delivered by transcutaneous immunostimulatory (IS) patches. Because transcutaneous delivery of adjuvants to the skin at the vaccination site has been shown to amplify the immune response to protein antigens, we hypothesized that the same IS patch when placed on the skin at the site of DNA injection could further enhance the immune response to a DNA influenza vaccine. We have combined an influenza DNA vaccine, hemagglutinin fused with three copies of complement C3d, to enhance uptake and antigen presentation, with an IS patch containing heat-labile enterotoxin from Escherichia coli. Coadministration of a potent adjuvant in IS patches placed on the skin at the site of DNA vaccination dramatically amplifies anti-influenza antibody immune response. Supplementing DNA vaccines with IS patches may be a particularly valuable strategy because DNA vaccines can be rapidly modified in response to mutations in pathogens, and individuals with compromised immune systems such as transplant patients and the elderly will benefit from the enhanced antibody response induced by the IS patches.

Changes in immunoreactivity of allergen-reduced peanuts due to post-enzyme treatment roasting.

The objective of this study was to evaluate the effect of enzyme treatment and post-enzyme treatment roasting on allergenicity of raw peanut kernels. Raw peanuts were treated by single- and two-enzyme treatments, respectively. Enzyme-treated raw peanuts were dry roasted. Reductions of four major allergens (Ara h 1, 2, 3, and 6) and in vitro allergenicity of peanuts were evaluated. Quantitative measurements show that enzyme treatment of raw peanuts reduced Ara h 1, 2, 3 and 6 in raw peanuts by 99-100%, 95-99%, 35-46% and 85-88%, respectively. Roasting of enzyme-treated peanuts significantly reduced the total soluble protein (P < 0.05), Ara h 3 and 6 (P < 0.0001), slightly increased Ara h 1 in the extracts (P < 0.05), but did not significantly affect Ara h 2. Immunoblot shows that the IgE-bindings of both soluble and insoluble proteins of enzyme-treated peanuts were slightly enhanced by roasting but still tremendously lower than that of untreated peanuts.