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Tofu has a high nutritional value, but it may also contain components that may have an antinutritional effect, such as trypsin inhibitors (TI), lectins and ureases. The aim of this study was to investigate the influence of the hydrothermal-cooking process of soybean in combination with commercial chymosin-pepsin rennet on the content and activity of TI, urease and lectins in tofu. High total TI content was found in tofu (5.00-16.87%). In addition, Kunitz (KTI = 3.52-4.32%) and Bowman-Birk (BBI = 5.00-12.53%) TI were registered, and BBI was detected in polymeric (1.38-2.71%) and monomeric (3.42-9.82%) forms. TI activity of tofu was very low (5.86-9.34%), corresponding to the very low activity of urease (0.51-3.07%). The percentage of lectin (2.62-4.63%) and urease (0.03-0.12%) in tofu was low. The results showed that the applied tofu production process is very effective in reducing the content and activity of TI, urease and lectin and provides the values without nutritional harmful effect. [ABSTRACT FROM AUTHOR]

Herein, we designed a series of guanidinobenzoic acid ester derivatives on the basis of approved AP drugs, such as nafamostat, gabexate and camostat, and evaluated their inhibitory effects on trypsin and anti-inflammatory activity. Among them, five compounds (6a, 6c–6e, 7j) showed excellent inhibitory effects on trypsin with IC50 values of 0.0756 μM to 0.1227 μM, which are more potent than nafamostat and gabexate. Moreover, compounds 6a, 6b and 6c also showed significant inhibitory potency against the pro-inflammatory molecule NO with IC50 values of 1.618 μM, 2.276 μM, and 3.022 μM, respectively. Consequently, the potential lead compounds simultaneously with anti-trypsin and anti-inflammatory activities were identified, which would profit further structural optimization for the treatment of AP. [ABSTRACT FROM AUTHOR]

BACKGROUND: The striped stem borer (SSB, Chilo suppressalis) is one of the most destructive insect pests on rice. As a chewing insect, SSB larval feeding causes a dramatic increase in rice defense responses. However, the effects of oral secretions (OSs) during SSB feeding on rice defense remain largely unexplored. RESULTS: In this study, based on transcriptome analysis results, treatment with SSB OSs regulated the expression of genes involved in the plant defense‐related pathways of calcium, mitogen‐activated protein kinases, reactive oxygen species, jasmonic acid (JA), herbivore‐induced plant volatiles (HIPVs), and protease inhibitors. Unsurprisingly, treatment with SSB OSs elicited the accumulation of JA and JA‐isoleucine in rice. The defense mechanisms activated by the cascade not only induced the expression of trypsin inhibitors, inhibiting the normal growth of SSB larvae but also induced HIPVs emission, rendering rice attractive to a common larval parasitoid. High‐throughput proteome sequencing of SSB OSs led to 534 proteins being identified and 343 proteins with two or more unique peptides being detected. CONCLUSION: The study demonstrates that SSB OSs trigger both direct and indirect defense mechanisms in rice, akin to the effects of SSB feeding. It identifies specific proteins in SSB OSs that may influence the interactions between SSB and rice during feeding, providing valuable insights for effectors research. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

AbstractSoybeans (Glycine max [L.] Merr.) are a globally significant crop, valued for their high protein content and nutritional versatility. However, they contain anti-nutritional factors (ANFs) that can interfere with nutrient absorption and pose health risks. This comprehensive review examines the presence and impact of key ANFs in soybeans, such as trypsin inhibitors, lectins, oxalates, phytates, tannins, and soybean polysaccharides, based on recent literature. The physiological roles, potential health hazards of the ANFs, and the detailed balance between their harmful and beneficial effects on human health, as well as the efficacy of deactivation or removal techniques in food processing, were discussed. The findings highlight the dual nature of ANFs in soybeans. Some ANFs have been found to offer health benefits include acting as antioxidants, potentially reducing the risk of cancer, and exhibiting anti-inflammatory effects. However, it is important to note that the same ANFs can also have negative impacts. For instance, trypsin inhibitors, lectins, and tannins may lead to gastrointestinal discomfort and contribute to mineral deficiencies when consumed in excess or without proper processing. This review will provide a clear understanding of the role of ANFs in soybean-based diets and to inform future research and food processing strategies. [ABSTRACT FROM AUTHOR]

The growing interest in plant-based proteins is driven by sustainability concerns and potential adverse reactions to animal protein consumption. Plant proteins, while promising, are known for their inferior digestibility compared to animal counterparts. Solid-state fermentation (SSF) offers an ancient, cost-effective approach to address this limitation. SSF enhances plant-based foods by augmenting protein content through microbial hydrolysis, thereby improving overall nutritional quality. This method not only boosts protein digestibility but also reduces larger polypeptides, generates bioactive peptides, refines amino acid profiles, and eliminates anti-nutritional factors such as trypsin inhibitors and tannins. Further investigation is essential to optimize the purification of protein isolates and examine their behaviour during digestion and absorption using in vitro, cellular, and in vivo models. In conclusion, SSF represents a promising, cost-effective, and high-yield processing method to produce nutritionally enhanced plant proteins, aligning with the demands of both the industry and consumers. [ABSTRACT FROM AUTHOR]

When insects harm plants, they activate relevant enzyme systems for defense, and changes in enzyme activity, to a certain extent, reflect the host plant's ability to resist insect damage. Alfalfa leaf weevils (Hypera postica Gyllenhal) are the main economic insect pest of alfalfa, which seriously affect its yield and quality. To clarify the effects of feeding induction by alfalfa leaf weevil larvae and adults on defense enzymes in alfalfa, 'Zhongmu No. 1' variety was used as the experimental material. Comprehensive correlation analysis and principal component analysis were used to evaluate the corresponding patterns of 12 physiological indicators of alfalfa induced by insect feeding of different densities. Results showed that after feeding induction by adult and larval alfalfa leaf weevils, total antioxidant capacity (T-AOC), malondialdehyde (MDA), phenylalanine ammonia-lyase (PAL), tyrosine ammonia lyase (TAL), lipoxygenase (LOX), chymotrypsin inhibitors (CI), trypsin inhibitor (TI), and jasmonic acid (JA) in the alfalfa leaves increased with increasing feeding time. However, activities of catalase (CAT), peroxidase (POD), superoxide dismutase (SOD) and polyphenolic oxidase (PPO) in alfalfa leaves first increased and then decreased, showing a downward trend. [ABSTRACT FROM AUTHOR]

In living systems, protein assemblies have essential functions, serving as structural supports, transport highways for molecular cargo, and containers of genetic material. The construction of protein assemblies, which involves control over space and time, remains a significant challenge in biotechnology. Here, we show that anionic boron clusters, 3,3′‐commo‐bis[closo‐1,2‐dicarba‐3‐cobaltadodecaborane] (COSAN−), and halogenated closo‐dodecarboranes (B12X122−, X=H, Cl, or I), described as super‐chaotropic nano‐ions, induce the formation of 2D assemblies of model proteins, myoglobin, carbonic anhydrase, and trypsin inhibitor. We found that the nano‐ion concentration reversibly controls the size of the protein assemblies. Furthermore, the secondary structures of the proteins are only slightly affected by assembly formation. For myoglobin, the formation of these assemblies even prevents temperature denaturation, highlighting a preservation effect of nano‐ions. Our study reveals that inorganic boron‐based nano‐ions act as a reversible molecular glue for proteins, providing a potential starting point for the further development of controlled protein assemblies. [ABSTRACT FROM AUTHOR]

In a bid to quantify the contribution of molecular structure to non-specific interactions leading to functionally important structural changes in cellular processes, the self-interaction of dextran-T70 (DT70) and its interaction with each of bovine serum albumin (BSA) and ovomucoid trypsin inhibitor (OVO) were studied at pH 7.4 between 5 and 37 °C. The dependences of the apparent molecular weight of each of BSA, OVO and DT70 on the concentration of DT70 were independent of temperature. The activity coefficient of the interaction of each species on DT70 concentration was also independent of temperature. The change in activity coefficient was however dependent on the molecular structure and size of the interacting species. The energy of insertion of each macromolecule in DT70 increased in the order DT70 > BSA > OVO. These findings show that although the enthalpic contribution is negligible, the extent of the entropic contribution to the macromolecular activity coefficient of interaction is chiefly the consequence of the exclusion volume of the interacting macromolecules. [ABSTRACT FROM AUTHOR]

Background: With the rapid development of animal husbandry, the demand for protein feed resources is increasing. Cottonseed meal (CSM) and soybean meal (SBM) are rich sources of protein. However, their application is limited due to the existence of anti-nutrients, which can be harmful to the digestion and absorption. A strain of Bacillus subtilis (Mafic-Y7) was isolated from digesta of intestines of Tibetan pigs. The strain showed high protease activity, which helps in degrading proteinic anti-nutritional factors in grain meal and in vitro degradation of free gossypol. In order to better understand this isolated strain, whole genome of Mafic-Y7 strain was sequenced and analyzed. Different effects on various grain meals were identified. Result: The GC-depth Poisson distributions showed no bias suggesting high-quality genome assembly of Mafic-Y7. The whole genome sequencing showed that one chromosome with 4,248,845 base pairs(bp)and the genes total length with 3,736,524 bp was predicted in Mafic-Y7. Additionally, Mafic-Y7 possessed 4,254 protein-coding genes, and several protease genes were annotated by aligning them with databases. There are 55 protease genes, one phytase gene and one laccase gene were annotated in the gene sequence of Mafic-Y7. The average nucleotide identity between Mafic-Y7 and the GCA-000009045.1 homologous genome was 0.9938, suggesting a close genetic relationship between them at the species level. Compared with the closest four whole genomes, Mafic-Y7 was annotated the most abundant of protease genes (55 genes). The fermentation supernatant of Mafic-Y7 could increase the content of small peptides, water-soluble proteins, and acid-soluble proteins in vitro by 411%, 281% and 317% in SBM and 420%, 257% and 338% in CSM. After fermentation in grain meal by Mafic-Y7, the degradation rate of anti-nutritional factors in SBM, such as trypsin inhibitor, glycinin, and β-conglycinin was greater than 70%, and lectin was greater than 30%. The degradation rates of anti-nutritional factors in CSM, such as gossypol and phytic acid, were 82% and 26%, respectively. [ABSTRACT FROM AUTHOR]

Background: Chickpea pod borer (CPB), Helicoverpa armigera (Hübner) is a pest of great economic importance in chickpea and it is the major limiting factor in chickpea cultivation. In severe cases it causes about 75 to 90 per cent losses in seed yield, despite the application of insecticides. Therefore, development of a cost effective and an environmentally friendly approach like improvement of cultivars resistant to H. armigera is necessary for management of the pest in chickpea. Methods: Morphological and chemical components of host plant resistance in chickpea germplasm was assessed under field and laboratory conditions against pod borer, Helicoverpa armigera at hotspot Pantnagar, during rabi, 2017-18 using standard protocols. Result: Observations recorded revealed that germplasm ICC4484 recorded highest phenol (4.73 mg/g) and flavonoid (0.19 mg/g) content, whereas the maximum tannin, protein and trypsin content were recorded in ICC6263 (1.33 mg/g), ICC3137 (19.41 g/100 g of seeds) and ICC372351 (31.83 IU/g), respectively. Germplasm with higher phenol, tannin, flavonoid and trypsin inhibitor content recorded minimum per cent pod damage. Phenol, tannin, flavonoid, trypsin content showed negative correlation, while protein content showed positive correlation with the per cent pod damage by H. armigera. [ABSTRACT FROM AUTHOR]

The fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), is a polyphagous insect of various agricultural crops. The methods used for its control have led to the selection of resistant insect populations, which justifies the search for new alternatives for the management of this insect. Accordingly, in the present study, trypsin inhibitors present in the leaf extract of Ricinus communis (Euphorbiaceae) were investigated for their activity against S. frugiperda. Chemometric optimization methods were developed for the extraction, purification, identification, and structural characterization of the inhibitors. In addition, the effect of R. communis extract on S. frugiperda development was evaluated. Inhibitor extraction was chemometrically optimized, yielding an extract with an antitryptic activity of 94,837.14 mUIT g−1. The R. communis extract was purified and found to contain two compounds (adenosine and ricinine) exhibiting trypsin inhibitor activity. However, after purification, only ricinine inhibited S. frugiperda trypsin in vitro (103.21 mUIT mg−1). The extract was added to the diet of S. frugiperda larvae, resulting in reduced digestion, increased protein in the feces (control = 12,571 μg protein/mg feces; 1818.2 g mL−1 = 16,867 μg protein/mg feces), and insect mortality. At the highest concentration, the treatment led to an LT50 of 15.9 days and a cumulative survival rate of 18.5%. Based on the results of this study, it is possible that ricinine binds to the catalytic site of trypsin, causing the mortality of S. frugiperda larvae. [ABSTRACT FROM AUTHOR]

BACKGROUND: Soybean meal (SBM) is used widely in animal feed but it contains anti‐nutritional factors (ANFs) such as protease inhibitors – immunogenic proteins that limit its utilization. Fermentative processes could help to reduce these ANFs. The aim of this study was to evaluate the nutritional attributes, bacterial community dynamics, and microbial metagenomic profile during the solid‐state fermentation of SBM using a strain of the bacterium Lactobacillus acidophilus with or without pre‐autoclaving treatment. RESULTS: Following fermentation, there was a reduction in the pH and a concurrent increase in the population of lactic acid bacteria. Fermentation also resulted in an increase in both crude and soluble protein levels. Trypsin inhibitor levels decreased after fermentation, particularly in fermented SBM that had not been pre‐autoclaved, with an inactivation rate higher than 90%. Moreover, high‐molecular‐weight peptides (44–158 kDa), specifically some polypeptides from the soybean immunogen glycinin and β‐conglycinin, underwent degradation during the fermentation process. Bacterial community analysis revealed the dominance of the Lactobacillus genus in all samples, regardless of the treatments applied. Metagenomic profiling identified L. acidophilus as the dominant species in inoculated SBM, irrespective of whether pre‐autoclaving was conducted or not. CONCLUSION: This study demonstrates the feasibility of solid‐state fermentation with L. acidophilus under non‐sterile conditions to inactivate trypsin inhibitor and increase protein concentration and hydrolysate immunogen proteins into low‐molecular‐weight peptides in SBM. Lactobacillus acidophilus inoculum also inhibited the growth of undesirable bacteria. This knowledge contributes to our understanding of the potential applications of solid‐state fermentation with L. acidophilus in improving the nutritional quality of SBM. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Global cultivation of the faba bean (Vicia faba L.), a nutritious member of the fabaceae family, is a popular coldtolerant crop. Leading faba bean-producing countries encompass China, Ethiopia, the United Kingdom, Australia, and France. Recently, the exploration of different foods enriched with bioactive components that offer enhanced functionality, nutritional value, and health benefits has sparked a growing interest in the health advantages of faba beans. Vicia faba is rich in derivatives of minerals, vitamins, carbohydrates, proteins abundant in lysine and numerous bioactive compounds. Furthermore, it serves as an excellent source of L-DOPA, a dopamine precursor with potential implications in Parkinson's disease treatment. The edible forms of faba bean seeds include dried, roasted, boiled, soaked, frozen and canned edibles. Yet, a number of antinutritional factors, such as phytic acid, trypsin inhibitors, saponins, vicine and convicine (substances associated with favism), lectins and condensed tannins, impede the best possible utilization of faba beans, thereby undermining their biological value. This comprehensive review encompasses global production, dietary characteristics and an array of antinutritional conditions. [ABSTRACT FROM AUTHOR]

Fava bean (Vicia faba L.) is a protein-rich pulse with high nutritional value, but its functional and sensory characteristics limit its application in foods. Solid-state fermentation (SSF) can modify the composition of plant proteins, modulate its functionality, and enhance the sensory aspects. In this study, fava bean flour (FB) was fermented with Aspergillus oryzae and Rhizopus oligosporus to produce FBA and FBR, respectively, ingredients with distinct nutritional, functional, and aroma characteristics. The protein content increased by 20% in FBA and 8% in FBR, while fat levels rose more significantly in FBR (+40%). The overall content of fermentable oligo-, di-, mono-saccharides, and polyols (FODMAPs) decreased by 47% (FBA) and 57% (FBR), although polyol production by A. oryzae was observed. SSF improved the nutritional profile of FBA and FBR, with a notable increase in the concentration of essential amino acids observed, and a reduction in most antinutrients, with the exception of trypsin inhibitors. SSF resulted in the formation of aggregates, which increased the particle size and reduced protein solubility. Emulsions prepared with the fermented ingredients separated faster, and the foaming capacity of both FBA and FBR was decreased, but an increase in water-holding capacity was observed. SSF resulted in the production of predominantly savoury-associated aroma compounds, with compounds characteristic of metallic and mouldy aromas reduced. These results indicate the potential of SSF to transform FB with enhanced nutritional value and improved sensory and functional properties. [ABSTRACT FROM AUTHOR]

Background Different methods for the extraction of trypsin inhibitors in beans (Phaseolus spp.) were investigated. Two randomised complete laboratory experiments were performed, one on the seeds and one on the pods. In the first, the seeds of common bean variety KIS Marcelijan, breeding line Ref_316 × 498 and runner bean variety Bonela were examined. In the second, the fresh pods of five common beans (three breeding lines, two varieties) were analysed. Four extraction methods were used, including ultrasonic-assisted extraction (UAE) for 15 and 30 min and shaking-assisted extraction for 60 and 180 min. Results The results showed a significant increase in trypsin inhibitor activity-related traits in UAE compared to shaking extraction, with the 15 min ultrasonic process showing better efficacy than the one with 30 min duration. In the seed experiment, the breeding line Ref_316 × 498 showed the highest Trypsin Units Inhibited (TUI) and TUI/mg sample after a 15 min UAE. In the pod experiment, the breeding line 228_4aa_ca also showed the highest TUI and TUI/mg sample after a 15 min extraction with UAE. These results underline the potential of UAE to maximise trypsin inhibitor content. In addition, remarkable correlations between TUI, TUI/mg sample and the percentage of trypsin inhibition (%TIn) were observed in both experiments. Conclusions These results provide valuable insights into the relationship between bean genetic resources, extraction methods and trypsin inhibitor content in bean pods and seeds and serve as a basis for refining extraction protocols. The study encourages further research on the practical implications of investigated protocols for breeding programmes and agricultural practices. [ABSTRACT FROM AUTHOR]

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Plant defense polypeptides play a crucial role in providing plants with constitutive immunity against various biotic and abiotic stressors. In this study, we explored a complex of proteins from wheatgrass (Elytrigia elongata) spikelets to estimate their role in the plant's tolerance to various environmental factors. The current research shows that in vitro protein extracts from E. elongata spikelets possess antifungal activity against certain Fusarium species, which are specific cereal pathogens, at concentrations of 1–2 mg/mL. In this study, we reproduced these antifungal activities using a 4 mg/mL extract in artificial fungal infection experiments on wheat grain (Triticum aestivum) under controlled laboratory conditions. Furthermore, the tested extract demonstrated a protective effect on Saccharomyces cerevisiae exposed to hyper-salinity stress at a concentration of 2 mg/mL. A combined scheme of fractionation and structural identification was applied for the estimation of the diversity of defense polypeptides. Defensins, lipid-transfer proteins, hydrolase inhibitors (cereal bifunctional trypsin/alpha-amylase inhibitors from a Bowman–Birk trypsin inhibitor), and high-molecular-weight disease resistance proteins were isolated from the extract. Thus, wheatgrass spikelets appear to be a reservoir of defense polypeptides. Our findings contribute to a deeper understanding of plant defense proteins and peptides and their involvement in the adaptation to various stress factors, and they reveal the regulatory effect at the ecosystem level. [ABSTRACT FROM AUTHOR]

Aeruginosins are common metabolites of cyanobacteria. In the course of re-isolation of the known aeruginosins KT608A and KT608B for bioassay studies, we isolated three new sulfated aeruginosins, named aeruginosins KT688 (1), KT718 (2), and KT575 (3), from the extract of a Microcystis cell mass collected during the 2016 spring bloom event in Lake Kinneret, Israel. The structures of the new compounds were established on the basis of analyses of the 1D and 2D NMR, as well as HRESIMS data. Marfey's method, coupled with HR ESI LCMS and chiral HPLC, was used to establish the absolute configuration of the amino acid and hydroxyphenyl lactic acid residues, respectively. Compounds 1–3 were tested for inhibition of the serine protease trypsin, and compounds 1 and 2 were found to exhibit IC50 values of 2.38 and 1.43 µM, respectively. [ABSTRACT FROM AUTHOR]

There is an unmet demand for plant‐based ingredients with desirable nutritional, techno‐functional properties, and health benefits. In this study, the composition, nutritional quality, and bioactives in industrial hempseed flours and protein isolate generated by milling, germination, isoelectric precipitation (IEP), and enzyme‐assisted extraction (EAE) were evaluated. Moisture, ash, fat, protein, phytic acid, tannin, and trypsin inhibitor content of the hempseed flours and protein isolate were 2.80%–6.46%, 5.07%–28.89%, 0.00%–31.44%, 22.71%–89.94%, 0.55%–1.05%, 274.24–1300.76 μg/g, and 0.00–42.66 U/g, respectively. IEP resulted in the highest protein content (89.94%), indicative of its effective to isolate hempseed protein. Germination and IEP significantly reduced phytic acid and tannin contents by 1.53‐ and 3.63‐fold, respectively. All processing methods improved in vitro protein digestibility (IVPD). SDS‐PAGE analysis revealed comparable band patterns in milled and protein isolate, with a strong 50 kDa band attributed as edestin. Amino acid analysis showed that EAE augmented total essential amino acids, particularly protease. Milling and pronase treatment yielded the highest and lowest IVPD‐corrected amino acid score (IVPDCAAS) of 76% and 47%, respectively. Milled and germinated flours contained varying amounts of γ‐tocopherol, lutein, zeaxanthin, α‐carotene, and β‐carotene. Germinated flour exhibited elevated levels of total phenolic (14.36 mg/g), and flavonoid (1.76 mg/g) contents, FRAP, TEAC, and DPPH compared to the other flours suggesting superior antioxidant capacity. Strong positive correlations (r >0.70) were found between IVPD and phytic acid for the protein isolate, total phenolic content, and FRAP for IEP and germinated flours. Overall, hempseed flours with diverse properties could be produced using non‐thermal processing. [ABSTRACT FROM AUTHOR]

Malva sylvestris L., Cynara cardunculus L., and Spinacia oleracea L. are spontaneous plants often consumed by humans and animals. Although, these plants possess nutritional and therapeutic value, they may also contain anti-nutritional factors in their carbohydrate fraction and trypsin inhibitors that interfere with protein digestion. This study aims to identify the antinutritional factors, and determine the trypsin inhibitory activity of Malva sylvestris L., Cynara cardunculus L., and Spinacia oleracea L. The sugar contents of the plants' ethanol extracts were determined using the Anthrone and Ortho-Toluidine colorimetric methods. Specific monosaccharides and oligosaccharides were identified by thin layer chromatography (TLC). The polysaccharides (starch and cellulose) contents were also assessed. The trypsin inhibitory activity was evaluated according to standard procedures. The results showed that the sugar contents of the plants ranged from 0.14 to 0.34 g/100 g dw. TLC analysis revealed the presence of sucrose in all the three plant extracts, while two oligosaccharides; raffinose and stachyose were identified in M. sylvestris and S. oleracea. The extracts exhibited significant trypsin inhibitory activity with C. cardunculus demonstrating the highest inhibitory effect with IC50 value of 4.92 mg/mL, while the IC50 values for M. sylvestris and S. oleracea where 9.26 mg/mL and 12.24 mg/mL, respectively. The presence of these anti-nutritional factors may interfere with digestion in both humans and animals, hence the need for caution in the use of these plants for medicinal purposes. [ABSTRACT FROM AUTHOR]

Severe alpha-1 antitrypsin deficiency (AATD) is associated with an increased risk of emphysema. However, the clinical manifestations are very heterogeneous, and an individual prognosis is very difficult to establish. Intravenous augmentation therapy with alpha-1 antitrypsin (AAT) from pooled blood donors is the only specific treatment available, but it requires weekly or biweekly administration for life. Several guidelines provide the indication criteria for the initiation of AAT augmentation therapy. However, in clinical practice, there are situations in which the decision as to when to start treatment becomes uncertain and some studies have shown great variability in the indication of this treatment even among specialists. The usual dilemma is between initiating augmentation therapy in individuals who may not develop significant lung disease or in whom disease will not progress or delaying it in patients who may otherwise rapidly and irreversibly progress. We illustrate this dilemma with five clinical cases: from the case of a patient with normal lung function who requests initiation of therapy to a moderately stable patient without augmentation or a mild patient who, after several years of remaining stable without treatment, deterioration in lung function initiated and, consequently, augmentation therapy was begun. All the nuances associated with the indication of augmentation justify a personalised approach and the decision about initiating augmentation therapy must be made after careful consideration of the pros and cons with the patient in reference centres with experience in treatment. These reference centres can work in collaboration with local hospitals where patients can be closely followed and augmentation therapy can be administered to avoid unnecessary travelling, making periodical administrations more comfortable for the patient. [ABSTRACT FROM AUTHOR]

Plant–soil interactions have bottom–up and top–down effects within a plant community. Heavy metal pollution can change plant–soil interactions, directly influence bottom–up effects and indirectly affect herbivores within the community. In turn, herbivores can affect plant–soil interactions through top–down effects. However, the combined effects of heavy metals and herbivores on soil enzymes, plants and herbivores have rarely been reported. Therefore, the effects of lead (Pb), Spodoptera litura and their combined effects on soil enzyme activities, pakchoi nutrition, defence compounds and S. litura fitness were examined here. Results showed that Pb, S. litura and their combined effects significantly affected soil enzymes, pakchoi and S. litura. Specifically, exposure to double stress (Pb and S. litura) decreased soil urease, phosphatase and sucrase activities compared with controls. Furthermore, the soluble protein and sugar contents of pakchoi decreased, and the trypsin inhibitor content and antioxidant enzyme activity increased. Finally, the S. litura development period was extended, and survival, emergence rates and body weight decreased after exposure to double stress. The combined stress of Pb and S. litura significantly decreased soil enzyme activities. Heavy metal accumulation in plants may create a superposition or synergistic effect with heavy metal-mediated plant chemical defence, further suppressing herbivore development. Pb, S. litura and their combined effects inhibited soil enzyme activities, improved pakchoi resistance and reduced S. litura development. The results reveal details of soil–plant–herbivore interactions and provide a reference for crop pest control management in the presence of heavy metal pollution. [ABSTRACT FROM AUTHOR]

Summary: Soybeans (Glycine max) are a preferred source of plant‐based proteins and oil in various applications in foods and feeds. When soybean is converted to soybean meal (SBM), the quality is determined by the protein content and its digestibility. Protein digestibility is reduced by anti‐nutritional factors like trypsin inhibitors (TI) and processing. The abundant genetic variations in soybean germplasm affect the quantitative trait of protein content. This study analysed the effect of location and planting dates on genetically identical soybeans grown in North Carolina (NC) and then compared the nutritional traits to similar commonly grown soybean varieties in other locations in the USA. The soybeans were analysed using NIRS and wet chemistry. It was found that soybean protein content ranged from 38.14% to 44.92% among the different varieties grown in three different locations in NC. Similarly, TI levels ranged between 28.22 and 33.11 mg g−1 among these varieties. The same varieties, when planted as either a full season or double crop, had different TI levels and were also impacted by planting location. However, there were no significant differences in the TI levels between the varieties and planting seasons. Furthermore, no significant difference in protein content among the varieties within the planting dates was found. Compared to soybeans grown in other parts of the USA, the average crude protein (CP) content was similar to those grown in Arkansas and Minnesota. The TI levels were significantly higher than those grown in Ohio, similar to that of Arkansas, and significantly lower than that of Minnesota. [ABSTRACT FROM AUTHOR]

Moringa oleifera Lam. (horseradish tree) leaves demonstrate high nutritional value, are rich in proteins, and are widely used in folk medicine and food. This study investigated the presence of secondary metabolites and antinutritional proteins in leaf extract (LE) and the protein-rich fraction (PRF) derived from M. oleifera leaves, as well as the cytotoxicity to human cells, hemolytic activity, and in vivo acute toxicity and genotoxicity in mice. The flavonoids rutin and vitexin as well as trypsin inhibitors and lectins were detected in LE and PRF. Neither sample demonstrated toxicity against human peripheral blood mononuclear cells and both showed low hemolytic action. In vivo, LE and PRF did not show antinutritional effects and caused no death. The hematological parameters of the animals in the treated group were similar to those of the control. A significant increase in the serum levels of alanine aminotransferase and a discrete leukocyte infiltration with cytoplasmic vacuolization of the hepatocytes in the liver were detected in LE-treated animals. The preparations were not genotoxic or mutagenic. This study shows that LE and PRF are not antinutritional agents and presented low acute toxicity and no genotoxicity or mutagenicity. The present study contributes to the determination of the safety of using M. oleifera leaf proteins. [ABSTRACT FROM AUTHOR]

Black soybeans with green cotyledon have long been widely consumed in the East due to their high content of health-promoting anthocyanins and lutein. However, major anti-nutritional and allergenic components such as lectin, Kunitz trypsin inhibitor (KTI), P34, lipoxygenase, and stachyose are contained in mature seed. The objective of this research is to breed a soybean line with a black seed coat, a green cotyledon, and the penta null genotype (lele-titi-p34p34-lox1lox1lox2lox2lox3lox3-rs2rs2) for all five components. The F2 plant strain with penta null genotype for lectin, KTI, P34, lipoxygenase, and stachyose components was developed. The breeding line has purple flower, determinate growth habit, brown pod, black seed coat and green cotyledon. The stem height of the breeding line was 57.0 cm and 100 seed weight was 31.5 g. This is the first soybean breeding line with black seed coat, green cotyledon and penta null genotypes for lectin, KTI, P34, lipoxygenase, and stachyose factors. This line will be used as parent to improve a black soybean cultivar with green cotyledon that have significantly reduced anti-nutritional and allergenic traits. [ABSTRACT FROM AUTHOR]

Trypsin inhibitors (TI) in raw soybean grain, mainly represented by the Kunitz trypsin inhibitor protein (KTI), prevent the normal activity of the digestive enzymes trypsin and chymotrypsin in humans and monogastric livestock. The inactivation of TI is achieved through costly and time-consuming heat treatment. Thermal processing also impairs the solubility and availability of the soybean grain protein. Therefore, the genetic elimination of KTI has been proposed as a suitable alternative to heat treatment. The aim of this study was to screen the collection of European soybean cultivars with six genetic markers (one SSR marker and five SNP markers) previously proposed as tightly linked to the KTI3 gene encoding the major Kunitz trypsin inhibitor seed protein of soybean and validate their usability for marker-assisted selection (MAS). The six markers were validated on a subset of 38 cultivars with wide variability in KTI content and in the F2 and F3:5 progenies of two crosses between the known high- and low-KTI cultivars. Three genetic markers (SSR Satt228 and two SNP markers, Gm08_45317135_T/G and Gm08_45541906_A/C) were significantly associated with KTI content in a subset of 38 cultivars. Low-KTI alleles were detected in both low- and high-KTI genotypes and vice versa, high-KTI alleles were found in both high- and low-KTI genotypes, indicating a tight but not perfect association of these markers with the KTI3 gene. The genetic marker SSR Satt228 showed a significant association with KTI content in the F2 progeny, while the SNP markers Gm08_45317135_T/G and Gm08_45541906_A/C allowed significant discrimination between progeny with high- vs. low-KTI progenies in the F3:5 generation. These three markers could be applied in MAS for low-KTI content but not without the additional phenotyping step to extract the desired low-KTI genotypes. [ABSTRACT FROM AUTHOR]

Knottins are very stable peptides (also called miniproteins) containing approximately 30 amino acid residues. Their structures are stabilized by three disulfide bonds forming a characteristic 'pseudo‐knotted' structure. This research was devoted to the design of potential inhibitors of human hepatitis C (HCV) NS3 protease by means of knottin fold using loop grafting, amino acid residues replacements and L/D amino acid epimerization. The initial structure used for design was a trypsin inhibitor from Momordica cochinchinensis seeds, MCOTI‐II. Modifications of the knottin template for optimization of the NS3 protease‐peptide interaction included deletion of several residues from the N‐terminus, replacement of residues in‐ and outside the inhibitor loop and replacement of certain L‐amino acids by their D‐stereoisomers. Stability of modified knottins and their complexes with HCV protease were evaluated by molecular dynamics simulation. Binding energy values for protein‐knottin complexes were calculated by MM‐GBSA method. The designed modified knottin characterized by the highest stability of the complex with HCV NS3 protease, was proposed for further experimental testing. [ABSTRACT FROM AUTHOR]

Background: Alpha-1 antitrypsin deficiency (AATD) is a genetic condition resulting from mutations in the alpha-1 antitrypsin (AAT) protein, a major systemic antiproteinase, resulting in reduced/no release of AAT, disrupting the proteinase/antiproteinase balance. A sustained imbalance can cause structural changes to the lung parenchyma, leading to emphysema. Predicting and assessing human responses to potential therapeutic candidates from preclinical animal studies have been challenging. Our aims were to develop a more physiologically relevant in vitro model of the proteinase/antiproteinase balance and assess whether the data generated could better predict the efficacy of pharmacological candidates to inform decisions on clinical trials, together with expected biomarker responses. Methods: We developed an in vitro model assessing the proteinase/antiproteinase balance by the changes in the fibrinogen cleavage products of neutrophil elastase (NE) and proteinase 3 (PR3). This allowed the assessment of physiological and pharmaceutical neutrophil serine proteinase (NSP) inhibitors to determine the putative threshold at which the maximal effect is achieved. Results: AAT significantly reduced NE and PR3 activity footprints, with themaximal reduction achieved at concentrations above 10 µM. The inhibitor MPH966 alone also significantly reduced NE footprint generation in a concentration-dependent manner, leveling out above 100 nM but had no effect on the PR3 footprint. At levels of AAT consistent with AATD, MPH966 had an additive effect, reducing the NE activity footprint more than either inhibitor alone. Conclusion: Our results support an inhibitor threshold above which the activity footprint generation appears resistant to increasing dosage. Our model can support the testing of inhibitors, confirming activity biomarkers as indicators of likely pharmaceutical efficacy, the assessment of NSP activity in the pathophysiology of emphysema, and the likely function of biological or pharmacological inhibitors in disease management. [ABSTRACT FROM AUTHOR]

Three sweet potato genotypes NSP-2, NSP-6 (white flesh) and NSP-12 (orange flesh), were subjected under different processing techniques (boiling, steaming, and roasting) to check the nutritional status. Sugars like glucose, fructose, sucrose, and maltose was increased under roasting treatment for all of the genotypes. Crude fiber, cellulose, hemicellulose and ash content were significantly deferred within different genotypes but different treatments remained at par with various processing techniques. Steaming was proved to be the best treatment to decrease loss of vitamin C and phenol content where as roasting was the best performing treatmant to decrease the antinutrients like phytic acid and trypsin inhibitor activity. Comparatively, higher antioxidant activity was observed in steaming treatmet. NSP-12 revealed comparatively lower Glycemic index (GI) indicating high amylase: amylopectin ratio may become a good source of good carbohydrate for diabetic friendly food for near future. [ABSTRACT FROM AUTHOR]

Kunitz trypsin inhibitor genes play important roles in stress resistance. In this study, we investigated RpKTI2 cloned from Robinia pseudoacacia and its effect on tobacco. RpKTI2 was introduced into the tobacco cultivar NC89 using Agrobacterium-mediated transformation. Six RpKTI2-overexpressing lines were obtained. Transgenic and wild-type tobacco plants were then compared for photosynthetic characteristics and endogenous hormone levels. Transgenic tobacco showed minor changes in chlorophyll content, fluorescence, and photosynthetic functions. However, the maximum photochemical efficiency (Fv/Fm) increased significantly while intercellular CO2 concentration (Ci) decreased significantly. Stomatal size and hormone content (indole-3-acetic acid, zeatin riboside, gibberellin, and indole-3-propionic acid) were reduced, while brassinosteroid content increased. Random forest regression revealed that RpKTI2 overexpression had the biggest impact on carotenoid content, initial fluorescence, Ci, stomatal area, and indole-3-acetic acid. Overall, RpKTI2 overexpression minimally affected chlorophyll synthesis and photosynthetic system characteristics but influenced stomatal development and likely enhanced the antioxidant capacity of tobacco. These findings provide a basis for future in-depth research on RpKTI2. [ABSTRACT FROM AUTHOR]

Copyright of Ziemniak Polski is the property of Instytutem Hodowli & Aklimatyzacji Roslin - Panstwowym Instytutem Badawczym and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The article presents a new method called no-laboratory epidermal cell paste (NLEP) for treating stable vitiligo, a skin disorder characterized by depigmentation. The method is simple, cost-effective, and does not require specialized equipment. The study involved three patients who experienced significant repigmentation and good color matching after 12 weeks of treatment. The article also discusses the psychological aspects of cosmetic procedures and the potential for excessive treatments to become a substance-related disorder. It emphasizes the importance of understanding patients' motivations in order to provide effective guidance during consultation and treatment. [Extracted from the article]

Pressure-induced perturbation of a protein structure leading to its folding–unfolding mechanism is an important yet not fully understood phenomenon. The key point here is the role of water and its coupling with protein conformations as a function of pressure. In the current work, using extensive molecular dynamics simulation at 298 K, we systematically examine the coupling between protein conformations and water structures of pressures of 0.001, 5, 10, 15, 20 kbar, starting from (partially) unfolded structures of the protein Bovine Pancreatic Trypsin Inhibitor (BPTI). We also calculate localized thermodynamics at those pressures as a function of protein–water distance. Our findings show that both protein-specific and generic effects of pressure are operating. In particular, we found that (1) the amount of increase in water density near the protein depends on the protein structural heterogeneity; (2) the intra-protein hydrogen bond decreases with pressure, while the water–water hydrogen bond per water in the first solvation shell (FSS) increases; protein–water hydrogen bonds also found to increase with pressure, (3) with pressure hydrogen bonds of waters in the FSS getting twisted; and (4) water's tetrahedrality in the FSS decreases with pressure, but it is dependent on the local environment. Thermodynamically, at higher pressure, the structural perturbation of BPTI is due to the pressure–volume work, while the entropy decreases with the increase of pressure due to the higher translational and rotational rigidity of waters in the FSS. The local and subtle effects of pressure, found in this work, are likely to be typical of pressure-induced protein structure perturbation. [ABSTRACT FROM AUTHOR]

Background: The PI*S variant is one of the most prevalent mutations within alpha-1 antitrypsin deficiency (AATD). The risk of developing AATD-related lung disease in individuals with the PI*SS genotype is poorly defined despite its substantial prevalence. Our study aimed to characterize this genotype and its risk for lung disease and compare it with the PI*ZZ and PI*SZ genotypes using data from the European Alpha-1 antitrypsin Deficiency Research Collaboration international registry. Method: Demographic, clinical, functional, and quality of life (QoL) parameters were assessed to compare the PI*SS characteristics with the PI*SZ and PI*ZZ controls. A propensity score with 1:3 nearest-neighbour matching was performed for the most important confounding variables. Results: The study included 1007 individuals, with PI*SS (n = 56; 5.6%), PI*ZZ (n = 578; 57.4%) and PI*SZ (n = 373; 37.0%). The PI*SS population consisted of 58.9% men, with a mean age of 59.2 years and a mean FEV1(% predicted) of 83.4%. Compared to PI*ZZ individuals they had less frequent lung disease (71.4% vs. 82.2%, p = 0.037), COPD (41.4% vs. 60%, p = 0.002), and emphysema (23.2% vs. 51.9%, p < 0.001) and better preserved lung function, fewer exacerbations, lower level of dyspnoea, and better QoL. In contrast, no significant differences were found in the prevalence of lung diseases between PI*SS and PI*SZ, or lung function parameters, exacerbations, dyspnoea, or QoL. Conclusions: We found that, as expected, the risk of lung disease associated with the PI*SS genotype is significantly lower compared with PI*ZZ, but does not differ from that observed in PI*SZ individuals, despite having higher serum AAT levels. Trial registration: www.clinicaltrials.gov (ID: NCT04180319). [ABSTRACT FROM AUTHOR]

Human induced pluripotent stem cell (iPSC) and CRISPR-Cas9 gene-editing technologies have become powerful tools in disease modeling and treatment. By harnessing recent biotechnological advancements, this review aims to equip researchers and clinicians with a comprehensive and updated understanding of the evolving treatment landscape for metabolic and genetic disorders, highlighting how iPSCs provide a unique platform for detailed pathological modeling and pharmacological testing, driving forward precision medicine and drug discovery. Concurrently, CRISPR-Cas9 offers unprecedented precision in gene correction, presenting potential curative therapies that move beyond symptomatic treatment. Therefore, this review examines the transformative role of iPSC technology and CRISPR-Cas9 gene editing in addressing metabolic and genetic disorders such as alpha-1 antitrypsin deficiency (A1AD) and glycogen storage disease (GSD), which significantly impact liver and pulmonary health and pose substantial challenges in clinical management. In addition, this review discusses significant achievements alongside persistent challenges such as technical limitations, ethical concerns, and regulatory hurdles. Future directions, including innovations in gene-editing accuracy and therapeutic delivery systems, are emphasized for next-generation therapies that leverage the full potential of iPSC and CRISPR-Cas9 technologies. [ABSTRACT FROM AUTHOR]

Background: Recombinant peptide production in Escherichia coli provides a sustainable alternative to environmentally harmful and size-limited chemical synthesis. However, in-vivo production of disulfide-bonded peptides at high yields remains challenging, due to degradation by host proteases/peptidases and the necessity of translocation into the periplasmic space for disulfide bond formation. Results: In this study, we established an expression system for efficient and soluble production of disulfide-bonded peptides in the periplasm of E. coli. We chose model peptides with varying complexity (size, structure, number of disulfide bonds), namely parathyroid hormone 1–84, somatostatin 1–28, plectasin, and bovine pancreatic trypsin inhibitor (aprotinin). All peptides were expressed without and with the N-terminal, low molecular weight CASPON™ tag (4.1 kDa), with the expression cassette being integrated into the host genome. During BioLector™ cultivations at microliter scale, we found that most of our model peptides can only be sufficiently expressed in combination with the CASPON™ tag, otherwise expression was only weak or undetectable on SDS-PAGE. Undesired degradation by host proteases/peptidases was evident even with the CASPON™ tag. Therefore, we investigated whether degradation happened before or after translocation by expressing the peptides in combination with either a co- or post-translational signal sequence. Our results suggest that degradation predominantly happened after the translocation, as degradation fragments appeared to be identical independent of the signal sequence, and expression was not enhanced with the co-translational signal sequence. Lastly, we expressed all CASPON™-tagged peptides in two industry-relevant host strains during C-limited fed-batch cultivations in bioreactors. We found that the process performance was highly dependent on the peptide-host-combination. The titers that were reached varied between 0.6–2.6 g L−1, and exceeded previously published data in E. coli. Moreover, all peptides were shown by mass spectrometry to be expressed to completion, including full formation of disulfide bonds. Conclusion: In this work, we demonstrated the potential of the CASPON™ technology as a highly efficient platform for the production of soluble peptides in the periplasm of E. coli. The titers we show here are unprecedented whenever parathyroid hormone, somatostatin, plectasin or bovine pancreatic trypsin inhibitor were produced in E. coli, thus making our proposed upstream platform favorable over previously published approaches and chemical synthesis. [ABSTRACT FROM AUTHOR]

Objective: The present work was conducted to evaluate suitable variables and develop prediction equations using chemical composition and physical characteristics for estimating standardized ileal digestibility (SID) of lysine (Lys) in full-fat soybeans (FFSB). Methods: The chemical composition and physical characteristics were determined including trypsin inhibitor activity (TIA), urease activity (UA), protein solubility in 0.2% potassium hydroxide (KOH), protein dispersibility index (PDI), lysine to crude protein ratio (Lys:CP), reactive Lys:CP ratio, neutral detergent fiber, neutral detergent insoluble nitrogen (NDIN), acid detergent insoluble nitrogen (ADIN), acid detergent fiber, L* (lightness), and a* (redness). Pearson's correlation (r) was computed, and the relationship between variables was determined by linear or quadratic regression. Stepwise multiple regression was performed to develop prediction equations for SID of Lys. Results: Negative correlations (p<0.01) between SID of Lys and protein quality indicators were observed for TIA (r = -0.80), PDI (r = -0.80), and UA (r = -0.76). The SID of Lys also showed a quadratic response (p<0.01) to UA, NDIN, TIA, L*, KOH, a* and Lys:CP. The best-fit model for predicting SID of Lys in FFSB included TIA, UA, NDIN, and ADIN, resulting in the highest coefficient of determination (R2 = 0.94). Conclusion: Quadratic regression with one variable indicated the high accuracy for UA, NDIN, TIA, and PDI. The multiple linear regression including TIA, UA, NDIN, and ADIN is an alternative model used to predict SID of Lys in FFSB to improve the accuracy. Therefore, multiple indicators are warranted to assess either insufficient or excessive heat treatment accurately, which can be employed by the feed industry as measures for quality control purposes to predict SID of Lys in FFSB. [ABSTRACT FROM AUTHOR]

Nonylphenol is an aromatic organic compound that has an estrogen-like effect and has a negative effect on the human endocrine system. A method has been developed for the competitive determination of nonylphenol using magnetic particles, rabbit antiserum, nonylphenol conjugate with soybean trypsin inhibitor (STI) and biotin. The principle of the analysis is the formation of immune complexes on the surface of magnetite particles due to covalent immobilization of protein G through the oriented immobilization of polyclonal antibodies from rabbit serum during a competitive reaction between the free analyte (nonylphenol) and the bound one (as part of the nonylphenol-STI-biotin conjugate) for the binding sites of specific antibodies. The detection of formed immune complexes is proposed to be carried out using a streptavidin-polyperoxidase conjugate, which makes it possible to achieve a nine-fold gain in the level of the analytical signal. The developed ELISA using magnetite particles allows us to achieve a detection limit of nonylphenol at the level of 3.8 ng/mL, which is 14.5 times lower in comparison with the classic competitive ELISA (55 ng/mL). Based on the results of the experimental work, the optimized volume of the test sample was 500 μL, which makes it possible to concentrate low-contaminated samples by 17 times. [ABSTRACT FROM AUTHOR]

Recovering valuable active substances from the by-products of agricultural processing is a crucial concern for scientific researchers. This paper focuses on the enrichment of soybean trypsin inhibitor (STI) from soybean whey wastewater using either ammonium sulfate salting or ethanol precipitation, and discusses their physicochemical properties. The results show that at a 60% ethanol content, the yield of STI was 3.983 mg/mL, whereas the yield was 3.833 mg/mL at 60% ammonium sulfate saturation. The inhibitory activity of STI obtained by ammonium sulfate salting out (A-STI) was higher than that obtained by ethanol precipitation (E-STI). A-STI exhibited better solubility than E-STI at specific temperatures and pH levels, as confirmed by turbidity and surface hydrophobicity measurements. Thermal characterization revealed that both A-STI and E-STI showed thermal transition temperatures above 90 °C. Scanning electron microscopy demonstrated that A-STI had a smooth surface with fewer pores, while E-STI had a rough surface with more pores. In conclusion, there was no significant difference in the yield of A-STI and E-STI (p < 0.05); however, the physicochemical properties of A-STI were superior to those of E-STI, making it more suitable for further processing and utilization. This study provides a theoretical reference for the enrichment of STI from soybean whey wastewater. [ABSTRACT FROM AUTHOR]

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a global pandemic. Known as COVID-19, it has affected billions of people worldwide, claiming millions of lives and posing a continuing threat to humanity. This is considered one of the most extensive pandemics ever recorded in human history, causing significant losses to both life and economies globally. However, the available evidence is currently insufficient to establish the effectiveness and safety of antiviral drugs or vaccines. The entry of the virus into host cells involves binding to angiotensin-converting enzyme 2 (ACE2), a cell surface receptor, via its spike protein. Meanwhile, transmembrane protease serine 2 (TMPRSS2), a host surface protease, cleaves and activates the virus's S protein, thus promoting viral infection. Plant protease inhibitors play a crucial role in protecting plants against insects and/or microorganisms. The major storage proteins in sweet potato roots include sweet potato trypsin inhibitor (SWTI), which accounts for approximately 60% of the total water-soluble protein and has been found to possess a variety of health-promoting properties, including antioxidant, anti-inflammatory, ACE-inhibitory, and anticancer functions. Our study found that SWTI caused a significant reduction in the expression of the ACE2 and TMPRSS2 proteins, without any adverse effects on cells. Therefore, our findings suggest that the ACE2 and TMPRSS2 axis can be targeted via SWTI to potentially inhibit SARS-CoV-2 infection. [ABSTRACT FROM AUTHOR]

Despite the high quality of soybean protein, raw soybeans and soybean meal cannot be directly included in animal feed mixtures due to the presence of Kunitz (KTi) and Bowman–Birk protease inhibitors (BBis), which reduces animal productivity. Heat treatment can substantially inactivate trypsin and chymotrypsin inhibitors (BBis), but such treatment is energy-intensive, adds expense, and negatively impacts the quality of seed proteins. As an alternative approach, we have employed CRISPR/Cas9 gene editing to create mutations in BBi genes to drastically lower the protease inhibitor content in soybean seed. Agrobacterium-mediated transformation was used to generate several stable transgenic soybean events. These independent CRISPR/Cas9 events were examined in comparison to wild-type plants using Sanger sequencing, proteomic analysis, trypsin/chymotrypsin inhibitor activity assays, and qRT-PCR. Collectively, our results demonstrate the creation of an allelic series of loss-of-function mutations affecting the major BBi gene in soybean. Mutations in two of the highly expressed seed-specific BBi genes lead to substantial reductions in both trypsin and chymotrypsin inhibitor activities. [ABSTRACT FROM AUTHOR]

Balaustium murorum (Acariformes: Erythraeidae) is a pollen feeder and predatory mite that occasionally causes dermatitis. We sought to determine the protease profile in the whole-mite extract of B. murorum using general and specific substrates as well as inhibitors. The pH profiles using two general substrates, azacasein and haemoglobin, revealed an optimal pH of 6 for both substrates. Specific trypsin inhibitor TLCK showed no influence activity towards BApNA; however, TPCK and AEBSF inhibited chymotrypsin and elastase, respectively. Specific cysteine protease inhibitors Cystatin and E-64 influenced both types of cathepsins B and L, while DTT as a specific activator increased the enzymatic activities. Only aminopeptidase showed significant activity and inhibition using negative control and phenanthroline. Chymotrypsin and elastase had the highest activity at pH of 9–10 and 10, respectively, by using both substrates alone and substrates along with specific inhibitors. Cathepsins B and L had pH optima of 6. Aminopeptidase revealed a broad pH optimum of 5–7 on both substrates HA and HPA alone and substrate along with inhibitor phenanthroline. Finally, different concentrations of each inhibitor were used to find IC50 values of TPCK, AEBSF, cystatin, E-64, DTT and phenanthroline required to inhibit half of enzyme activity. [ABSTRACT FROM AUTHOR]

Typical Kunitz proteins (I2 family of the MEROPS database, Kunitz-A family) are metazoan competitive inhibitors of serine peptidases that form tight complexes of 1:1 stoichiometry, mimicking substrates. The cestode Echinococcus granulosus, the dog tapeworm causing cystic echinococcosis in humans and livestock, encodes an expanded family of monodomain Kunitz proteins, some of which are secreted to the dog host interface. The Kunitz protein EgKU-7 contains, in addition to the Kunitz domain with the anti-peptidase loop comprising a critical arginine, a C-terminal extension of ~20 amino acids. Kinetic, electrophoretic, and mass spectrometry studies using EgKU-7, a C-terminally truncated variant, and a mutant in which the critical arginine was substituted by alanine, show that EgKU-7 is a tight inhibitor of bovine and canine trypsins with the unusual property of possessing two instead of one site of interaction with the peptidases. One site resides in the anti-peptidase loop and is partially hydrolyzed by bovine but not canine trypsins, suggesting specificity for the target enzymes. The other site is located in the C-terminal extension. This extension can be hydrolyzed in a particular arginine by cationic bovine and canine trypsins but not by anionic canine trypsin. This is the first time to our knowledge that a monodomain Kunitz-A protein is reported to have two interaction sites with its target. Considering that putative orthologs of EgKU-7 are present in other cestodes, our finding unveils a novel piece in the repertoire of peptidase-inhibitor interactions and adds new notes to the evolutionary host-parasite concerto. [ABSTRACT FROM AUTHOR]

Although peach kernels are rich in oil, there is a lack of information about its chemical and biological properties. Therefore, the purpose of this study was to determine the lipid profile, antioxidant capacity, and trypsin inhibitory propriety of peach oil extracted from two varieties (sweet cap and O'Henry) cultivated in Tunisia. The investigated peach kernel oil contains significant amount of unsaponifiable (2.1±0.5-2.8±0.2% of oil) and phenolic compounds (45.8±0.92-74.6±1.3 mg GAE/g of oil). Its n-alkane profile was characterized by the predominance of tetracosane n-C24 (47.24%) followed by tricosane n-C23 (34.43%). An important total tocopherol content (1192.83±3.1 mg/kg oil) has been found in sweet cap cultivar. Although rich in polyphenols and tocopherols, the tested oil did not display an inhibitory effect on trypsin. However, all peach oil samples showed effective antioxidant capacity and the highest values (86.34±1.3% and 603.50±2.6 µmol TE/g oil for DPPH test and ORAC assay, respectively) were observed for sweet cap oil. Peach oil has an excellent potential for application in the food and pharmaceutical industries as source of naturally-occurring bioactive substances. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Soil Sciences & Agricultural Engineering is the property of Egyptian National Agricultural Library (ENAL) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Soy remains the legume protein of excellence for plant-based meat alternatives due to its fiber-forming potential. In this study, protein-rich powders from soy protein isolate (SPI), concentrate (SPC), and their mixture (SPM) were thoroughly characterized for their proximate composition, nutritional quality, and physicochemical properties to understand their structuring behavior during high-moisture extrusion. SPI presented higher degrees of protein denaturation and aggregation, least gelation concentration and lower essential amino acid contents. Thus, an SPI:SPC combination (1:9 ratio, 70% protein) was extruded at three different screw speeds (300, 350, and 400 rpm) and two temperature profiles (120 and 140 °C maximum temperature). The effects of the processing parameters on the extrudates were evaluated for their appearance (fibrousness), texture (TPA, cutting force, and anisotropy), color, protein structure (FTIR), and trypsin inhibitors. Higher temperatures resulted in softer and darker extrudates, with increased visual and instrumental anisotropy. Increasing screw speeds led to softer and lighter extrudates, without a clear fibrousness effect. β-sheet structures decreased and intermolecular aggregates (A1) increased after extrusion, especially at 140 °C, together with the formation of intramolecular aggregates (A2). Extrusion also significantly decreased the amount of trypsin inhibitors (>90%). This study demonstrates that extrusion parameters need to be carefully selected to achieve meat analogs with optimal textural and nutritional characteristics. [ABSTRACT FROM AUTHOR]

The peptide derived from E. contortisiliquum trypsin inhibitor (Pep-3-EcTI), peptide derived from kallikrein inhibitor isolated from B. bauhinioides (Pep-BbKI), and B. rufa peptide modified from B. bauhinioides (Pep-BrTI) peptides exhibit anti-inflammatory and antioxidant activities, suggesting their potential for treating asthma--chronic obstructive pulmonary disease (COPD) overlap (ACO). We compared the effects of these peptides with dexamethasone (DX) treatment in an ACO model. In this study, 11 groups of male BALB/c mice were pre-treated under different conditions, including sensitization with intraperitoneal injection and inhalation of ovalbumin (OVA), intratracheal instillation of porcine pancreatic elastase (ELA), sensitization with intraperitoneal injection, and various combinations of peptide treatments with Pep-3-EcTI, Pep-BbKI, Pep-BrTI, dexamethasone, and non-treated controls (SAL-saline). Respiratory system resistance, airway resistance, lung tissue resistance, exhaled nitric oxide, linear mean intercept, immune cell counts in the bronchoalveolar lavage fluid, cytokine expression, extracellular matrix remodeling, and oxidative stress in the airways and alveolar septa were evaluated on day 28. Results showed increased respiratory parameters, inflammatory markers, and tissue remodeling in the ACO group compared to controls. Treatment with the peptides or DX attenuated or reversed these responses, with the peptides showing effectiveness in controlling hyperresponsiveness, inflammation, remodeling, and oxidative stress markers. These peptides demonstrated an efficacy comparable to that of corticosteroids in the ACO model. However, this study highlights the need for further research to assess their safety, mechanisms of action, and potential translation to clinical studies before considering these peptides for human use. [ABSTRACT FROM AUTHOR]