The identification and expression analysis of walnut Acyl-ACP thioesterases.

Walnuts (Juglans regia L.), renowned for their nutritional potency, are a rich source of unsaturated fatty acids. Their regular intake plays a pivotal role in health maintenance and recuperation from a myriad of ailments. Fatty acyl-acyl carrier protein thioesterases, which orchestrate the hydrolysis of acyl-ACP thioester bonds, thereby yielding fatty acids of varying chain lengths, are instrumental in augmenting plant fatty acid content and modulating the balance between saturated and unsaturated fatty acids. Despite some investigative efforts into the synthesis and metabolic pathways of fatty acids in walnuts, our comprehension of Fat in walnuts remains rudimentary. This research undertook a comprehensive characterization of the JrFat family, predicated on the complete genome sequence of walnuts, leading to the identification of 8 JrFat genes and an exploration of their protein physicochemical properties. Utilizing Arabidopsis and soybean Fat genes as outgroups, JrFat genes can be categorized into 5 distinct subgroups, three of which encompass a pair of homologous gene pairs. These genes have demonstrated remarkable conservation throughout the evolutionary process, with highly analogous conserved base sequences. The promoter region of JrFats genes predominantly harbors light response and plant hormone response regulatory elements, with no discernible disparity in promoter elements among different JrFats. Predictive analyses indicate that JrFats proteins engage extensively with walnut fatty acid synthesis and metabolism-associated proteins. qRT-PCR analysis reveals an initial surge in the expression of JrFats during the development of walnut kernels, which either stabilizes or diminishes following the hard core period. Homologous gene pairs exhibit analogous expression patterns, and the expression trajectory of JrFats aligns with the dynamic accumulation of fatty acids in kernels. The expression of JrFatA2 exhibits a strong correlation with the content of Alpha-linolenic acid, while the expression of JrFatB2 is inversely correlated with the content of two saturated fatty acids. Collectively, these findings enrich our understanding of fatty acid synthesis and metabolism in walnuts and furnish gene resources for enhancing the content and ratio of fatty acids in walnuts.

JrPPO1/2 play distinct roles in regulating walnut fruit browning by different spatiotemporal expression and enzymatic characteristics.

Polyphenol oxidase (PPO) activity drives walnut fruit browning, but the roles of its only two-family genes, JrPPO1 and JrPPO2, remain unclear. This study explores the spatiotemporal expression and enzymatic characteristics of JrPPO1 and JrPPO2 in walnut. Treatment with the PPO activator CuSO4 and H2O2 accelerated fruit browning and up-regulated JrPPO1/2 expression, whereas treatment with the PPO inhibitor ascorbic acid delayed browning, down-regulating JrPPO1 and up-regulating JrPPO2 expression. Compared to mJrPPO1, mJrPPO2 can exhibited better enzyme activity at higher temperatures (47 °C) and in more acidic environments (pH 4.25). mJrPPO2 exhibited a higher substrate specificity over mJrPPO1, and the preferred substrates are catechol, chlorogenic acid, and epicatechin. Additionally, mJrPPO2 adapted better to low concentration of oxygen (as low as 1.0% O2) and slightly elevated CO2 levels compared to mJrPPO1. Subcellular localization and spatiotemporal expression patterns showed that JrPPO1 is only expressed in green tissues and located in chloroplasts, while JrPPO2 is also located in chloroplasts, partly associated with membranes, and is expressed in both green and non-green tissues. Silencing JrPPO1/2 with virus-induced gene silencing (VIGS) reduced fruit browning, maintained higher total phenols, and decreased MDA production. Notably, silencing JrPPO1 had a greater impact on browning than JrPPO2, indicating JrPPO1's greater contribution to PPO activity and fruit browning in walnut fruits. Consequently, JrPPO1 can be effectively regulated both at the molecular level and by manipulating environmental conditions, to achieve the objective of controlling fruit browning.

Preparation, characterization and microencapsulation of walnut (Juglans regia L.) peptides-zinc chelate.

In this research, a novel kind of walnut (Juglans regia L.) peptides-zinc (Zn-WPs) chelate was obtained using the mass ratio of the walnut peptides (WPs) to ZnSO4.7H2O of 3.5:1 at pH 8.5 and 50°C for 84 min, with the chelation rate of 84.5%. In comparison to walnut peptides (WPs), the contents of aspartic acid and glutamic acid in Zn-WPs chelate are approximately 27%, indicating that hydrophilic amino acids predominantly bind with walnut peptides. Following chelation with zinc ions, the ultraviolet-visible (UV) characteristic absorption peak shifted from 213 nm to 210 nm, while the average particle size of the chelate increased to 8.0 ± 0.14 µm, presenting a loose spherical structure under scanning electron microscopy. These findings suggest the formation of new substances. Fourier-transform infrared spectroscopy (FTIR) revealed carboxyl, amino, and peptide bonds as the chelation sites of WPs and zinc. The IC50 of walnut peptides-zinc (Zn-WPs) chelate is 2.91 mg/mL, indicative of a favorable DPPH radical scavenging rate. Furthermore, Zn-WPs chelate microcapsules were produced via the spray drying method, achieving an encapsulation rate of 75.67 ± 0.83% under optimal conditions. These microcapsules demonstrate robust stability across diverse environmental conditions. This study underscores the potential of Zn-WPs and its chelate microcapsules to enhance stability and bioactivity under varying circumstances. PRACTICAL APPLICATION: In this study, a new walnut peptide-zinc (Zn-WPs) chelate was prepared. The presence of zinc ions changes the structure and properties of walnut peptides and improves its stability. The production of Zn-WPs chelate microcapsules enables Zn-WPs to have strong in vitro stability under different pH and simulated gastrointestinal digestion conditions. These results provide novel insights for developing the walnut peptides as bioactive ingredients in functional foods.

Preparation and Properties of Walnut Protein Isolate-Whey Protein Isolate Nanoparticles Stabilizing High Internal Phase Pickering Emulsions.

To enhance the functional properties of walnut protein isolate (WalPI), hydrophilic whey protein isolate (WPI) was selected to formulate WalPI-WPI nanoparticles (nano-WalPI-WPI) via a pH cycling technique. These nano-WalPI-WPI particles were subsequently employed to stabilize high internal phase Pickering emulsions (HIPEs). By adjusting the mass ratio of WalPI to WPI from 9:1 to 1:1, the resultant nano-WalPI-WPI exhibited sizes ranging from 70.98 to 124.57 nm, with a polydispersity index of less than 0.326. When the mass ratio of WalPI to WPI was 7:3, there were significant enhancements in various functional properties: the solubility, denaturation peak temperature, emulsifying activity index, and emulsifying stability index increased by 6.09 times, 0.54 °C, 318.94 m2/g, and 552.95 min, respectively, and the surface hydrophobicity decreased by 59.23%, compared with that of WalPI nanoparticles (nano-WalPI), with the best overall performance. The nano-WalPI-WPI were held together by hydrophobic interactions, hydrogen bonding, and electrostatic forces, which preserved the intact primary structure and improved resistance to structural changes during the neutralization process. The HIPEs stabilized by nano-WalPI-WPI exhibited an average droplet size of less than 30 µm, with droplets uniformly dispersed and maintaining an intact spherical structure, demonstrating superior storage stability. All HIPEs exhibited pseudoplastic behavior with good thixotropic properties. This study provides a theoretical foundation for enhancing the functional properties of hydrophobic proteins and introduces a novel approach for constructing emulsion systems stabilized by composite proteins as emulsifiers.

A new approach for artificial pollination in walnut trees: AirPoll.

Pollination is the first step in the plant's fruit development. Therefore, fruit setting does not occur without pollination. Some problems encountered in natural pollination cause pollination not to be achieved as desired and cause significant losses in yield and fruit quality. Artificial pollination applications with drones are the best way to solve these problems. In this study, the AirPoll artificial pollination machine, which performs artificial pollination through the air using drone technology, was developed and the operating success of the machine was tested in walnut gardens. In the experiment gardens, female flowers on 18 branches of 5 trees each in the artificially pollinated area with a drone and in the control area were marked with colored strings. Control trees were selected from a distance that would not be possible to transport pollen with a drone. As a result of the study carried out in 2020 and 2021, the average fruit setting rate in trees pollinated by drone was determined as 94.61%. In control trees, 32.33% fruit setting was achieved. Thus, it was determined that the productivity increase in artificial pollination with AirPoll was 62.28%. In addition, in the study, Computational Fluid Dynamics (CFD) simulation analysis was performed using ANSYS Fluent 2024 R1 software to predict the downward air flow and pollen distribution in the walnut tree crown. The analysis was carried out in 680 iterations using drone propellers at a rotation speed of 4500 rpm, 4 m/s airflow and a k-w viscous model. In the analysis, it was observed that the pollen was distributed homogeneously with the determined height and the created artificial pollination environment. Based on the results obtained from the simulations, a convergence criterion of 5e-3 for continuity and 1e-6 for speed, k, w was determined. Considering all the results, the ease of use of the developed AirPoll artificial pollination machine and the successful results obtained in field trials reveal the effectiveness of the AirPoll artificial pollination machine.

Robust peroxidase from Bacillus mojavensis TH309: Immobilization on walnut shell hydrochar and evaluation of its potential in dye decolorization.

Peroxidases have received considerable attention as a cost-effective and environmentally friendly catalyst for bioremediation. Their rapid activity loss under harsh environmental conditions and inability to be used repetitively limit their exploitation in real-world wastewater treatment. First, a peroxidase was produced extracellularly by Bacillus mojavensis TH309 and purified 8.12-fold with a final yield of 47.10 % using Sephadex G-100 superfine resin. The pure peroxidase (BmPer) possessed a relatively low molecular weight of ~21 kDa and was active against L-DOPA on acrylamide gel after electrophoresis. BmPer was immobilized by adsorption functionalized walnut shell hydrochar (WsH) with 61.99 ±â€¯1.34 % efficiency and 37.07 ±â€¯4.16 % activity loss. BmPer and its immobilized form (WsH-BmPer) exhibited maximum activity at 50 °C and pH 9. WsH-BmPer exhibited 3.23-, 2.37-, 1.65-, and 2.25-fold longer half-life than BmPer at 50, 60, 70, and 80 °C, respectively. Immobilization significantly enhanced the stability of the enzyme under acidic conditions. BmPer and WsH-BmPer showed maximal activity in the presence of 1 % salt and retained more than 85 % of their activity even after pre-incubation with 2.5 M salt for 60 min at 50 °C. Their catalytic efficiency was significantly stimulated by pre-incubation with Triton X-100 (1 mM), Tween20 (1 mM), and Mg2+ (1 and 10 mM). Immobilization strongly reduced the loss of activity caused by inhibitors including Ba2+, Hg2+, and Cu2+. Moreover, both forms of the enzyme were compatible with solvents. The Michaelis constant (Km) values of BmPer and WsH-BmPer were 0.88 and 2.66 mM for 2,4 DCP, respectively. WsH-BmPer peroxidase maintained about 82 % and 85 % of its activity when stored at 4 °C for 30 days and reused for up to 10 cycles, respectively. Furthermore, it decolorized Cibacron red (CR), Poly R-478 (PR), Remazol Brilliant Blue R (RBBR), and Methyl red (MR) dyes by 60.13 %, 91.34 %, 86.41 %, and 50.51 % within 60 min, respectively.

Unveiling the "hidden quality" of the walnut pellicle: a precious source of bioactive lipids.

Tree nut consumption has been widely associated with various health benefits, with walnuts, in particular, being linked with improved cardiovascular and neurological health. These benefits have been attributed to walnuts' vast array of phenolic antioxidants and abundant polyunsaturated fatty acids. However, recent studies have revealed unexpected clinical outcomes related to walnut consumption, which cannot be explained simply with the aforementioned molecular hallmarks. With the goal of discovering potential molecular sources of these unexplained clinical outcomes, an exploratory untargeted metabolomics analysis of the isolated walnut pellicle was conducted. This analysis revealed a myriad of unusual lipids, including oxylipins and endocannabinoids. These lipid classes, which are likely present in the pellicle to enhance the seeds' defenses due to their antimicrobial properties, also have known potent bioactivities as mammalian signaling molecules and homeostatic regulators. Given the potential value of this tissue for human health, with respect to its "bioactive" lipid fraction, we sought to quantify the amounts of these compounds in pellicle-enriched waste by-products of mechanized walnut processing in California. An impressive repertoire of these compounds was revealed in these matrices, and in notably significant concentrations. This discovery establishes these low-value agriculture wastes promising candidates for valorization and translation into high-value, health-promoting products; as these molecules represent a potential explanation for the unexpected clinical outcomes of walnut consumption. This "hidden quality" of the walnut pellicle may encourage further consumption of walnuts, and walnut industries may benefit from a revaluation of abundant pellicle-enriched waste streams, leading to increased sustainability and profitability through waste upcycling.

Walnut oil: a promising nutraceutical in reducing oxidative stress and improving cholinergic activity in an in vitro Alzheimer's disease model.

Improving the quality of life in elderly patients and finding new treatment options for neurological diseases such as Alzheimer's has become one of the priorities in the scientific world. In recent years, the beneficial effects and therapeutic properties of natural foods on neurological health have become a very remarkable issue. Walnut oil (WO) is a promising nutraceutical, with many phytochemicals and polyunsaturated fatty acids and is thought to be promising in the treatment of many neurological diseases and cognitive deficits, such as Alzheimer's disease (AD). Polyphenolic compounds found in WO enhance intraneuronal signaling and neurogenesis and improve the sequestration of insoluble toxic protein aggregates. The objective of this study was to investigate the potential protective and therapeutic effects of WO in a model of AD induced by retinoic acid (RA) and brain-derived neurotrophic factor (BDNF). In order to achieve this, the experimental groups were formed as follows: Control group, WO group, Alzheimer's disease (AD) group, AD + WO applied group (AD + WO). WO supplementation almost significantly reduced oxidative stress in the ad model, providing 2-fold protection against protein oxidation. Additionally, WO showed a significant reduction in tau protein levels (2-fold), increased acetylcholine (ACh) levels (12%), and decreased acetylcholine esterase (AChE) activity (~50%). Since it has been known for centuries that WO does show any adverse effects on human health and has neuroprotective properties, it may be used in the treatment of AD as an additional nutraceutical to drug treatments.

A dual RNA-seq analyses revealed dynamic arms race during the invasion of walnut by Colletotrichum gloeosporioides.

BACKGROUND: Walnut anthracnose caused by Colletotrichum gloeosporioides seriously endangers the yield and quality of walnut, and has now become a catastrophic disease in the walnut industry. Therefore, understanding both pathogen invasion mechanisms and host response processes is crucial to defense against C. gloeosporioides infection. RESULTS: Here, we investigated the mechanisms of interaction between walnut fruits (anthracnose-resistant F26 fruit bracts and anthracnose-susceptible F423 fruit bracts) and C. gloeosporioides at three infection time points (24hpi, 48hpi, and 72hpi) using a high-resolution time series dual transcriptomic analysis, characterizing the arms race between walnut and C. gloeosporioides. A total of 20,780 and 6670 differentially expressed genes (DEGs) were identified in walnut and C. gloeosporioides against 24hpi, respectively. Generous DEGs in walnut exhibited opposite expression patterns between F26 and F423, which indicated that different resistant materials exhibited different transcriptional responses to C. gloeosporioides during the infection process. KEGG functional enrichment analysis indicated that F26 displayed a broader response to C. gloeosporioides than F423. Meanwhile, the functional analysis of the C. gloeosporioides transcriptome was conducted and found that PHI, SignalP, CAZy, TCDB genes, the Fungal Zn (2)-Cys (6) binuclear cluster domain (PF00172.19) and the Cytochrome P450 (PF00067.23) were largely prominent in F26 fruit. These results suggested that C. gloeosporioides secreted some type of effector proteins in walnut fruit and appeared a different behavior based on the developmental stage of the walnut. CONCLUSIONS: Our present results shed light on the arms race process by which C. gloeosporioides attacked host and walnut against pathogen infection, laying the foundation for the green prevention of walnut anthracnose.

Tolerance of peanuts and tree nuts in Spanish children with exclusive sensitization to lipid transfer proteins.

BACKGROUND: Allergy to peanuts and tree nuts is a common cause of food allergy in Spain, with lipid transfer proteins (LTP) being the most frequently recognized panallergen. LTP sensitization often leads to multiple food group sensitivities, resulting in overly restrictive diets that hinder patient's quality of life. This study aimed to assess the tolerance of peanuts and tree nuts (hazelnuts and walnuts) in children sensitized to LTP, potentially mitigating the need for such diets. METHODS: This prospective study enrolled individuals diagnosed with allergy to peanuts, hazelnuts, or walnuts. Data were collected from medical records, including demographics and clinical history. Allergological assessment comprised skin prick tests using commercial extracts and the nuts in question, alongside measurements of total and specific IgE to nuts and their primary molecular components. Participants showing positive LTP sensitization without sensitization to seed storage proteins underwent open oral nut challenges. RESULTS: A total of 75 individuals labeled as allergic to peanuts, 44 to hazelnuts, and 51 to walnuts were included. All of them underwent an open oral provocation test with the incriminated nut, showing a high tolerance rate. Peanut was tolerated by 98.6% of patients, 97.72% tolerated hazelnut, and 84.3% tolerated walnut. CONCLUSION: The findings suggest that the majority of patients allergic to peanuts, hazelnuts, or walnuts, due to LTP sensitization and lacking IgE reactivity to seed storage proteins, can tolerate these nuts. This supports the need for personalized nut tolerance assessments to avoid unnecessary dietary restrictions.

Dietary Walnuts Prevented Indomethacin-Induced Gastric Damage via AP-1 Transcribed 15-PGDH, Nrf2-Mediated HO-1, and n-3 PUFA-Derived Resolvin E1.

Non-steroidal anti-inflammatory drugs (NSAIDs), the most highly prescribed drugs in the world for the treatment of pain, inflammation, and fever, cause gastric mucosal damage, including ulcers, directly or indirectly, by which the development of GI-safer (-sparing) NSAIDs relates to unmet medical needs. This study aimed to document the preventive effects of walnut polyphenol extracts (WPEs) against NSAID-induced gastric damage along with the molecular mechanisms. RGM-1 gastric mucosal cells were administered with indomethacin, and the expressions of the inflammatory mediators between indomethacin alone or a combination with WPEs were compared. The expressions of the inflammatory mediators, including COX-1 and COX-2, prostaglandin E2, 15-hydroxyprostaglandin dehydrogenase (15-PGDH), and antioxidant capacity, were analyzed by Western blot analysis, RT-PCR, and ELISA, respectively. HO-1, Nrf-2, and keap1 were investigated. The in vivo animal models were followed with in vitro investigations. The NSAIDs increased the expression of COX-2 and decreased COX-1 and 15-PGDH, but the WPEs significantly attenuated the NSAID-induced COX-2 expression. Interestingly, the WPEs induced the expression of 15-PGDH. By using the deletion constructs of the 15-PGDH promoter, we found that c-Jun is the most essential determinant of the WPE-induced up-regulation of 15-PGDH expression. We confirmed that the knockdown of c-Jun abolished the ability of the WPEs to up-regulate the 15-PGDH expression. In addition, the WPEs significantly increased the HO-1 expression. The WPEs increased the nuclear translocation of Nrf2 by Keap-1 degradation, and silencing Nrf2 markedly reduced the WPE-induced HO-1 expression. We found that the WPE-induced HO-1 up-regulation was attenuated in the cells harboring the mutant Keap1, in which the cysteine 151 residue was replaced by serine. These in vitro findings were exactly validated in indomethacin-induced gastric rat models. Daily walnut intake can be a promising nutritional supplement providing potent anti-inflammatory, antioxidative, and mucosa-protective effects against NSAID-induced GI damage.

Utilization of low-grade walnut kernels for oil extraction using eco-friendly methods: a comparative analysis of oil composition, antioxidant and antimicrobial activity.

Walnut oil was extracted using three different eco-friendly extraction methods, solvent extraction (using ethyl acetate [EA] and ethanol [ET]), aqueous enzymatic extraction (AEE), and ultrasound-assisted enzymatic extraction (UAEE), and their lipid yield, lipid composition, physicochemical analysis, mineral composition, total phenols, antioxidant capacity, and antimicrobial activity were analyzed and compared. The AEE technique offered a greater yield (50.6%) than the other extraction methods and gave comparatively higher linoleic acid (66.12%) content. Palmitic, oleic, linoleic, linolenic, and stearic acids were the principal components that GC/MS detected in all the oil samples. UAEE produced the most polyphenols (0.49 mgGAE/g), followed by AEE (0.46 mgGAE/g), EA (0.45 mgGAE/g), and ET (0.35 mgGAE/g). The DPPH assay results were in the order of UAEE (191 µmolTE/kg) > AEE (186 µmolTE/kg) > EA (153 µmolTE/kg) > ET (130 µmolTE/kg). The FRAP assay findings showed a similar pattern: UAEE (112 molTE/kg) > AEE (102 molTE/kg) > EA (96 molTE/kg) > ET (82 molTE/kg). Results suggested that for a higher extraction yield, AEE is the better technique and UAEE is the recommended method for enhancing walnut oil antioxidant capacity. Additionally, it was found that polyphenols considerably increased the antioxidant capacity of walnut oil and are thought to be health-promoting. The results demonstrated the antibacterial effectiveness of the extracted oil against Bacillus subtilis, Bacillus licheniformis, and Staphylococcus aureus. This study provides information about low-cost and ecofriendly technologies of walnut oil extraction for food, cosmetic, and medical uses.

Lipidomics and metabolomics reveal the molecular mechanisms underlying the effect of thermal treatment on composition and oxidative stability of walnut oil.

Roasting walnut kernel significantly improves the oxidative stability and sensory properties of its oil. However, the effect of roasting temperatures on the molecular change of main components and micronutrients in walnut oil is still unclear. Herein, lipidomics and metabolomics were integrated to comprehensively profile the walnut oil obtained at different roasting temperatures (30 °C, 120 °C, 140 °C, 160 °C, and 180 °C). Lipidomics showed that the content of glycerolipids, sphingolipids, and glycerophospholipids decreased with roasting temperatures, while the oxidized fatty acids and triglycerides increased. Ratios of linoleic acid and linolenic acid varied with roasting temperatures and were most close to 4-6:1 at 140 °C, 160 °C, and 180 °C. Major classes of micronutrients showed a tendency to increase at the roasting temperature of 120 °C and 140 °C, then decrease at 160 °C and 180 °C. Liposoluble amino acids identified for the first time in walnut oil varied with roasting temperatures. Correlation analysis demonstrated that the higher contents of liposoluble amino acids and phenolics are positively associated with enhanced oxidative stability of walnut oil obtained at 140 °C. Furthermore, glutamine and 5-oxo-D-proline were expected to be potential biomarkers to differentiate the fresh and roasted walnut oil. The study is expected to provide new insight into the change mechanism of both major lipids and micronutrients in walnut oil during the roasting process.

Comparative Proteomic Analysis of Floral Buds before and after Opening in Walnut (Juglans regia L.).

The walnut (Juglans regia L.) is a typical and an economically important tree species for nut production with heterodichogamy. The absence of female and male flowering periods seriously affects both the pollination and fruit setting rates of walnuts, thereby affecting the yield and quality. Therefore, studying the characteristics and processes of flower bud differentiation helps in gaining a deeper understanding of the regularity of the mechanism of heterodichogamy in walnuts. In this study, a total of 3540 proteins were detected in walnut and 885 unique differentially expressed proteins (DEPs) were identified using the isobaric tags for the relative and absolute quantitation (iTRAQ)-labeling method. Among all DEPs, 12 common proteins were detected in all four of the obtained contrasts. GO and KEGG analyses of 12 common DEPs showed that their functions are distributed in the cytoplasm metabolic pathways, photosynthesis, glyoxylate and dicarboxylate metabolism, and the biosynthesis of secondary metabolites, which are involved in energy production and conversion, synthesis, and the breakdown of proteomes. In addition, a function analysis was performed, whereby the DEPs were classified as involved in photosynthesis, morphogenesis, metabolism, or the stress response. A total of eight proteins were identified as associated with the morphogenesis of stamen development, such as stamen-specific protein FIL1-like (XP_018830780.1), putative leucine-rich repeat receptor-like serine/threonine-protein kinase At2g24130 (XP_018822513.1), cytochrome P450 704B1-like isoform X2 (XP_018845266.1), ervatamin-B-like (XP_018824181.1), probable glucan endo-1,3-beta-glucosidase A6 (XP_018844051.1), pathogenesis-related protein 5-like (XP_018835774.1), GDSL esterase/lipase At5g22810-like (XP_018833146.1), and fatty acyl-CoA reductase 2 (XP_018848853.1). Our results predict several crucial proteins and deepen the understanding of the biochemical mechanism that regulates the formation of male and female flower buds in walnuts.

Interaction between the Neuroprotective and Hyperglycemia Mitigation Effects of Walnut-Derived Peptide LVRL via the Wnt3a/ß-Catenin/GSK-3ß Pathway in a Type 2 Diabetes Mellitus Model.

The term type 3 diabetes mellitus (T3DM) has been considered for Alzheimer's disease (AD) due to the common molecular and cellular characteristics found between type 2 diabetes mellitus (T2DM) and cognitive deficits. However, the specific mechanism of T3DM remains elusive, especially the neuroprotective effects of dietary components in hyperglycemic individuals. In this study, a peptide, Leu-Val-Arg-Leu (LVRL), found in walnuts significantly improved memory decline in streptozotocin (STZ)- and high-fat-diet (HFD)-stimulated T2DM mouse models (p < 0.05). The LVRL peptide also mitigated hyperglycemia, enhanced synaptic plasticity, and ameliorated mitochondrial dysfunction, as demonstrated by Morris water maze tests, immunoblotting, immunofluorescence, immunohistochemistry, transmission electron microscopy, and cellular staining. A Wnt3a inhibitor, DKK1, was subsequently used to verify the possible role of the Wnt3a/ß-Catenin/GSK-3ß pathway in glucose-induced insulin resistance in PC12 cells. In vitro LVRL treatment dramatically modulated the protein expression of p-Tau (Ser404), Synapsin-1, and PSD95, elevated the insulin level, increased glucose consumption, and relieved the mitochondrial membrane potential, and MitoSOX (p < 0.05). These data suggested that peptides like LVRL could modulate the relationship between brain insulin and altered cognition status via the Wnt3a/ß-Catenin/GSK-3ß pathway.

Comparison of chemical compositions and aroma characteristics of walnut oil prepared by different roasting processes.

Walnut oil is an edible oil with high nutritional value, and the roasting process influences its quality and flavor. This study aimed to investigate the effects of roasting on the fatty acid composition, bioactive compounds (tocopherols, polyphenols, and phytosterols), and antioxidant capacity of walnut oil. Additionally, the aroma compounds and sensory characteristics were evaluated to comprehensively assess the variations in walnut oil after roasting. Roasting resulted in no notable impact on the fatty acid composition of walnut oil but increased the content of tocopherols and polyphenols in walnut oil, increasing its antioxidant capacity. Heavy roasting (160°C/20 min) reduced the phytosterol content in walnut oil by 2.3%. In total, 146 volatile compounds were detected in both cold-pressed and roasted walnut oil using headspace solid-phase microextraction-gas chromatography-mass spectrometry, and 32 key aroma compounds were identified. Aromatic aldehydes, aliphatic aldehydes, and heterocyclic compounds significantly contributed to fragrant walnut oil. Furthermore, the principal component analysis based on quality characteristics and sensory evaluation indicated that moderate roasting (130°C/20 min, 130°C/30 min, and 160°C/10 min) provided walnut oil with a sweet, nutty, and roasted aroma, as well as high levels of linoleic acid, phytosterols, and γ-tocopherol. Although heavy roasting (160°C/15 min and 160°C/20 min) enhanced the antioxidant capacities of walnut oils due to high levels of polyphenols, the oils exhibited an unpleasant burnt aroma. This study showed that roasting promoted the quality and flavor of walnut oil, and moderate conditions endowed walnut oil with a characteristic-rich flavor while maintaining excellent quality.

External browning mechanism in walnut kernel pellicles under different drying conditions based on the combination of widely-targeted and anthocyanin-targeted metabolomics.

There has been limited research on external browning (EB) of walnut. This work discovered 1888 metabolites and 34 anthocyanins in walnut pellicles (WPs) after three drying methods using widely-targeted and anthocyanin-targeted metabolomics. Based on OPLS-DA and correlation analysis, 64 temperature-responsive metabolites (TRMs; 13 anthocyanins and 51 flavonoids) were identified as critical components in relation to EB. Notably, 14 flavonoids exhibited a strong positive correlation (r > 0.9) with the browning index (BI), with upregulation of >60% after browning. Most of the identified anthocyanins were negatively linked with BI because of degradation (>45%), with correlation coefficients ranging from 0.75 to 0.97. Furthermore, anthocyanidin reductase and laccase were the two key enzymes involved in the EB of WPs, with their activities increasing by 10.57-fold and 1.32-fold, respectively, with increasing drying temperature. A metabolic pathway network of the TRM was built to provide insights into the potential mechanisms underlying EB in WPs.

Construction and antibacterial activities of walnut green husk polysaccharide based silver nanoparticles (AgNPs).

In this paper, the size-controllable nano­silver particles (AgNPs) were synthesized from walnut green husk polysaccharide, and its cytotoxicity and antibacterial activity were evaluated. Firstly, acidic polysaccharide WGHP2 was extracted from walnut green husk, and then the silver ion in AgNO3 was reduced in WGHP2 aqueous solution using NaBH4, so as to synthesize the nano­silver composite. The nano­silver composite was characterized by transmission electron microscope, Fourier infrared spectroscopy, ultraviolet-visible spectrometer, scanning electron microscope, inductively coupled plasma mass spectrometry and X-ray photoelectron spectroscopy. The results show that AgNPs stabilized by WGHP2 are mainly regular spheres with an average particle size distribution of 15.04-19.23 nm. The particle size distribution and morphology of AgNPs changed with the concentration of silver precursor, which is related to the dispersion of silver precursor in polysaccharide aqueous solution and the formation of AgO coordination bond between silver precursor and polysaccharide molecules. These coordination bonds changed the ability of nanoparticles to produce and release Ag+, and thus regulated their antibacterial activity and cytotoxicity, as evidenced by the experimental result of the cytotoxicity of the nano­silver particle against PC12 cells and the bacteriostatic effect on E.coli and S.aureus. Conclusively, WGHP2-Ag has good stability, antibacterial activity and low cytotoxicity.

Walnut protein isolate based emulsion as a promising delivery system enhanced lutein bioaccessibility.

Lutein, a natural pigment with multiple beneficial bioactivities, faces limitations in food processing due to its instability. In this study, we constructed four modified walnut protein isolate (WNPI) based emulsions as emulsion-based delivery systems (EBDS) for lutein fortification. The modification treatments enhanced the encapsulation efficiency of the WNPI-based EBDS on lutein. The modified WNPI-based EBDS exhibited improved storage and digestive stability, as well as increased lutein delivery capability in simulated gastrointestinal conditions. After in vitro digestion, the lutein retention in the modified WNPI-based EBDS was higher than in the untreated WNPI-based EBDS, with a maximum retention of 49.67 ± 1.10 % achieved after ultrasonic modification. Furthermore, the modified WNPI-based EBDS exhibited an elevated lutein bioaccessibility, reaching a maximum value of 40.49 ± 1.29 % after ultrasonic modification, nearly twice as high as the untreated WNPI-based EBDS. Molecular docking analysis indicated a robust affinity between WNPI and lutein, involving hydrogen bonds and hydrophobic interactions. Collectively, this study broadens WNPI's application and provides a foundation for fortifying other fat-soluble bioactive substances.