Search Results (5,475)

Common wheat (Triticum aestivum L.) is a staple food crop for humans, yet it primarily accumulates the non-provitamin A carotenoid lutein and exhibits limited natural variation in provitamin A β-carotene among its various accessions. This characteristic necessitates the development of alternative strategies for provitamin A biofortification. To address this challenge, we targeted key control points in the carotenoid biosynthetic pathway using the CRISPR-Cas9 system in a wheat cultivar Fielder. Specifically, we knocked out the gene encoding lycopene ε-cyclase (LCYE), an enzyme that acts as a gatekeeper opposing the production of β-branch carotenoids. Biochemical analysis of homozygous transgene-free mutant endosperms at 20 days post-anthesis (DPA) revealed marked metabolic rerouting of carotenoid biosynthesis, characterized by differential, line-specific accumulation patterns. Provitamin A carotenoids—specifically β-carotene—increased by 26.1–34.5% relative to wild-type controls, concomitant with elevated 22.9–125.4% for zeaxanthin, 41.6–73.9% for violaxanthin, and 26.2–186.5% for antheraxanthin. However, these gains were offset by drastic lutein reduction in lines 1–4 and 5–1. Consequently, total carotenoid levels displayed non-uniform responses, with line 5–1 exhibiting a modest decrease relative to wild-type. Moreover, the mutant lines exhibited elevated levels of amylose and soluble sugar, and the seed coats and endosperms of the triple homozygous transgene-free mutant lines exhibited an orange-yellow hue. In conclusion, we have successfully developed novel carotenoids biofortified wheat lines through a gene-editing approach. This study demonstrates targeted redirection of carotenoid biosynthesis via gene editing as an effective strategy to enhance the nutritional value of commercial wheat and mitigate micronutrient deficiencies in modern food systems. Full article

When cultivating a selected field crop (alfalfa), we aimed to examine its positive effects on the variability of soil infiltration capacity. A total of 21 monitoring points were proposed for investigating soil hydraulic conductivity on the targeted plot with a total area of 47.64 ha, divided between the irrigated and non-irrigated areas. The plot is located outside the village of Oponice (Slovak Republic) and is managed by VPP Kolíňany. The study of hydraulic conductivity has been ongoing on the selected plot for several years. The presented results come from a two-year experiment, during which work operations related to the cultivation of alfalfa were carried out on the plot. The unsaturated hydraulic conductivity of the soil was assessed several times a year using a Mini Disk Infiltrometer, while soil moisture at monitoring points and the dependence of the measurement date (work operations, weather conditions) were also monitored. The average soil moisture content in the pilot measurements reached 18.77% vol. (CV = 1.44%), in the secondary measurements 17.21% vol. (CV 20.49%), in tertiary measurements 15.27% vol. (CV = 10.38%), and in the last measurements 15.26% vol. (CV = 10%), which ultimately represents a positive result of soil moisture balance. To test the significance of the differences between measurements taken across the entire surveyed plot, a one-factor ANOVA analysis was used to compare the measurement dates. The results showed a statistically significant difference when examining the effect of the time period of soil infiltration capacity monitoring between all measurements (p = 0.004). The mutual combinations of individual measurement dates were mostly significant (p = 0.03 for IDM1, IDM2; p = 0.003 for IDM2, IDM3), except for one case without a significant difference (IDM3, IDM4; p = 0.52). The second hypothesis was confirmed only at some monitoring points, and it can be stated that the irrigated area had a more significant effect on the soil infiltration capacity. The results obtained by the Shapiro–Wilk test and Welch’s test in irrigated and non-irrigated areas at individual dates showed statistically insignificant differences in three cases (IDM1, p = 0.123; IDM3, p = 0.382; IDM4, p = 0.445) and statistically significant in one case (IDM2, p = 0.0175). Based on the hypotheses and the results obtained, it can be said that the work tasks performed have a decisive influence on the infiltration capacity of the soil. The phenomenon of “water resistance” did not manifest itself in our research on soil infiltration capacity. The results were also evaluated using ArcGIS software 10.0 to display the spatial variability of soil hydraulic conductivity. The last application used to evaluate the results was Orange software 3.40.0, using clustering maps and hierarchical clustering. The results also pointed to variability depending on the dates of monitoring. Full article

The biochars obtained by pyrolyzing tomato stems at temperatures of 400, 500, 600, and 700 °C were characterized, and their ability to absorb anionic (Direct Orange 26, DO26) and cationic (Rhodamine B, RhB) dyes from aqueous solutions was investigated. The effects of solution pH and ionic strength were studied. It was found that the adsorption process of both dyes was pH-dependent, but no effect of ionic strength was observed. The kinetics of dye adsorption on biochars were well described by the pseudo-second-order model. The equilibrium adsorption data were analyzed using the Freundlich, Langmuir, and Temkin isotherms. All three equations described dye adsorption on biochars quite well, although a slightly better fit was observed for the Freundlich model. The maximum adsorption capacities of BCs ranged from 54.44 mg/g (BC400) to 108.1 mg/g (BC700) for DO26 and from 4.483 mg/g (BC700) to 8.887 mg/g (BC400) for RhB. The study reveals that biochars derived from tomato stems can be used as efficient, low-cost adsorbents for the removal of anionic and cationic dyes from water. Full article

Laccases are environmentally friendly biocatalysts capable of oxidizing a broad range of organic pollutants by using molecular oxygen as the sole electron acceptor, producing water as the only by-product. The cofactor-independent activity makes them attractive for sustainable wastewater treatment, particularly for the removal of synthetic dyes. In this study, laccase from Trametes versicolor was evaluated for the decolorization of the azo dye Methyl Orange, with emphasis on the effects of redox mediators and metal ions. Laccase alone exhibited maximum activity at pH 3.0, achieving 40.5% decolorization after 24 h. The addition of redox mediators markedly enhanced dye removal. Both synthetic mediators (ABTS, HBT, and TEMPO) and natural mediators (p-coumaric acid, vanillin, gallic acid, ascorbic acid, and syringaldehyde) improved decolorization in a concentration-dependent manner. Among them, ABTS and syringaldehyde were the most effective, achieving 98.8% and 96.9% decolorization, respectively, at 0.2 mM after 24 h, with syringaldehyde offering the advantage of natural origin. Metal ions also modulated laccase activity, with several ions enhancing decolorization at 0.5 mM, whereas higher concentrations were generally inhibitory; potassium ions showed the strongest enhancement (80.3%). Molecular docking analysis suggested favorable binding of Methyl Orange within the laccase active pocket, supported by predicted hydrogen-bonding and hydrophobic interactions. The molecular docking analysis was performed using a representative T. versicolor laccase structure and provides supportive computational insight into plausible enzyme–dye interactions rather than isoform-specific mechanistic validation. Overall, these findings demonstrate that optimization of mediators and metal ion concentrations significantly improve laccase-mediated dye decolorization, while docking provides supportive computational insight into possible enzyme–dye interactions relevant to sustainable wastewater treatment. Full article

During the examination of a bicolor angelfish (Centropyge bicolor) imported from the Philippines and intended for sale on the Polish market, clinical signs of weakness and respiratory distress were observed. Mild hyperemia was noted along the lateral sides of the body and around the mouth. Necropsy revealed the presence of five orange-colored trematodes in the intestinal lumen, with an average body length of 3.12 mm. Based on morphological features and molecular analyses, the parasite was identified as Paragyiauchen sp. The following gene loci were amplified: the gene-encoding component of the large ribosomal subunit (28S rRNA), the gene-encoding NADH dehydrogenase subunit 1 (ND1), the gene cluster 5.8S rRNA-ITS2-28S rRNA and the gene cluster ITS2-28S rRNA. Bacterial species identification using MALDI-TOF MS revealed the presence of three species: Shewanella putrefaciens and Brevundimonas diminuta isolated from the head kidney, and Aeromonas caviae isolated from the liver. This study documents the first detection of representatives of Paragyliauchen genus in C. bicolor imported to Europe and highlights the potential risk of introducing new parasites and opportunistic bacterial pathogens through the ornamental fish trade. These findings emphasize the need for parasitological and microbiological screening of imported ornamental fish. Full article

Background: The eco-friendly synthesis of silver nanoparticles (AgNPs) utilizing medicinal flora presents a viable strategy for the development of multifunctional agents exhibiting antimicrobial, antioxidant, anti-inflammatory, and anticancer properties. This investigation aims to elucidate the phytochemical composition of Calotropis gigantea and its contribution to the synthesis of CG-AgNPs that demonstrate efficacy against Helicobacter pylori and gastric cancer cell lines. Methods: The aqueous plant leaf extract of C. gigantea underwent comprehensive analysis via gas chromatography-mass spectrometry (GC-MS), identifying a total of 25 bioactive constituents, including oleic and oxalic acid derivatives. The fabrication and analysis of silver nanoparticles (AgNPs) were performed utilizing methodologies including ultraviolet-visible (UV–Vis) spectroscopy, X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), high-resolution transmission electron microscopy (HR-TEM), dynamic light scattering (DLS), and assessments of zeta potential. Antibacterial efficacy was evaluated through methods including agar well diffusion, time-kill kinetics, and biofilm assays. The cytotoxic impact on AGS gastric cancer cells was investigated using MTT assays, DAPI staining, and acridine orange/ethidium bromide (AO/EtBr) staining techniques. The assessment of antioxidant potential was performed utilizing DPPH and ABTS assays. The anti-inflammatory properties were analyzed through protein denaturation and membrane stabilization tests. Results: CG-AgNPs exhibited a spherical morphology (11–17 nm) with commendable stability, denoted by using zeta potential analysis measurement of −30.2 mV. The antibacterial activity showed a significant inhibition zone of 16.00 ± 0.17 mm at a concentration of 50 µg/mL against H. pylori, in addition to notable biofilm disruption. The viability of AGS cells was reduced by 61% at a concentration of 100 micrograms per milliliter, with apoptosis being confirmed through relevant assays. The antioxidant potential varied from 18% to 83% (DPPH) and reached 74% (ABTS) at a concentration of 100 µg/mL. The anti-inflammatory assays indicated a BSA denaturation inhibition ranging from 45% to 80% and a membrane stabilization effect between 54% and 85%. Conclusions: CG-AgNPs exhibit substantial antibacterial, antioxidant, anti-inflammatory, and anticancer activities, underscoring their pharmaceutical potential, particularly for combating antibiotic-resistant pathogens and gastric malignancies. Full article

Importance: Uncomplicated urinary tract infections (uUTIs) are among the most common bacterial infections and are typically treated with existing oral antibiotics. In 2024–2025, the FDA approved two new oral agents, sulopenem etzadroxil/probenecid and gepotidacin, via expedited review pathways, for the treatment of uUTIs. Objective: To assess the clinical significance and regulatory evidence supporting FDA approval of sulopenem/probenecid and gepotidacin for uUTI, and to analyze the adherence of pivotal phase III trials to regulatory standards for approval and FDA guidelines. Design, Setting, and Participants: Comparative descriptive analysis of publicly available data from phase 3 randomized, double-blind, active-controlled clinical trials submitted to the FDA for approval. Pivotal phase III clinical trial data and FDA integrated reviews, guidance for the industry, and approved drug labels. Adult women with uUTI enrolled in pivotal phase III clinical trials, with subgroup analysis stratified by pathogen susceptibility to comparator antibiotics. Interventions: Sulopenem/probenecid was compared to ciprofloxacin and to amoxicillin/clavulanate and gepotidacin was compared to nitrofurantoin. Main Outcomes and Measures: Primary endpoints were clinical and microbiological responses assessed in the microbiologic modified intention-to-treat (micro-MITT) subjects. Safety outcomes and FDA regulatory determinations were also assessed. Results: Sulopenem/probenecid was inferior to ciprofloxacin and noninferior to amoxicillin/clavulanate in subjects with susceptible pathogens, and superior in subjects with resistant pathogens. Gepotidacin showed noninferiority to nitrofurantoin in one trial and superiority in another. Trials excluded randomized subjects, included post-randomization subgroup analyses, and enrolled control-arm subjects with resistant pathogens. Clinical cure rates were lower than historical comparators. Both new antibiotics had higher adverse event rates than controls. Conclusions and Relevance: Pivotal clinical trials for sulopenem/probenecid and gepotidacin for uUTI had significant design limitations and relied on surrogate endpoints of limited clinical interpretability, undermining reliability and clinical relevance. Future antibiotic development for uUTI should prioritize representative populations, standard-of-care comparators, clinically meaningful outcomes and robust, well-controlled trial designs to ensure meaningful clinical evidence of safety and efficacy. Full article

This study aimed to investigate the flavor quality and starch physicochemical properties of three orange-fleshed sweet potato varieties commonly cultivated in northeastern China. Fresh and baked samples were evaluated using sensory analysis, electronic nose and tongue, gas chromatography-mass spectrometry for volatile compound profiling, and chemical methods for starch characterization. Liankaoshu 1 exhibited the highest sensory score (88.6), reflecting superior taste and aroma. A total of 70 volatile organic compounds were identified, including β-damascenone, maltol, and β-ionone, as key contributors to baked flavor. Significant varietal differences were found in starch content, particle size, and crystalline structures, with Pushu 32 showing CA-type crystals, Yanshu 25 A-type, and Liankaoshu 1 B-type. Baking increased maltose and soluble sugar levels, which were strongly correlated with sensory attributes. Spearman correlation analysis revealed that sweetness and overall sensory scores were significantly and positively correlated with maltose, soluble sugar, and reducing sugar contents, as well as starch particle size parameters (p ≤ 0.05). These results indicate that starch structural characteristics and saccharification efficiency play critical roles in regulating flavor formation during baking, providing a theoretical basis for sweet potato breeding and processing optimization. Full article

This study investigates the potential of pure chitosan powder as an effective, sustainable, and low-cost adsorbent for the removal of synthetic dyes from aqueous media. The work demonstrates the potential of pristine chitosan for practical wastewater treatment applications by adsorbing two commonly used textile dyes, methyl orange (MO) and methylene blue (MB). To elucidate the adsorption mechanism, chitosan was comprehensively characterized using zeta potential analysis, Fourier Transform Infrared Spectroscopy (FTIR), X-ray Diffraction (XRD), Scanning Electron Microscopy coupled with Energy-Dispersive X-ray Spectroscopy (SEM–EDX), Thermogravimetric Analysis (TGA), Brunauer–Emmett–Teller (BET) surface area analysis, and point of zero charge (pHpzc) determination. FTIR analysis revealed notable shifts in –NH₂ and –OH functional groups after dye adsorption, confirming their involvement in electrostatic interactions and hydrogen bonding with MO and MB. SEM images demonstrated significant surface morphological changes following adsorption, while EDX spectra confirmed successful dye uptake through the appearance of sulfur and nitrogen signals characteristic of MO and MB, respectively. Zeta potential and pHpzc results explained the strong pH-dependent adsorption behavior, highlighting favorable electrostatic attraction between chitosan and the ionic dyes. The optimum adsorption conditions were achieved at adsorbent dosages of 0.5 g for MO and 1.0 g for MB, a contact time of 30 min, initial dye concentrations of 20 and 100 mg/L, and solution pH values of 3 for MO and 9 for MB at room temperature. The adsorption data fit the Langmuir isotherm model, indicating monolayer adsorption on a homogeneous chitosan surface, with maximum adsorption capacities of 7.843 mg/g for MO and 7.605 mg/g for MB. Kinetic studies showed that adsorption followed the pseudo-second-order model, suggesting chemisorption as the dominant mechanism. Thermodynamic analysis indicated that the adsorption process was endothermic and non-spontaneous under the investigated conditions. In conclusion, these findings demonstrate that unmodified chitosan is a practical, eco-friendly adsorbent for dye removal, achieving removal efficiencies comparable to many modified chitosan composites, and represents a promising candidate for sustainable wastewater treatment. Full article

Background: Dietary guidance for type 2 diabetes mellitus (T2DM) frequently discourages fruit consumption due to intrinsic sugars, despite extensive evidence supporting the anti-diabetic properties of fruit-derived polyphenols. This reductionist, carbohydrate-only model inadequately reflects the complex bioactive matrices of whole fruits. Objective: To develop an integrated analytical framework that quantitatively balances the predicted anti-diabetic bioactivity of fruit polyphenols against their glycemic burden, and to apply this model to fruits commonly consumed in the Korean market. Methods: Nutritional and phytochemical composition data for five fruits sourced from Korea and India were obtained from national food databases to ensure broader phytochemical diversity. Six representative polyphenols were selected based on abundance and reported significance. Molecular docking was conducted against α-glucosidase and peroxisome proliferator-activated receptor gamma (PPAR-γ) to estimate target-specific affinity, and a “Total Predicted Anti-Diabetic Score” (TPAS) was computed by weighting docking potency by compound concentration. A risk–benefit matrix integrating TPAS and sugar content was applied to classify fruits, followed by a cultivar-level comparison of major grape varieties. Results: Hesperidin exhibited the strongest predicted PPAR-γ binding (−9.3 kcal/mol). Among whole fruits, grapes and oranges showed the highest TPAS (593.19 and 448.77, respectively), placing them in the “redemptive choice” category (high benefit/high glycemic risk). Comparative cultivar analysis identified the Campbell Early grape as the most advantageous option, with a Holistic Value Score (HVS) of 9.5, notably higher than Shine Muscat (3.9). Conclusions: This study presents a structured, computation-driven framework capable of integrating phytochemical potency and nutritional risk into a unified metric for dietary evaluation. Despite higher sugar content, fruits rich in potent polyphenols may confer substantial metabolic benefit when consumed judiciously. These findings challenge sugar-centric dietary models and provide an evidence-based tool for consumer-level guidance in T2DM dietary management. Full article

Bismuth oxychloride (BiOCl)– copper oxide (CuO) heterostructures were synthesized via a solvothermal route and assessed as visible-light-driven photocatalysts for methyl orange (MO) degradation. Different CuO loadings deposited on BiOCl microspheres were investigated to identify the optimal composition. Structural and physicochemical characterization revealed that low CuO content (0.6 wt. %) promoted uniform dispersion and enhanced surface area, whereas higher loadings led to nonuniform coverage and reduced photocatalytic efficiency. Operating conditions were optimized using response surface methodology based on a central composite design, considering catalyst dosage (0.1–0.8 g L⁻¹) and pH (4–9). The highest degradation efficiency (~50% after 60 min irradiation) was achieved at pH = 4 and a catalyst dosage of 0.8 g L⁻¹ using the BiOCl surface modified with 0.6% CuO. Kinetic analysis followed a pseudo-first-order model. Mass spectrometry identified transient intermediates associated with demethylation and desulfonation pathways, while radical scavenger experiments confirmed hydroxyl radicals (^•OH) as the dominant oxidizing species, with a secondary contribution from superoxide radicals (^•O₂⁻). These results highlight the critical role of CuO dispersion and interfacial quality in enhancing charge separation and photocatalytic performance, providing practical guidelines for the rational design of BiOX-CuO heterostructures for water remediation applications. Full article

Blackberry (Rubus spp.) is a highly perishable fruit rich in bioactive compounds, particularly anthocyanins, which are associated with significant health benefits. This study investigated the application of nanofiltration using a pilot-scale spiral-wound module (DOW^® NF90-2540) as a mild technology to concentrate phenolic compounds, especially anthocyanins, in blackberry juice. The process achieved concentration factors (CF) of 2.2 for monomeric anthocyanins and 1.9 for total phenolic content (TPC), reaching values of 54.3 mg C3G·100 mL⁻¹ and 326.85 mg GAE·100 mL⁻¹, respectively. The antioxidant capacity (ABTS⁺ and DPPH methods) also increased significantly in the concentrated fraction (CF 1.9 and 1.7, respectively). Stability of the concentrated juice was evaluated during 90 days of frozen storage, showing that low temperatures effectively preserved anthocyanin levels and visual quality, with only minor variations in color parameters (L*, a*, b*). Furthermore, the concentrated blackberry juice was successfully incorporated into apple–orange juice blends, generating formulations with progressively increased phenolic content, antioxidant activity, and red color intensity as the proportion of blackberry concentrate increased. Anthocyanin bioaccessibility in these juice blends was also evaluated and was not proportional to the increase in anthocyanin content. Strong correlations between anthocyanin concentration, antioxidant capacity, and CIELAB color parameters highlight the dual functional and technological role of blackberry compounds. In conclusion, this study demonstrates the feasibility of nanofiltration as a mild and efficient strategy for concentrating anthocyanins and phenolic compounds from blackberry juice while preserving physicochemical quality and color attributes. Full article

Clementine, mandarin, and orange peels, which are usually discarded, can serve as promising, sustainable dietary supplements with beneficial compositions, as demonstrated in this study. Citrus peels are low in ash, fat, and protein, but high in moisture, fibre, sugar, and polyunsaturated fatty acids (PUFAs) (up to 60%). They contain high levels of omega-3 and omega-6 fatty acids, up to 30% each, making them a good health-promoting source, as shown by the values of nutritional indices as follows: PUFA/saturated fatty acid (SFA) (1.94 to 2.30), monounsaturated fatty acid (MUFA)/SFA (0.39 to 0.84), and PUFA/MUFA (2.37 to 5.82). Essential macro elements (K > Ca > Mg > S > P > Na) and trace elements (Fe > Zn > Mn > Cu > Cr > Mo > Co > Se) are unevenly distributed among the peels, along with non-essential elements, with Al (37 to 51 mg/kg) and Sr (17 to 30 mg/kg) predominating. Rare elements in food, such as V and W, are found up to 41 and 79 µg/kg respectively, followed by Nb > Ga > Y > Ge (5 to 11 µg/kg). Although citrus peels have a nutrient-dense composition, their monitoring must be ensured before inclusion in the common diet, particularly regarding non-essential elements, as for most of them the reference doses are not established and they could be harmful to human health. Full article

Background/Objectives: Cardiomyopathy (CM) is a leading cause of morbidity and mortality in Duchenne muscular dystrophy (DMD) patients. Ferroptosis, an iron-dependent form of cell death characterized by lipid peroxidation, is implicated in various cardiovascular diseases. However, the role of ferroptosis in DMD-CM remains unexplored. Methods: Here, we used dystrophin and utrophin double-knockout (mdx:utr^−/−) mice as a model that exhibits cardiac pathological phenotypes similar to those seen in DMD patients to investigate the potential role of ferroptosis. Results: We observed an increased level of iron deposition and lipid peroxidation in the hearts of mdx:utr^−/− mice. Live/Dead viability assays revealed that mdx:utr^−/− cardiomyocytes exhibited greater susceptibility to ferroptosis than WT cardiomyocytes both at baseline and upon exposure to ferroptosis inducers. We also used mdx:utr^−/− mice with a heterozygous sarcolipin (SLN) knockout background (sln^+/−) to investigate the effect of SLN reduction on ferroptosis susceptibility in DMD-CM. Notably, ferroptosis was significantly suppressed in cardiomyocytes from mdx:utr^−/−:sln^+/− mice (p < 0.01). Western blot analysis confirmed the upregulation of transferrin receptor 1 (TfR1) and 15-lipoxygenase-1 (15LOX1), along with the downregulation of heme oxygenase-1 (HMOX-1) and ferroptosis suppressor protein 1 (FSP1) in mdx:utr^−/− hearts, while glutathione peroxidase 4 (GPX4) levels remained unchanged. A similar pattern of alterations in ferroptosis-related biomarkers was observed in human heart samples from DMD patients compared to healthy controls. Conclusions: Our results provide direct evidence that ferroptosis contributes to the pathology of DMD-CM and suggest that reducing SLN expression and inhibiting ferroptosis may represent potential therapeutic strategies for this condition. Full article

Background/Objectives: The physiological disorder known as “orange spot” in Juglans regia L. cv. Chandler is an emerging kernel quality issue of increasing concern for the Chilean walnut industry. Characterized by a localized orange discoloration of the embryo, its etiology remains unknown. This study aimed to determine whether orange spot is associated with bacterial infection and to elucidate the metabolic alterations underlying its development, testing the hypothesis that it results from oxidative imbalance and lipid disorganization. Methods: Untargeted metabolomic profiling and fatty acid methyl ester (FAME) analysis were applied to compare affected (orange) and unaffected (white) walnut kernels collected from a commercial orchard in Melipilla, Chile, during the 2018 and 2022 seasons. Bacterial DNA was screened by PCR amplification of the 16S rRNA gene (799F/1193R). Results: Orange spot incidence reached 31–34% across seasons. No bacterial-sized amplicons were detected in either kernel type, indicating the absence of detectable bacterial DNA under the analyzed conditions. Metabolomic analyses revealed significant alterations in glyoxylate and dicarboxylate metabolism, glutamate-related pathways, and glycerophospholipid metabolism. Affected kernels exhibited reduced levels of L-glutamic acid and antioxidant-associated metabolites, alongside increased gluconic, citric and quinic acid, consistent with the redox imbalance and membrane oxidative degradation. FAME analysis showed higher total fatty acid content in affected kernels, suggesting intensified lipid turnover associated under oxidative stress. Conclusions: Orange spot is a physiological, non-infectious disorder linked to premature oxidative stress and lipid disorganization, negatively affecting walnut kernel quality. Full article