Long-chain metabolites produced through hepatic and microbiota-associated ω-oxidation of vitamin E are increasingly recognized as bioactive regulators of lipid metabolism and inflammatory pathways. These properties suggest interesting opportunities in drug development and garcinoic acid (GA) - a δ-tocotrienol-derived natural product and a chemically accessible analogue of these metabolites - is a useful probe for investigating their molecular and pharmacological properties. GA has been identified as an agonist of pregnane X receptor, a modulator of peroxisome proliferator-activated receptor γ, and an inhibitor of enzymes involved in biosynthesis of inflammatory lipid mediators, including 5-lipoxygenase and microsomal prostaglandin E₂ synthase-1, while its effects on cyclooxygenase pathways are context-dependent. Through these activities, GA functionally links xenobiotic sensing, lipid metabolism, and inflammatory regulation across selected tissues, including the intestine, liver and brain. GA can be viewed within the broader framework of metabolite-inspired pharmacology, highlighting how plant-derived natural products that mimic endogenous or microbiota-associated metabolites may carry privileged recognition motifs for pharmacological targets. These aspects, together with the biological effects of GA identified in preclinical models, suggest therapeutic potential. However, direct applications are constrained by unfavorable pharmacokinetic properties, supporting its use as a biomimetic scaffold for the design of improved modulators inspired by vitamin E metabolite biology.

Eggplant (Solanum melongena L.) processing produces large amounts of mostly unexploited by-products, despite their richness in bioactive compounds. These residues represent promising sources of functional ingredients for nutraceutical, food, and pharmaceutical applications. Among eggplant metabolites, phenolic compounds, particularly chlorogenic acids (caffeoylquinic acids, CQAs), stand out for their health-promoting properties. This study provides a comprehensive characterization of eggplant pulp, peel, calyx, and leaves as sources of phenolic compounds, with emphasis on CQAs. Non-conventional extraction techniques, including microwave- and ultrasound (bath- and probe)-assisted extraction were compared and optimized through a Design of Experiments to maximize 5-CQA (chlorogenic acid) recovery while minimizing isomer degradation. Antioxidant capacity was evaluated through spectrophotometric assays, and cytotoxicity was assessed in Caco-2 cells to determine extract functionality and safety. Polyphenol profiles were determined by HPLC-PDA and tentatively identified by UHPLC-HRMS/MS based on HRMS/MS databases. The stability of 5-CQA and its main isomers was monitored over four weeks, supporting the valorization of eggplant by-products.

This study aimed to assess the effects of rearing system on egg quality traits, eggshell characteristics including eggshell breaking strength (EBS), yolk vitamin D-3 (YD3) content and keel bone damage (KBD) occurrence. Sixty White Leghorn hens (18-week-old) were raised in either indoor barn or free-range system (BA and FR, respectively; 3 replicates of 10 hens) and fed a basal diet for 52 weeks. Egg production was monitored daily, while feed intake was registered weekly. In both autumn-winter and spring-summer periods, 204 eggs/rearing system were collected to assess egg and eggshell quality traits. The YD3 content was determined on six eggs/rearing system/period, while KBD occurrence was evaluated on all hens. FR hens presented a lower feed consumption than BA ones in both periods, resulting in a reduced Ca and vitamin D-3 supply. During the autumn-winter, FR eggs showed similar YD3 content than BA ones (19.63 vs. 17.83 mu g/100 g DM yolk; p = 0.33), while EBS tended to be lower (4.29 vs. 4.38 kg, respectively; p = 0.08). In spring-summer, FR eggs had identical EBS (3.63 vs. 3.63 kg; p = 0.97) and higher YD3 content than BA (25.39 vs. 20.81 mu g/100 g DM yolk; p < 0.01). The estimated endogenous vitamin D-3 synthesis accounted for 22% and 43% of the total YD3 content in autumn-winter and spring-summer, respectively. KBD occurrence was similar in both periods. These findings suggest that sunlight exposure in free-range system can positively affect eggshell quality traits (particularly EBS) and yolk vitamin D-3 content likely by stimulating vitamin D-3 synthesis, although with seasonal variations.

Autoreactive lymphocytes are thought to contribute to tissue injury in the pathogenesis of multiple sclerosis (MS). However, reliable biomarkers reflecting myelin-specific immune responses and disease activity remain limited. In HLA-DRB1*15:01-positive individuals, the myelin basic protein (MBP) epitope MBP85-99 represents a well-characterized immunodominant target of CD4+ T cells. Using classical spectratyping, we characterized the peripheral T-cell receptor (TCR) β-chain repertoire reactive to MBP85-99 in individuals with MS, according to their HLA-DRB1 haplotypes. Selected rearrangements were further evaluated in cerebrospinal fluid (CSF), cytokine-defined T-cell subsets, and longitudinal samples collected during interferon beta-1a treatment. A restricted set of shared ("public") MBP85-99-reactive TCR rearrangements was identified in individuals with MS and differed from the repertoire observed in healthy controls. These TCRs were more frequently detected during periods of active disease than during remission. Two disease-associated public rearrangements were enriched in active MS, whereas one rearrangement was preferentially observed in healthy controls. HLA-DRB1*15:01-restricted MBP85-99-specific public TCR signatures were associated with inflammatory disease activity in MS and may help distinguish pathogenic from non-pathogenic autoreactive responses. In a longitudinal sub-cohort (n = 15, ~10-year follow-up), the RSI of the principal disease-associated rearrangement TRBV19-TRBJ2.4 correlated positively with cumulative relapse burden (Spearman ρ = 0.782, p = 0.002), providing preliminary evidence that these TCR signatures reflect long-term inflammatory disease activity. These findings support further validation of antigen-specific TCR profiling as a biomarker strategy in MS using contemporary high-throughput immune repertoire technologies.

Chronic obstructive pulmonary disease (COPD) is a lung disease with an incidence expected to rise in the coming years. Beyond their hypolipidemic effects, statins, including atorvastatin (ATV), exhibit pleiotropic activities, and several studies have reported their efficacy in treating COPD and its exacerbations. This study aims to develop and characterize a spray-dried high-content (>90%) ATV powder for inhalation, also evaluating its effects in BEAS-2B lung epithelial cells. ATV was co-spray-dried with leucine resulting in a production yield of approximately 50%. SEM images revealed small (similar to 1-2 mu m) and spherical particles with an irregular surface. Aerodynamic evaluation using a multi-stage liquid impinger (MSLI) showed a respirable fraction between 41 and 52%, emitted fractions of 58-66%, and mass median aerodynamic diameters (MMAD) of 2.6-3.0 mu m. To facilitate the dosing for in vitro studies, a blend of the spray-dried ATV with lactose and magnesium stearate was prepared. LPS-stimulated BEAS-2B cells were used to investigate the antioxidant and anti-inflammatory potential of ATV. ATV solution and, more efficiently, the spray-dried ATV blend reduced the extracellular H2O2 and IL-6 levels. Excipients alone showed no anti-inflammatory effect. Real-time PCR showed that both the ATV solution and ATV blend downregulated IL-6 expression whereas TGF-beta, PPAR-gamma, and PLIN-2 were upregulated. In conclusion, an inhalable ATV dry powder was developed, demonstrating strong anti-inflammatory and antioxidant effects in LPS-treated BEAS-2 cells.

Over the past century, research on vitamin E has evolved from its initial identification as a fertility factor to the recognition of α-tocopherol (α-TOH) as an essential micronutrient with antioxidant and gene regulatory functions. Despite extensive investigation, controversies persist regarding its precise biological role, clinical efficacy, and classification as a vitamin.This review summarizes scientific milestones in vitamin E (tocochromanol) research. It highlights recent advances in understanding the absorption, metabolism, and molecular mechanisms of vitamin E. The review also outlines unresolved questions and methodological challenges. The focus is on α-TOH and its metabolites, which exhibit biological activities beyond classical antioxidant effects, such as the regulation of inflammation, lipid metabolism, and immune responses.Evidence from genetic, biochemical, and clinical studies supports the essentiality of α-TOH in humans, as demonstrated by ataxia with vitamin E deficiency (AVED). Other tocochromanols and their metabolites exhibit promising biological activities that suggest potential therapeutic applications. However, their physiological relevance must be confirmed. Advances in metabolomic profiling and molecular modeling now allow for a more comprehensive characterization of vitamin E metabolism and function.Understanding the molecular pathways and biological roles of vitamin E and its derivatives is crucial for refining its definition, establishing evidence-based dietary recommendations, and evaluating its potential in disease prevention and therapy.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a highly prevalent liver disorder deriving from the chronic exposure to hepatocyte steatosis and its lipotoxicity effects, as mitochondrial dysfunction, overactivation of stress response and inflammatory genes, and cell death signaling. Genomics and post-genomic disciplines are now offering unprecedent opportunities in disease mechanisms characterization, precision diagnostics, and treatment. These disciplines include nutrigenomics, i.e. the use of genomics techniques to study the health effects of the interaction between diet/nutrients and the genome. Its applications can be particularly useful to study lifestyle and dietary modifications, and to assess the efficacy of nutritional interventions through the criteria of precision medicine. Specific examples include interventions with fat-soluble vitamins and other lipid nutrients as omega-3 fatty acids, which have shown cytoprotective properties and molecular effects useful in modulating key steps of the hepatocellular lipotoxicity process, such as lipid biosynthesis, lipid peroxidation inflammatory gene activation and cell death signaling. Other applications of nutrigenomics concern drug discovery and preclinical studies to explore therapeutic mechanisms, efficacy and safety of new vitamin products and nutraceuticals. This review discusses current evidence on hepatocyte lipotoxicity and its nutrigenomic exploration to identify disease mechanisms, nutritional defects and intervention strategies with these lipid nutrients. Their properties, limitations, and potential for translation in the prevention and clinical management of MASLD are critically evaluated.

Naturally occurring vitamin E is a lipophilic plant-derived molecule corresponding to the 2 R forms of alpha-tocopherol. A series of natural analogs or tocochromanols are present in nature, including β-, γ- and δ-tocopherol (βT, γT, δT), the corresponding tocotrienols (αTE, βTE, γTE, δTE) and tocomonoenols. Differences between these analogs as lipophilic antioxidants and modulators of molecular processes suggest specific therapeutic properties against various disorders associated with acute and chronic inflammation. However, hepatic metabolism of these compounds via cytochrome P450-initiated side chain ω-oxidation involves the production of long-chain metabolites (LCMs) followed by intermediate (ICMs) and short-chain metabolites (SCMs), respectively. Despite the initial studies indicating these metabolites as catabolic-end products, recent findings identify their importance in providing biological functions. In this scope, LCMs, especially 13'-carboxychromanols (13'-COOHs), have been reported to hold stronger anti-inflammatory capacity than their unmetabolized precursors due to their ability to inhibit 5-lipoxygenase and cyclooxygenase-catalyzed eicosanoid formation, as well as their modulation of the pro-inflammatory transcriptional protein nuclear factor κB (NF-κB). Also, these LCMs have been reported to enhance detoxification and lipid metabolism pathways associated with cellular inflammation by modulating the nuclear receptors peroxisome proliferator-activated receptor-γ (PPARγ) and pregnane x receptor (PXR). These properties of LCMs will be described in this narrative review article focusing on recent information regarding their bioavailability, anti-inflammatory effects, and mechanisms of action in acute and chronic inflammatory disorders.

Storage conditions significantly impact the quality and functional properties of extra-virgin olive oil (EVOO). This study investigated the impact of light and dark storage on the nutritional quality of Umbrian EVOO and its effectiveness in tissue repair. The research aimed to simulate real-world conditions occurring during transport, retail, and domestic storage. Light exposure accelerated EVOO oxidation, significantly affecting peroxide levels (ranging from 5.19 to 24.30 meq O2/kg of oil), total antioxidant capacity (measured spectrophotometrically, collectively ranging from 399.47 to 684.63 mg TE/kg of oil), and phenolic compound concentrations, particularly secoiridoids, lignans, and flavonoids (measured by HPLC, collectively ranging from 41.92 to 169.74 mg/kg of oil). Statistically significant differences (p < 0.01) were recorded between the control sample and the others in almost all cases, after storage. For instance, both light and dark exposure over a 24-month period resulted in a marked reduction (p < 0.01) in oleocanthal, pinoresinol, luteolin, and apigenin. Pigment levels were also affected, representing a rapid and cost-effective indicator of product oxidative degradation. The loss of phenolic compounds (especially oleacein and oleocanthal, which showed the most significant reductions of approximately 75% and 60%, respectively), impaired the EVOO’s wound-healing properties, affecting key tissue regeneration processes such as keratinocyte migration, hyaluronic acid synthesis, and angiogenesis. Notably, oleocanthal and oleacein, present at higher concentrations in fresh extracts, emerged as the primary contributors to the observed dermal effects and wound-healing processes, demonstrating a significant highest efficacy (p < 0.0001) in promoting wound closure. These findings underscore the critical role of EVOO storage in preserving its sensory properties and labile components with tissue repair and regeneration functions.

[This corrects the article DOI: 10.3389/fendo.2023.1063916.].

The aim of this study was to analyze how recombinant rabbit NGF (Nerve Growth Factor) encapsulated in chitosan (rrβNGFch) affects sperm viability, motility, capacitation, acrosome reaction (AR), kinetic traits, and apoptosis after 30 min and 2 h of storage. Specific intracellular signaling pathways associated with either cell survival, such as protein kinase B (AKT) and extracellular signal-regulated kinases 1/2 (ERK1/2), or programmed cell death, such as c-Jun N-terminal kinase (JNK), were also analyzed. The results confirmed the effect of rrβNGFch on capacitation and AR, whereas a longer storage time (2 h) decreased all qualitative sperm traits. AKT and JNK did not show treatment-dependent activation and lacked a correlation with functional traits, as shown by ERK1/2. These findings suggest that rrβNGFch may promote the functional activation of sperm cells, particularly during early incubation. The increase in capacitation and AR was not linked to significant changes in pathways related to cell survival or death, indicating a specific action of the treatment. In contrast, prolonged storage negatively affected all sperm parameters. ERK1/2 activation correlated with capacitation, AR, and apoptosis, supporting its role as an NGF downstream mediator. Further studies should analyze other molecular mechanisms of sperm and the potential applications of NGF in assisted reproduction.

In response to Yoshiyasu Takefuji's critique regarding the use of Principal Component Analysis (PCA) and Partial Least Squares Discriminant Analysis (PLS-DA) in the study "Melatonin Repairs the Lipidome of Human Hepatocytes Exposed to Cd and Free Fatty Acid-Induced Lipotoxicity," we provide a methodological clarification. PCA and PLS-DA are well-established, widely validated tools for exploratory analysis of high-dimensional omics data, including lipidomics data. Although these methods are linear, they are appropriate for capturing systematic and directional variations in complex biological systems, particularly in controlled in vitro models like ours. Our analytical approach integrates PCA and PLS-DA with rigorous statistical testing, data transformations, and biological validation, ensuring robustness and biological relevance of the findings. We reaffirm that these methods represent a standard, reliable practice in lipidomics, and the potential of nonlinear techniques does not diminish the appropriateness or utility of linear multivariate models when applied with scientific rigor.

The aim of this study was to evaluate the influence of various agronomic factors on plant growth and the accumulation of secondary metabolites with antioxidant properties. The three Mediterranean Officinal Wild Plants selected for this investigation were Silybum marianum, Achillea millefolium, and Trifolium pratense whose extracts, enriched in phenolic compounds, are well documented for their potential therapeutic effects. Three agronomic factors were evaluated, each with two treatment options, resulting in eight experimental combinations: (1) inoculation with plant growth-promoting rhizobacteria (PGPR) versus control (no inoculation); (2) high versus low fertilization rates of K₂O and P₂O₅ to modulate nutrient availability; (3) water stress at 40% of field capacity compared to the control with full field capacity. Plant growth was monitored using the BBCH (Biologische Bundesanstalt, Bundessortenamt and CHemical industry) scale to delineate key phenological phases, with treatments applied until the flowering stage was reached. Only the leaves of the plants were collected, and hydroalcoholic extracts were prepared for the evaluation of total antioxidant capacity (TAC) using the FRAP, DPPH, and ABTS assays. These assays were selected due to their complementary insights into the chemical mechanisms underlying TAC, as well as their ability to assess the physicochemical characteristics of the phytochemical constituents.

Natural sources of vitamin D (VD), have been proposed to represent an alternative to synthetic vitamin D in nutritional interventions, also holding therapeutic potential in non-alcoholic fatty liver disease (NAFLD). In this study lipidomics was used to comparatively investigate the molecular mechanisms behind the therapeutic effects of a natural VD formulation consisting of a Shitake mushroom extracts (NVD) and a synthetic cholecalciferol formulation (SVD) in HepaRG human hepatocytes exposed to free fatty acid (FFA)-induced lipotoxicity. The results demonstrate that the two VD formulations prevent lipotoxicity with similar efficacy, but different lipidomic fingerprints. Differentially expressed lipids in NVD' in vitro therapeutic effect indicated a reduced synthesis of cellular triglycerides; combined with a marked reshaping of glycerophospholipid metabolism and characteristic changes of the chain length and number of double bonds in the phosphatidylcholine pool that were absent in SVD treatment. Bioinformatics interpretation of lipidomics data associated NVD therapeutic properties to an enhanced insulin function and glycerophospholipid metabolism, whereas SVD was primarily associated with the inflammatory signaling and death pathways of the liver cell. These differences between the two VD formulations were further highlighted matching lipidomics data with gene microarray (transcriptomics) data available from previous studies on this experimental model; the resulting multiomics data identified lipid metabolism nodes specific for the multimolecular mechanisms of the two formulations which may deserve further pre-clinical investigation in the treatment of hepatocyte lipotoxicity.

ABSTRACTThe objective of this study is to develop an HPLC‐UV method for the cost‐effective and quantitative determination of vitamin D3 in food, even in the presence of vitamin D2, with a specific focus on egg yolk. During method development, the performance of three stationary phases in resolving the peak of vitamin D2 from that of vitamin D3 was investigated. The physicochemical properties of these phases differed particularly in the extent of hydrophobicity and silanophilic activity, including a GraceSmart RP C18 column without silanol endcapping, a Robusta RP C18 column with silanol endcapping, and a Waters Xbridge RP C18 column with ethylene‐bridged hybrid (BEH) particle technology. The Xbridge C18 stationary phase exhibited the most favorable performance, leading to an RS of 1.6 under the following nonaqueous reversed‐phase (NARP) experimental conditions: mobile phase, acetonitrile, methanol, and trifluoroacetic acid in a (99/1/0.1, v/v/v) ratio; column temperature, 15°C. The developed chromatographic method does not require preanalytical purification steps and is also compatible with mass spectrometry. The identity of the vitamin D3 peak observed in the HPLC analysis was verified via GC–MS. The NARP‐HPLC‐UV method was partially validated, demonstrating satisfactory linearity, precision, accuracy, limit of quantification, and robustness. The HPLC method was then successfully applied to the analysis of real egg yolk samples, revealing average concentrations of vitamin D3 of 4–5 µg/g of wet weight sample.