Bioactive Compounds in Health and Disease

The gut–lung axis represents a bidirectional communication network linking intestinal and pulmonary immunity through microbial metabolites, immune cell trafficking, and systemic signaling. Short-chain fatty acids (SCFAs) — acetate, propionate, and butyrate — produced through colonic bacterial fermentation of dietary fiber, have emerged as critical immunomodulatory mediators in this axis with significant therapeutic potential for respiratory diseases. SCFAs exert anti-inflammatory effects in the lungs through two principal mechanisms: activation of G protein-coupled receptors FFAR2 (GPR43) and FFAR3 (GPR41) in alveolar macrophages, dendritic cells, and epithelial cells, suppressing pro-inflammatory cytokine production (TNF-α, IL-6, IL-1β) and modulating neutrophil trafficking; and inhibition of histone deacetylases (HDACs), driving epigenetic reprogramming that promotes regulatory T cell (Treg) differentiation, enhances epithelial barrier integrity, and reduces Th2-driven eosinophilia. Preclinical evidence shows that SCFA supplementation or high-fiber diets reduce airway hyperresponsiveness, eosinophilia, and mucus production in murine asthma models, with propionate acting via GPR41 signaling. In acute respiratory distress syndrome (ARDS) models, acetate and butyrate reduce neutrophil infiltration and alveolar-capillary barrier disruption. In chronic obstructive pulmonary disease (COPD), butyrate inhibits IL-13-producing innate lymphoid cells (ILC2) and modulates the NLRP3 inflammasome via GPR43. Limited clinical data associate reduced fecal SCFA levels with increased asthma risk in children, while early-life propionate exposure correlates with protection against atopy. Therapeutic strategies include dietary fiber supplementation, prebiotics (inulin, fructooligosaccharides), probiotics (Faecalibacterium prausnitzii, Bifidobacterium spp., Roseburia spp.), and direct SCFA administration. Postbiotics — bioactive compounds produced during microbial fermentation — represent an emerging approach that bypasses live microbial colonization. Context-dependent effects related to dose, route, disease stage, and baseline microbiome composition necessitate personalized approaches. Translation to clinical practice requires well-designed human trials addressing optimal dosing, delivery routes, patient stratification, and long-term safety. Novelty of the Study: This review highlights short-chain fatty acids as central mediators of the gut–lung axis, integrating receptor-mediated and epigenetic mechanisms with emerging translational strategies such as postbiotics. It emphasizes the role of SCFAs in modulating pulmonary inflammation across multiple respiratory diseases and identifies key gaps for clinical application, particularly in personalized nutrition, dosage, and targeted delivery approaches. Keywords: Short-chain fatty acids; gut–lung axis; pulmonary inflammation; microbiota; FFAR2; FFAR3; histone deacetylase; asthma; COPD; ARDS

Background: To improve nutrient availability from serpentinite, the raw material was subjected to acid treatment, resulting in the production of a novel combined fertilizer. Objective: The aim of this study was to investigate the effect of the obtained serpentinite fertilizer, applied both separately and in combination with mineral fertilizers, on soil fertility improvement, as well as on the yield and grain nutritional quality of winter wheat. Methods: Laboratory studies of fertilizer solubility were conducted to determine the rate at which nutrients pass into the soil solution. The climatic conditions of the experimental sites were described, and the agrochemical and agrophysical characteristics of the experimental plots were presented. Field and pot experiments with the fertilizer were conducted, along with soil and crop analyses, observations, and calculations using standard methods. Results: Fertilizer application had a significant positive effect on the field germination, germination energy, and yield of winter wheat, resulting in an increase in plant survival, grain number per spike, and grain weight. The contents of nutrients, protein, and lysine in the grains increased, together with the quantity and quality of gluten. Fertilizer application also reduced fungal infection of the root system. These effects contributed to improved nutritional value of the grain and enhanced bread-making quality. Conclusion: The application of serpentinite fertilizer improves several important agronomic parameters; however, its greatest effectiveness was observed when applied in combination with mineral fertilizers (NPK). Novelty: A new type of combined fertilizer was produced through acid treatment of serpentinites. The fertilizer contains Mg, Fe, Ca, P, K, Na, active SiO₂, and other nutrients. This study is the first to evaluate the agronomic and nutritional effects of phosphoric acid-activated serpentinite fertilizer under both field and pot conditions, and the effectiveness of the developed fertilizer was investigated. Keywords: serpentinite, combined fertilizer, winter wheat, grain yield capacity, proteins, lysine, gluten content and quality, bread-making quality.

Background: Post-weaning stress in piglets is characterized by the development of diarrhea, intestinal dysbiosis, systemic inflammation, and significant economic losses in pig farming. Limiting the use of feed antibiotics stimulates the search for safe functional products capable of modulating intestinal homeostasis. The historical protocol of E. Moreau (1908) for long-boiled carrots has demonstrated anti-adhesive and prebiotic potential but has not been considered as an element of systemic prevention in industrial pig farming. Objectives: To develop and scientifically substantiate the use of a functional carrot-based food product (Moro soup) in the prevention of post-weaning stress and associated diarrhea in piglets. Methods: The experiment was conducted on 60 piglets of a large white breed (28-35 days old, live weight 7.2 ± 0.5 kg). The animals are divided into 4 groups: 1st – control (standard diet); 2nd – antibiotic (colistin); 3rd – probiotic (Bacillus subtilis); 4th – experimental (Moro soup at a dose of 5-7 ml/kg 3 times a day, 5 days before, and 7 days after weaning). The Moro soup's main ingredients of 1 kg carrots, 2 liters of water, and 1.5 g of salt were cooked for 2 hours, after which 5% oat broth was added. The frequency and severity of diarrhea, body weight gain, large intestinal microbiota (PCR-RV), IL-6, secretory IgA (ELISA) levels, and jejunum histology were evaluated. Results: Moro soup supplementation resulted in a 67% decrease in the incidence of diarrhea (p<0.01) compared with the control group and reduced the duration of diarrhea syndrome to 1.6 ± 0.3 days. The average daily weight gain during the 14 days after weaning in the experimental group exceeded that of the control group by 18.4% (p<0.05) and was comparable to the antibiotic group. Microbiological analysis revealed a 2.3-fold increase in Lactobacillus spp. and a 70% decrease in enteropathogenic E. coli. The level of the pro-inflammatory cytokine IL-6 decreased by 40% (p<0.05), and the concentration of sIgA in the intestinal contents increased by 1.7 times. Morphometry of jejunum villi showed an increase in villi height of 22% and crypt depth by 12%, indicating a regenerative effect. Conclusion։ Moro soup is an effective functional food product that provides systemic prevention of post-weaning stress in piglets due to its combined anti-adhesive, prebiotic, anti-inflammatory, and reparative effects. Including the product in standard feeding protocols reduces reliance on antibiotics, increases livestock safety, and improves productivity. Next steps should focus on developing a free-dried form of Moro soup for industrial production and on incorporating it into commercial pre-starter feeds. Novelty of study: This is the first study to develop a functional carrot-based food product (Moro soup) and examine efficacy in the prevention of post-weaning stress in piglets. Additionally, this study is the first to obtain comprehensive data on the effects of Moro soup on the microbiota, immune response, and morphofunctional state of piglets' intestines, compared with antibiotics and probiotics. Keywords: post-weaning stress, functional animal food product, Moro soup, oligogalacturonides, intestinal microbiota, alternatives to antibiotics.

Background: Tomato (Solanum lycopersicum L.) is not only one of the key vegetable crops worldwide but also an important source of bioactive compounds, including lycopene, vitamin C, and polyphenols, which are associated with antioxidant and health-promoting properties. In the Republic of Armenia, the intensive development of greenhouse vegetable production contributes to increased crop productivity; however, it simultaneously increases the risk of bacterial diseases. One of the most problematic diseases is Bacterial canker of tomato, caused by Clavibacter michiganensis subsp. michiganensis (Cmm), which can lead to massive plant losses and significant yield reduction. Early diagnosis using quantitative polymerase chain reaction (qPCR) is crucial for effective disease control and the prevention of epiphytotics in greenhouses. Objective: To study the manifestation of bacterial canker of tomato under commercial greenhouse conditions and to evaluate the effectiveness of a qPCR method for the early and latent detection of the Cmm pathogen. Methods: The studies were conducted in commercial hydroponic greenhouses of the Kotayk and Ararat regions of Armenia. DNA was extracted from infected tomato stems and leaves using the CTAB method with Genetic Analysis Strategies S.L. kits and was spectrophotometrically analysed (A260/280 =1.78-1.92). Identification of Cmm was performed by qPCR using species-specific primers, and quantitative assessment of the pathogen was carried out using a standard curve based on serial tenfold dilutions. The reliability of the method was confirmed by analysis of sensitivity, linearity, repeatability, and reproducibility (Efficiency 98.1%, R² =1.0, LOD 3.1±0.4 × 10¹ copies/µl). Results: In commercial greenhouses, the first manifestations of bacterial canker in tomato include unilateral leaf wilting, darkening and necrosis of the stem vascular system, as well as alterations in fruit structure. The qPCR method enabled the detection of Cmm in all 12 examined samples with high amplification efficiency (97.5%). Samples with low Ct values (12.5-17.5) corresponded to a high number of pathogen DNA copies and exhibited pronounced visual disease symptoms. In contrast, samples with higher Ct values (26.7-36.8) contained low pathogen concentrations and, in most cases, showed no symptoms, indicating a latent stage of infection. Application of qPCR for early diagnosis reduced the proportion of infected plants from 13.2-14.9% to 4.8-5.1%, and yield losses from 9.4-10.1% to 3.5-3.9%. In addition to yield reduction, bacterial infection may affect the physiological processes of fruit development. In particular, the present study revealed a decrease in average fruit weight from 165.4 to 134.2 g, corresponding to a reduction of 18.9%. At the same time, accelerated fruit ripening was observed. The obtained results show that qPCR ensures effective early detection of infection and limits its spread under protected cultivation conditions, leading to reduced production losses and contributing to the production of healthier and high-quality plant products. Novelty: This study presents, for the first time, a comprehensive diagnosis of Cmm in commercial greenhouses in Armenia using qPCR to detect both symptomatic and latent tomato infections. The data demonstrates a direct correlation between bacterial load and symptom severity and confirms that early molecular detection of the pathogen can significantly reduce yield losses. The results highlight the practical value of implementing qPCR in greenhouse monitoring systems and provide a basis for developing a rapid phytosanitary control strategy, which has not previously been documented for the study region. Conclusion: The qPCR method demonstrated high efficiency for the early diagnosis of bacterial canker in tomato, enabling the detection of the pathogen at latent stages and preventing widespread infection. Early detection of Cmm allows timely implementation of protective measures, minimizes yield losses, and enhances the resilience of greenhouse tomato production. Thus, early molecular detection of Cmm is important not only for plant disease management but also for preserving the bioactive compounds and functional food value of tomato. Keywords: tomato, Clavibacter michiganensis subsp. michiganensis, bacterial canker, qPCR, greenhouse, early detection

Background: Ischemic heart disease (IHD) is associated with mitochondrial dysfunction, oxidative stress, impaired cellular bioenergetics, and persistent vascular injury. These disturbances are reflected by reduced ATP production, altered NAD/NADH redox balance, and lower coenzyme Q10 (CoQ10) levels, together with adverse anthropometric and hemodynamic indicators such as elevated body mass index (BMI), systolic blood pressure (SBP), and diastolic blood pressure (DBP). Squalene, a natural triterpene with antioxidant properties and relevance to sterol and ubiquinone biosynthesis, may support mitochondrial function and cellular energy metabolism. Objective: This study evaluated the effects of oral squalene supplementation (300, 600, or 900 mg/day) on BMI, SBP, DBP, and cellular energy biomarkers (ATP, NAD/NADH, and CoQ10) in patients with stable IHD over 84 days. Methods: A total of 180 participants were allocated into six groups (n = 30 each): healthy controls, healthy volunteers receiving 600 mg/day squalene, cardiac patients without squalene, and cardiac patients receiving 300, 600, or 900 mg/day squalene. Blood samples were collected after overnight fasting on days 1, 14, 28, 56, and 84. ATP and NAD/NADH were measured using colorimetric assays, and CoQ10 was assessed by ELISA. Data was analyzed using repeated-measures ANOVA, one-way ANOVA with Tukey post hoc testing, and independent t-tests, with p < 0.05 considered significant. Results: At baseline, patients with stable IHD showed significantly elevated BMI, SBP, and DBP compared with healthy controls, together with reduced ATP, NAD/NADH ratio, and CoQ10 levels (p < 0.05). Oral squalene supplementation produced dose-dependent improvements in blood pressure and cellular energy biomarkers over 84 days. ATP increased from baseline values of 3.89–4.54 to 4.62, 5.40, and 6.14 in the 300, 600, and 900 mg/day groups, respectively, by day 84, corresponding to an approximate 19–35% increase across treated groups, while untreated cardiac controls declined from 3.67 to 2.73, corresponding to an approximate 25% decrease. NAD/NADH ratio and CoQ10 also increased progressively across treated groups, with the strongest responses observed at 600 and 900 mg/day, while untreated cardiac controls showed continued declines in these biomarkers. BMI declined modestly in treated groups, while untreated controls remained largely unchanged. Conclusion: Oral squalene supplementation for 84 days improved blood pressure and enhanced cellular energy biomarkers in patients with stable IHD. The 300 mg/day dose showed beneficial effects on blood pressure, while the 600–900 mg/day doses produced stronger improvements in cellular energy biomarkers, including ATP, NAD/NADH ratio, and CoQ10. These findings suggest that squalene may serve as a promising adjunctive nutritional strategy to support hemodynamic regulation, mitochondrial bioenergetics, and cardiovascular health. Novelty of Study: This study investigated the dose-dependent effects of oral squalene supplementation on blood pressure, BMI, and key cellular energy biomarkers, including ATP, NAD/NADH ratio, and CoQ10, in patients with stable ischemic heart disease. The findings suggest a dose-related response, with blood pressure improvements observed even at 300 mg/day and stronger cellular energy biomarker responses observed at 600–900 mg/day. This highlights the potential of squalene as an adjunctive nutritional strategy for supporting hemodynamic status and mitochondrial bioenergetics in IHD. Keywords: Squalene, ischemic heart disease, ATP, NAD/NADH, CoQ10, mitochondrial function, body mass index, systolic blood pressure, diastolic blood pressure, oxidative stress

Background: Breeding pigs under free-range forest conditions is considered one of the most effective ways to ensure the production of high-quality, environmentally clean meat with relatively low costs. However, free-range pig farming also carries certain risks, particularly the likelihood of infection with invasive diseases, among which metastrongylosis is the most common. Milk from cows is mainly processed into semi-dried salty cheese, and the whey produced in this process is traditionally used as a fresh feed additive for pigs. Additionally, the cheese whey may improve the chemical composition and biochemical parameters of muscle tissues in pigs infected with metastrongylosis. Objectives: To study the effect of cheese whey on the hematological composition of blood and biochemical indicators of muscle tissues in healthy and metastrongylosis-infected pigs, and to evaluate the compliance of the obtained pork with the requirements of functional food. Methods: The research was conducted in 2025–2026 in the northeastern mountainous-forest zone of Armenia, in abattoirs, and at the Research Center of Veterinary Medicine and Veterinary Sanitary Expert Examination of the Armenian National Agrarian University. The studies were conducted on 28 pigs aged 3 months to 1.5 years kept on the farm. To improve metabolic processes, fresh whey obtained during semi-dry Chanakh cheese production was used as a feed supplement. Hematological and biochemical analyses were performed using automated analyzers. Results: In porcine metastrongylosis, the organism is under stress and exposed to toxic metabolic by-products. Given these conditions, the use of cheese whey helps to alleviate pathological processes. As a result, protein restoration and anabolic processes are activated, and the organism begins to provide the structural materials necessary for maintaining and restoring muscle tissue. Conclusion։ The inclusion of cheese whey in the diet of healthy and metastrongylosis-infected pigs is an effective feed supplement. It promotes muscle growth, strengthens the muscular system, and improves the quality of meat obtained. Novelty of study: For the first time, the effectiveness of cheese whey as a natural, biologically active feed additive in pigs with metastrongylosis has been studied. The research results have shown that the inclusion of cheese whey in the pig diet improves blood parameters in both healthy pigs and those with pig metastrongylosis, as well as chemical and biochemical parameters of muscle tissue. Keywords: cheese whey, bioactive compounds, porcine metastrongylosis, functional food production, hematological and biochemical biomarkers, muscle tissue quality, livestock nutrition

Background: Shiitake mushrooms (Lentinula edodes) are valued for their nutritional and medicinal properties, including antioxidant and anti-inflammatory effects. However, their impact on skeletal muscle health at the cellular level remains underexplored. Investigating their bioactive potential may reveal natural therapeutic strategies for reducing muscle inflammation and oxidative stress while promoting cell viability. Objective: This study aims to investigate the nutritional composition, bioactive compounds, and therapeutic potential of shiitake extract on muscle cells. Methods: Shiitake mushrooms were freeze-dried, powdered, and extracted with 70% ethanol. Nutritional and amino acid profiles were analyzed using standard methods and LC-MS/MS. identified by HPLC, and antioxidant activity was evaluated by measuring the radical scavenging activity using the DPPH assay. C2C12 myoblast cells were treated with the extract at concentrations of 125–1000 µg/mL. Cellular responses, including cell viability (MTS assay), IL-6 secretion (ELISA), ROS production, apoptosis, and cell cycle distribution (flow cytometry), were subsequently evaluated. Results: Nutritional analysis revealed that shiitake mushrooms provide 356 calories per 100 grams, with a high protein content (27.43 g), low fat (1.5 g), and abundant carbohydrates (58.2 g), including significant dietary fiber (34.98 g). The amino acid profile was dominated by glycine, L-Aspartic acid, and L-Leucine, along with various essential and non-essential amino acids. Bioactive compounds, such as gallic acid, p-Coumaric acid, and methyl gallate, were identified in the extract, which also exhibited potent antioxidant activity (314.04 mg Trolox/100g). In vitro experiments demonstrated that the extract reduced oxidative stress and inflammation in LPS-stimulated myoblast cells, as evidenced by decreased IL-6 secretion and ROS production. Moreover, the extract enhanced cell viability and reduced apoptosis at concentrations of 125–500 µg/mL, while concentrations above 1000 µg/mL showed toxicity. Conclusion: The findings suggest that shiitake extract has favorable potential as a therapeutic agent for promoting muscle health, and cell survival in muscle cells, as well as reducing inflammation and oxidative stress. Novelty of the Study: This study is unique in that investigates the therapeutic potential of Lentinula edodes (shiitake) extract on muscle cells, focusing on its protective effects against inflammation and oxidative stress in LPS-stimulated myoblasts. Unlike prior studies, it highlights specific bioactive compounds and demonstrates improved cell viability and reduced apoptosis, suggesting its potential as a natural agent for muscle health support. Keywords: Anti-inflammation, Antioxidant activity, Bioactive compounds, C2C12 cells, Nutritional composition, Shiitake mushroom

Hereditary cancer syndromes, accounting for approximately 5–10% of all malignancies, are caused by germline mutations in high-penetrance genes such as BRCA1/2, TP53, and mismatch repair genes, leading to an increased risk of early-onset and aggressive cancers. Accurate prognostic biomarkers are essential for risk stratification and personalized prevention in individuals with genetic predisposition. This review focuses on the prognostic relevance of oxidative stress and redox-dependent mechanisms in hereditary cancers. Redox imbalance, characterized by excessive reactive oxygen species (ROS) and impaired antioxidant defense, promotes genomic instability, tumor progression, and therapy resistance. Key redox-sensitive pathways, including transcription factors nuclear factor erythroid 2–related factor 2 (NRF2), nuclear factor-κB (NF-κB), and hypoxia-inducible factor-1α (HIF-1α), are discussed alongside emerging molecular and liquid biopsy biomarkers. Importantly, several dietary polyphenols, such as resveratrol, epigallocatechin gallate (EGCG), and curcumin, exhibit antioxidant, anti-inflammatory, and epigenetic effects that may help counteract mutation-associated oxidative damage. Integrating prognostic biomarker profiling with polyphenol-based, biomarker-guided functional food strategies represents a promising, non-invasive approach to personalized prevention in hereditary cancer. Novelty of Study: This review is novel in that it integrates oxidative-stress–related prognostic biomarkers with polyphenol-based dietary interventions within the structured framework of functional food science, specifically applying the Functional Food Centre’s 17-step development model to hereditary cancer prevention, thereby proposing a biomarker-guided, personalized nutrition strategy that bridges molecular oncology and functional food applications. Keywords: Hereditary cancer syndromes; Prognostic biomarkers; Oxidative stress; Redox signaling; dietary polyphenols; Personalized cancer prevention; Functional food science

Background: The primary raw material used to form the structure of ice cream waffle cones is bakery-grade wheat flour, which possesses specific technological characteristics. High levels of simple carbohydrates, primarily starch (up to 76.22 g/100 g of product), significantly increase the caloric content of finished waffle products. As a result, the product cannot be considered dietary, and its application is limited to certain consumer groups. The nutritional value and organoleptic characteristics of waffle cones can be enhanced while maintaining the required technological properties by partially replacing wheat flour with various fortifying ingredients in the recipe. Therefore, the production of waffle products with specified structural, mechanical, and technological characteristics is a modern and relevant area of research and development. Objective: To investigate the technological feasibility of using freeze-dried fruit, berry and vegetable powders in the recipe for ice cream waffle cones as a partial replacement for wheat flour. Methods: The moisture content of the powders was determined thermogravimetrically. The content of water-soluble pectin substances was determined using the calcium-pectate method, and crude fiber was determined using the intermediate filtration method. Ashing and determination of the amount of organic residue were carried out gravimetrically. The organoleptic, physicochemical (moisture content, alkalinity, wettability), and structural-mechanical (product strength) parameters of model samples and finished wafer products were evaluated. The moisture content was determined by drying the product sample for 30 minutes at the temperature of 130°C to the constant dry mass. Alkalinity was determined by titration. The strength of wafer products was determined using the Structurometer-ST2 device with microprocessor control using the ST2-07 method. The ST2-07 method is based on determining the ultimate load using the Knife No. 1 indenter. Results: The addition of vegetable, fruit, and berry powders to the waffle batter at levels of 4% to 6% resulted in a more flexible, less viscous, denser, and more structured batter. As a result, the fragility of the finished waffle cones decreased. These changes in the structural and mechanical properties of the finished waffle products can be seen as a positive effect of partially replacing traditional wheat flour with freeze-dried vegetable, fruit, and berry powders, which may ultimately make the product more appealing to consumers. Conclusions: Replacing some of the wheat flour (4 to 6% of the dry matter) with freeze-dried fruit, berry, and vegetable powders had no negative impact on the technological characteristics of the waffle cones. The resulting product exhibited acceptable organoleptic, physicochemical, and structural-mechanical properties. The vitamin-enriched formulation, due to its natural plant-based components, may contribute to the nutritional value of the product. The advantage of the developed technology is that it does not require retooling the waffle cone production line. Novelty of the Study: The results of the study can be used in the design of waffle cones with targeted functionality and specified structural and mechanical characteristics, including those with an extended shelf life. Keywords: waffle cones; plant resources; vegetable and fruit and berry powders; organoleptic properties; biochemical properties; rheological properties; quality assessment; quality products; ecological and hygienic safety; health preservation.

Aging is associated with progressive cellular dysfunction, mitochondrial damage, impaired proteostasis, and chronic low-grade inflammation, commonly referred to as inflammaging. Autophagy and mitophagy, the cellular quality control mechanisms responsible for degrading dysfunctional proteins and mitochondria, decline with age and contribute to age-related pathologiesFood-derived bioactive compounds modulate these processes through key signaling pathways, including AMP-activated protein kinase (AMPK), mechanistic target of rapamycin (mTOR), sirtuin 1 (SIRT1), nuclear factor erythroid 2-related factor 2 (Nrf2), nuclear factor kappa B (NF-κB), and the PTEN-induced kinase 1 (PINK1)/Parkin pathway. This mini review synthesizes current mechanistic, preclinical, and clinical evidence on how specific bioactives—resveratrol, curcumin, quercetin, spermidine, urolithin A, carotenoids, omega-3 fatty acids, sulforaphane, and microbiota-derived metabolites— may influence pathways relevant to healthy aging, including autophagy and mitophagy, based primarily on mechanistic and preclinical evidence. Particular attention is given to bioavailability, microbiome-dependent metabolism, biomarker limitations, and functional food development, where translation from mechanistic findings to clinically validated applications remains incomplete. Overall, dietary bioactives represent a scientifically grounded but clinically developing approach for supporting cellular quality control and reducing inflammatory mechanisms associated with biological aging. Novelty of the study: This review provides an integrated mechanistic framework linking food-derived bioactive compounds with autophagy, mitophagy, mitochondrial quality control, and inflammaging. Rather than presenting bioactives as general antioxidants or anti-inflammatory agents, it emphasizes their effects on specific regulatory pathways—AMPK, mTOR, SIRT1, Nrf2, NF-κB, and PINK1/Parkin— and their reported capacity to influence cellular quality-control pathways that decline with age. The review further distinguishes compounds by the strength of evidence across mechanistic, preclinical, and clinical levels. It also highlights microbiota-derived metabolites and the translation of functional foods as priority areas that the field must advance before dietary bioactives can be responsibly recommended for healthy aging applications. Keywords: autophagy; mitophagy; inflammaging; healthy aging; food bioactives; AMPK; mTOR; SIRT1; Nrf2; PINK1/Parkin; resveratrol; curcumin; spermidine; urolithin A; functional food

Background: Wheat is a major dietary staple and an important source of phenolic bioactive compounds with antioxidant and cytoprotective properties. However, environmental stress and conventional farming methods can limit the accumulation of these beneficial compounds and affect their functional quality. A promising strategy to enhance the natural synthesis of bioactive compounds is still under investigation. The functional relevance of laser-induced phenolic enrichment in human cell systems remains largely unexplored. Objective: To determine the effect of CO₂ laser seed priming on wheat phenolic content and functional (antioxidant and cytoprotective) activity. Methods: This study assessed the effects of continuous-wave CO₂ laser seed priming in the wheat cultivar “Red Doly” at three developmental stages: grain, germination, and green sprouts. Total free and bound phenolics, as well as phenolic profiles, were determined using High-Performance Liquid Chromatography (HPLC). Antioxidant capacity was evaluated through radical-scavenging assays. Cytoprotective activity was examined in Jurkat T lymphocytes under oxidative stress induced by hydrogen peroxide. Results: Laser priming enhanced germination and early growth (5–25% increase vs. control). Total free phenolic content increased significantly, particularly in green sprouts (p < 0.01). The antioxidant capacity increased at all developmental stages and correlated strongly with phenolic content (r = 0.81, p = 0.0004). Caffeic acid was most abundant in the free fractions, while ferulic acid dominated in the bound fractions. Phenolic extracts from laser-treated sprouts significantly improoved Jurkat cell viability and reduced oxidative cytotoxicity (p < 0.001), with more pronounced effects observed for free-phenolic–enriched fractions. Conclusions: This study establishes a functional connection between laser-induced metabolic activation and the protection of human cells by integrating compound identification, biomarker validation, and preclinical cellular efficacy testing (Steps 1, 2, 5, 6, and 8 of the Functional Food Development Model). CO₂ laser priming offers a scalable, chemical-free approach to producing phenolic-enriched functional wheat ingredients. Notably, this research is one of the first to demonstrate that phenolic enrichment induced by CO₂ laser priming results in measurable cytoprotective effects in a human cell-based model. Novelty: This study is one of the first to establish a connection between CO₂ laser priming-induced phenolic enrichment in wheat and a functional biological outcome. It demonstrates significant cytoprotection of human Jurkat T lymphocytes under oxidative stress. The research also includes detailed phenolic profiling using HPLC and validation of antioxidant biomarkers. Keywords: CO₂ laser seed priming; Wheat phenolics; Antioxidant capacity; Cytoprotective activity; Functional foods; Wheat sprouts; Oxidative stress

Background: Natural substances derived from dill (Anethum graveolens) are characterized by broad antimicrobial activity against numerous human pathogens, attributable to their rich content of bioactive compounds and strong antioxidant and antimicrobial properties. Consequently, dill may be considered both a valuable nutritional supplement and a promising source of herbal medicinal agents. Objective: This research aimed to develop high-fiber biscuits through partial replacement of wheat flour with dill powder levels at 5% and 10%, and to evaluate the effects on chemical composition, antioxidant capacity, and antibacterial activity against selected pathogenic microorganisms. Results: Chemical analysis of dill revealed carbohydrate, protein, fat, crude fiber, and ash contents of 9.02%, 3.26%, 2.10%, 3.12%, and 0.96%, respectively. The moisture content was 81.34%, indicting a high water-retention capacity that may positively influence both sensory and functional properties. Phytochemical analysis demonstrated a diverse profile of phenolic and aromatic compounds, as well as amino acids. Microbiological analysis confirmed that all biscuit samples were free of coliform bacteria. Additionally, the alcohol extract of dill exhibited strong inhibitory activity against the tested bacteria, including E. coli, Salmonella, and Staphylococcus. Sensory evaluation indicated that dill incorporation improved taste, aroma, texture, and overall acceptability of the biscuits compared with the control formulation. Novelty: This study is the first to systematically incorporate dill powder into high-fiber biscuits while simultaneously evaluating its nutritional enhancement, antioxidant capacity, and antimicrobial activity against foodborne pathogens. Unlike previous studies that have focused on dill extracts or on bakery product fortification in isolation, the present work demonstrates the dual functionality of dill as both a natural preservative and a functional food ingredient within a ready-to-eat bakery product. Conclusion: Dill exhibits valuable nutritional properties, including appreciable protein and lipid contents, in addition to serving as a rich natural source of phytonutrients. These findings highlight the multifaceted potential of dill as a source of bioactive compounds, supporting its application in the functional food industry as a natural antioxidant and preservative. Consequently, dill may serve as a safe and effective ingredient for enhancing both the nutritional value and sensory quality of food products. Keywords: Dill powder; Functional food; Natural preservative; Plant-based fortification; High-fiber biscuits; Antimicrobial activity; Antioxidant capacity

Background: Proteins in food are degraded by proteases in the digestive tract, and their products may exhibit physiological activities. In vitro protease digestion mimics digestion of food protein in the body. When crude herbal drugs used in Japanese Kampo medicines are subjected to in vitro protease digestion, which mimics in vivo processes, decarboxylated amino acids (biogenic amines) are detected in the digest. The protease-resistant dipeptide, pyroglutamyl leucine (pyroGlu-Leu), is present in fermented foods. In vitro protease digestion of functional foods and ingredients has not yet been studied. Objective: To identify protein-derived compounds by in vitro protease digestion of three functional ingredients that possess anti-inflammatory effects: a standardized extract of Asparagus officinalis stem (EAS), a standardized extract of cultured Lentinula edodes mycelia (ECLM), and a standardized oligomerized-polyphenol from Litchi chinensis fruit extract (OPLFE). Methods: Functional ingredients were treated with endoproteinases and exopeptidases. To detect decarboxylated amino acids, amino groups were derivatized with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC), and a precursor ion scan was performed using liquid chromatography separation–tandem mass spectrometry (LC–MS/MS) to target the product ions. AQC-derivatized amines were identified, and decarboxylated amino acids and pyroGlu-Leu in the digests were analyzed and quantified using LC–MS/MS in the multiple reaction monitoring (MRM) mode. Undigested functional ingredients were analyzed by LC–MS/MS and used as the negative control. Results: Decarboxylated amino acids with anti-inflammatory activity were detected in both undigested functional ingredients and their protease digestion. Tyramine was abundant in OPLFE, but its content was significantly decreased after in vitro protease digestion. 2-Methylbutylamine and isoamylamine were present in both undigested ECLM and its digest, whereas trace amounts of isobutylamine were also detected. In addition, pyroGlu-Leu, which also exhibits anti-inflammatory effects, was highly abundant in undigested ECLM, and its content was significantly increased after in vitro protease digestion. Conclusion: We first reported that decarboxylated amino acids and pyroGlu-Leu are present in undigested functional ingredients and/or formed from their proteins by in vitro protease digestion. When functional ingredients are ingested, these protein-derived compounds may exert anti-inflammatory and other beneficial effects. This methodology may be applied to other functional foods and ingredients, leading to understanding of their effects in the body. Keywords: Functional food, bioactive compound, decarboxylated amino acid, trace amine, in vitro protease digestion, LC–MS/MS.

Background: Intestinal barrier function is critically important for human health. Despite the promising biological properties of herbal extracts, limited research has addressed the effects of Astragalus membranaceus and Panax notoginseng saponin (APS) extracts on intestinal barrier function and probiotic adhesion, particularly under conditions of inflammatory stress. In the present study, we investigated the effects of APS on Caco-2 cell monolayers challenged with LPS as an in vitro model of intestinal inflammation. Objective: This study aimed to evaluate the anti-inflammatory effects of Astragalus membranaceus and Panax notoginseng saponin (APS) extracts in preserving the intestinal epithelial barrier in a lipopolysaccharide (LPS)-stimulated human Caco-2 cell monolayer model. Methods: Caco-2 monolayers were exposed to LPS to induce barrier dysfunction. Subsequently, the monolayers were treated with APS. Barrier function was evaluated by measuring transepithelial electrical resistance (TEER), FITC-dextran flux permeability, tight junction protein (Claudin-1, Occludin, ZO-1) expression, and cellular ATP levels. In addition, the growth and adhesion of Lactobacillus rhamnosus to Caco-2 cell monolayers were assessed. Results: APS treatment significantly increased TEER values, decreased FITC-dextran permeability, restored the expression of tight junction proteins (Claudin-1, Occludin, and ZO-1), and recovered intracellular ATP levels compared to the LPS group. Furthermore, APS significantly enhanced the growth and adhesion of Lactobacillus rhamnosus to Caco-2 cell monolayers, even under inflammatory conditions. Novelty: This study identifies a previously uncharacterized synergistic role of Astragalus membranaceus and Panax notoginseng saponins in coordinating intestinal barrier protection and probiotic adhesion under inflammatory stress. These findings provide new mechanistic insight into the integrated actions of combined botanical saponins in maintaining intestinal homeostasis. Conclusion: These findings suggest that APS exerts protective effects on intestinal barrier function and promotes probiotic adherence against LPS-induced intestinal injury in human epithelial cells. APS may be considered a valuable nutritional and therapeutic supplement for maintaining gastrointestinal health. Keywords: Astragalus membranaceus and Panax notoginseng extracts; Inflammatory bowel disease; Intestinal probiotics; Tight junction proteins.

Background: Lipid oxidation is a major factor affecting the chemical stability and shelf life of processed meat products during frozen storage. Natural plant oils rich in bioactive compounds have gained attention as effective alternatives to Mentha longifolia antioxidants. These compounds can reduce oxidative deterioration and improve product quality. Objective: This study aimed to evaluate the effect of different concentrations of Mentha longifolia oil on the chemical properties and oxidative stability of processed burger patties during frozen storage. Materials and Methods: Crude Mentha longifolia oil was extracted using a Clevenger apparatus. Burger samples were divided into four treatments: control (T1) without oil addition, and T2, T3, and T4, each supplemented with 1, 2, and 3 g of Mentha longifolia oil, respectively. Chemical analyses were conducted before freezing and during 60 days of frozen storage at −18 °C, with testing performed twice per month. Fatty acid composition and flavonoid profiles were determined using GC–MS and HPLC, while oxidative stability was evaluated by measuring free fatty acids (FFA), peroxide value (PV), and thiobarbituric acid (TBA). Results: Mentha longifolia oil exhibited a high content of essential fatty acids and 38 identified flavonoid compounds, dominated by caffeic acid, apigenin, quercetin, and 4-hydroxy derivatives. No significant differences (P ≥ 0.05) were observed among treatments before freezing. During frozen storage, oxidative indices increased in all samples; however, oil-treated burgers showed significantly lower FFA, PV, and TBA values, with treatments T3 and T4 recording the lowest values compared to the control. Keywords: antioxidant activity; natural antioxidants; flavonoids; Mentha longifolia aromatic oils; beef burger; shelf-life extension

Helicobacter pylori is a Gram-negative bacterium that colonizes the human gastric mucosa and affects more than half of the world´s population. It is associated with gastritis, peptic ulcers, and gastric cancers. Despite antibiotics being long effective, increasing multi-resistance to key drugs has led to lower eradication rates, highlighting the need for alternative approaches. Natural bioactive compounds from microalgae have increased attention due to their antioxidant, antibacterial, and anti-inflammatory properties. Microalgae produce a wide variety of metabolites, such as photosynthetic pigments, fatty acids, peptides, and carbohydrates, many of which show promising activity against H. pylori. Antioxidants like astaxanthin and polyphenols help neutralize reactive oxygen species, protect gastric epithelial cells, and modulate inflammatory signaling. Sulfated polysaccharides and antibacterial peptides can interfere with bacterial adhesion, damage the cell membrane, and inhibit virulence factors such as cagA and vacA. Additionally, omega-3 fatty acids (EPA and DHA) and phycocyanin have strong immunomodulatory effects, reducing NF-κB activation and lowering pro-inflammatory cytokines like IL-6 and TNF-α. In contrast to previous reviews that discuss microalgal bioactivities separately, this article takes an integrated view, connecting specific microalgal compounds with H. pylori virulence mechanisms and host inflammatory responses. Using different approaches at once, microalgal metabolites can help limit bacterial growth, reduce oxidative stress, and mitigate chronic inflammation. Overall, microalgae represent a sustainable and innovative source of bioactive compounds that could complement current therapies for H. pylori infection. Incorporating these compounds into functional foods or nutraceuticals may improve treatment outcomes, protect the gastric mucosa, and reduce the use of antibiotics. Future work should focus on improving extraction methods, enhancing bioavailability, and confirming their efficacy and safety through preclinical and clinical studies. Microalgal compounds offer a promising, safe, and environmentally friendly strategy for preventing and managing H. pylori-related diseases. Keywords: H. pylori, microalgae, bioactive compounds, antibacterial activity, antioxidant activity, anti-inflammatory activity

Background: The Areni grape (Vitis vinifera L.) is an indigenous Armenian variety renowned for its adaptation to local terroir and historical significance in winemaking. Its genetic diversity, particularly among clones, offers valuable resources for enhancing wine quality through variations in phenolic compounds, chemical composition, and technological traits. Understanding these differences is crucial for selecting optimal genotypes in viticulture and enology, especially in regions like Armenia, where traditional varieties contribute to unique wine profiles. Objective: To assess the chemical, polyphenolic, and color variations among three Areni grape genotypes and their impact on wine quality indicators. Methods: Analyses of grapes and wines produced under microvinification conditions were performed according to OIV standards, using spectrophotometry and HPLC to determine organic acids, physicochemical parameters, total phenolics, anthocyanins, flavonoids, and color indices. Results: Nosr Areni displayed the highest pigment content in the fruit, Areni clone 15 produced wines with the highest total phenolics, and Sev Areni wines exhibited the greatest color intensity. Chemical parameters varied depending on the specific genotype characteristics. Conclusion: Areni genotypes exhibit pronounced biochemical and technological diversity, which significantly influences wine composition and color development. Areni clone 15 stands out for its high phenolic potential, making it a promising raw material to produce high-quality red wines. Novelty of the Study։ This study provides the first comparative evaluation of Sev Areni and its clones in terms of polyphenolic composition and wine quality, identifying clones with high phenolic potential and technological suitability for winemaking. Keywords: Sev Areni grape, Vitis vinifera, polyphenols, anthocyanins, flavonoids, organic acids, wine color, clonal selection

Background: The neem tree (Azadirachta indica) is an evergreen species native to regions such as India and Pakistan and is also cultivated in parts of the Americas. Products derived from neem have numerous applications, particularly in skin, hair, and nail care, as well as in food preservation, due to their ability to inhibit the growth of various microorganisms. Objective: Three organic solvents, hexane, ethanol, and petroleum ether (80-‎‎100°C), were used to extract neem oil from dried, ground seeds using the Soxhlet extraction method. High-performance liquid chromatography (HPLC) was employed to determine the fatty acid composition of the extracted oil. The antibacterial activity of neem seed oil was evaluated against four bacterial strains (Staphylococcus aureus, Bacillus spp., Pseudomonas aeruginosa, and Escherichia coli) and one yeast (Candida spp.) at three different concentrations. Results: The extraction yields were 44.89% using petroleum ether, 42.63% using hexane, and 39.17% using ethanol. Analysis revealed that long-chain fatty acids accounted for 19.77% of the neem seed oil composition, including palmitoleic acid (14.23%), oleic acid (2.49%), palmitic acid (1.50%), stearic acid (0.90%), linoleic acid (1.50%), and linolenic acid (0.05%). Neem seed oil exhibited strong antibacterial activity. Staphylococcus aureus showed inhibition zones of 36, 30, and 28 mm at concentrations of 1000, 500, and 250 µg/mL, respectively. Bacillus spp. demonstrated inhibition zones of 28 , 26 , and 24 mm at the same concentrations. Escherichia coli exhibited inhibition zones of 26 , 22 , and 10 mm, respectively. Lower inhibitory effects were observed against Pseudomonas aeruginosa, with inhibition zones of 18 , 19 , and 3 mm. Novelty: Neem seed oil represents an effective, sustainable, and environmentally friendly alternative to synthetic antimicrobial agents. Conclusion: Neem seeds are a valuable source of oil rich in long-chain fatty acids with significant therapeutic potential. The oil demonstrated notable antibacterial and antifungal activity, emphasizing its potential importance for medical and pharmaceutical applications. Keywords: Neem tree, Oil extraction, Fatty acid and Antimicrobial, antifungal activity, long-chain fatty acids.

Background: Beyond dietary modification, bioactive compounds have emerged as modulators of glycemic regulation. Silico analysis has identified N-trans-caffeoyltyramine (NCT) and N-trans-feruloyltyramine (NFT) as potent modulators of hepatocyte nuclear factor 4α (HNF4α), thereby favorably regulating blood glucose metabolism. Objective: To evaluate the effects of a proprietary dietary supplement containing novel bioactives, NCT and NFT, on parameters of continuous glucose monitoring in individuals with prediabetes. Methods: A 4-week randomized, double-blind, placebo-controlled parallel arm trial was conducted in adults (Age: 18-50 years; BMI: 25-30 kg/m2) with prediabetes (fasting blood glucose >100 mg/dL) and abdominal adiposity (waist circumference >80cm for females and >90 cm for males). A total of 126 participants were randomized to receive either 120 mg/d NCT/NFT (n=63) or placebo (n=63). Glycemic control measures were assessed by continuous glucose monitoring (CGM) and included total 24-hour glucose AUC, mean 24-hour glucose, maximum 24-hour glucose, minimum nighttime glucose, MAGE, MoDD, and basal hyperglycemia AUC. Homeostatic model assessments of insulin resistance (HOMA-IR) and

Stress and sleep disturbances have emerged as significant global health challenges, intricately linked through the hypothalamic–pituitary–adrenal (HPA) axis. Cortisol, the primary stress hormone, serves as a key biomarker of HPA axis activity, influencing metabolism, immune response, and sleep regulation. Chronic elevation of cortisol disrupts circadian rhythm, inhibits melatonin synthesis, and contributes to insomnia, anxiety, and fatigue. Therefore, identifying bioactive compounds capable of modulating cortisol and restoring HPA axis balance offers a natural strategy for improving stress resilience and sleep quality. This review consolidates scientific evidence on several plant-derived and microbial bioactive compounds with cortisol-lowering and sleep-promoting effects. Bioactive compounds that modulate the HPA axis, therefore, represent a multidimensional therapeutic opportunity, acting not only on endocrine regulation but also on neurochemical balance and immune modulation. Integrating these compounds into dietary or nutraceutical interventions could provide a preventive and restorative approach for individuals experiencing chronic stress or sleep disorders. Withania somnifera (ashwagandha) exhibits adaptogenic properties via withanolides that downregulate cortisol and improve sleep onset. Panax ginseng contains ginsenosides that modulate stress hormone release and enhance neural adaptability. Ganoderma lucidum (reishi mushroom) triterpenoids exert anxiolytic and sedative effects, promoting restorative sleep. Centella asiatica triterpenes, including asiaticoside and madecassoside, contribute to cortisol regulation and neuroprotection, improving REM sleep architecture. Matricaria chamomilla flavonoids, notably apigenin, interact with GABAergic receptors and reduce HPA hyperactivation. Emerging evidence also highlights postbiotics, such as those from Limosilactobacillus fermentum PS150 and Bifidobacterium breve BB091109, which influence the gut–brain axis by enhancing neurotransmitter balance, reducing systemic inflammation, and lowering cortisol. The novelty of this study lies in presenting an integrated linkage between plant-derived bioactive compounds and postbiotics in the regulation of stress and sleep through modulation of the HPA axis. This framework highlights their potential influence on cortisol dynamics, providing valuable insights for the future development of functional products aimed at stress reduction and the promotion of mental wellness. Future research should focus on clinical validation, pharmacokinetic profiling, and formulation optimization to translate these natural agents into evidence-based functional foods and nutraceuticals capable of safely supporting stress regulation and sleep health. Keyword: Cortisol modulation, Sleep quality, Hypothalamic-pituitary-adrenal (HPA) axis regulation, Adaptogenic bioactive compounds, Gut-brain axis, Neuroendocrine regulation