Background: Linking metabolite composition to biological activities remains challenging in herbal medicines, where diverse metabolites may collectively contribute to pharmacological effects. Although metabolite profiling is fundamental to quality assessment, conventional quality evaluation often provides limited information on how variation in metabolite composition relates to biological activities. Objectives: This study aimed to develop a multi-component–multi-bioactivity framework integrating metabolite profiling and bioactivity assessment, enabling bidirectional mapping between metabolite composition and biological responses and providing a functional perspective for quality characterization of complex herbal medicines. Salvia miltiorrhiza Bunge (Danshen), a widely used medicinal plant with a chemically diverse metabolome and well-documented cardiovascular activities, was selected as a representative model system. Methods: Quantitative 1H NMR spectroscopy was employed to profile 28 metabolites in Danshen, while zebrafish-based bioassays were used to assess three cardiovascular-related bioactivities: pro-angiogenic, anti-thrombotic, and vasodilatory activities. A lifecycle-oriented approach was introduced for bioassay development to improve the robustness and reproducibility of biological measurements. Multiple machine learning approaches were integrated within a two-level stacking ensemble framework to construct quantitative metabolite–bioactivity relationship models and capture nonlinear associations between metabolite composition and biological activities. Results: The stacked models consistently outperformed the individual learners across the three cardiovascular bioactivities, achieving R2 values of 0.895–0.931 and RMSE values of 0.116–0.130. External validation using 10 independent Danshen samples further supported the predictive reliability of the stacked models. Based on model-derived metabolite contributions, a quantitative and interpretable metabolite–bioactivity network linking 28 metabolites with three cardiovascular-related activities was constructed, revealing both shared and activity-specific metabolic contributors. Conclusions: This study establishes an interpretable framework integrating NMR-based metabolite profiling, robust multi-endpoint bioassays, and ensemble learning to connect metabolite composition with functional bioactivity. The framework provides a complementary bioactivity-informed perspective for the quality characterization of medicinal plants and supports systems-level evaluation of complex herbal medicines.
Background: Global expansion of Litopenaeus vannamei aquaculture has intensified the reliance on antibiotics for disease control, raising concerns about hepatotoxicity, microbiota dysbiosis and antimicrobial resistance. Synthesized scorpion venom-derived antimicrobial peptide BmKn2-7 is a promising alternative, but its efficacy and mechanisms as a dietary additive in shrimp remain unclear. Methods: A 56-day feeding trial was conducted with six diets: a basal diet (FM), four diets supplemented with BmKn2-7 (0.8, 1.6, 2.4 and 3.2 mg/kg), and a florfenicol diet (FL, 3.2 mg/kg). Results: Growth performance was unaffected by any treatment. However, FL group significantly reduced condition factor and hepatosomatic index and markedly increased serum ALT and AST. The A2.4 diet increased whole-body crude lipid contents and significantly elevated serum ACP, AKP, HDL-C, and TG. 16S rRNA gene sequencing showed that intestinal microbial diversity indices were numerically highest in the A2.4 group, although pairwise differences were not statistically significant. A2.4 was also associated with reduced Vibrio abundance and enrichment of genera such as Acinetobacter, while FL induced pronounced community shifts. Hepatopancreatic transcriptomics and pathway analysis indicated that A2.4 was associated with shifts in amino acid, organic acid, and carbohydrate metabolism, together with enhanced endoplasmic reticulum protein processing, whereas FL was mainly associated with reduced V-ATPase-related proton transport. Key BmKn2-7 responsive genes were positively correlated with phosphatase activities and lipid parameters, and Hsp90b1 was negatively correlated with ALT and AST. Following V. parahaemolyticus challenge, A2.4 showed the lowest cumulative mortality, significantly outperforming FL and high-dose BmKn2-7. Conclusions: Dietary supplementation with BmKn2-7 at 2.4 mg/kg improves intestinal microbial homeostasis, metabolic and proteostatic functions, and resistance to V. parahaemolyticus. These findings support the further evaluation of BmKn2-7 as a potential feed additive for reducing antibiotic dependence in shrimp aquaculture.
Background: This research aimed to examine the associations between the total “Nutrition Knowledge Scale” score and metabolic profile indicators among fitness center members, and to explore whether selected nutrition knowledge items differed according to daily meal consumption. Methods: The research, conducted using a cross-sectional and relational screening model, included a total of 230 volunteers, consisting of 67 women and 163 men. The participants’ HDL, LDL, fasting glucose, and triglyceride levels were obtained from biochemical analysis results via the e-Nabız system, while their nutrition knowledge levels were assessed using the “Nutrition Knowledge Scale”. Descriptive statistics, simple and multiple linear regression analyses, and one-way ANOVA were used for data analysis. Results: The total “Nutrition Knowledge Scale” score was significantly associated with LDL levels after adjustment for BMI and gender (β = −0.136, p = 0.037), whereas no statistically significant associations were observed with HDL, fasting glucose, or triglyceride levels. In secondary exploratory analyses, selected nutrition knowledge items showed significant associations with LDL levels, while the vitamin B12-related item was positively but not significantly associated with triglyceride levels (β = 0.099, p = 0.136). Additionally, responses to three selected nutrition knowledge items differed significantly according to daily meal consumption, with small effect sizes (η2 = 0.029–0.046). Conclusions: Overall nutrition knowledge was associated with LDL levels after adjustment for BMI and gender, but not with HDL, fasting glucose, or triglyceride levels. Secondary exploratory findings indicated associations between selected nutrition knowledge items, LDL levels, and daily meal consumption. Causal inference is limited by the cross-sectional design.
Background/Objectives: Evidence suggests that plant-based diets are associated with reduced risk of chronic inflammatory diseases and lower circulating C-reactive protein (CRP) levels. However, associations with other inflammatory biomarkers, including interleukins (ILs), tumor necrosis factors (TNFs), interferons (IFNs), and chemokines, remain limited and inconclusive. This study compared multiple serum cytokine and chemokine levels between healthy vegetarians and omnivores. Methods: 160 apparently healthy, normal-weight participants aged 25–50 years, without acute and chronic illnesses or allergies, were included: 80 omnivores and 80 vegetarians (67 vegans and 13 lacto-ovo vegetarians). Serum concentrations of 13 cytokines/chemokines were measured using flow cytometry with a LEGENDplex™ bead-based multiplex immunoassay kit. Anthropometry, standard clinical hematological and biochemistry parameters, CRP, and dietary intakes were also analyzed. Results: The groups did not differ by sex (50% women), age, smoking, physical activity, or anthropometrics. Vegetarians had lower levels of TNF-α, IL-6, IL-8, IL-10, IL-12p70, IL-17A, IFN-γ, and IFN-α2, as well as lower leukocyte and lymphocyte counts, with no difference in IL-1β, IL-18, IL-33, IL-23, MCP-1, or CRP. The differences persisted after adjustment for potential confounders (although fewer remained significant after multiple-testing correction), and were principally present in both vegetarian subgroups, alongside substantial differences in multiple dietary factors. Regression analyses further confirmed the association of vegetarian dietary patterns with cytokine levels that differed. Conclusions: Vegetarian dietary patterns were associated with lower circulating concentrations of selected cytokines/chemokines in this cohort. However, the number of associations remaining after multiple-testing correction was limited; there was high multicollinearity between dietary factors, and causality could not be established. Further research is needed to elucidate causal relationships, specific dietary factors, underlying mechanisms, and clinical implications.
