Search Results (768)

Lameness is a major welfare and economic disorder in dairy production and a clinical sign with multiple causes. This review focuses on claw horn disruption lesions (CHDL), a major cause of periparturient lameness. Although hoof-centered prevention and treatment remain essential, preclinical systemic alterations, recurrence, and clustering with other transition-period disorders suggest that CHDL may sometimes represent the local expression of broader pathophysiology. We propose the Lameness Triad, in which three mechanisms converge: sustained endotoxemic pressure and TLR4-mediated innate immune activation, endothelial glycocalyx degradation and vascular dysfunction, and metabolic triage that reallocates resources from production toward defense. Their combined pathogenic pressure is proposed to produce clinical disease when it exceeds a cow-specific Susceptibility–Resilience Gate comprising mucosal barrier integrity, hepatic endotoxin clearance, immune calibration, redox reserve, hoof structural reserve, and prior damage. Ruminal dysbiosis, mammary involution, and postpartum uterine contamination may contribute to inflammatory and endotoxemic pressure, whereas the confined, continuously loaded digital corium may become a principal site of clinical expression. Longitudinal multi-omics studies identified inflammatory, urinary, and milk-metabolite alterations before diagnosis, with within-cohort serum and urine models discriminating cows that later became lame as early as eight weeks prepartum. These findings require prospective validation and do not establish the complete causal sequence. The framework extends, rather than replaces, established hoof-health programs and provides testable hypotheses for earlier detection, prevention, and selection for resilience. Full article

Agathosma betulina (P.J. Bergius) Pillans is an aromatic medicinal shrub endemic to South Africa’s Cape Floristic Region, widely used in traditional medicine to treat urinary tract infections, gastrointestinal disorders, colds, and wounds. Despite its commercial importance, limited information is available on how different growing environments influence its phytochemical composition. This study profiled the secondary metabolite composition of A. betulina leaves collected from wild, open-field cultivation, and glasshouse-grown environments using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) combined with chemometric analyses. Methanolic extracts prepared from nine samples were analysed, resulting in the confident annotation of 21 secondary metabolites (MSI Level 2), comprising phenolic acids, flavanones, flavones, and flavonol glycosides. Rutin was the predominant flavonoid, with concentrations of 194.62, 185.71, and 58.82 mg CE kg⁻¹ DW in wild, cultivated, and glasshouse-grown plants, respectively. Neochlorogenic acid accumulated predominantly in cultivated plants (51.84 mg CE kg⁻¹ DW) compared with wild plants (3.31 mg CE kg⁻¹ DW), while coumaroylquinic acid (125.93 mg CE kg⁻¹ DW) and feruloylquinic acid (30.74 mg CE kg⁻¹ DW) were most abundant under glasshouse conditions. Tricin-7-O-glucoside (62.93 mg CE kg⁻¹ DW) and isorhamnetin-3-O-glucoside (31.26 mg CE kg⁻¹ DW) also accumulated significantly in cultivated plants. Partial Least Squares Discriminant Analysis (PLS-DA) score explained 92.1% of the total variation (C1 = 63.8%; C2 = 28.3%), together with hierarchical heat map clustering and VIP, clearly separating samples according to growing environment. Wild plants exhibited the greatest metabolite diversity, cultivated plants showed intermediate metabolite profiles, and glasshouse-grown plants displayed reduced flavonoid accumulation and overall chemical diversity. These findings demonstrate that growing environment strongly influences secondary metabolite accumulation in A. betulina and provide valuable information for optimizing cultivation practices, quality control, and conservation strategies for this economically important medicinal species. Full article

Urologic malignancies (UMs), including prostate cancer (PCa), bladder cancer (BCa), renal cell carcinoma (RCC), and testicular germ cell tumors (TGCT), are governed by interconnected molecular pathways that regulate proliferation, angiogenesis, metabolism, invasion, immune escape, and treatment resistance. Key pathways include PI3K/AKT/mTOR, VEGF/VEGFR, EGFR, FGFR, c-MET, androgen receptor signaling, DNA damage response, inflammatory transcriptional programs, and regulation of the tumor microenvironment. This structured expert review evaluates mechanistic, translational, epidemiological, and clinical evidence on food- and botanical-derived nutraceuticals that may influence cancer-related signaling, redox balance, inflammation, metabolic adaptation, and host–tumor interactions. Nutraceuticals are considered investigational adjunctive exposures rather than anticancer treatments or alternatives to standard care. Relevant literature was identified through a structured, non-systematic search of PubMed/MEDLINE, Scopus, Web of Science Core Collection, and Embase from database inception to 17 June 2026, supplemented by backward and forward citation searches. Eligible evidence comprised preclinical, observational, interventional, pharmacokinetic, formulation, safety, and drug–nutraceutical interaction studies. Evidence was evaluated by cancer type, compound class, molecular context, formulation, exposure, translational maturity, and safety, and was classified into five stages: prevention signal, mechanistic plausibility, bioavailability and exposure feasibility, exposure-linked biomarker activity, and clinical benefit. An exposure–species–compartment framework was used to assess whether parent compounds and relevant metabolites reached systemic, urinary, or target-tissue concentrations compatible with the proposed effects. Findings based solely on supraphysiological concentrations of unconjugated parent compounds were considered hypothesis-generating unless supported by human exposure or tissue-distribution data. Curcumin, green tea catechins, isoflavones, carotenoids, flavonols, stilbenes, and triterpenoids affect several cancer-related pathways, primarily in experimental models. Translation to clinical practice is constrained by inconsistent formulations, limited bioavailability, inadequate target-tissue exposure data, few biomarker-linked studies, and possible interactions with anticancer therapies. PCa and BCa provide the most suitable settings for exposure-verified mechanistic studies. In RCC, safety and treatment interactions should be prioritized, whereas in TGCTs, non-interference with curative cisplatin-based therapy must be demonstrated. Future studies should use chemically defined formulations, verify clinically relevant exposure, incorporate mechanism-matched biological-response endpoints, and confirm compatibility with established treatment. Current evidence does not support nutraceuticals as treatments for urologic malignancies. Full article

Neonicotinoid insecticides (NEOs) are widely used neuroactive pesticides, but biomonitoring evidence among older adults remains limited. This multicenter cross-sectional study characterized urinary exposure profiles of NEOs and their metabolites among 419 older adults from South China. Ten urinary NEO biomarkers, including six parent compounds and four metabolites, were measured using high-performance liquid chromatography coupled with tandem mass spectrometry. NEOs were frequently detected in urine samples; except for thiacloprid and acetamiprid, with detection frequencies of 81.67% and 65.48%, respectively, all other compounds were detected in more than 90% of participants. Among the parent NEOs, clothianidin had the highest median concentration (1.402 μg/L), followed by dinotefuran (1.240 μg/L) and thiamethoxam (0.888 μg/L). The main metabolite was N-desmethyl-acetamiprid (1.643 μg/L). Looking at the overall composition, dinotefuran, clothianidin, thiamethoxam, and N-desmethyl-acetamiprid were the biggest contributors to the total urinary NEO burden, though their shares varied by region and recruitment center. Correlation analysis pointed to common co-exposure patterns, especially between thiamethoxam and clothianidin, and between imidacloprid and its metabolites. In exploratory analyses, N-desmethyl-thiamethoxam showed a positive correlation pattern, with significant positive correlations with glucose, glycated hemoglobin, triglycerides, and alanine aminotransferase (all q < 0.01). CLO was positively correlated with high-density lipoprotein cholesterol (q < 0.05). These findings indicate widespread co-exposure to multiple NEOs among older adults in South China and highlight the need for longitudinal studies to clarify exposure sources, temporal variability, and potential health implications. Full article

Background/Objectives: Chronic kidney disease (CKD) is a major global public health problem, and conventional biomarkers such as serum creatinine primarily reflect excretory function with limited sensitivity for early detection and for predicting progression. Amino acids (AAs) and acylcarnitines (ACs) reflect metabolic processes related to nitrogen metabolism and mitochondrial fatty-acid oxidation that are influenced by renal function. This study aimed to characterize CKD-associated alterations in AAs and ACs profiles using an integrated analytical framework combining differential analysis, supervised multivariate modeling, differential network enrichment analysis (DNEA), and cross-compartment plasma–urine profiling, moving beyond individual metabolite associations toward a multi-level characterization of CKD-associated metabolic reprogramming. Methods: Plasma and urine samples from 78 patients with CKD and plasma samples from 70 healthy controls were analyzed using flow-injection tandem mass spectrometry (FIA-MS/MS). An integrated targeted metabolomics framework combining differential, multivariate and network-based analyses including DNEA was applied to plasma and paired urine samples to characterize systemic and urinary metabolic alterations in CKD, focusing on AAs and ACs. Results: CKD was characterized by significant elevated short-chain dicarboxylic acylcarnitines, increased methylhistidine (MetHis) and argininosuccinic acid (ASA), together with reduced tryptophan, serine, methionine, and tyrosine in plasma. These metabolites were consistently identified across different analyses and correlated with kidney function markers. DNEA revealed coherent network-level reorganization, with acylcarnitine pathways gaining connectivity and centrality while amino acid modules lost integration in CKD. Cross-compartment analysis identified both systemic and compartment-specific patterns of metabolite distribution. An exploratory clustering-guided biomarker panel combining MetHis, C3DC, and Trp achieved an area under the curve (AUC) of 0.881 for discriminating patients with CKD from controls. Moreover, C6, C8, and C10 remained significantly associated with CKD after additional adjustment for estimated glomerular filtration rate (eGFR). Conclusions: Targeted metabolomic profiling revealed a coordinated metabolic signature in CKD suggesting disturbances in pathways related to fatty-acid oxidation, nitrogen imbalance, and altered amino acid metabolism. Network-level analysis provided evidence of systemic metabolic reorganization beyond individual metabolite changes. As findings derive from an observational cohort with high comorbidity prevalence, the identified signatures should be considered CKD-associated rather than CKD-specific. Full article

Cannabis use among breastfeeding women is increasing. While studies have shown that cannabinoids transfer into human milk, whether breastfed infants of mothers who use cannabis are exposed to volatile organic compounds (VOCs) is not known. Here, we characterized VOC metabolite concentrations in urine produced by exclusively breastfed infants whose mothers used cannabis. Relationships between infant urinary VOC metabolite concentrations and milk delta-9-tetrahydrocannabinol (∆⁹-THC) concentrations and cannabis use modality (smoking/vaping) were documented. Twenty mother–infant dyads (<6 months postpartum) living in Washington and Oregon were enrolled in the Lactation and Cannabis (LAC) Study, and nineteen were included in the current study. Mothers collected baseline infant urine and milk samples (following ≥12 h of cannabis abstention) and three infant urine and five milk samples over 8–12 h following cannabis use. Sixty-five urine samples were analyzed for VOC metabolite concentrations, and 111 milk samples were previously analyzed for ∆⁹-THC. Results indicate detectable levels of 24 VOC metabolites in infant urine. Non-baseline infant urine had greater concentrations of 15 VOC metabolites than matched baseline samples (p < 0.05). Urinary 2CyEMA (acrylonitrile metabolite) and 2CaEMA (acrylamide metabolite) concentrations were positively correlated with ∆⁹-THC concentrations in milk. Urinary 2CyEMA concentrations were higher in infants whose mothers primarily smoked (including those who smoked and vaped) compared to those who exclusively vaped (B = −0.637, SE = 0.317, p = 0.044). 2CaEMA concentrations were also greater in the urine produced by infants whose mothers reported any cannabis smoking versus only vaping on the study day (B = −4.06, SE = 1.42, p = 0.004). Results indicate that infants whose mothers use cannabis have measurable VOC exposure. Further research is needed to assess the potential health implications, if any. Full article

Background: Prostaglandin E₂ (PGE₂) is a key lipid mediator involved in inflammation and bone homeostasis. Its systemic production is reliably reflected by 24 h urinary excretion of PGE₂ (U-PGE₂) and its major metabolite (U-PGEM). However, the physiological association between systemic PGE₂ production, calcium-phosphorus homeostasis and bone turnover markers remains unclear. This study aims to elucidate these relationships in a general Chinese adult population. Methods: In this multicenter, cross-sectional study, 737 Chinese adults underwent standardized 24 h urine collection. Multivariable linear regression was used to assess independent associations with bone metabolism markers. Restricted cubic spline models were further employed to examine nonlinear relationships. Results: The median 24 h U-PGE₂ and U-PGEM excretion levels were 133.87 and 246.76 pg/mmol creatinine, respectively, with no significant sex differences (both p > 0.05). Multivariable regression analyses revealed that both 24 h U-PGE₂ and U-PGEM were independently and positively associated with advancing age. Notably, both 24 h U-PGE₂ and U-PGEM maintained a significant inverse association with serum calcium (Overall p < 0.05). Restricted cubic spline analyses further demonstrated a significant non-linear association between both 24 h U-PGE₂ and U-PGEM and total procollagen type 1 N-propeptide (P1NP; both Overall p < 0.05). This relationship was characterized by a steep decline in U-PGE₂ and U-PGEM excretion at lower P1NP concentrations, which subsequently plateaued at higher concentrations (Overall p < 0.05). Additionally, U-PGEM exhibited a significant inverse linear association with intact parathyroid hormone (PTH; Overall p < 0.05). Conclusions: This study provided valuable insights into the clinical determinants of 24 h U-PGE₂ and U-PGEM in Chinese adults and their independent associations with calcium-phosphorus homeostasis and bone turnover markers. Full article

Nicotine is the main psychoactive component of tobacco and is considered to be the main substance responsible for the development of tobacco addiction. The main enzyme responsible for nicotine metabolism, CYP2A6, catalyzes the conversion of nicotine to cotinine and the subsequent metabolism of cotinine to trans-3′-hydroxycotinine. CYP2A6 activity is known to be modulated by various compounds, such as quercetin. This repeated-measures study examined the effects of short-term quercetin supplementation on urinary nicotine metabolism biomarkers in adult users of conventional and alternative nicotine products. Seventy-two participants completed a two-week study protocol involving first-morning urine collection at four time points: baseline, immediately after three days of quercetin supplementation (500 mg/day), seven days after supplementation, and ten days after supplementation. Urinary nicotine, cotinine, and trans-3′-hydroxycotinine concentrations were measured, and the nicotine metabolite ratio was calculated as trans-3′-hydroxycotinine/cotinine. Repeated-measures analysis of variance was used to evaluate biomarker changes over time according to sex, nicotine product type, and self-reported nicotine consumption intensity. Quercetin supplementation did not consistently alter nicotine metabolism biomarkers, while a descriptive increase in median urinary nicotine concentration after supplementation was observed in participants reporting lower daily nicotine consumption compared with other groups. These findings suggest that further studies are warranted to better clarify the effects of quercetin on nicotine metabolism across different levels of nicotine exposure. Full article

Background: A high-fat diet (HFD) contributes to cardiometabolic disease. Gut microbiota-derived metabolites may participate in this process, but their contribution to lipid regulation is not well defined. Indoxyl sulfate (IS), a microbiota-derived metabolite, has been linked to vascular and metabolic dysfunction. Its role in lipid metabolism remains unclear. Methods: In Part A, plasma and urinary concentrations of IS were measured in plasma and urine from HFD-fed rats in which dyslipidemia had developed, together with controls. In Part B, HepG2 cells were exposed to IS, and cell viability and selected cholesterol metabolism-related transcripts and proteins were assessed. In Part C, 10-week-old, male Sprague–Dawley rats maintained on a standard diet received vehicle or IS at two doses for 8 weeks. Hepatic expression of LDLR, SREBP-2, HMG-CoA reductase, and related cholesterol metabolism markers were measured by quantitative real-time PCR and Western blotting. Results: In Part A, higher plasma IS concentrations and higher daily urinary IS excretion were found in samples collected from HFD-fed rats compared to controls. In HepG2 cells, IS reduced cell viability at higher concentrations and increased LDLR mRNA and protein expression. In IS-treated rats, total cholesterol, LDL-cholesterol, and triglycerides increased in a dose-dependent manner. Hepatic SREBP-2 and HMG-CoA reductase protein levels were increased at both IS doses, whereas LDLR protein abundance was increased at the higher dose. Moreover, serum PCSK9 levels were reduced in IS-treated rats. Conclusion: IS increased in HFD-fed rats. IS altered cholesterol metabolism-related pathways in HepG2 cells and in rats. In vivo IS administration increased circulating lipids and hepatic proteins involved in cholesterol synthesis and uptake. These findings indicate that IS may contribute to disturbed lipid homeostasis, although its role in HFD-induced dyslipidemia requires further mechanistic confirmation. Full article

This in vitro study examined the influence of supersaturation level on the efficacy of different inhibitors in preventing the crystallization of calcium oxalate, uric acid, and cystine. Kinetic–turbidimetric assays were used to determine the induction time of crystallization (t_i) in the absence and presence of different inhibitors. For calcium oxalate, the inhibitory effects of citrate, hydroxycitrate, tartronate, and phytate decreased significantly as the Ca²⁺ concentration increased from 220 mg/L to 240 mg/L. The polyhydroxycarboxylic compounds (citrate, hydroxycitrate, tartronate) can form complexes with calcium ions, decreasing overall supersaturation. Even though phytate also forms complexes with calcium, its urinary concentration is much lower than that of calcium, so the decrease in supersaturation can be considered negligible. For uric acid, theobromine and two of its metabolites (3-methylxanthine and 7-methylxanthine) inhibited the formation of these crystals. For each compound, the efficacy of inhibition decreased as the supersaturation of uric acid increased. For cystine, the inhibitory effect of N-acetylcysteine was lower at pH 4.5 and 5.0 than at pH 6.0. These studies of the crystallization of calcium oxalate, uric acid, and cystine showed that the efficacy of crystallization inhibitors decreased as the level of supersaturation increased. The results demonstrate that, in patients prone to kidney stone formation, the effectiveness of crystallization inhibitors can be improved by reducing the supersaturation of the specific stone-forming compound. Full article

Apical pelvic organ prolapse (POP) is characterized by descent of the uterus or post-hysterectomy vaginal vault resulting from failure of level I pelvic support and represents a major contributor to pelvic floor dysfunction and recurrent prolapse surgery. Loss of apical support is frequently associated with anterior and posterior compartment defects, leading to vaginal bulge symptoms, pelvic pressure, urinary and bowel dysfunction, sexual dysfunction, and reduced quality of life. This narrative review summarizes current knowledge on POP, from molecular mechanisms and emerging biomarkers to contemporary surgical management, with particular emphasis on sacrocolpopexy and robotic-assisted approaches. A literature search of PubMed, Scopus, Google Scholar, and Consensus identified peer-reviewed studies published up to February 2026. Evidence demonstrates that POP has a multifactorial and polygenic background involving extracellular matrix remodeling, connective tissue integrity, smooth muscle dysfunction, and altered level of protein expression. Several candidate biomarkers, including single-nucleotide polymorphisms, circulating proteins, metabolites, and imaging-based parameters, show potential for risk prediction and earlier diagnosis, although routine clinical implementation remains limited. Sacrocolpopexy remains the gold standard for apical prolapse repair because of superior anatomical outcomes, low recurrence, and significant quality-of-life improvement. Laparoscopic and robotic-assisted sacrocolpopexy provide comparable efficacy with reduced blood loss, shorter hospitalization, and faster recovery. The objective success rate is usually over 90%. Complications are very rare and typically include mesh erosion in 2–4% of cases and the need for reoperation in 6% of cases. Our own experience shows that, for a group of surgeons, the learning curve for the laparoscopic approach reached a plateau after a total of 30 operations. Robotic platforms may facilitate complex pelvic dissection and shorten the learning curve, although higher procedural costs remain a major limitation. Full article

Patients presenting with conditions attributed to environmental exposures face complex, multi-toxicant burdens, yet the stability of multivariate toxicant patterns under demographic adjustment remains poorly evaluated. This study assessed the sensitivity of principal component analysis (PCA) structures to demographic confounding and variable composition in 551 patients (aged <1–86 years) with environmentally related conditions. Nine urinary biomarkers were analysed using PCA with Varimax and Promax rotation on both raw data and residuals adjusted for age and sex. Among the identified patterns, the solvent/industrial cluster (xylene and styrene metabolites) was the most stable, persisting across both raw and residual analyses regardless of rotation method. However, overall component structures were sensitive to preprocessing: the clustering pattern of the herbicide 2,4-dichlorophenoxyacetic acid shifted markedly after demographic adjustment, illustrating indirect confounding whereby demographic effects on co-variables altered apparent biomarker associations. Notably, inclusion of an endogenous mitochondrial marker (tiglylglycine) alongside exogenous toxicant biomarkers produced Heywood cases (loadings > 1), violating factor analysis assumptions and indicating that mixing exposure and response variables destabilises the model. Cumulative variance explained was modest, consistent with the weak inter-biomarker correlations observed (KMO ≈ 0.52). These findings do not support the identification of robust, demographically stable clustering patterns in this cohort. Instead, they demonstrate that PCA-derived structures in heterogeneous clinical data are vulnerable to demographic confounding, variable selection and marker type, and caution against interpreting transient clustering patterns as definitive exposure signatures without rigorous validation. It should be noted that the absence of a non-clinical matched control group means that whether the identified co-exposure signatures are distinctive to symptomatic patients or reflective of general population exposure patterns cannot be determined; future studies should incorporate matched controls to address this directly. Full article

Background/Objectives: Bile acid synthesis disorders (BASDs) represent a distinct category of progressive familiar cholestatic liver disease. A novel targeted mass spectrometry assay was developed for the accurate measurement of the major urinary atypical bile acids and bile alcohols that are biomarkers for HSD3B7, AKR1D1, CYP7B1 and CYP27A1 deficiencies, the four most common BASDs. Methods: Stable-isotope dilution UPLC tandem mass spectrometry was used for the simultaneous quantification of 12 key atypical bile acid biomarkers in urine from patients with BASD. Typical concentration ranges for these metabolites were established from urine samples from patients with biochemically and/or genetically confirmed BASD and compared with non-cholestatic and cholestatic controls. Results: The separation of major 3β-hydroxy-Δ⁵-bile acid sulfates, taurine- and glycine-conjugated 3-oxo-Δ⁴-bile acids, and bile alcohol glucuronides was achieved in a 20 min chromatographic run with intra- and inter-batch imprecisions of <15% for all metabolites. The mean ± SEM urinary concentration of total 3β-sulfated-Δ⁵-cholenoic acids in patients with HSD3B7 deficiency was 704 ± 204 µmol/L (n = 22), approximately 2000-fold higher than in cholestastic patients (n = 168) or non-cholestatic controls (n = 127). Similarly, the concentration of 5β-cholestane-3α,7α,12α,24,25-pentol-glucuronide, the major bile alcohol, in patients with CYP27A1 deficiency was 95 ± 17 µmol/L (n = 12). For CYP7B1 deficiency, two confirmed cases showed elevated levels (average, 7.5 µmol/L) of the glycine conjugate of 3β-sulfooxy-Δ⁵-bile acid. In AKR1D1 deficiency, total 3-oxo-Δ⁴-bile acids in urine were elevated (81 ± 16 µmol/L, n = 48), but concentrations showed overlap with cholestatic and non-cholestatic controls. Conclusions: A novel quantitative tandem mass spectrometry assay is described for the measurement of the major atypical metabolites and biomarkers in urine applicable to the accurate monitoring of treatment responses, and for the first time typical concentration ranges are established for each of these BASDs. Full article

Ashwagandha (Withania somnifera (L.) Dunal; WS) is an adaptogenic herb, widely used in botanical dietary supplements. Plasma pharmacokinetics of selected withanolides, an important group of bioactive phytochemicals found in WS, have been described but no previous reports were found on their urinary excretion following WS ingestion. Healthy, older adults were administered oral doses of 240 or 480 mg of Shoden^®, a commercial extract of WS roots and leaves, and urine was collected over the subsequent 12 h. Using validated methods employing liquid chromatography coupled to multiple reaction monitoring mass spectrometry (LC-MRM-MS), we analyzed Shoden^® and participant urine samples for the presence of 11 withanolides for which standards were available. Only withaferin A (0.09 µmol/mg Shoden^®), withanoside IV (0.03 µmol/mg Shoden^®), and a mixture of 2,3-didehydrosomnifericin and 3R-viscosalactone B (0.19 µmol/mg Shoden^®) were present in Shoden^® powder in amounts greater than 0.01 µmol/mg Shoden^®. We detected four withanolides in participant urine: withaferin A; sominone, the aglycone of withanoside IV; 3R-viscosalactone B; and an abundant unidentified metabolite (WFNX), isomeric with 4-oxo withaferin A, and not detected in Shoden^®. Urinary recovery of the other 3 detected compounds was generally low (<3% of the amount in Shoden^®). These results provide new insights into the oral absorption, metabolism, and urinary excretion of withanolides found in WS and highlight additional withanolides to consider in future human pharmacokinetic and bioactivity studies of WS products. Full article

Background/Objectives: We aimed to evaluate the efficacy and safety of a short-term dietary intervention using trigonelline-rich Sakurajima radish on vascular endothelial function in patients with metabolic syndrome (MetS). Methods: In this multicenter, open-label, randomized, three-period crossover phase IIb trial, 21 patients with MetS were assigned to three 14-day sequences (Sakurajima radish powder, Aokubi radish powder, and a usual diet), separated by 14-day washouts. The primary outcome was flow-mediated dilation (FMD). Key Secondary outcomes included blood pressure (BP), urinary nitric oxide metabolites (NOx), and the oxidative stress marker 8-hydroxy-2′-deoxyguanosine (8-OHdG). Results: Sakurajima radish did not improve FMD versus the usual diet (p = 0.58) or Aokubi radish (p = 0.59), although a significant negative carryover effect following the Aokubi period likely confounded this estimation. Despite successfully stimulating NO production (elevated urinary NOx, p = 0.03), the intervention paradoxically increased oxidative stress (elevated 8-OHdG/creatinine, p = 0.02) and significantly elevated systolic BP compared with the usual diet (+9.67 mmHg, p = 0.03) and Aokubi radish (+8.86 mmHg, p = 0.04). Conclusions: Sakurajima radish does not appear to improve endothelial function in patients with MetS within the constraints of this short-term crossover design. Importantly, the unexpected negative carryover effect inherently limits the interpretability of this primary FMD outcome, as it may have masked potential physiological benefits. Despite boosting NO production, the intervention paradoxically exacerbated systemic oxidative stress and elevated systolic BP. These findings suggest that in the pro-oxidant environment of MetS, NO-boosting functional foods may induce unintended adverse hemodynamic responses, underscoring the need for careful risk–benefit evaluation and parallel-group trial designs in this specific population. Full article