Metabolites

Background/Objectives: Studies have shown that Atractylenolide I (AT-I) can exert anti-inflammatory and anti-oxidative effects, protecting against the development of various kinds of cardiovascular diseases. However, whether AT-I prevents nicotine-induced atherogenesis is unknown. This study was designed to explore the effects of AT-I on nicotine-induced macrophage pyroptosis and the progression of atherosclerosis. Methods: RT-qPCR and Western blot were used to detect the mRNA and protein levels of TXNIP and pyroptosis-related factors in THP-1-derived macrophages. ELISA was used to detect the secretion of pro-inflammatory cytokines. Hoechst/PI double-staining assay was used to assess plasma membrane integrity. The ROS assay kit, LDH release assay kit, and caspase-1 activity assay kit were used to detect ROS production, LDH release, and caspase-1 activity. Oil Red O, HE, and Masson staining were used to evaluate lipid accumulation, lesion size, and plaque stability in HFD-fed apoE−/− mice. Results: AT-I treatment significantly decreased pyroptosis-related factors expression, disrupted plasma membrane integrity, and down-regulated pro-inflammatory cytokines secretion, thereby inhibiting nicotine-induced pyroptosis of THP-1-derived macrophages. In addition, AT-I decreased ROS production and the expression of TLR4 and TXNIP. Lentivirus overexpression of TLR4 or TXNIP, or pre-treatment with ROS agonist, mainly reversed the anti-pyroptotic effects of AT-I in nicotine-treated THP-1-derived macrophages. Additionally, administering AT-I to HFD-fed apoE−/− mice markedly decreased nicotine-induced up-regulation of pyroptosis-related proteins in the aortas. Enzymatic methods and ELISA assay suggested that AT-I improved dyslipidemia and inflammation in vivo. Oil Red O, HE, and Masson staining showed that AT-I alleviated lipid accumulation, decreased plaque size, and increased plaque stability. Conclusions: Taken together, AT-I can be regarded as a potential phytomedicine that protects against macrophage pyroptosis and atherosclerosis triggered by nicotine.

Background/Objectives: Strawberries (Fragaria × ananassa) are among the most commonly consumed fruits due to their taste and nutritional benefits. However, their high rate of spoilage poses a major problem during the period from harvest and transport to further processing or marketing. The aim of this study was, therefore, to investigate the effects of passive modified atmosphere packaging on the metabolome and shelf life of strawberries as a more sustainable alternative compared to standard market storage conditions. Methods: A total of 99 strawberry samples were analyzed for microbial viable counts, water content, and metabolomic changes using non-targeted low-resolution near-infrared spectroscopy, high-resolution mass spectrometry, and microbial culture-based methods. Results: Using near-infrared spectroscopy as a rapid screening method, the linear regression model indicated that strawberries stored under modified atmosphere packaging conditions had a longer shelf life. Furthermore, lipidomic analysis using mass spectrometry showed that the levels of spoilage biomarkers, such as oxidized phosphatidylcholines and lysophosphatidylcholines, were increased under common market storage conditions without a controlled atmosphere. In contrast, the levels of these metabolites were reduced when strawberries were stored in modified atmosphere packaging. Moreover, the strawberries stored under modified atmosphere packaging had a lower number of bacteria, yeasts, and molds as well as a lower water loss throughout the entire storage period. Conclusions: Overall, the study highlights the potential of passive modified atmosphere packaging films to extend the shelf life and thus maintain the edibility of strawberries over a longer period.

Background: Infective endocarditis continues to represent a challenge for healthcare systems, requiring careful management and resources. Recent studies have indicated a shift in the predominant pathogens of concern, with Streptococcus sp. a being superseded by Staphylococcus sp. and Enterococcus sp. as the leading causes of concern. This shift is of concern as it is associated with Staphylococcus Aureus which has a high virulence rate and a tendency to form a biofilm, meaning that non-surgical therapy may not be effective. It is imperative to deliberate on the likelihood of platelet blood clot formation, which may be accompanied by bacterial infestation and the development of a biofilm. Methods: MEDLINE, Embase, and Pubmed were searched using terms relating to ‘endocarditis’ and ‘Staphilococcus aureus’, along with ‘epidemiology’, ‘pathogenesis’, ‘coagulation’, ‘platelet’, ‘aggregation’, and ‘immunity’. The search focused on publications from the past 15 years, but excluded older, highly regarded articles. We also searched the reference lists of relevant articles. Recommended review articles are cited for more details. Results: An endocarditis lesion is believed to be a blood clot infected with bacteria that adheres to the heart valves. Infective endocarditis is a good example of immunothrombosis, where the coagulation system, innate immunity and the function of coagulation in isolating and eliminating pathogens interact. However, in the context of infective endocarditis, immunothrombosis unintentionally establishes an environment conducive to bacterial proliferation. The process of immunothrombosis impedes the immune system, enabling bacterial proliferation. The coagulation system plays a pivotal role in the progression of this condition. Conclusion: The coagulation system is key to how bacteria attach to the heart valves, how vegetations develop, and how complications like embolisation and valve dysfunction occur. Staphylococcus aureus, the main cause of infective endocarditis, can change blood clotting, growing well in the fibrin-rich environment of vegetation. The coagulation system is a good target for treating infective endocarditis because of its central role in the disease. But we must be careful, as using blood-thinning medicines in patients with endocarditis can often lead to an increased risk of bleeding.

Background: Sugarcane is an important sugar crop. Sugarcane stems are mainly used for sugar extraction, while leaves can only be burned as waste. However, sugarcane leaves can also produce a large number of secondary metabolites, and these metabolites have significant nutritional and pharmacological value. At present, there are few studies on sugarcane compounds. Methods: Therefore, the stems and leaves of three sugarcane varieties (Yacheng 89-159, Dianzhe 01-58, ROC22) were selected as experimental materials, and the compounds of stems and leaves of different sugarcane were studied using high-performance liquid chromatography. Results: Metabolomics analysis detected 1197 metabolites that could be broadly divided into 11 categories. Orthogonal partial least squares discriminant analysis identified metabolites that were differentially abundant across groups (stems and leaves within and across the three varieties). Flavonoids, phenolic acids, and lipids were the main differential metabolites. Notably, tricin-4′-O-(guaiacylglycerol)ether-7-O-glucoside, quercetin-3,4-O-di-glucoside, cyanidin-3-O-(6′′-O-malony)glucoside were significantly higher in the stems than in the leaves across all three varieties. The content of methylenesuccinic acid was higher in the leaves of Dianzhe 01-58 and ROC22. In the comparative analysis of the top 20 differential metabolites among different varieties, it was found that the metabolite content of stems and leaves of Yacheng 89-9 and ROC22 was significantly higher than that of Dianzhe 01-58. Next, KEGG analysis showed that these differential metabolites were mainly enriched in pathways related to flavonoid, phenylpropanoid, and isoflavonoid biosynthesis, as well as starch and sucrose metabolism. Leaves also had significantly fewer metabolites involved in starch and sucrose metabolism than stems did. Conclusion: In conclusion, this study provides a scientific basis for utilization of sugarcane compounds, laying a theoretical foundation for further processing of sugarcane by-products into higher-value materials.

Background: Rose processing faces critical challenges in preserving bioactive compounds and aroma profiles during thermal treatments, particularly given the growing demand for natural ingredients in the food and cosmetic industries. Methods: Using widely targeted metabolomics, we first characterized volatile profiles of four major commercial cultivars (Hetian, Damask, Bulgarian, and Fenghua; n = 6 replicates per cultivar), identifying terpenoids as dominant components (p < 0.05). Subsequent thermal optimization focused on Hetian rose, where WGCNA and K-means analyses revealed temperature-dependent dynamics (40–55 °C, triplicate drying trials per temperature). Results: Hetian rose exhibited significantly higher accumulation (p < 0.05) of a unique sesquiterpene marker, 4-(1,5-dimethyl-1,4-hexadienyl)-1-methyl-cyclohexene. Systematic drying optimization identified 50 °C as the thermal threshold for optimal color, bioactive retention, and sensory quality. Mechanistic analysis identified 193 temperature-responsive metabolites (VIP > 1, FC < 0.25 or >4, p < 0.01), with terpenoid biosynthesis (MVA/MEP pathways) and esterification dynamics emerging as critical control points. Conclusions: This study establishes the first cultivar-specific processing framework for roses, demonstrating that metabolic signature-guided drying improves product quality. The findings advance our understanding of thermal impacts on aroma biochemistry while providing actionable protocols for natural product industries.

Background: Ammonia (NH3), a harmful gas, reduces livestock productivity, threatens their health, and causes economic losses. Luteolin (Lut), an anti-inflammatory flavonoid, may counteract these effects. Methods: Our study explored luteolin’s protective mechanisms on chicken splenic lymphocytes under ammonia stress using a simulation model and four-dimensional fast data-independent acquisition (4D-FastDIA) proteomics. We identified 316 proteins, with 69 related to ammonia’s negative effects and 247 to Lut’s protection. Thirty differentially expressed proteins (DEPs) were common to both groups, with 27 showing counter-regulation with Lut. Results: Gene Ontology (GO) analysis showed DEPs enriched in molecular responses to interferons and the negative regulation of immune responses, mainly located extracellularly. Molecular function analysis revealed DEPs in antigen binding and synthase activity. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis linked DEPs to pathways like estrogen signaling, NOD-like receptor signaling, cytokine–cytokine receptor interaction, and JAK-STAT signaling. The quantitative real-time polymerase chain reaction (qRT-PCR) results indicated that the mRNA levels of Interferon Alpha and Beta Receptor subunit 2 (IFNAR2) and Signal Transducer and Activator of Transcription 1 (STAT1) were trending downward. This observation was in strong agreement with the downregulation noted in the proteomics analysis. Conclusions: Lut’s protective role against ammonia’s adverse effects on chicken splenic lymphocytes is linked to the modulation of key signaling pathways, offering insights for further research on treating ammonia exposure with Lut.

Background: Whether intestinal epithelial cells can regulate distant adipose tissue remains a mystery. Methods: Cold-stimulated intestinal epithelial cell-derived exosomes (Cold IEC-Exo) play a pivotal role in enhancing adipose thermogenesis and metabolic homeostasis, as demonstrated in this study. Results: IEC-Exo can accumulate in adipose tissue. Compared with IEC-Exo derived from room temperature mice (RT IEC-Exo), Cold IEC-Exo significantly enhanced the thermogenesis of adipose. In vitro, Cold IEC-Exo directly stimulated thermogenesis in primary adipocytes by elevating oxygen consumption rate, proton leak, and fatty acid uptake, with no effect on glucose uptake. Small RNA sequencing identified miR-674-3p as a key mediator enriched in Cold IEC-Exo. miR-674-3p mimicry replicated Cold IEC-Exo effects, augmenting Ucp1 expression, mitochondrial uncoupling, and fatty acid utilization in adipocytes. Local overexpression of miR-674-3p in BAT and sWAT via AAV in vivo enhanced thermogenesis and attenuated diet-induced glucose intolerance. Conclusions: These findings establish that Cold IEC-Exo, via miR-674-3p transfer, drive adipose thermogenic activation and mitigate metabolic dysfunction, highlighting their therapeutic potential in obesity-related disorders.

Background: Muscle loss during sarcopenia and atrophy is also commonly associated with age-related insulin resistance. Interestingly, branched-chain amino acids (BCAA) which are known for stimulating muscle protein synthesis are commonly elevated during insulin resistance and sarcopenic obesity. Objectives: This study investigated the effects of the interplay between atrophy and insulin resistance on insulin sensitivity, mitochondrial metabolism, and BCAA catabolic capacity in a myotube model of skeletal muscle insulin resistance. Methods: C2C12 myotubes were treated with dexamethasone to induce atrophy. Insulin resistance was induced via hyperinsulinemia. Gene and expression were measured using qRT-PCR and Western blot, while mitochondrial and lipid content were assessed using fluorescent staining. Cell metabolism was analyzed via Seahorse metabolic assays. Results: Both dexamethasone-induced atrophy and insulin resistance independently reduced insulin-stimulated pAkt levels, as well as mitochondrial function and content. However, neither treatment affected gene or protein expression associated with mitochondrial biogenesis or content. Although dexamethasone independently reduced insulin sensitivity in otherwise previously insulin-sensitive cells, dexamethasone had no significant effect on extracellular BCAA content. Conclusions: Our findings indicate the metabolic interplay between atrophy and insulin resistance and demonstrate that both can reduce mitochondrial function, though only limited effects were observed on indicators of BCAA catabolism and utilization. This emphasizes the need for future studies to investigate the mechanisms that underlie atrophy and other metabolic disorders to develop new interventions.

Background/Objectives: Strawberries are highly appreciated for their rich phytochemical composition, but rapid postharvest deterioration limits their shelf life and nutritional quality. This study aimed to investigate the metabolic changes occurring in both strawberry fruits and leaves during storage and to evaluate the NADPH oxidase 2 (NOX2) inhibitory potential of strawberry-derived metabolites. Methods: Untargeted LC-MS/MS analysis was conducted on fruit and leaf tissues stored at 8 ± 0.5 °C. A total of 37 metabolites were identified, including organic acids, phenolic acids, flavonoids, and hydroxycinnamic acid derivatives. Multivariate statistical analyses (ANOVA, PLS-DA, and volcano plots) were used to assess temporal and tissue-specific metabolic shifts. Additionally, a machine learning-based predictive model was applied to evaluate the NOX2 inhibitory potential of 24 structurally characterized metabolites. Results: Storage induced significant and tissue-specific metabolic changes. In fruits, malic acid, caffeic acid, and quercetin-3-glucuronide showed notable variations, while ellagic acid aglycone and galloylquinic acid emerged as prominent markers in leaves. The predictive model identified 21 out of 24 metabolites as likely NOX2 inhibitors, suggesting potential antioxidant and anti-inflammatory bioactivity. Conclusions: These findings provide new insights into postharvest biochemical dynamics in both strawberry fruits and leaves. The results highlight the value of leaves as a source of bioactive compounds and support their potential valorization in functional food and nutraceutical applications.

Background/Objectives: Metabolomics, in combination with genetic data, is a powerful approach to study the biochemical consequences of genetic variation. We assessed the impact of human gene knockouts (KOs) on the metabolite levels of Estonia Biobank (EstBB) participants and integrated the results with electronic health record data. Methods: In 150,000 EstBB genotyped participants, we identified 723 KOs with 152 different predicted loss of function (pLoF) variants in 115 genes. For those KOs and 258 controls, 1387 metabolites were profiled using ultra-high-performance liquid chromatography–tandem mass spectrometry. Results: We identified 48 associations linking rare pLoF variants in 22 genes to 43 metabolites. Out of 48 associations, 27 (56%) were found in genes that cause inborn errors of metabolism. The top associations identified in our analysis included genes and metabolites involved in the degradation pathway of the pyrimidine bases uracil and thymine (DPYD and UPB1). We found DPYD gene KOs to be associated with elevated levels of Uracil, confirming that DPD-deficiency is a leading cause of severe 5-Fluorouracil toxicity. Overall, 54% of reported associations are gene targets of approved drugs or bioactive drug-like compounds. Conclusions: Our findings contribute to assessing the impact of human KOs on metabolite levels and offer insights into gene functions, disease mechanism, and drug target validation.

Background/Objectives: Maternal free fatty acids (FFAs) play a critical role in maternal metabolism, fetal growth, and pregnancy outcomes. However, their relationship with maternal metabolic status in early pregnancy and the subsequent development of gestational diabetes mellitus (GDM) remains unclear. Aim: Assess the trajectory of FFA concentrations during pregnancy, considering first-trimester metabolic status (obesity, insulin resistance—IR) and the development of GDM, and evaluate whether first-trimester FFA is a relevant risk factor for GDM. Methods: A case–control study nested within the OBESO cohort (Mexico City, pregnant women and their children), classified women according to first-trimester metabolic status (pregestational body mass index—pBMI, insulin resistance homeostasis model assessment—HOMA-IR > 1.6), as well as the presence of GDM: Group 1 (normal weight without IR, n = 60), Group 2 (obesity without IR, no GDM, n = 20), Group 3 (obesity with IR, no GDM, n = 20), and Group 4 (obesity with IR, with GDM, n = 9). FFA concentrations were measured each trimester. Statistical analyses included repeated measures ANOVA and logistic regression models. Results: FFA concentrations were the highest in Group 4 across all trimesters (p < 0.05). FFAs decreased throughout pregnancy in all groups (p = 0.023), with the most significant decline from the first to the third trimester (p < 0.001). The greatest reduction occurred in Group 4 (p < 0.001), followed by Group 3. Multivariate logistic regression showed no association between first-trimester FFAs and the development of GDM. Higher gestational weight gain was associated with a higher GDM risk (OR: 1.22, 95%CI: 1.01–1.48), when the FFAs difference was accounted for. Conclusions: FFA levels are higher in women with GDM compared with women with obesity or a normal weight. However, FFAs progressively decline from the first to the third trimester, with the most pronounced decrease in women with obesity, IR, and GDM.

Background: Small RNA, defined as RNA molecules of less than 200 nucleotides in length, play pivotal regulatory roles in plant growth, development, and environmental stress responses. However, research on modifications in plant small RNA remains limited. Methods: In this study, we developed a liquid chromatography–tandem mass spectrometry (LC-MS/MS) method for the simultaneous detection of 41 RNA modifications, facilitating the systematic qualification and quantification of modifications in plant small RNA. Results: We identified a total of nine modifications, among which N6,N6-dimethyladenosine (m6,6A) is a newly identified modification in plant small RNA. Furthermore, we conducted a quantitative analysis of these modifications in Arabidopsis thaliana during the germination process and observed significant dynamic changes in their abundance from 1 to 5 days post-germination. Notably, the trends in the contents of these modifications exhibited a strong correlation with the reported gene expression levels of the relevant modifying enzymes and demodifying enzymes, suggesting that these modifications may play essential roles during seed germination and are tightly regulated by the genes of the corresponding enzymes. Conclusion: The discovery of these modifications in plant small RNA, coupled with the dynamic changes in their levels during germination, holds great promise for a further understanding of the physiological functions of small RNA modifications and their associated regulatory mechanisms in plant seed germination.

Background/Objectives: Fatty liver and hyperuricemia are growing public health concerns linked to unhealthy lifestyles, yet their combined effects in working populations remain underexplored. This study investigates their associations with metabolic risk factors, inflammation, and liver dysfunction to inform workplace health strategies. Methods: The participants were employees aged 20 or older from four industrial enterprises located in central Taiwan. A total of 3089 participants (2571 males, 518 females) were analyzed. Lifestyle factors were assessed via a self-administered questionnaire, fatty liver was diagnosed using ultrasound, and serum uric acid levels, metabolic parameters, inflammatory markers, and liver function were measured. Results: The prevalence of fatty liver (43.2%) exceeded that of hyperuricemia (25.5%), with a higher burden among males. Fatty liver was associated with lower physical activity, while alcohol consumption was significantly higher in individuals with both conditions. Both conditions correlated with increased metabolic risk factors, liver dysfunction, and inflammation. Health-related risk factors were compared across four groups, using Group A (no hyperuricemia/no fatty liver, OR = 1.00) as the reference. The risk of metabolic syndrome increased progressively: 2.90 (Group B: hyperuricemia/no fatty liver), 6.15 (Group C: no hyperuricemia/fatty liver), and 11.52 (Group D: hyperuricemia/fatty liver), following the trend A < B < C < D. Notably, Group D had the highest risk, with exacerbated inflammation and liver dysfunction. Conclusions: Fatty liver and hyperuricemia synergistically worsen metabolic disorders, inflammation, and liver dysfunction. Early detection and lifestyle interventions are crucial to mitigating long-term health risks.

Background: There is a continuous demand to create new, superior sensory food experiences. In the food industry, yeast-derived flavor products (YPs) are often used as ingredients in foods to create new aromas and taste qualities that are appreciated by consumers. Methods: Chicken bouillon samples containing diverse YPs were chemically and sensorially characterized using statistical multivariate analyses. The sensory evaluation was performed using quantitative descriptive analysis (QDA) by trained panelists. Thirty-four sensory attributes were scored, including odor, flavor, mouthfeel, aftertaste and afterfeel. Untargeted metabolomic profiles were obtained using stir bar sorptive extraction (SBSE) coupled to GC-MS, RPLC-MS and targeted HILIC-MS. Results: In total, 261 volatiles were detected using GC-MS, from chemical groups of predominantly aldehydes, esters, pyrazines and ketones. Random Forest (RF) modeling revealed volatiles associated with roast odor (2-ethyl-5-methyl pyrazine, 2,3,5-trimethyl-6-isopentyl pyrazine) and chicken odor (2,4-nonadienal, 2,4-decadienal, 2-acetyl furan), which could be predicted by our combined model with R2 > 0.5. In total, 2305 non-volatiles were detected for RPLC-MS and 34 for targeted HILIC-MS, where fructose-isoleucine and cyclo-leucine-proline were found to correlate with roast flavor and odor. Furthermore, a list of metabolites (glutamate, monophosphates, methionyl-leucine) was linked to umami-related flavor. This study describes a straightforward data-driven approach for studying foods with added YPs to identify flavor-impacting correlations between molecular composition and sensory perception. It also highlights limitations and preconditions for good prediction models. Overall, this study emphasizes a matrix-based approach for the prediction of food taste, which can be used to analyze foods for targeted flavor design or quality control.

Background: Maillard reaction products (MRPs) are known for their antioxidant properties; however, their effects on muscle cells remain unclear. This study aims to elucidate the effects of MRPs on muscle hypertrophy and atrophy in C2C12 myotubes. Methods: MRPs were prepared by heating L-lysine and D-glucose, and their antioxidant activity was assessed using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. Subsequently, mouse C2C12 myoblasts were cultured with MRPs until myotubes formed, and their diameters were measured to assess hypertrophic and atrophic changes. Akt phosphorylation was evaluated by Western blotting, and gene expression levels were analyzed via quantitative PCR. Results: The prepared MRPs exhibited high antioxidant activity in the DPPH radical scavenging assay. MRP treatment significantly increased the average myotube diameter by approximately 40% and enlarged the largest myotube diameter by up to 80%, potentially mediated by enhanced Akt phosphorylation. Under dexamethasone-induced atrophy, MRPs modestly attenuated the reduction in myotube diameter by approximately 20%, although the effect was not statistically significant, and did not significantly alter the fusion index either. Quantitative PCR analysis revealed that MRP treatment significantly reduced the mRNA expression of Nfe2l2, a key regulator of antioxidant response, whereas it had no notable effects on the expression of genes related to myoblast proliferation (Myod1), differentiation (Myog), hypertrophy (Igf1), atrophy (Foxo1 and Trim63), and oxidative stress (Cat, Gclc, and Nqo1). Conclusions: Our findings suggested that MRPs possess antioxidant activity and promote myotube hypertrophy via Akt signaling. This study highlighted the potential of MRPs as functional ingredients for promoting muscle health, though further in vivo studies are required to validate their physiological relevance.

Background/Objectives: Carvacrol is a naturally occurring phenolic monoterpene that is one of the main constituents of the essential oils of oregano (Origanum vulgare) and other herbs. Carvacrol has anti-inflammatory and antinociceptive effects. Carvacrol can activate and inhibit several second messengers and ionic channels at the systemic level. However, there is no evidence of the peripheral antinociception of carvacrol and its mechanism of action. This study was designed to determine whether the opioid receptor-nitric oxide (NO)-cyclic guanosine monophosphate (cGMP)-K+ channel pathway is involved in the local antinociception of carvacrol. Methods: Wistar rats were injected with 1% formalin subcutaneously on the dorsal surface of the right hind paw with the vehicle or carvacrol (100–300 µg/paw). To determine whether the opioid receptor-NO-cGMP-K+ channel pathway and a biguanide-dependent mechanism are responsible for the local antinociception induced by carvacrol, the effect of the injection (10 min before the 1% formalin injection) with the corresponding vehicles, metformin, naltrexone, NG-L-nitro-arginine methyl ester (L-NAME), 1 H-(1,2,4)-oxadiazolo (4,2-a) quinoxalin-1-one (ODQ), and K+ channel blockers on the antinociception induced by local carvacrol (300 µg/paw) was determined. Results: In both phases of the formalin test, carvacrol produced antinociception. Naltrexone, metformin, L-NAME, ODQ, glibenclamide and glipizide (both ATP-sensitive K+ channel blockers), tetraethylammonium and 4-aminopyridine (voltage-gated K+ channel blockers), and apamin and charybdotoxin (Ca2+-activated K+ channel blockers) reversed the carvacrol-induced peripheral antinociception. Conclusions: The local peripheral administration of carvacrol produced significant antinociception and activated the opioid receptor-NO-cGMP-K+ channel pathway.

Objectives: This study aims to elucidate the metabolic differences between obese adolescents categorized into low-weight-loss (LWL) and high-weight-loss (HWL) groups. Methods: The objective of this study is to investigate the metabolic characteristics of obese adolescents, with a focus on the statistically significant individual differences observed in weight loss outcomes after the same dietary and exercise training intervention. A four-week exercise and dietary intervention was administered to the participants. Obese adolescents were categorized into LWL (with a weight loss percentage of 5–10%) and HWL (with a weight loss percentage of >10%) groups on the basis of their weight loss outcomes. Post-intervention changes in body morphology and body composition between the two groups were compared using Analysis of Covariance (ANCOVA), with gender as a covariate. Additionally, metabolic changes were analyzed in depth; differential metabolites between the groups were identified through ANCOVA adjusted for gender, followed by pathway analysis. Results: After the four-week exercise intervention, the body morphology and composition of the obese adolescents showed significant improvements compared with those before the intervention (p < 0.001). For example, weight decreased from 80.65 kg to 72.35 kg, BMI decreased from 30.57 kg/m2 to 27.26 kg/m2, waist circumference decreased from 103.64 cm to 94.72 cm, and body fat percentage decreased from 32.68% to 28.54%. Prior to the exercise intervention, no significant differences in body morphology and composition were observed between the HWL and LWL groups (p > 0.05). After the intervention, the HWL group demonstrated significant improvements in weight, body mass index, waist circumference, body fat percentage, fat mass, fat-free mass, body water amount, and skeletal muscle mass compared with the LWL group (p < 0.001). After controlling for the levels of pre-intervention metabolites, 27 differential metabolites were identified between the HWL and LWL groups. These metabolites were categorized into fatty acids, amino acids, organic acids, carnitines, indoles, benzoic acids, and carbohydrates. Notably, they were significantly enriched in the eight metabolic pathways involved in amino acid metabolism, fatty acid biosynthesis, and coenzyme A biosynthesis. Conclusions: A four-week exercise intervention enhanced the body morphology and physical fitness of obese adolescents, although the degree of weight loss varied among individuals. Considerable weight reduction was significantly correlated with metabolites involved in lipid, amino acid, organic acid, carbohydrate, and gut microbiota metabolism and with the enrichment of pathways involved in amino acid metabolism, fatty acid biosynthesis, and coenzyme A biosynthesis. These findings indicate that intrinsic metabolic characteristics considerably influence individual responsiveness to exercise-based weight-loss interventions.

Background: Premenstrual syndrome is characterized by emotional changes, including anxiety and depression symptoms, which may be treated with anxiolytic and antidepressant drugs, as well as estrogen therapy. However, steroidal estrogen therapy is contraindicated for patients with a potential risk of developing estrogen-dependent cancers through interactions with estrogen receptor α (ERα). Alternatively, genistein produces estrogenic effects in animals and humans at dietary dosages that act on the nuclear and membrane ERα, estrogen receptor β (ERβ), and the G-protein-coupled estrogen receptor (GPER). These receptors are likely involved in the anxiety symptoms observed in premenstrual disorders. The objective of this study was to evaluate the effects of genistein and 17β-estradiol on anxiety-like behavior and the plasma concentrations of estradiol and progesterone throughout the ovarian cycle of Wistar rats. Methods: The effect of the administration of 0.09 mg/kg of genistein or 17β-estradiol was evaluated using the elevated plus maze (EPM) test, locomotor activity test (LAT), and light/dark box (LDB) test, as well as by assessing the plasma concentrations of estradiol and progesterone, while considering the ovarian cycle phases. Results: Higher levels of anxiety-like behavior were detected in the metestrus–diestrus phase compared to the proestrus–estrus phase, which was associated with low concentrations of estradiol. Genistein, similarly to 17β-estradiol, significantly reduced anxiety-like behaviors in the EPM and LDB; however, 17β-estradiol, but not genistein, significantly increased the plasma estradiol concentration. No significant changes were found in locomotor activity or the plasma progesterone concentrations due to the treatments. Conclusions: These findings suggest that genistein may be useful in the development of alternative therapies to reduce the anxiety associated with low steroid hormone concentrations, which occur in premenstrual syndrome. Genistein could be an alternative to steroidal estrogen therapy to avoid potential side effects due to estradiol or antidepressant treatments, although it still requires medical care.

Background: The advent of tandem mass spectrometry (MS/MS) had an essential role in the expansion of newborn screening (NBS) for different inborn errors of metabolism (IEMs). Nowadays, almost 50 IEMs are screened in Italy. The use of second-tier tests (2-TTs) in NBS minimizes the false positive rate; nevertheless, the metabolic profile is influenced not only by the genome but also by environmental factors and clinical variables. We reviewed the MS/MS NBS data from over 37,000 newborns (of which 8% required 2-TTs) screened in the Italian Abruzzo region to evaluate the impact of neonatal and maternal variables on propionate-related primary biomarker levels. Methods: Expanded NBS and 2-TT analyses were performed using MS/MS and liquid chromatography–MS/MS methods. We set up layered cut-offs dividing all 37,000 newborns into categories. Statistical analysis was used to create alarm thresholds for NBS-positive samples. Statistically significant differences were found in both neonatal and maternal conditions based on the 2-TTs carried out. According to the stratified cut-offs, only 1.47% of the newborns would have required a 2-TT while still retaining the ability to recognize the true-positive case of methylmalonic acidemia with homocystinuria, which has been identified by NBS. To further support the clinical applicability, we performed an external evaluation considering nine positive cases from an extra-regional neonatal population, confirming the potential of our model. Interestingly, the setting of alarm thresholds and their application would allow for establishing the degree of priority/urgency for 2-TTs. Conclusions: Tailoring NBS by customized cut-offs may enhance the application of precision medicine, focusing on true-positive cases and also reducing analysis costs and times.

Background/Objectives: The prevalence of obesity worldwide has rapidly increased. Numerous studies showed a beneficial effect of probiotics in obese individuals, and changes in hematological parameters are observed in obesity. Therefore, the aim of this study was to investigate the effect of a novel probiotic approach on the red blood cells (RBCs) and platelets. Methods: Twenty-five obese women participated in a randomized, placebo-controlled study and were divided into the experimental group (one capsule daily containing Lactiplantibacillus plantarum 299v (DSM9843), Saccharomyces cerevisiae var. boulardii, and 40 mg octacosanol; n = 13) and the placebo group (n = 12). Blood samples were collected for light microscopic examination, morphometric analysis, and an automated hematology analyzer. A possible relationship between hematological parameters and body mass index (BMI), a common indicator of obesity, was investigated using Spearman correlation. The plasma concentration of soluble P-selectin and fibrinogen were determined using an ELISA assay. All measurements were performed before (T0) and after 12 weeks of supplementation (T1). Results: The three-month supplementation of probiotics improved hemoglobin levels, chromic status, and red blood cell morphology. The mean platelet volume (MPV), a measure of platelet size, was restored to normal levels, platelet morphology was improved, and the number of activated platelets was significantly reduced (p < 0.05). A strong negative correlation (r = −0.5904, p < 0.05) was found between BMI and platelet distribution width (PDW), a measure of variation in platelet size and shape. Conclusions: The results show that the probiotic approach improves morphology and normalizes the values of disturbed hematological parameters of RBCs and platelets in obese women.

Background: Microbesas chemical factories provide an alternative sustainable approach for producing platform chemicals. Until recently, most efforts have involved engineering heterologous pathways into a single microbial chassis to maximise its production of a target chemical. More recently, cohorts of microbes have been used to engineer microbial communities to achieve higher yields than achieved in a single chassis.

Background: This study presents a sensitive method for the simultaneous determination of organic acids, flavonoids, and amino acids in Mume Fructus (MF) using ultra-performance liquid chromatography coupled with triple-quadrupole linear ion-trap tandem mass spectrometry (UPLC-QTRAP-MS/MS). Methods: Analysis was performed on a UPLC system (Shimadzu, Kyoto, Japan) equipped with a quaternary pump solvent management system, an online degasser, a triple-quadrupole mass detector, and an autosampler. An Agilent ZORBAX SB-C18 column (3.0 mm × 100 mm, 1.8 µm) was used for chromatographic analyses. The mobile phase was distributed between 0.2% aqueous formic acid (A) and 0.2% formic acid acetonitrile (B) at a velocity of 0.2 mL/min. The gradient evolution protocol was 0–2 min at 90–70% B; 3–7 min at 70–50% B; 7–10 min at 50–20% B; 10–14.5 min at 20–90% B; and 14.5–17 min at 10% B. Results: The method was validated for matrix effects, linearity, limits of detection/quantification, precision, repeatability, stability, and recovery of target components. It effectively determined all target compounds in 12 MF batches from different drying methods. Conclusions: Principal component analysis (PCA) of 47 active components was conducted to evaluate MF quality comprehensively. The proposed method serves as a reliable approach for assessing the consistency of MF’s quality and therapeutic efficacy.

Background: Parkinson's disorder (PD) affects around 1:500 individuals and is associated with enlarged ventricles and symptoms of normal pressure hydrocephalus (NPH). These features suggest disrupted cerebrospinal fluid (CSF) dynamics and folate metabolism. With L-DOPA treatment showing diminishing benefits over time, there is an urgent need to investigate upstream metabolic disruptions, including folate and tetrahydro-biopterin (BH4) pathways, in post-mortem CSF and brain tissue to understand their roles in PD pathogenesis. Methods: CSF and brain tissue from 20 PD patients (mean age 84 years; 55% male; disease duration 10-30 years) and 20 controls (mean age 82 years; 50% male) were analysed. Western and Dot Blots measured proteins and metabolites, spectroscopic assays assessed enzyme activities, BH4 and Neopterin levels were measured using ELISA, and levels of hydrogen peroxide, used as a proxy for reactive oxygen species, and calcium were quantified using horseradish peroxidase and flame photometry assays, respectively. ClinVar genetic data were analysed for variants in genes encoding key enzymes. Statistical significance was assessed using unpaired t-tests (p < 0.05). Results: All enzymes were significantly reduced in PD compared to controls (p < 0.01) except for methyltetrahydrofolate reductase (MTHFR), which was elevated (p < 0.0001). Enzymes were functional in control but undetectable in PD CSF except tyrosine hydroxylase (TH). BH4 and Neopterin were elevated in PD CSF (p < 0.0001, p < 0.001) but significantly reduced (p < 0.001) or unchanged in tissue. Peroxide was increased in both PD CSF (p < 0.001) and tissue (p < 0.0001) selectively inhibiting TH. Calcium was 40% higher in PD than controls (p < 0.05). No pathogenic variants in enzyme genes were found in ClinVar data searches, suggesting the observed deficiencies are physiological. Conclusions: We identified significant disruptions in folate and BH4 pathways in PD, with enzyme deficiencies, oxidative stress and calcium dysregu-lation pointing to choroid plexus dysfunction. These findings highlight the choroid plexus and CSF as key players in cerebral metabolism and promote further exploration of these as therapeutic targets to address dopaminergic dysfunction and ventricular enlargement in PD.

Background/Objectives: Plant-derived secondary metabolites have long contributed to the discovery of novel therapeutic agents, especially in the treatment of parasitic and infectious diseases in developing countries. Metabolomics provides a systems-level approach to understanding plant biochemistry, enabling the discovery of secondary metabolites with pharmacological relevance. Taraxacum kok-saghyz (TKS), widely known for its rubber-producing capabilities, remains underexplored as a medicinal plant. Given the well-established therapeutic properties of Taraxacum officinale and the emerging pharmacological profiles of related species, this study investigates the metabolic composition of TKS roots and leaves to uncover bioactive compounds with antioxidant, anti-inflammatory, or hepatoprotective potential. Methods: Widely targeted metabolomics was conducted on 10-month-old field-grown Kultevar™ TKS plants using ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS). Samples were hand-harvested and preserved on dry ice to maintain biochemical integrity. Metabolite identification and classification were performed using the MWDB and KEGG databases. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to evaluate metabolic variation between tissues. Results: A total of 1813 metabolites were identified, including flavonoids, alkaloids, lipids, amino acids, and phenolic compounds. Differential analysis revealed 964 significantly altered metabolites—609 downregulated and 355 upregulated in roots relative to leaves. Multivariate analysis confirmed clear tissue-specific metabolic profiles. KEGG pathway enrichment highlighted the involvement of flavonoid biosynthesis, amino acid metabolism, and lipid metabolism pathways, suggesting bioactive potential. This study presents the first comprehensive metabolic profile of TKS, highlighting its potential value beyond rubber production. The detection of numerous therapeutic secondary metabolites supports its promise as a pharmaceutical and nutraceutical resource. Further functional validation of identified compounds is warranted.