Search Results (278)

This study evaluated the effects of including spray-dried porcine plasma (SDP) in nursery diets and enzymatically hydrolyzed plasma (EHP) in drinking water on piglet growth performance and post-weaning diarrhea (PWD). Four treatments were tested: CONTROL (soy protein concentrate, SPC), P1SDP (5% SDP in phase 1), P1 + P2SDP (5% SDP and 2% SDP in phases 1 and 2), and EHP (0.88% in water during phases 1 and 2). No significant differences among treatments were observed during phase 1. During phase 2 (14–28 days) pigs fed SDP or pigs provided EHP in water had higher average daily gain (ADG; p = 0.001) and feed conversion (GFR; p = 0.013) versus the other groups. Pigs fed SDP in the first two phases had an average d-42 body weight that was 1.54 kg heavier than controls. Post-weaning diarrhea was not observed at any time during the study. These results support the use of SDP and EHP as effective nutritional strategies to enhance the growth and resilience of pigs during the post-weaning period. Both ingredients contribute to sustainable pig production by improving efficiency and promoting circular economy practices through the valorization of animal by-products. Full article

Saskatoon berry (SB), a traditional food of Indigenous people, has been associated with cardiometabolic benefits in animal models; however, its effects on humans remain unclear. This study investigated the effects of dried SB consumption on cardiometabolic outcomes, gut microbiota, and short-chain fatty acids (SCFAs) profiles in healthy adults. In a 10-week, single-arm, and open-label trial, 20 healthy adults consumed 40 g/day of freeze-dried whole SB. Biochemical measures, physical exams, dietary records, participant feedback, and fecal samples were collected before and after the intervention. Gut microbiota composition and fecal SCFAs were profiled using 16S-rRNA sequencing and gas chromatography–mass spectrometry, respectively. SB intake significantly reduced fasting plasma glucose, total cholesterol (TC), low-density lipoprotein-cholesterol (LDL-c), non-high-density lipoprotein-cholesterol (non-HDL-c), systolic blood pressure, and high-sensitivity C-reactive protein, while increasing dietary fiber intake. Fiber intake was negatively correlated with TC, LDL-c and non-HDL-c (p < 0.05). The relative abundance of fecal Prevotellaceae increased after SB consumption and was positively correlated with multiple fecal SCFAs (p < 0.05–0.0001), while being negatively associated with lipid profiles and blood pressure. No adverse cardiovascular, hepatic, or renal dysfunction were observed; however, the significant increase in sugar intake may pose a risk for elevated blood glucose. Therefore, limiting other high-sugar foods during SB supplementation may be advisable for individuals with glucose intolerance. Overall, SB intake improved glucose and lipid metabolism and lowered blood pressure and inflammatory markers in healthy adults. These cardiometabolic benefits may be mediated by fiber and anthocyanins in SB and through modulation of gut microbiota and SCFA production; however, further confirmation is needed in subsequent randomized controlled trials. Full article

Background: Metabolomic studies have generated extensive data on metabolic changes in aging and disease, yet translating this data into practical nutrition guidelines remains challenging. Recent analysis identified pathways common to both processes, termed the metapathway. As a network, it features key central metabolites that most representatively reflect its state. The manageable number of these key metabolites provides a practical basis for translating complex metabolomic data into actionable nutritional information. Methods: We developed a conceptual framework for precision nutrition approach involving: (1) selecting an initial (baseline) diet with minimal impact on key metapathway metabolites, (2) defining dietary modifications using foods and supplements that are capable of elevating them, and (3) implementing mass spectrometry-based metabolome fingerprinting to assess individual responses. This capability was evaluated using blood plasma and dried blood spot samples. Results: A promising precision nutrition was created, consisting of a selected baseline diet and its metabolomics-guided modification. The metabolic fingerprinting demonstrated the possibility of determining the diet outcome by identifying biological age change with an accuracy of 1 month. Conclusions: The fully metabolomics-guided nutrition strategy has been developed and is ready for further human testing to validate its translational potential and health benefits. Full article

Hyperlipidemia is a major global risk factor for cardiovascular disease, underscoring the need for safe, multi-target preventive strategies. In this study, two novel dietary supplements were developed by blending freeze-dried aqueous extracts of chamomile (CDS) or thyme (TDS) with linseed oil (1:1, w/w) and evaluated for their phytochemical composition, antioxidant capacity, and hypolipidemic efficacy. Total phenolics, total flavonoids, fatty acid composition, volatile constituents, and individual phenolic profiles were characterized, while antioxidant activity was assessed using DPPH· radical-scavenging and FRAP assays. Hypolipidemic activity was investigated in a Triton X-100-induced hyperlipidemia rat model through an assessment of plasma lipid parameters, oxidative stress and inflammatory markers, and liver and kidney function indices, supported by hepatic histopathology. Molecular docking was performed to explore the interactions of major bioactive compounds with AMP-activated protein kinase (AMPK) and HMG-CoA reductase. Both CDS and TDS exhibited strong antioxidant activity and high polyphenol content, with kaempferol and chlorogenic acid identified as the predominant phenolics in CDS and TDS, respectively. β-Farnesene and carvacrol were the main volatile constituents. In vivo, both formulations significantly reduced total cholesterol, triglycerides, LDL-C, lipid peroxidation markers, and TNF-α, while increasing HDL-C and improving cardiac risk indices, with more pronounced effects observed for TDS. Histopathological analyses confirmed marked hepatoprotection, particularly in the TDS-treated group. Docking analyses identified ellagic acid as the strongest dual binder to both AMPK and HMG-CoA reductase. Overall, these findings demonstrate that chamomile-linseed and thyme-linseed formulations exert synergistic, multi-target antioxidant and hypolipidemic effects, supporting their potential as nutraceutical strategies for the early prevention and management of hyperlipidemia and cardiometabolic risk. Full article

Epilepsy remains one of the most prevalent neurological disorders, characterised by complex aetiology encompassing genetic, structural, metabolic, and inflammatory factors. Despite advances in neuroimaging and neurophysiological diagnostics, there is a persistent lack of sensitive and specific biomarkers to enable early diagnosis, risk stratification, and monitoring of therapeutic efficacy. Key epilepsy biomarkers include neurotransmitters, energy–related compounds, tryptophan pathway metabolites, and choline derivatives. Their determination employs liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS), high–performance liquid chromatography (HPLC) with electrochemical or fluorescence detection, gas chromatography with tandem mass spectrometry (GC–MS/MS), high–resolution mass spectrometry (HRMS), and proton nuclear magnetic resonance (¹H–NMR) spectroscopy, revealing metabolic disturbances in neurotransmission, energy metabolism, and oxidative stress associated with epileptogenesis. Among these techniques, LC–MS/MS currently provides the highest analytical sensitivity and specificity for quantifying low–abundance epilepsy–related metabolites, while HPLC with conventional detection remains a simpler and more cost–effective alternative for routine clinical laboratories. This review presents the current state of knowledge regarding chromatographic techniques applied to the analysis of mentioned metabolites, as well as therapeutic drug monitoring of antiepileptic drugs. Key sample preparation stages are also discussed. Various biological matrices–plasma, serum, urine, cerebrospinal fluid (CSF), dried blood spots (DBSs), and brain tissue—are evaluated. Novel approaches are also presented, including hair samples, microsampling techniques, and headspace analysis of volatile metabolites. Chromatographic techniques constitute the foundation of contemporary metabolomic research in epileptology, enabling biomarker identification and supporting personalised medicine. Further standardisation and translational validation remain necessary, as current evidence is insufficient for routine clinical implementation. Full article

Background/Objectives: Dried blood spot (DBS) sampling, a technique for collecting capillary blood samples, is widely used in therapeutic drug monitoring, pharmacokinetic and toxicology research, newborn screening, and population health because it enables simple, non-invasive sampling across large cohorts. However, it presents several challenges, mainly due to the effect of hematocrit (HCT), which can influence the quantification of analytes. Methods: A combination of methods was developed to estimate the HCT and blood volume in DBS samples. Image analysis and hemoglobin (Hb) quantification using UV-VIS spectrometry were used for HCT estimation, and conductivity was used to determine blood volume. DBS samples from five donors were prepared with HCT between 0.2 and 0.6 and were used to prepare calibrators and quality control samples. The developed methods were applied to 23 samples obtained from ten adult patients with inflammatory bowel disease. Results: The methods for HCT determination using image analysis or Hb measurements were linear (r² > 0.994), with acceptable accuracy (90.3–102.2%) and precision (<7.4%). Moreover, the conductivity method was linear (r² = 0.999) and enabled accurate (96.8–100%) and precise (<5.65%) determination of blood volume in DBS samples. All three methods were in good agreement with the reference values in patient samples. Finally, strategies to correct HCT- and volume-related bias in DBS samples were proposed for analytes with different blood cell-to-plasma partition coefficients. Conclusions: We accurately and precisely estimated HCT in DBS samples using image analysis and Hb determination, and the volume of blood in DBS using conductivity measurement. We evaluated different approaches and derived an optimal procedure for HCT-bias correction. Full article

Gaucher disease (GD) is a lysosomal storage disorder caused by mutations in the glucocerebrosidase gene (GBA1), leading to acid β-glucosidase deficiency and the accumulation of glucosylceramide-derived glycosphingolipids. Its three phenotypes (non-neuronopathic, acute neuronopathic, and chronic neuronopathic) have variable clinical presentations including hepatosplenomegaly, cytopenia, bone disease, and neurological involvement. Early diagnosis and treatment are critical for improving outcomes, but GD is under-recognized due to non-specific symptoms and limited access to appropriate diagnostic testing. Glucosylsphingosine (lyso-Gb1), a deacylated metabolite of glucosylceramide, has been identified as a candidate biomarker for diagnosis and monitoring. This narrative review examines the role of biomarkers in GD, focusing on lyso-Gb1 as a potential diagnostic and prognostic biomarker. Lyso-Gb1 is markedly elevated in GD patients and correlates with disease burden, severity, and response to therapy. It is detectable in plasma and dried blood spots, making it suitable for newborn screening, diagnosis, and monitoring. Lyso-Gb1 is a sensitive and specific biomarker for GD, facilitating early detection, guiding treatment decisions, and enabling personalized disease management. Lyso-Gb1 levels reflect substrate accumulation and therapeutic response more reliably than other biomarkers such as chitotriosidase or CCL18. Ongoing research aims to refine diagnostic thresholds and integrate lyso-Gb1 monitoring into routine clinical practice for optimal patient outcomes. Full article

Low-processed ready-to-eat (RTE) meat products are highly vulnerable to microbial contamination, yet data on in-process dynamics remain limited. This study investigated microbial dynamics and environmental hygiene during the production of vacuum-packed RTE pork bars containing dried plasma, with a focus on identifying process-inherent contamination risks. Samples were collected at successive processing stages and from food-contact and non-food-contact surfaces. Process hygiene was assessed using indicator organisms (Aerobic Plate Count, Enterobacteriaceae, lactic acid bacteria, yeast and mold, E. coli, S. aureus counts), while food safety relevance was addressed by monitoring Listeria monocytogenes and Salmonella spp. Microbial counts increased by approximately 1.5–2.3 log CFU/g between early processing steps, indicating that these operations are critical contamination-prone steps. Environmental monitoring revealed contamination hotspots on frequently handled surfaces, highlighting the vulnerability of pre- and post-lethality stages. Despite the baking achieving a mean microbial reduction of ~3 log CFU/g, consistent with effective thermal processing, low-level microbial reappearance during packaging and maturation indicated the potential for post-process contamination. The results demonstrate that production-inherent factors largely drive microbial contamination patterns and may persist even in facilities operating under implemented GHP, GMP, and HACCP-based procedures, highlighting step-specific limitations rather than system failure. By providing empirical data on in-process microbial dynamics, this study supports both scientifically based and risk-based approaches within Food Safety Management Systems, offering transferable insights applicable to similar RTE meat production environments. The findings may assist food business operators in optimising targeted control measures and strengthening risk-based decision-making in low-processed RTE meat production. Full article

Around eight million people—mainly in cities—die prematurely from pollution-related diseases; thus, studies of urban dust have become increasingly relevant over the last two decades. In this study, an assessment of heavy metal and metalloid contamination in urban dust was conducted in downtown Murcia, Spain. The objectives were to evaluate the level of contamination and the associated health risks, both with a spatially explicit focus. One hundred and twenty-eight urban dust samples were collected, each from a 1-square-meter area, using plastic tools to prevent contamination. The dust was dried and weighed, then acid-digested before analysis via inductively coupled plasma mass spectrometry. Corresponding maps were then generated using a geographic information system. The elements analyzed in the urban dust (with their median concentrations, given in mg/kg) were As (2.14), Bi (14.06), Cd (0.38), Co (1.88), Cr (71.17), Cu (142.60), Fe (13,752), Mn (316.64), Mo (3.90), Ni (21.94), Pb (106.27), Sb (6.54), Se (4.34), Sr (488.08), V (28.05), and Zn (357.33). The sequence of median concentrations for the analyzed elements was Fe > Sr > Zn > Mn > Cu > Pb > Cr > V > Ni > Bi > Sb > Se > Mo > As > Co > Cd. The pollution assessment reveals that the city is moderately polluted. Using local background levels, the elements with median values exceeding the threshold for considerable contamination were As, Cu, Pb, Sb, Se, and Zn. Using the global background level, the elements with median values exceeding the threshold for considerable contamination were Bi, Cu, Mo, Pb, Sb, Se, and Zn. The median value of the sum of the hazard index (1.82) indicates a risk to children’s health. The hazard index revealed that 43% of the sites pose a relative risk to children. In contrast to previous global studies, the present research provides a multi-scale assessment of urban pollution and health risks. Pollution is evaluated by metal, city, zone, and site, while health risks are assessed by metal, city, and site. We recommend a strategy for both local authorities and residents. Full article

Background: Metabolomics is recognized as crucial technology for advancing our ability to diagnose, characterize, and monitor treatment of disease. Yet, metabolomics-based diagnostic testing has not been widely adapted into clinical practice because its technical requirements make it generally incompatible with operation at the point of care. One way to expand the reach of metabolomics-based testing, and its clinical benefits, is to utilize dried blood spots (DBS) as a testing sample type. Their easy collection, ambient storage capability, and cost-effective shipment make DBSs ideal for diagnostic tests that require the use of a centralized technology. Methods: To date, relatively few studies have investigated the performance of DBSs at capturing the global metabolome and reporting changes associated with physiological processes. In this study, we investigated those factors by performing global metabolomic profiling on DBSs collected from study volunteers under fasted and postprandial states, with and without dietary fish oil supplementation. Results: DBSs demonstrated broad coverage of metabolic pathways and captured numerous metabolic changes associated with feeding, fasting, and fish oil supplementation that have been reported in plasma and serum. Conclusions: Our findings support the hypothesis that DBSs are a viable sample type for metabolomics-based diagnostic testing and justify follow-up validation studies. Full article

Purpose: Fetal bovine serum (FBS) is widely used in cell culture due to its rich nutrient and growth factor content, but it poses ethical concerns, biosafety risks, and cost limitations. This study investigates plasma-depleted lyophilized (freeze-dried) porcine platelet lysate (pPL) as a potential alternative to FBS for use in cell-based research and biomanufacturing. Materials and Methods: Fresh porcine blood was processed to obtain plasma-depleted pPL using double centrifugation and repeated freeze–thaw cycles. The lysate was analyzed for growth factor content via ELISA, then freeze-dried and sterilized with gamma irradiation. Endotoxin levels and cytotoxicity were evaluated. The ability of plasma-depleted lyophilized pPL to promote cell proliferation was assessed using L929 fibroblast cultures and compared with FBS. Results: Plasma-depleted lyophilized pPL contained significantly higher levels of TGF-β1 than FBS. The freeze-dried product remained stable for at least three months at room temperature. Gamma irradiation effectively sterilized the lysate without degrading key growth factors. Plasma-depleted lyophilized pPL showed no cytotoxicity and promoted greater proliferation of L929 cells compared to FBS, indicating enhanced mitogenic activity. Conclusions: Plasma-depleted lyophilized pPL is a stable, safe, and growth factor-rich alternative to FBS. It supports fibroblast proliferation, retains bioactivity after sterilization and storage, and may provide a scalable, ethical option for cell culture in biomedical research, regenerative medicine, and therapeutic product development. Full article

3-Methylcrotonyl-CoA carboxylase deficiency (3-MCCD) is a metabolic disorder with a wide clinical spectrum ranging from asymptomatic individuals to severe metabolic decompensation. Following the introduction of expanded newborn screening, a high number of asymptomatic individuals with 3-MCCD were identified, prompting debates about its inclusion in screening panels. In order to inform policy and healthcare decisions regarding the inclusion of 3-MCCD in newborn screening programs, we evaluated the long-term outcomes for newborns with positive results over a decade of screening experience in North-East Italy, as well as the psychological impact on their parents. Of the 336,668 newborns screened between 2014 and 2025, 9 were confirmed to be affected. These infants underwent annual clinical and biochemical assessments, including dried blood spot acylcarnitine profile, plasma free carnitine, and urinary organic acids assays. An emergency protocol was provided to all affected children to manage intercurrent illnesses. An ad hoc survey was developed to assess the psychological impact of the disease on parents. During follow-up (mean age at last visit: 4.2 years), one patient experienced metabolic decompensation during an intercurrent illness, which was promptly treated. One patient presented with growth retardation and another with transient psychomotor delay. Five patients developed carnitine deficiency, requiring supplementation. Psychological assessments revealed an initial high level of parental psychological impact, which decreased over time. All parents strongly supported the screening program. Newborn screening for 3-MCCD enabled the early identification and management of affected individuals, thereby avoiding severe metabolic decompensation. Although there is an initial psychological burden on parents, it significantly decreases over time. Therefore, the long-term benefits of newborn screening for 3-MCCD seem to outweigh the psychological drawbacks. Full article

Background/Objectives: Blueberries are an important nutraceutical due to their excellent nutritional profile, with particularly high levels of polyphenols and anthocyanins. These compounds can improve mood, cognition, and health. As such, blueberry consumption can potentially benefit those coping with depression and anxiety. In this regard, there is an unmet need for novel, effective, and accessible treatments for these conditions, particularly in rural communities, where specialized health care is often limited. Methods: Therefore, we conducted a double-blind, randomized pilot study in a rural population to assess whether daily blueberry supplementation affected symptoms of anxiety and depression. We employed a crossover design to test the effects of 12 weeks daily ingestion of 24 g of whole freeze-dried blueberry powder versus placebo on symptoms of depression and anxiety in those diagnosed with a depressive or anxiety disorder including major depressive disorder (MDD) and generalized anxiety disorder (GAD). We collected behavioral data (HDRS, GAD-7, MDI) at baseline, mid-, and post-treatment timepoints. We collected blood, serum, plasma, and behavioral data (HDRS, GAD-7, MDI) at baseline, mid-, and post-treatment timepoints. We measured inflammatory cytokines IL-1β, IL-6, TNF-α, IFN-γ, and IL-10 in serum, CRP in whole blood, and performed global metabolomics in plasma. Results: Blueberries significantly reduced symptoms of depression and anxiety compared to placebo. CRP and inflammatory cytokine levels were unaffected. Our global metabolomic measures suggested that different metabolites were differentially affected at the middle and post-intervention timepoints in the study. Conclusions: Overall, this study found potential improvements in symptoms of depression and anxiety following daily blueberry supplementation, although the biochemical mechanisms underlying these behavioral improvements remain unresolved. Full article

Background: Mycophenolic acid (MPA) is frequently used in pediatric renal transplantation as part of immunosuppressive therapy, yet therapeutic drug monitoring (TDM) remains challenging. Accurate monitoring is essential due to MPA’s narrow therapeutic window, variable pharmacokinetics, and high protein binding. This study examined whether saliva could serve as a non-invasive alternative to plasma for measuring MPA exposure. Methods and Results: Concentrations of MPA and its primary glucuronide metabolite (MPAG) were determined in plasma, capillary blood, plasma ultrafiltrate, wet saliva, and dried saliva collected using volumetric absorptive microsampling (VAMS). A novel LC–MS/MS method for quantifying MPA and MPAG in dried saliva collected with the Mitra™ device was developed and validated within a 1–700 μg/L calibration range, demonstrating robust analytical performance. Dried and wet saliva showed high correlation (r = 0.99 and 0.98 for MPA and MPAG, respectively). However, both salivary matrices—dried saliva collected with Mitra™ (vsMPA, vsMPAG) and wet saliva (sMPA, sMPAG)—exhibited poor correlation with unbound (fMPA, fMPAG) and total plasma concentrations (tMPA, tMPAG). A modest, yet positive, correlation was observed between the measured concentrations for the following pairs: sMPA versus fMPA (r = 0.376, p = 0.1036), sMPA versus tMPA (r = 0.305, p = 0.1904), sMPAG versus fMPAG (r = 0.205, p = 0.3851), and sMPAG versus tMPAG (r = 0.472, p = 0.0012). Pharmacokinetic parameters supported these findings, highlighting discrepancies between saliva and plasma. Conclusions: From a clinical perspective, saliva sampling—although minimally invasive and patient-friendly—does not offer a reliable substitute for plasma in routine TDM of MPA and MPAG. Capillary blood collected through VAMS remains a promising alternative for long-term monitoring of pediatric patients; however, several considerations still need to be addressed. Full article

Dried blood spots (DBSs) are a practical tool for diagnosing infectious diseases, especially in remote or resource-limited settings. This study assessed the efficacy of DBS-based serological assays for syphilis screening. EDTA blood samples from 171 syphilis-seropositive and 122 seronegative individuals were used to prepare DBSs by spotting whole blood onto filter paper. After drying, 12 mm disks were punched, incubated overnight in buffered solution, and centrifuged. Syphilis serological screening was conducted using the Liaison^® Treponema Screen assay, Macro-Vue™ Reagin Plasma Rapid (RPR) card test, and Dual Path Platform (DPP) Syphilis Screen and Confirm test. The Liaison^® assay demonstrated 100% sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) with an optimized cut-off. The nontreponemal RPR test showed very low sensitivity (2.9%) on DBS but perfect specificity (100%). The DPP test for treponemal antibodies achieved high sensitivity (92.1%) and specificity (98.2%) with microreader adjustment. Visual reading of the DPP test had variable accuracy, with sensitivity reaching 100% but lower specificity (42.1%). Nontreponemal antibody detection by DPP showed moderate sensitivity and specificity. Although nontreponemal testing requires refinement, DBS testing combined with point-of-care tests like DPP holds promise for expanding syphilis screening accessibility and decentralization globally, particularly in resource-constrained environments. Full article