Search Results (3,843)

Background/Objectives: Natural multi-component nutraceutical formulations may modulate interconnected pathways involved in metabolic and bone health. This study evaluated, using in vitro models, the effects of a standardized botanical–vitamin formulation on intestinal barrier integrity, osteoblastic activity, and osteoclast differentiation, focusing on intestinal-bone crosstalk, redox-inflammatory signalling, and potential synergistic interactions among components. Methods: A combined in vitro approach using intestinal, osteoblastic, and osteoclastic cell models was applied to assess a formulation containing characterized plant extracts and vitamin D₃. The study evaluated cytocompatibility, intestinal barrier function, cellular uptake, and the modulation of markers related to osteogenesis and osteoclastogenesis, using biochemical, molecular, and enzymatic assays, as well as oxidative stress measurements and synergy analysis. Results: The formulation maintained intestinal barrier integrity and bioavailability without cytotoxicity, promoted osteoblastic differentiation and reduced oxidative stress, while inhibiting osteoclast differentiation and resorptive activity. These effects were associated with modulation of inflammatory and redox-related signalling pathways and showed additive to synergistic interactions among components. Conclusions: These findings support a multi-target nutraceutical approach that can concurrently influence intestinal barrier and bone remodelling in vitro, offering mechanistic evidence for its role in modulating the gut–bone axis, and highlight the need for further studies in advanced models and clinical trials. Full article

Neuroblastoma is characterized by noticeable resistance to chemotherapy, largely driven by the ability of tumour cells to reorganize stress-adaptive signalling networks rather than relying on single oncogenic drivers. We conducted a study to investigate the pharmacological mode of action of doxorubicin in modifying adaptive signalling pathways in SH-SY5Y neuroblastoma cells, and whether the capacity of plant metabolites can exploit emergent biochemical vulnerabilities. Transcriptomic profiling through RNA sequencing conducted 48 h post-doxorubicin exposure unveiled the organized disruption of pathways linked with amyloidogenic processes, oncogenic signalling pathways, oxidative stress, and DNA repair. The protein–protein interactions, coupled with Kyoto Encyclopedia of Genes and Genomes pathway evaluations, revealed five network-central-hubs: BRAF, GSK3β, PARP1, BACE1, and MAOB. Structural docking integrated with 200 ns molecular dynamics simulations illustrated binding stability across multiple targets driven by three metabolites, Lactol binding to BRAF (−54.13 kcal/mol) and MAOB (−39.08 kcal/mol), Amino(1H-indol-2-yl)acetic acid to BACE1 (−41.07 kcal/mol) and GSK3β (−47.38 kcal/mol), and Quercetin-3-(6″-malonyl-glucoside) binding to PARP1 (−46.03 kcal/mol). In vitro Cell Counting Kit-8 proliferation assays validated the significant anti-neuroblastoma efficacy, with the lowest IC₅₀ (0.2397 µM) being exhibited by Amino(1H-indol-2-yl)acetic acid, followed by Lactol (1.226 µM) and Quercetin-3-(6″-malonyl-glucoside) (1.301 µM), which mirrored the cytotoxic action of doxorubicin (1.306 µM). These results suggest that plant-derived metabolites may interact with stress-adaptive signalling pathways connected with neuroblastoma. However, direct experimental validation of target engagement and pathway modulation will be required to confirm these predicted interactions. Full article

Pheochromocytoma (PhC) and Paraganglioma (PG) are rare neuroendocrine tumors characterized by uncontrolled catecholamine secretion. Although rare, much attention is still devoted to identifying a unique biochemical signature of these diseases to reduce false positives, thus improving patient outcomes, and customizing clinical laboratory practices to available resources and specific diagnostic needs. Emerging knowledge into catecholamine metabolism has greatly improved diagnostic strategies, with current international guidelines recognizing plasma free or urinary fractionated metanephrine measurements as the recommended first-line biochemical tests. This narrative review highlights the clinical utility of measuring plasma free metanephrines compared to urinary catecholamines in the diagnosis of these conditions. Plasma free metanephrines offer superior sensitivity and specificity compared to catecholamines due to their continuous secretion, which is independent from tumor size and catecholamine fluctuations. This review also addresses preanalytical and methodological challenges, emphasizing patient preparation, sample stability and advanced analytical techniques currently available. Methodologies such as LC-MS/MS have demonstrated improved diagnostic accuracy compared to traditional immunoassays, offering enhanced analytical performance in terms of sensitivity, specificity, and reduced susceptibility to interferences. Full article

Feline hepatic lipidosis (FHL) is a life-threatening, common hepatobiliary disease characterized by massive triglyceride accumulation in the liver, often triggered by anorexia and negative energy balance in cats. This condition causes severe metabolic stress that may secondarily impact myocardial integrity. This observational clinical study evaluated serum homocysteine (Hcy) and high-sensitivity cardiac troponins (hs-cTnI, hs-cTnT) to assess secondary myocardial injury and their prognostic value in FHL. Fifty cats, comprising 30 with naturally occurring FHL and 20 healthy controls, were included. Serum Hcy, hs-cTnI, and hs-cTnT concentrations were measured using feline-specific ELISA kits, and routine biochemical parameters, alongside hospitalization times, were recorded. Results indicated that Hcy, total bilirubin, and liver enzymes were significantly elevated in the FHL group compared to controls (p < 0.001). However, hs-cTnI and hs-cTnT levels did not differ significantly between the groups. Notably, Hcy exhibited a strong positive correlation with hospitalization time (r = 0.89, p < 0.001). The absence of significant hs-cTn elevations suggests the feline myocardium remains largely resistant to acute necrosis during FHL. Nevertheless, relative hyperhomocysteinemia is a powerful prognostic biomarker for prolonged hospitalization in affected cats. Full article

Innovation in the field of bioinspired therapeutic anticoagulants, as an alternative to heparin and its derivatives, is increasingly focused toward the discovery of new molecules from natural sources. Inspired by the compelling observation that marine organisms possess a wide array of acidic polysaccharides (APs) within their extracellular matrix (ECM) with promising anticoagulant potential, this study investigates AP topographic distribution, content, and bioactivity in the body of seven phylogenetically distant Mediterranean sponge species. Tissue architecture was evidenced by Alcian Blue histological staining whereas biochemical analyses were carried out on APs purified from papain-digested tissue extracts using anion-exchange chromatography. Four polysaccharide fractions were obtained and assessed for hexuronic acid content. The two most abundant ones, up to 98% of the total, were characterized by electrophoretic analyses, and assessed in vitro for inhibitory activity on blood coagulation. Histology evidenced a heterogeneous distribution of APs within the sponge’s ECM, along with marked interspecific variability in both concentration and electrophoretic profiles. Neither Tethya aurantium nor Crambe crambe showed any significant in vitro effects on coagulation, whereas the other species exhibited a strong inhibitory effect on both activated partial thromboplastin time (aPTT) and thrombin time (TT), comparable to standard heparin (Hep). Concerning aPTT, both I. retidermata and H. communis reached the endpoint of 300 s at 5 µgUA/mL, whereas S. spinosulus, O. lobularis, A. aerophoba reached it at 10 µgUA/mL. With reference to TT, A. aerophoba, I. retidermata, S. spinosulus and H. communis had comparable effects on coagulation time with respect to Hep (endpoint at 5 µgUA/mL), whereas O. lobularis was less effective (endpoint at 100 µgUA/mL). These findings show that Porifera, one of the most basal Metazoa, have an ECM rich in APs with anticoagulant properties towards both intrinsic and common pathways of coagulation, consistent with known inhibitory mechanisms reported for certain marine sulfated polysaccharides derived from other invertebrate taxa. These molecules, obtainable through sustainable blue technology, represent compelling candidates for bioinspired next-generation anticoagulant therapeutics, with broader applications in regenerative medicine and tissue engineering. Full article

Pyranose oxidases (POXs, EC 1.1.3.10) are a class of fungal FAD-dependent oxidoreductases with potential for lignocellulosic bioconversion because they generate H₂O₂ during sugar oxidation. Despite their known catalytic properties, the role of these enzymes in promoting lignocellulose enzymatic saccharification remains largely unexplored. In this study, POXs from Phanerochaete chrysosporium (PcPOX) and Trametes versicolor (TvPOX) were comparatively evaluated through biochemical characterization, kinetic analysis, molecular simulation, and supplementation for lignocellulose hydrolysis. PcPOX exhibited a broader substrate spectrum and a slightly higher optimum temperature, whereas TvPOX demonstrated greater stability under acidic and hydrolysis-relevant conditions and a longer half-life at 50 °C. TvPOX also showed a numerically lower apparent K_m toward D-glucose, while the apparent catalytic efficiencies were comparable between the two enzymes. Molecular simulation results suggested more stable glucose binding in TvPOX. Accordingly, TvPOX was selected for hydrolysis experiments and was shown to increase the measured glucan conversion of phosphoric acid-swollen cellulose, Avicel, and corncob residue. Mixture design analysis further indicated that this positive effect depended on balanced peroxide regulation, with low catalase supplementation providing better performance. These results identify TvPOX as a promising auxiliary enzyme for cellulase-based lignocellulosic saccharification. Full article

Background: Diabetes mellitus is a metabolic disturbance characterized by chronic hyperglycemia, which stems from defective secretion and/or action of insulin. D-Limonene has been studied for the confirmation of its antidiabetic and antioxidant effects. This paper aims to investigate the antidiabetic and antioxidants effects of D-Limonene in an experimental model of DM1. Methods: Female Wistar rats (180–250g) received streptozotocin (STZ, 45 mg/kg) intraperitoneally. Animals with capillary glycemia ≥ 250 mg/dL were considered diabetic. D-Limonene at oral doses of 12.5 mg/kg, 25 mg/kg and 50 mg/kg was administered during 28-day treatment. Water and food intake, weight gain and capillary glycemia were evaluated. At the end of the treatment, the following biochemical parameters were assessed: serum glucose, HbA1c, urea, creatinine, AST, ALT, GGT, ALP and albumin. The oxidative stress markers were determined in plasma, erythrocytes, and aortic homogenates: malondialdehyde, nitrite, myeloperoxidase, superoxide dismutase and catalase. Results: D-Limonene (25 and 50 mg/kg) significantly reduced serum glucose, HbA1c, AST, ALT, GGT and ALP when compared to DC, as well as plasma MDA and nitrite concentrations. Interestingly, D-Limonene (25 and 50 mg/kg) decreased both plasma and aortic myeloperoxidase activities, as well as increased both erythrocytic and aortic catalase activities. Conclusions: These findings, besides a marked D-Limonene-induced hypoglycemic effect, pave the way for further studies comprising a multi-target treatment by providing benefits on hepatic and vascular complications related to the diabetic condition. Full article

Growing evidence implicates neuroinflammation, gut-derived endotoxemia, and dysregulated lipid metabolism in the pathogenesis of Parkinson’s disease (PD). However, the relationships among circulating lipopolysaccharide (LPS), LPS-handling proteins, systemic inflammatory activation, and lipid fractions remain insufficiently characterized. The aim of this study was to compare LPS levels, LPS-related inflammatory mediators, and plasma lipid parameters between PD patients and matched controls, and to explore correlations among these biomarkers. Twenty PD patients and twenty matched controls underwent fasting venous sampling. Circulating LPS, lipopolysaccharide binding protein (LBP), soluble cluster of differentiation 14 (sCD14), high-sensitivity C-reactive protein (hsCRP), and phospholipid transfer protein (PLTP) were quantified via LAL assay and ELISAs. Serum cholesterol, HDL cholesterol (HDL-C), phospholipids (PLs), HDL-PLs and triacylglycerols (TAGs) were assessed using validated biochemical techniques. LPS concentrations did not differ between groups. However, PD patients showed elevated sCD14 and hsCRP levels, reduced LBP, and increased PLTP. Lipid profiling revealed lower total cholesterol and reduced HDL-associated cholesterol and phospholipids in PD, while TAG levels remained unchanged. Correlation analyses indicated coordinated associations between inflammatory markers and lipid fractions, with distinct interaction patterns in PD compared with controls. These findings support a mechanistic interplay among endotoxemia, innate immune activation, and lipid dysregulation in the pathophysiology of PD. Full article

Background: Cataract is a progressive lens opacity. According to the World Health Organization, about 45 million people in the world are blind, with about half of these cases attributable to cataracts. Due to the complexity of cataract disease, current research on cataracts is far from sufficient, so it is especially important to understand the development process and the pathogenic factors of cataracts. Myodes rufocanus (M. rufocanus) is an animal of the M. rufocanus of the hamster family Volinae. In developing M. rufocanus, we found an individual of M. rufocanus with a congenital cataract phenotype. We confirmed the symptoms of cataract under natural light and using a slit lamp. Methods: Therefore, we analyzed the mechanism of congenital cataract in M. rufocanus from the aspects of pathological histology, physiology and biochemistry, and gene level, aiming to explore the feasibility of its development into an animal model of cataract. Cataract is a progressive lens opacity and a leading cause of visual impairment. Understanding its pathogenesis requires appropriate animal models. In a laboratory-bred colony of M. rufocanus, we identified individuals with a spontaneous congenital cataract phenotype, confirmed by gross observation and slit lamp examination. To characterize this phenotype and explore its potential as an animal model, we performed pathological, physiological, biochemical, and transcriptomic analyses using three cataract-affected and three normal age-matched male individuals (8 weeks old per group). Results: Blood tests revealed significantly lower white blood cell and lymphocyte counts in the cataract group, while blood glucose and other biochemical parameters showed no significant differences. Histologically, cataractous lenses exhibited eosinophilic aggregation in the nuclear region with disorganized fiber cells. Transcriptome analysis identified 6544 differentially expressed genes, including downregulation of crystallin genes (CRYBB2, CRYBA4, CRYGS) known to be associated with congenital cataract. KEGG pathway enrichment analysis highlighted retinol metabolism, tyrosine metabolism, and cytochrome P450-related pathways. RT-qPCR confirmed reduced CRYBB2 expression in cataractous eyes. Conclusions: This study provides the first transcriptome dataset for M. rufocanus ocular tissues and offers preliminary evidence that this naturally occurring cataract phenotype may serve as a potential model for congenital cataract research. Full article

Fermented foods and probiotics represent complementary yet distinct components of human nutrition. Fermented foods are shaped by biochemical transformations driven by microbial metabolism, whereas probiotics are live microorganisms that may confer health benefits to the host. In both cases, bacteria, yeasts, and filamentous fungi mediate key metabolic activities that generate bioactive compounds and modulate host–microbiota interactions. During fermentation, microbial communities synthesize organic acids, peptides, exopolysaccharides, vitamins, and other metabolites that enhance food safety, sensory attributes, and potential health-promoting properties. Several microbial products, such as bacteriocins, reuterin, hydroxylated fatty acids, and exopolysaccharides, exhibit antimicrobial, immunomodulatory, antioxidant, and cholesterol-lowering activities. Advancing our understanding of microbial metabolism in fermented foods is essential for developing next-generation functional foods and nutraceuticals that leverage microbial biotransformations to support human health. Nonetheless, multiple challenges limit the translation of these advances into commercial products. Inadequately controlled fermentation may introduce microbiological or chemical hazards, regulatory frameworks governing microbial use in foods remain insufficiently defined, and standardized procedures for microbial strain handling and characterization are still lacking. This narrative review integrates current evidence on the nutraceutical properties of fermented foods and probiotics, while also examining the associated safety considerations and the technological factors that influence fermentation processes. Full article

Agri-food processing in Europe generates large quantities of organic residues that remain insufficiently valorized despite their significant biochemical potential. Among these, wastes derived from root vegetables and anthocyanin-rich crops represent a distinct category of non-lignocellulosic biomass characterized by high moisture content, low lignin levels, and substantial concentrations of fermentable carbohydrates and bioactive compounds. This review provides a systematic overview of the origin, composition, and valorization potential of these residues, as well as extraction methods, with particular emphasis on root vegetable processing wastes and pigment-rich agri-food by-products. Valorization options are discussed within an integrated biorefinery perspective, particularly for specific compositional characteristics of the investigated waste streams related to suitable recovery strategies, followed by the conversion of post-extraction residues into secondary products and bioenergy. These options are evaluated in relation to the origin, biochemical profile, and valorization potential of each waste stream, as detailed in the dedicated sections of the review. Cascading utilization strategies are highlighted as a means to improve resource efficiency and reduce environmental burdens compared to single-route treatment options. By integrating information on feedstock characteristics and processing pathways, this review contributes to a better understanding of non-lignocellulosic agri-food wastes and supports the development of sustainable valorization strategies in the European circular bioeconomy. Full article

Background/Objectives: Obesity is characterized by chronic low-grade inflammation, and bariatric surgery promotes substantial metabolic and inflammatory improvement. However, residual obesity and microvascular complications may persist in some individuals, suggesting potential genetic influences on postoperative outcomes. This exploratory pilot study investigated the association between inflammatory cytokine gene polymorphisms and clinical, metabolic, and inflammatory outcomes in older women one year after bariatric surgery. Methods: This cross-sectional, hypothesis-generating pilot study included 21 women aged ≥50 years (mean 61.6 ± 5.0) who underwent Roux-en-Y gastric bypass at a public bariatric center in Brazil. Anthropometry, body composition, biochemical markers, and serum levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were assessed 12 months postoperatively. Genotyping for IL6-174G/C (rs1800795) and TNFA-308G/A (rs1800629) was performed using PCR-RFLP. Associations were analyzed using non-parametric statistical tests. Results: Notably, the IL6-174CC genotype was associated with persistent obesity, whereas carriers of the TNFA-308A allele showed a higher prevalence of diabetic retinopathy. These results highlight genotype-specific postoperative outcomes. No significant genotype-related differences were observed for most anthropometric, biochemical, or inflammatory parameters, indicating substantial overall metabolic improvement after surgery regardless of genetic background. However, the observed associations were based on a small sample and should be interpreted cautiously. Conclusions: This exploratory pilot study revealed associations between inflammatory cytokine gene polymorphisms and selected postoperative outcomes, particularly persistent obesity and diabetic retinopathy, in older women one year after bariatric surgery. These hypothesis-generating findings emphasize the need for larger, longitudinal studies to clarify the role of genetic factors in postoperative heterogeneity after bariatric surgery. Full article

Betaine-homocysteine methyltransferase (BHMT) is an enzyme involved in one-carbon metabolism and plays a crucial role in maintaining liver health. In this study, we investigated the impact of liver-specific deletion of BHMT on liver dysfunction using a mouse model. We generated BHMT floxed mice and bred them with albumin Cre to generate liver-specific BHMT knockout (BHMT LKO) mice. Liver tissues harvested from six-month-old chow-fed BHMT floxed and LKO mice were characterized through histological, biochemical, and molecular analyses. BHMT LKO mice displayed a complete loss of hepatic expression of BHMT mRNA, protein and enzyme activity. Histopathological analysis revealed the development of hepatic steatosis in BHMT LKO mice compared to the floxed mice. These morphological changes were supported by biochemical analysis showing elevated levels of hepatic triglycerides in conjunction with a profound decrease in the methylation potential (i.e., reduced S-adenosylmethionine (SAM): S-adenosylhomocysteine (SAH) ratio), which was mainly driven by a six- to sevenfold increase in SAH levels. BHMT LKO mice also exhibited increased lipid peroxidation and lysosomal dysfunction compared to floxed mice. Early signs of inflammation were seen in the livers of BHMT LKO mice of both sexes, as evident from significant increase in CD68-positive cells and interleukin 1β levels. Additionally, there was a moderate increase in fibrosis, as evidenced by the upregulated expression of α-smooth muscle actin and collagen II levels and the histological assessment of picrosirius red-stained liver sections of BHMT LKO mice of both sexes compared to their respective counterparts. These findings demonstrate that hepatic BHMT deficiency promotes lipid accumulation, lysosomal/proteasomal dysfunction, and early inflammatory and fibrotic changes in the liver by reducing the methylation potential. Collectively, our results underscore BHMT as a critical regulator of liver homeostasis and a potential therapeutic target in liver-related disorders. Full article

Hypovolemia is a systemic state characterized by severe reduction in the effective blood volume with subsequent tissue hypoperfusion. It may be due to fluid loss, decreased water intake, fluid redistribution, or systemic disease. The present study aimed to evaluate the diagnostic significance of the renal artery resistive index (RRI), caudal vena cava (CVC) diameter, and aorta (Ao) diameter in dogs with hypovolemia. For this purpose, 30 dogs (hypovolemic, n = 15; control, n = 15) were investigated. Clinical variables and hematological findings were investigated for each dog. Pulsed-wave Doppler ultrasound was performed to measure the RRI and diameters of the CVC and Ao. Ultrasound examination revealed a markedly elevated RRI (p < 0.001) and significantly reduced CVC (p < 0.001) and Ao (p < 0.001) diameters in hypovolemic dogs compared to controls, reflecting increased vascular resistance and impaired venous return. Biochemical analysis showed significant increases in blood urea nitrogen (BUN) and BUN:Cr ratio, while Cr remained unchanged. Hematological variables demonstrated limited diagnostic value, with only mild increases in packed cell volume (PCV%). Correlation analysis confirmed a strong positive correlation between RRI and BUN (r = 0.917; p < 0.01), RRI and BUN:Cr ratio (r = 0.664; p < 0.01), and CVC and Ao diameters (r = 0.832; p < 0.05). Receiver operating characteristic (ROC) analysis with area under the curve (AUC) identified RRI (AUC = 0.99), CVC diameter (AUC = 0.93), and Ao diameter (AUC = 0.88) as highly accurate markers of hypovolemia, whereas the CVC:Ao ratio and hematological markers provided poor discrimination. Logistic regression confirmed significant diagnostic value for RRI, CVC diameter, Ao diameter, and BUN, but final multivariate analysis revealed RRI as the sole independent early diagnostic marker (p < 0.001; OR: 196.0; 95% CI: 11.12–34.72). In conclusion, RRI measured by Doppler ultrasound is the most reliable and sensitive early diagnostic marker for hypovolemia in dogs, outperforming conventional biochemical and hematological markers. Full article

Background: Primary biliary cholangitis (PBC) is a rare autoimmune liver disease characterized by marked clinical and serological heterogeneity. Although diagnosis is mainly based on antimitochondrial antibodies (AMAs) and alkaline phosphatase (ALP), non-classical presentations remain a relevant cause of diagnostic delay. In this context, laboratory medicine plays a pivotal role in both diagnosis and long-term disease management. Methods: This manuscript represents a structured clinical review of laboratory biomarkers relevant to the diagnosis, monitoring, and prognostic stratification of PBC, integrated with a representative atypical case with long-term follow-up to illustrate the practical application of laboratory-driven diagnostic. Results: The analysis confirms the central role of immunological and biochemical markers in treatment monitoring and prognostic assessment, while highlighting their limitations in selected clinical scenarios. The reported case, characterized by persistent AMA negativity and consistently normal ALP levels, illustrates how expanded laboratory testing can support the identification of non-standard disease phenotypes. In this setting, parallel testing for AMA- and PBC-specific autoantibodies was essential to achieve a correct diagnosis. Moreover, alternative biomarkers, including gamma-glutamyl transferase (GGT) and selected immunological markers, provided clinically meaningful information when conventional markers were not informative. Conclusions: By integrating current evidence with a long-term clinical case, this work moves beyond a descriptive overview and proposes a practical, laboratory-driven diagnostic and follow-up framework for PBC. It highlights laboratory opportunities to facilitate timely diagnosis, appropriate prognostic stratification, and disease monitoring, including the assessment of associated comorbidities. Full article