Search Results (47)

Background: Neuropathic pain is a complex condition often resistant to current therapies due to limited efficacy and adverse effects. Nutraceuticals offer promising alternatives, combining antioxidant and anti-inflammatory properties with good tolerability. This study aimed to compare the effects of a commercial nutraceutical formulation, SUPERALA CARNITINE^® (Pharma Suisse Laboratories SpA, Milan, Italy), containing Alpha-Lipoic Acid (ALA), with a novel formulation, called SUPERALA CARNITINE^® Forte, where ALA and vitamin B6 were replaced by N-acetylcysteine (NAC), Glutathione (GSH), and Uridine monophosphate (UMP). Methods: An indirect gut–peripheral nerve axis was employed to simulate oral absorption, metabolism, and effect on nervous tissues using 3D in vitro models. Both formulations and their individual components were assessed for cytotoxicity and permeability in the gut model (Caco-2 cells in Transwell^®) and, after gut metabolism, for antioxidant capacity, anti-inflammatory activity, and neuroprotective potential in the peripheral nerve model. Results: SUPERALA CARNITINE^® Forte improved cell viability and favoured the maintenance of intestinal integrity, showing enhanced permeability, and significantly reduced oxidative stress (OS) and pro-inflammatory cytokines (TNF-α, IL-2) at the peripheral nervous system. In addition, it increased levels of neuronal markers (p75, MPZ, NRG1, ERβ) and decreased NaV1.7 and NaV1.8 activity, indicating greater neuroprotection and analgesic modulation than the ALA-based formula. Conclusions: The replacement of ALA and vitamin B6 with NAC, GSH, and UMP produced favorable responses in vitro on neuronal cells, supporting a hypothetical potential interest in this nutraceutical combination and justifying further future in vivo investigations. Full article

Gyrification, the folding of the cerebral cortex, plays a crucial role in brain development and function. Perinatal hypoxia–ischemia (HI) is a leading cause of neonatal brain injury, affecting cortical folding that can be measured by the gyrification index (GI). Using a late-preterm ferret model, our objective was to explore the relationships between HI injury, GI changes, and behavior, as well as the potential moderating effects of sex and treatment. Animals received 3 mg/kg E. coli lipopolysaccharide and underwent bilateral carotid artery ligation followed by alternating hypoxia and hyperoxia (HIH) and were randomized to saline vehicle (n = 25), erythropoietin (n = 20), therapeutic hypothermia (6 h at 33.5 °C, n = 20), and uridine monophosphate (n = 6), with n = 20 unexposed littermates serving as controls. Early reflex testing, CatWalk gait analysis, open-field behavior, and an open-water swim test were performed. Average, peak, motor, and somatosensory strip GIs were then assessed using ex vivo MRI. In control animals, males had lower GI than females; however, HIH exposure resulted in male GI being more similar to females, where HIH animals had significantly higher average GI than controls (p = 0.02). Adjusting for brain volume and injury, GIs in motor and somatosensory areas were associated with faster reflex outcomes in males but not females. In female controls, motor and somatosensory GIs were associated with increased anxiety-like behaviors, such as spending less time in open water during the swim test. By comparison, in male controls, higher GI was associated with decreased anxiety-like behaviors, including higher exploration index in the swim test. These sex-specific relationships between GI and behavior were lost with HIH injury. Treatment did not meaningfully restore the relationship between GI and behavior after HIH, but targeting this outcome may be an important measure for use in future neuroprotection studies in the ferret. Full article

Background/Objectives: Neuropathic pain, resulting from damage or pathology affecting the somatosensory nervous system, is a prevalent form of chronic pain that significantly impacts quality of life. Combined therapies are often utilised to manage this condition. Three-dimensional printing (3DP) offers a promising approach for personalising medication doses and dosage forms to meet individual patient needs. Methods: In this study, a formulation suitable for 3D printing was developed using magnesium citrate, uridine monophosphate, vitamins B₃ (niacin), B₆ (pyridoxine), B₁₂ (cobalamin), B₉ (folic acid), and spermidine to create a novel gel-based oral tablet for the targeted treatment of neurological pain. The antioxidant potential of the active pharmaceutical ingredients (APIs) was assessed using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) methods. The physical properties of the tablets were evaluated using a texture analyser, while the in vitro release profiles were determined by high-performance liquid chromatography (HPLC). Results: Results demonstrated that pectin–gelatin tablets hardened over time, with higher citric acid concentrations further enhancing this effect. Formulation AVII exhibited good hardness and low stickiness. Formulation AV, however, showed poor performance across all physical parameters and lacked sufficient structural integrity for practical application. While uridine monophosphate, B₁₂, and B₉ showed no significant differences in the release profiles of the tablets, spermidine, B₆, and B₃ displayed statistically significant variations. Specifically, AVII outperformed AV in terms of spermidine and B₆ release, and AV showed a higher release of B₃ compared to AV. Conclusions: The AVII tablet demonstrates potential for use in combined therapy targeting neurological pain disorders. Full article

Background/Objectives: Constipation can be induced in animal models through various factors such as loperamide (Lop) or complement component 3 (C3) deficiency. The effectiveness of therapeutic agents in the clinical management of constipation has been primarily evaluated within only one model, but between-model comparisons have not been performed so far. Therefore, we investigated whether the effectiveness of the laxative drugs for the clinical management is related to etiological factors. Methods: The changes in the key parameters for defecation were compared between C3 knockout (KO) mice with C3-deficiency-induced constipation and ICR mice with Lop-induced constipation after the oral administration of Uridine (Urd) and aqueous extract of Liriope platyphylla L. (AELP). Results: Similar effectiveness of Urd and AELP were detected on the stool frequency, intestinal epithelial barrier structure, and mucin secretion in both models. However, other parameters (namely gastrointestinal (GI) transit, water retention, and enteric nervous system (ENS) structure and function) showed higher effectiveness in C3 KO mice than in the Lop-induced model. Only the effectiveness of the two therapeutic agents on the histological structure of the mid-colon was greater in the Lop-induced mice model compared to the C3 KO mice model. Furthermore, these differences in the therapeutic effectiveness of Urd and AELP were partially reflected in alterations in the cyclic adenosine monophosphate (cAMP) downstream signaling pathway. Conclusions: The results suggest that the therapeutic effectiveness of Urd and AELP is sensitive to C3-deficiency-induced constipation and these differences may be linked to the alternative regulation of the cAMP downstream signaling. Full article

This study used a targeted metabolomics approach to examine changes in metabolites within green tea infusions fermented by G. lucidum (TFG) and evaluate the in vitro antioxidant and lipid-lowering properties of TFG. Fermentation decreased tea polyphenols, flavonoids, caffeine, soluble sugars, theaflavins, and catechins, while increasing free amino acids and theabrownins. The microbial bioconversion process led to the generation of decorated flavonoids, phenolic acids, terpenoids, alkaloids, nucleotides, and amino acids. This process shifted the tea’s taste from bitter and astringent to mellow, primarily due to the transformation of flavonoid glycosides, caffeine, catechins, 5′-guanosine monophosphate, 5′-uridine monophosphate, and theabrownins. Volatile metabolites added woody, floral, sweet, and fruity aromas. Reduced gallic acid and catechins lowered antioxidant activity, whereas increased theabrownins enhanced lipid-lowering activity and imparted a reddish-brown color. These findings indicate that fermentation significantly affects the flavor, aroma, and lipid-lowering ability of green tea infusion. Full article

This study used NMR-based metabolomics to investigate the mode of action (MoA) of 6-hydroxydopamine (6-OHDA) toxicity in the SH-SY5Y neuroblastoma cell model. 6-OHDA, a structural analogue of dopamine, has been used to create a Parkinson’s disease model since 1968. Its selective uptake via catecholaminergic transporters leads to intracellular oxidative stress and mitochondrial dysfunction. SH-SY5Y cells were treated with 6-OHDA at its IC₅₀ concentration of 60 μM, and samples of treated and untreated groups were collected after 24 h. The endo metabolome was extracted using a methanol–water mixture, while the exo metabolome was represented by the culture media. Further, endo- and exo metabolomes of treated and untreated cells were analysed for metabolic changes. Our results demonstrated significantly high levels of glutathione, acetate, propionate, and NAD⁺, which are oxidative stress markers, enhanced due to ROS production in the system. In addition, alteration of myoinositol, taurine, and o-phosphocholine could be due to oxidative stress-induced membrane potential disturbance. Mitochondrial complex I inhibition causes electron transport chain (ETC) dysfunction. Changes in key metabolites of glycolysis and energy metabolism, such as glucose, pyruvate, lactate, creatine, creatine phosphate, glycine, and methionine, respectively, demonstrated ETC dysfunction. We also identified changes in amino acids such as glutamine, glutamate, and proline, followed by nucleotide metabolism such as uridine and uridine monophosphate levels, which were decreased in the treated group. Full article

Background: Verticillium wilt (VW), caused by the fungal pathogen Verticillium dahliae, is a destructive disease that severely compromises cotton yield and fiber quality. Pyrimidine nucleotides, as essential metabolites and nucleic acid components, play critical roles in plant development and stress responses. However, genes involved in pyrimidine metabolism, especially their roles in disease resistance, remain largely uncharacterized in plants. Methods: Ghir_D05G039120, a gene encoding uridine kinase, shown to be associated with VW resistance in our previous study, was cloned and named as GhUKL4. The differential expression of GhUKL4 between the resistant and susceptible cultivars at multiple time points post-inoculation with V. dahliae was analyzed by quantitative real-time PCR (qRT-PCR), and the uracil phosphoribosyl transferase (UPRT) and uridine 5′-monophosphate kinase (UMPK) domains were verified by analyzing the amino acid sequences of GhUKL4. The role of GhUKL4 in the defense against VW infection was estimated by silencing GhUKL4 in the resistant and susceptible cultivars using virus-induced gene silencing (VIGS) analysis. Results: There were significant differences in the expression level of Ghir_D05G039120/ GhUKL4 among resistant and susceptible cotton lines. GhUKL4 contains UPRTase and UMPK domains, and there was one SNP between the resistant and susceptible cultivars in its 3′-UTR region. The silencing of GhUKL4 reduced cotton’s resistance to VW through mediating hormone signaling (JA) and oxidative stress (ROS) pathways. Conclusions: GhUKL4, encoding UMPK and UPRTase domain proteins, is a new regulatory factor associated with VW resistance in Gossypium hirsutum through fine-tuning JA-signalling and ROS bursting. Full article

Tendons are connective tissues that join muscles and bones and are rich in glycosaminoglycans (GAGs). Decorin is a proteoglycan with one dermatan sulfate (DS) or chondroitin sulfate (CS) chain (a type of GAG) attached to its core protein and is involved in regulating the assembly of collagen fibrils in the tendon extracellular matrix (ECM). Calcium-activated nucleotidase 1 (CANT1), a nucleotidase that hydrolyzes uridine diphosphate into uridine monophosphate and phosphate, plays an important role in GAG synthesis in cartilage. In the present study, we performed detailed histological and biochemical analyses of the tendons from Cant1 knockout (Cant1^−/−) mice. No abnormalities were observed in the tendons on postnatal day 1 (P1); however, remarkable hypoplasia was observed on P30 and P180. The collagen fibrils were more angular and larger in the Cant1^−/− tendons than in the control (Ctrl) tendons. In the Cant1^−/− tendons, the DS/CS content was significantly reduced, and the DC/CS chains attached to the decorin core protein became shorter than those in the Ctrl tendons. No abnormalities were observed in the proliferation and differentiation of tendon fibroblasts (tenocytes) in the Cant1^−/− mice. These results strongly suggest that CANT1 dysfunction causes defective DS/CS synthesis, followed by impairment of decorin function, which regulates collagen fibrogenesis in the tendon ECM. Multiple joint dislocations are a clinical feature of Desbuquois dysplasia type 1 caused by human CANT1 mutations. The multiple joint dislocations associated with this genetic disorder may be attributed to tendon fragility resulting from CANT1 dysfunction. Full article

To study the differences in meat quality and nutritional components between the local Shanghai pig breeds Meishan pig (MS), Shawutou pig (SWT), Fengjing pig (FJ), and the commercial Duroc × Landrace × Yorkshire (DLY) crossbred pigs, and to provide data support for the selection and breeding of superior pig breeds, this study selected 30 piglets each of three local pig breeds and DLY with similar birth ages and weights around 25 kg, fed them the same daily ration with uniform nutritional components, and slaughtered ten of them at around 100 kg weight for evaluation of differences in meat quality indicators (primarily intramuscular fat content, tenderness value, texture, etc.) and amino acid content among the varieties. The results indicated significant differences among the four pig breeds in intramuscular fat content, with MS having the highest content and significant differences in tenderness value compared to the other three breeds (p < 0.05). In terms of texture indicators, MS and SWT differed significantly from FJ and DLY in terms of hardness and fracturability indicators (p < 0.05), with certain differences existing among the four breeds in other indicators. In amino acid content, the differences in total amino acid content among the three Shanghai local pig breeds were not significant (p > 0.05), but all were significantly higher than DLY. Further analysis revealed significant differences in amino acid content between Shanghai local pig breeds and DLY, with Shanghai local pigs showing markedly higher levels of serine, proline, isoleucine, leucine, and histidine compared to DLY (p < 0.05). Regarding nucleotides, the cytidine monophosphate (CMP) indicator of MS differed significantly from the other three breeds (p < 0.05), SWT’s uridine monophosphate (UMP) indicator differed significantly from FJ and DLY, and FJ and DLY’s inosine monophosphate (IMP) indicator was significantly higher than MS and SWT (p < 0.05), while SWT’s adenosine monophosphate (AMP) indicator was significantly higher than the other three breeds (p < 0.05). The results of this study suggest that the meat quality and nutritional composition of Shanghai local pigs are significantly superior to DLY, with MS exhibiting significantly better meat quality and nutrition compared to SWT and FJ among the three local pig breeds. Full article

Dihydroorotase (DHOase; EC 3.5.2.3) is a zinc-dependent metalloenzyme that plays a key role in the de novo pyrimidine biosynthesis pathway, catalyzing the reversible cyclization of N-carbamoyl aspartate to dihydroorotate. This reaction is essential for the production of uridine monophosphate, the precursor of all pyrimidine nucleotides required for DNA and RNA synthesis. Despite its conserved enzymatic function, DHOase exhibits significant structural diversity across species, particularly in its oligomeric states, gene fusion patterns, and active site architecture. A crucial structural feature of DHOase is its flexible active site loop, which undergoes dynamic conformational changes during catalysis. Previously, the loop-in conformation was associated with substrate binding, whereas the loop-out conformation was linked to product release and non-substrate ligand binding. However, recent crystallographic studies challenge this paradigm, revealing that certain non-substrate ligands and inhibitors, including malate, 5-fluoroorotate, plumbagin, 5-aminouracil, and 5-fluorouracil, interact with DHOase via a loop-in binding mechanism rather than the previously assumed loop-out mode. These findings necessitate a reassessment of the catalytic mechanism of DHOase and underscore the active site loop as a potential target for drug development. This review revisits the structural and biochemical mechanisms of DHOase, with a focus on recent crystallographic insights that redefine the loop-in binding mode for ligand interaction. By leveraging the unique conformational dynamics of the active site loop, novel inhibitors may be developed to selectively target pyrimidine biosynthesis in cancer cells and microbial pathogens. These insights emphasize the crucial role of structural biology in therapeutic design and highlight DHOase as a promising drug target. Full article

Salivary biomarkers are promising molecules for diagnosing systemic diseases. Cytidine/uridine monophosphate kinase 2 (CMPK2) is associated with various systemic diseases. However, little is known about the role of the CMPK2 gene in saliva and dyslipidemia. This study investigated the relationship between serum lipid levels and Cmpk2 mRNA expression in the saliva of dyslipidemic mice. Additionally, immunofluorescence staining was employed to assess the localization of the CMPK2 protein in the submandibular gland. Two types of dyslipidemic mice were utilized: mice fed a high-fat and high-cholesterol (HFHC) diet and genetically dyslipidemic ApoE-deficient mice. The mice at 9 to 46 weeks were analyzed for serum lipid levels, Cmpk2 mRNA expression in saliva, and CMPK2 protein localization in the submandibular glands. Both dyslipidemic mice displayed elevated low-density lipoprotein cholesterol and total cholesterol in serum. ApoE-deficient mice apparently exhibited increased Cmpk2 expression in saliva. Immunofluorescence staining indicated that CMPK2 proteins were primarily localized in the serous acini, potentially associated with the secretion of Cmpk2 mRNA in saliva. These findings suggest that Cmpk2 mRNA increases and is detectable in the saliva of dyslipidemic mice, providing a viable experimental model to assess the potential use of CMPK2 as a biomarker for dyslipidemia. Full article

Genetic disorders are caused by a hereditary change in the structure of DNA that may hurt the health and life of animals. Several recessive haplotypes and a few causative mutations are known in Holstein Friesian cattle: CDH (Holstein cholesterol deficiency), haplotypes with a homozygous deficiency in Holstein (HH1, HH3, HH4, HH5, HH6, HH7), BLAD (bovine leukocyte adhesion deficiency), DUMPS (deficiency of uridine monophosphate synthase), FXI (factor XI deficiency), HHM (mule foot, syndactyly), and BC (citrullinaemia). From a breeding point of view, these genetic diseases have highly negative effects and are a significant problem for breeders, exposing them to economic losses and hurting animal welfare. This study aimed to characterize the Polish population of Holstein Friesian dairy cattle, considering the carrier status of twelve selected genetic defects. This study was based on genotype data collected from 78,884 cows and 691 bulls of the Holstein Friesian variety. The studies were performed using Illumina Infinium microarrays. Among both bulls and cows, the highest numbers of carriers were detected for HH5 (appropriately 6.7% and 5.4%). The lowest numbers of carriers were detected for DUMPS, factor XI, and HHM. The study revealed one calf suffering from cholesterol deficiency. Full article

Genetic disorders arise from alterations in the hereditary information encoded in DNA, leading to potential detrimental effects on the well-being and vitality of organisms. Within the bovine population, genetic conditions inherited in an autosomal recessive manner are frequently associated with particular breeds. In recent years, several recessive haplotypes and a few causative mutations have been discovered in Holstein cattle: CDH (Holstein cholesterol deficiency), haplotypes with a homozygous deficiency in Holstein (HH1, HH3, HH4, HH5, HH6 and HH7), BLAD (bovine leukocyte adhesion deficiency) and DUMPS (deficiency of uridine monophosphate synthase). All of these diseases are inherited in an autosomal recessive manner. From a breeding perspective, recessive mutations specifically exhibit considerable detrimental effects and are a significant problem for breeders, exposing them to economic losses. Individual mutations can cause embryo death at any stage of pregnancy. Only genetic research and conscious selection of animals for mating will lead to a reduction in the number of carriers and elimination of mutations from the population. Full article

Latilactobacillus (L.) sakei is a species of lactic acid bacteria (LAB) mostly studied according to its application in food fermentation. Previously, L. sakei L3 was isolated by our laboratory and possessed the capability of high exopolysaccharide (EPS) yield during sucrose-added fermentation. However, the understanding of sucrose promoting EPS production is still limited. Here, we analyzed the growth characteristics of L. sakei L3 and alterations of its transcriptional profiles during sucrose-added fermentation. The results showed that L. sakei L3 could survive between pH 4.0 and pH 9.0, tolerant to NaCl (<10%, w/v) and urea (<6%, w/v). Meanwhile, transcriptomic analysis showed that a total of 426 differentially expressed genes and eight non-coding RNAs were identified. Genes associated with sucrose metabolism were significantly induced, so L. sakei L3 increased the utilization of sucrose to produce EPS, while genes related to uridine monophosphate (UMP), fatty acids and folate synthetic pathways were significantly inhibited, indicating that L. sakei L3 decreased self-growth, substance and energy metabolism to satisfy EPS production. Overall, transcriptome analysis provided valuable insights into the mechanisms by which L. sakei L3 utilizes sucrose for EPS biosynthesis. The study provided a theoretical foundation for the further application of functional EPS in the food industry. Full article

Currently, an important group of biomaterials used in the research in the field of tissue engineering is thermosensitive chitosan hydrogels. Their main advantage is the possibility of introducing their precursors (sols) into the implantation site using a minimally invasive method—by injection. In this publication, the results of studies on the new chitosan structures in the form of thermosensitive hydrogels containing graphene oxide as a nanofiller are presented. These systems were prepared from chitosan lactate and chitosan chloride solutions with the use of a salt of pyrimidine nucleotide—uridine 5′-monophosphate disodium salt—as the cross-linking agent. In order to perform the characterization of the developed hydrogels, the sol–gel transition temperature of the colloidal systems was first determined based on rheological measurements. The hydrogels were also analyzed using FTIR spectroscopy and SEM. Biological studies assessed the cytotoxicity (resazurin assay) and genotoxicity (alkaline version of the comet assay) of the nanocomposite chitosan hydrogels against normal human BJ fibroblasts. The conducted research allowed us to conclude that the developed hydrogels containing graphene oxide are an attractive material for potential use as scaffolds for the regeneration of damaged tissues. Full article