Search Results (287)

To understand glycan–protein interactions at biological interfaces, designing surfaces modified with structurally controlled glycans is highly important. In particular, naturally occurring glycosaminoglycans (GAGs) possess periodic sugar arrangements that play important roles in protein recognition, highlighting the need for the development of periodic glycopolymer model systems that can serve as GAG mimics for quantitative interaction analysis. In this study, sequence-controlled periodic glycopolymers were synthesized by reversible addition–fragmentation chain-transfer (RAFT) polymerization and immobilized onto gold surfaces to construct glycan-modified interfaces. The synthesized material was a terminally functionalized periodic glycopolymer with the most basic structure, consisting of alternating maltose-containing vinyl ether (MalVE) units and ethyl maleimide (EtMI) units, with a trithiocarbonate group at the ω-terminal. This trithiocarbonate group was converted to a thiol group for immobilization through Au–S bond formation. Structural characterization by ¹H NMR spectroscopy, size exclusion chromatography (SEC), MALDI-TOF mass spectrometry, and UV–vis spectroscopy confirmed the structure as designed. Quartz crystal microbalance (QCM) measurements verified the stable immobilization of thiol-terminated periodic glycopolymers on the gold surface, and allowed for estimation of graft density and quantitative analysis of glycan-protein interactions at the modified interface. The periodic glycopolymer-modified surfaces exhibited selective binding behavior toward concanavalin A (ConA) compared to bovine serum albumin (BSA), with apparent binding constants on the order of 10⁶–10⁷ L mol⁻¹. This enhanced binding behavior indicated that specific and multivalent interactions with proteins also occurred at periodic pendant maltose residues along the main chain. These results demonstrate that the gold surface modified with end-functional periodic glycopolymers synthesized by RAFT polymerization provides a versatile platform for quantitative analysis of glycan-protein interactions and suggests potential applications for periodic glycopolymers as functional materials. Full article

Atherosclerosis is a progressive vascular disease characterized by lipid-rich plaque accumulation, oxidative stress, and chronic inflammation, contributing to coronary heart disease, stroke, and peripheral arterial disease. This study investigated the impact of inflammation, vascular calcification, and statin therapy on redox balance in blood and carotid artery plaques, aiming to identify potential biomarkers for disease assessment. Thirty-two patients undergoing carotid endarterectomy provided 34 plaque samples. Enzyme activities in plaque/erythrocytes and –SH group concentration in plasma/plaque were measured. Pathological analysis was performed to determine inflammation/calcification grade, the presence of mast cells and plaque composition. The results showed that mast cells were associated with reduced non-protein –SH groups, indicating selective thiol consumption and serving as a qualitative marker of oxidative burden. Reduced catalase activity in erythrocytes was associated with advanced calcification, pointing to long-standing systemic oxidative stress. Statin therapy enhanced systemic superoxide-dismutase 1 activity, increased –SH groups, and modulated plaque-specific glutathione reductase activity, attenuating sex-related differences in redox regulation. These findings highlight the complex interplay between systemic and local oxidative processes in atherosclerosis through alterations in redox-related biomarkers such as plasma –SH group concentrations and catalase activity. Full article

The exact mechanisms of skin involvement in type 1 diabetes (DM1) remain poorly understood. This study aimed to evaluate the relationship between antioxidants, oxidative stress, protein glycation, and glycoxidation, as well as matrix metalloproteinase (MMP) activity, in the skin of rats with DM1, while investigating whether insulin administration improves skin homeostasis. Male Wistar rats were assigned to three groups: control, diabetes, and diabetes treated with insulin. Significantly higher expression of GSH (gluthatione) and GSH-Px (glutathione peroxidase), elevated levels of AGE (Advanced Glycation End products), DT (dityrosine), KN (kynurenine), NFKN (N-formylkynurenine) and ONOO- (peroxynitrite), as well as increased activity of GLU (β-D-glucuronidase), NADPH oxidase (NOX) and MMP-1, -2, -3, -7, -9, -11 and -13 were observed in the skin of rats with DM1. Insulin treatment normalizes the skin’s antioxidant barrier and eliminates oxidative stress. It also reduces the intensity of protein glycation and glycoxidation, though not to the levels observed in the control group. Summarizing, in diabetic skin there is a complex interaction between the thiol antioxidant barrier, oxidative damage, protein glycation and glycoxidation as well as MMP expression. Insulin restores physiological balance in skin cells; however, glycation and ECM remodeling are still more pronounced than in healthy skin. Full article

This study examined alterations in serum redox biomarkers before and one month after administration of the coronavirus disease 2019 (COVID-19) booster (third) doses across four vaccine regimens. A longitudinal cohort of 410 adults was analyzed following homologous Pfizer-BioNTech, Sinopharm [Vero Cell]-Inactivated, Sputnik V, or heterologous Sinopharm/Pfizer vaccination. Serum total proteins, albumin, total thiols, nitrites, ferric-reducing antioxidant power (FRAP), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity were measured, with DPPH interpreted as an ex vivo surrogate of serum radical-scavenging capacity. Additional analyses included stratification by prior severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, multivariable regression, correlation analysis, effect-size estimation, and sensitivity testing. Booster vaccination was associated with modest but consistent decreases in DPPH activity, albumin, and total proteins, whereas FRAP, nitrite, and total thiol levels remained stable. This pattern supports a transient shift in antioxidant buffering capacity but, by itself, does not exclude oxidative stress, as direct oxidative damage markers were not assessed. The most pronounced changes were observed in Sinopharm-based regimens, particularly in the heterologous Sinopharm/Pfizer group. Prior SARS-CoV-2 infection did not materially alter the qualitative response pattern, whereas older age and comorbidities were associated with greater declines in DPPH activity and albumin. Overall, the findings indicate a modest, transient redox-associated response following booster-induced immune activation and suggest that host-related factors, such as age and comorbidity burden, may accentuate short-term changes in antioxidant buffering capacity. Full article

This study focused on the inhibitory effects of wheat bran feruloyl oligosaccharides (FOs) on the formation of AGEs in three bovine serum albumin (BSA)-based non-enzymatic glycation models, namely BSA-fructose, BSA-methylglyoxal (MGO), and BSA-glyoxal (GO). In the BSA-fructose model, FOs at 0.25 mg/mL achieved a 62% inhibition rate of fructosamine, equivalent to approximately 78% of the activity of the positive control aminoguanidine (AG), and reduced fluorescent AGEs by over 50% on day 12. Additionally, FOs suppressed the accumulation of α-dicarbonyl compounds, key intermediates in the glycation pathway. In the BSA-MGO and BSA-GO system, the decreased fluorescence intensity of tryptophan residues indicated that FOs bound to BSA, inducing conformational changes in the protein microenvironment; this binding also inhibited protein carbonyl formation and the loss of thiol groups, thereby modulating the protein glycation process. Compared with their precursors (ferulic acid, FA; xylooligosaccharides, XOS), FOs exhibited comparable or even superior inhibitory activity against specific AGE subtypes, suggesting a synergistic effect between the feruloyl and oligosaccharide moieties. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed that FOs reduced the band intensity of 90 kDa AGEs in the glycation system, indicating the inhibition of protein-fructose cross-linking. Fluorescence spectroscopy confirmed that FOs dynamically quenched BSA with a single binding site, and thermodynamic calculations demonstrated that the binding was spontaneous (ΔG < 0), primarily driven by hydrogen bonds and van der Waals forces (ΔH < 0, ΔS < 0). This study systematically investigated the anti-glycation activities of FOs and their precursors. The findings demonstrate that FOs are promising natural glycation inhibitors and provide important theoretical and experimental support for related research. Furthermore, this study establish a basis for the green and high-value utilization of agricultural by-products like wheat bran. Full article

Mussel foot proteins (Mfps) are renowned for their underwater adhesion, whereas their biotechnological potential for cutaneous wound repair remains largely underexplored. In this study, we identified and characterized a cysteine-rich mussel foot protein, Mfp6-1, from Mytilus coruscus and investigated its therapeutic potential for wound healing. Sequence analysis showed that Mfp6-1 is enriched in cysteine (11.0%) and tyrosine (~16.5%). We successfully expressed recombinant Mfp6-1 (rMfp6-1) in E. coli. Structural prediction based on the mature peptide sequence suggested that rMfp6-1 adopts a relatively compact fold containing several short β-structural elements. In vitro assays demonstrated that rMfp6-1 possesses antioxidant activity in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, and alkylation experiments suggested that cysteine residues contribute importantly to this activity. Dithio-bis-nitrobenzoic acid (DTNB)-based thiol quantification further demonstrated that rMfp6-1 contained abundant accessible free sulfhydryl groups, supporting an important contribution of cysteine-derived thiols to its antioxidant activity. Experiments on a full-thickness mouse wound model showed that rMfp6-1 treatment resulted in significantly faster wound contraction. Morphological analysis further revealed that rMfp6-1 optimizes the healing microenvironment by promoting collagen accumulation and re-epithelialization. Additionally, the treatment was found to trigger vascular endothelial growth factor (VEGF)-mediated angiogenesis, thereby improving the overall quality of the regenerated tissue. Furthermore, rMfp6-1 treatment significantly reduced interleukin-6 (IL-6) expression, suggesting that its antioxidant capacity creates a permissive microenvironment for tissue regeneration by suppressing excessive inflammation. These findings indicate that recombinant rMfp6-1 is a promising bioactive candidate for wound-healing applications. Full article

Background: Metabolic syndrome (MetS) is a cluster of pathologies (obesity, dyslipidemia, insulin resistance, hypertension) that affects over one quarter of old adults. MetS is a condition that markedly increases the susceptibility of various organs to dysfunctionality and is associated with the development of oxidative stress. The existing guidelines point out that exercise is highly advantageous for patients with MetS. However, there is a need for specific guidance and clinical evidence. Objective: This study aimed to investigate the effects of a moderate aerobic exercise program on older women without and with MetS. Methods: A total of 120 women aged 60–70 years old were recruited and divided into two groups: healthy old women (HOW, N = 60) and old women with MetS (OW-MetS, N = 60). Anthropometric values, biochemical parameters and markers of oxidative damage were evaluated before and after moderate aerobic exercise. Exercise was performed five days per week for three months (64 sessions). Each exercise session consisted of 40 min and included the following: (a) five minutes of warm-up exercise; (b) ten minutes of flexibility exercise with resistance using own weight and coordination; (c) twenty minutes of moderate-intensity aerobic exercise (heart rate max between 60% and 70%); and (d) five minutes to cool down/stretching with respiratory techniques. Results: A significant decrease in anthropometric variables was generated by the exercise program [waist circumference 4.35 cm (p < 0.05) in OW-MetS, body fat −1.55, −1.39% (p < 0.05) and muscle mass 0.8, 1.1% (p < 0.05) in HOW and OW-MetS, respectively]. The exercise program resulted in beneficial changes in all biochemical parameters in both groups. Importantly, HOMA values showed a significant decline of −0.85 and −6.17 in HOW and OW-MetS, respectively. Furthermore, oxidative stress was present in the OW-MetS group, which was reduced by the exercise program, resulting in a decrease in protein damage [formazan 45% and 42% in HOW and OW-MetS respectively] and an increase in antioxidant defenses (thiol groups 36%, 99% and GPx 55%, 20% in HOW and OW-MetS, respectively). Conclusions: The data of this study show that moderate aerobic exercise may be potentially useful in treating and preventing MetS in older patients. Full article

The dose–response relationship for fluoride (F⁻) exposure remains largely unexplored. Hence, the current study assessed the hepatotoxic and nephrotoxic effects of subacute exposure (28 days) to increasing F⁻ concentrations in Wistar rats via the benchmark dose (BMD5) method. Thirty male rats were assigned to six groups (n = 5): a control group (tap water) along with five groups that received F⁻ via drinking water at increasing concentrations (10, 25, 50, 100, and 150 mg/L). F⁻ toxicity was determined via water intake, weight gain, histological analyses, redox status, and essential element levels. PROASTweb 70.1 software was utilized to investigate the external and internal F⁻ dose–response relationships. Specified major cytoarchitecture damage and superoxide anion (O₂·⁻), total oxidative status (TOS), superoxide dismutase (SOD) activity, total thiol groups (SH), and advanced oxidation protein product (AOPP) level alterations were detected in both sets of tissues. Moreover, F⁻ caused an imbalance in copper (Cu), zinc (Zn), iron (Fe), and manganese (Mn). The most sensitive parameters were O₂·⁻ (0.06 mg F⁻/kg) in the liver and AOPP (6.5 × 10⁻⁶ mg F⁻/L) in the kidneys. These findings contribute to the limited risk assessment of fluorides and highlight the dose-dependent relationship between redox status parameters and bioelements in the liver and kidneys. Full article

Cyclophosphamide (CYP) is an effective chemotherapeutic, but its use is limited by hemorrhagic cystitis caused by its toxic metabolite acrolein. Acrolein, when concentrated in the urine, triggers oxidative stress, leading to urothelial inflammation and cell death. Given that albumin is the most abundant plasma protein that contains free thiol groups capable of neutralizing electrophiles and oxidants, we, therefore, hypothesized that albumin could mitigate CYP-induced bladder injury. Here, we tested this hypothesis. In CYP-induced mouse cystitis, albumin administration markedly reduced bladder enlargement, edema, and hemorrhage, effectively normalizing the bladder weight. Albumin also reduced bladder oxidative injury and preserved the expression of anti-ferroptotic proteins, including the cystine/glutamate antiporter xCT and glutathione peroxidase 4 (GPX4). In addition, albumin-treated mice showed less leakage of inflammatory protein into bladder tissue. In vitro, albumin protected urothelial cells from acrolein-induced cell death. It also significantly prevented H₂O₂-induced cytotoxicity. Mechanistically, albumin acted as an extracellular scavenger that preferentially reacted with acrolein and H₂O₂, thereby sparing cellular components from oxidative damage. Notably, oral albumin supplementation similarly attenuated CYP-induced cystitis. Furthermore, albumin administration improved survival in a high-dose CYP toxicity model. These findings establish albumin as a potent protector against CYP-induced toxicity by sequestering acrolein and scavenging reactive oxygen species. Albumin supplementation could be a practical strategy to mitigate chemotherapy-associated bladder and systemic injury. Full article

Background/Objectives: Syringa vulgaris L. (common lilac) is one of the most popular ornamental plant species. Through the ages, many parts of S. vulgaris, including fruits, flowers, leaves, and branches, have been used in folk medicine due to their beneficial biological activity. Lilac flowers are the basis of many supplements available on the market. Moreover, its petals and flowers are edible and are an aromatic ingredient in preserves and desserts. However, the data about the antioxidant properties of various parts of S. vulgaris is limited only to the in vitro antioxidant capacity of the extracts—so far, the effect of S. vulgaris flower extract on the parameters of oxidative stress in biological materials, including plasma, has not been demonstrated. Therefore, the aim of our study was to investigate the protective effects of the extract from S. vulgaris L. flowers against oxidative stress in human plasma, and its influence on the coagulation process in vitro. Methods: We measured the levels of three parameters of oxidative stress in human plasma treated with H₂O₂/Fe²⁺ (the donor of hydroxyl radicals): lipid peroxidation (based on the level of thiobarbituric acid reactive substances (TBARS)), protein carbonylation, and thiol oxidation. Ascorbic acid (vitamin C) was used as a reference antioxidant. In addition, we studied the effect of the extract on three coagulation parameters of human plasma-activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT). We also compared the biological properties of the extract from S. vulgaris flowers with the properties of a phenolic extract from Taraxacum officinalis (dandelion) flowers, as they have proven antioxidant activity in both in vitro and in vivo models and can modulate hemostasis in vitro. Results: Our UHPLC-HRMS analyses of S. vulgaris extract led to a tentative identification of 50 compounds, mainly phenolics and secoiridoids. For the first time, the present study demonstrated that the extract from S. vulgaris flowers (at the concentrations of 1–50 µg/mL) significantly reduced plasma lipid peroxidation and protein carbonylation induced by H₂O₂/Fe²⁺. Moreover, the concentrations of 1–25 µg/mL significantly reduced the oxidation of thiol groups in plasma treated with H₂O₂/Fe²⁺. The anticoagulant tests also demonstrated that S. vulgaris flowers extract, at physiologically relevant concentrations (1–50 µg/mL), did not affect blood clotting times in vitro, suggesting that it is hemostatically safe. Conclusions: Despite the differences in composition, the extracts from lilac flowers and dandelion flowers exhibited similar protective effects against oxidative damage to human plasma components. However, the extract from S. vulgaris flowers had a stronger inhibitory effect on lipid peroxidation than the extract from dandelion flowers. Full article

Circadian rhythm regulates several physiological functions, and influences endocrine and metabolic responses in mammals, with cortisol acting as important modulator of this mechanism. Cortisol secretion is affected by both internal and external factors and is intensified under stress conditions. The response to surgical stress is consistently observed after surgical procedures, such as ovariohysterectomy (OVH). Therefore, this study aimed to evaluate the influence of the circadian rhythm on the surgical stress response following elective OVH in healthy bitches. Twenty patients weighing between 10 and 20 kg were hospitalized 48 h before surgery and remained hospitalized for 48 h postoperative. The animals were randomly allocated into two groups and underwent OVH either in the morning (6–8 h—a.m., GAM) or at night (18–20 h—p.m., GPM). Surgical procedures were standardized with respect to the surgical team, technique applied and duration; this was carried out in order to induce a comparable level of surgical stress. Physical parameters (systolic blood pressure, heart rate, respiratory rate and rectal temperature) and laboratorial analyses (cortisol, leukogram, protein thiols, no protein thiols, vitamin C, ferric reducing ability of plasma and thiobarbituric acid reactive substances) were assessed immediately before surgery and at 2, 4, 6, 12, 24 and 48 h, as well as 14 days postoperatively. No significant changes in cortisol profile were detected. However, significant alteration in the respiratory rate, rectal temperature, time to first urine, and lipid peroxidation were observed in the GPM group, suggesting that surgeries performed at night induce greater disturbances in homeostasis than those performed in the morning. Full article

Due to the non-polar and chemically inert nature of carbon fiber surfaces, the interfacial bonding strength between carbon fibers and norbornene-polyimide (PI-NA) resin matrix is relatively weak. To address this issue, this study constructed a composite coating on the carbon fiber surface and proposed a novel method to build robust interfaces based on multiple interfacial interactions, thereby effectively enhancing the interfacial properties between carbon fibers and PI-NA resin. Inspired by mussel adhesive proteins, this study established a multi-level synergistic interfacial reinforcement system by sequentially constructing a C-PEI@OPDA coating, in situ growing zinc oxide nanorods (ZW) arrays, and grafting 3-mercaptopropyltrimethoxysilane (MPS) onto carbon fiber surfaces. The C-PEI@OPDA coating, rich in amino (–NH₂) and hydroxyl groups (–OH), enhanced adhesion to carbon fibers and adsorbed Zn²⁺ via coordination interactions to provide nucleation sites for ZW growth. Meanwhile, the active hydrogen in the coating promoted the crosslinking of PI-NA resin, thereby increasing the resin crosslinking density in the interfacial region. The vertically aligned ZW significantly increased surface roughness, enhanced mechanical interlocking effects, and provided secondary reaction sites for MPS grafting. The thiol groups (–SH) in MPS formed covalent bonds with PI-NA resin through thiol-ene click reactions, further strengthening interfacial bonding. The results showed that the ILSS, IFSS, and flexural strength of C-PEI@OPDA/ZW/MPS modified carbon fiber composites reached 75.15 MPa, 102.93 MPa, and 1735.56 MPa, representing improvements of 39.09%, 48.79%, and 31.16%, respectively. This study effectively enhanced the carbon fiber-reinforced polymer composites interfacial bonding strength through the synergistic effects of hydrogen bonding, mechanical interlocking, chemical bonding, and increased resin crosslinking density. Full article

Introduction: cigarette smoking is a major source of systemic oxidative stress and a well-established risk factor for cardiovascular disease. Heated tobacco products (HTPs) are increasingly promoted as reduced-risk alternatives, yet their cellular effects remain incompletely understood. Methods: this study compared the oxidative stress-mediated effects of conventional cigarette smoking and HTP use on human erythrocytes. Erythrocytes from healthy non-smokers, conventional smokers, and HTP users were analyzed using biochemical, functional, and cytological approaches to assess redox status, membrane and cytoskeletal organization, anion exchanger 1 (AE1) function, antioxidant response, and redox-sensitive signaling pathways. Results: conventional smokers exhibited higher intracellular reactive oxygen species (ROS) levels, thiol depletion, methemoglobin and hemichrome formation, whereas HTP users showed marked lipid peroxidation despite lower ROS availability. Both groups instead displayed altered expression and distribution of key membrane and cytoskeletal proteins, including glycophorin A, AE1, spectrin, ankyrin, and band 4.1, indicating impaired membrane–cytoskeleton interactions. Functional analyses revealed an accelerated AE1-mediated anion exchange in erythrocytes from conventional smokers, whereas cells from HTP users exhibited a reduced sulfate accumulation, indicating altered transport capacity. In both groups, G6PDH activity was significantly increased, and redox-sensitive signaling pathways involving ERK, AKT, and eNOS were activated, accompanied by sex-dependent alterations in estrogen receptor expression and distribution. Conclusions: collectively, these findings identify erythrocytes as sensitive biomarkers of tobacco-related systemic damage and indicate that smoking-induced erythrocyte dysfunction, including that associated with HTP use, may actively contribute to vascular impairment. This evidence challenges the assumption that heated tobacco products confer a substantially reduced cardiovascular risk compared with conventional cigarettes. Full article

Silver nanoparticles were biosynthesized using the culture supernatant of Staphylococcus sp. YRA, a strain isolated from Colombian mining sediments. Synthesis was optimized at 1 mM AgNO₃, pH 7, 40 °C and 7 μg/mL extract, producing spherical, protein-capped AgNPs with primary sizes in the tens-of-nanometers range (~35–90 nm by SEM), while DLS indicated larger hydrodynamic diameters (~250–320 nm) consistent with aggregation in suspension (ζ-potential −16.6 mV). These nanoparticles remained stable over 6 months. Characterization by UV–Vis, SEM, AFM, EDS and FTIR confirmed extracellular protein-mediated reduction and capping. The AgNPs showed antibacterial activity against multidrug-resistant clinical isolates (Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Salmonella bongori, Enterococcus spp.), with inhibition zones of 8–16 mm at 400–1000 μg/mL. Biofilm formation was reduced by >50% at 700 μg/mL in both Gram-positive and Gram-negative strains. In Phaseolus vulgaris (P. vulgaris), low concentrations (5–100 μg/mL) increased growth and chlorophyll content, while 500 μg/mL caused moderate inhibition. FTIR analysis identified amide and thiol groups from bacterial enzymes as capping agents. These results suggest Staphylococcus sp. YRA as a bacterial platform for AgNPs production with antibiofilm activity against MDR pathogens and acceptable phytotoxicity profile for potential applications. Full article

Coated tofu is prone to spoilage and degradation during processing, storage, and transportation. As the material basis for gel of coated tofu, proteins determine coated tofu’s unique qualities, such as its colour, flavour, and texture. This study aimed to investigate the changes in the quality of coated tofu and the physicochemical properties of its proteins during cold storage (4 °C and 10 °C, 14 days), as well as the intrinsic correlations between these variables. Quality deterioration and protein structural changes were significantly slower at 4 °C than at 10 °C, with lower temperature effectively delaying quality loss. The results indicated that as storage time increased, the freshness of coated tofu declined, its textural properties significantly deteriorated, and the protein gel network structure became impaired. Meanwhile, the proteins underwent significant oxidative denaturation, characterized by a decrease in the free thiol group content and an increase in surface hydrophobicity. The tertiary structure exhibited unfolding and disruption, while the secondary structure transitioned from an ordered to a disordered state. Specifically, the contents of α-helixes and β-sheets decreased significantly, reaching 34.96% and 8.68%, respectively, after 14 days of storage at 4 °C. In contrast, the contents of β-turns and random coils increased to 30.11% and 26.25%, respectively, under the same storage conditions. The subunit bands of the 11S and 7S proteins gradually weakened, and the protein structure tended to loosen. Correlation analysis revealed that the oxidative denaturation, structural depolymerization, and reaggregation of proteins were highly significantly correlated with the textural breakdown and colour deterioration of coated tofu, which together contributed to the quality degradation of coated tofu during cold storage. The findings of this study provide fundamental data and technical support for the development of cold storage methods for coated tofu. Full article