Search Results (1,998)

This study investigates the antioxidant properties of several synthetic peptides derived from milk proteins previously identified in milk permeate, a by-product of the dairy industry. The aim of the research is to identify which peptides present in milk permeate are responsible for its antioxidant activity. A comprehensive experimental strategy was employed to evaluate their antioxidant potential, including in silico selection, in vitro free radical scavenging assays and cellular models using Caco-2 and HCT116 cell lines. The peptides were screened using a molecular docking approach for their potential interaction with the Kelch-like ECH-associated protein 1/nuclear factor erythroid 2-related factor 2 (Keap1/Nrf2) pathway, and eight out of twenty-eight were selected and synthesized for further analyses. In vitro, six of the selected peptides demonstrated significant direct antioxidant activity in the DPPH scavenging assay, and two in the ABTS scavenging test. In cellular environments, three peptides (LPAPELGPRQA, LPIIQKLEPQI and NGQVWEESLKRL) effectively protect cells from oxidative stress induced by tert-butyl hydroperoxide, reducing reactive oxygen species production and partially mitigating lipid peroxidation. Further investigation showed that two of them (LPAPELGPRQA and LPIIQKLEPQI) effectively induce the Keap1/Nrf2 pathway, as evidenced by a ∼1.5-fold increase in Nrf2 levels and overexpression of downstream proteins. Permeability studies revealed that these peptides can cross the intestinal monolayer (2–3% in 2 h), suggesting potential systemic effects. Overall, these findings highlight the multifunctional antioxidant properties of the investigated peptides and support their potential application as nutraceuticals or therapeutic agents for oxidative stress-related conditions. Full article

Cigarette smoke (CS) is the primary risk factor for the development of chronic obstructive pulmonary disease (COPD). Investigating the impact of CS on human airway epithelium is important for understanding COPD development and combating its effects. While some studies show that long exposure to CS activates inflammasome formation in airway epithelium, leading to cytokines’ maturation and release, its acute effect on inflammation regulation requires further elucidation. Due to the importance of acute cellular responses in modulating cell survival and controlling inflammatory outcomes, we examined the effect of acute cigarette smoke extract exposure on human bronchial epithelial cells. Due to the high reactive oxygen species content in CS, we hypothesize that acute CS exposure activates the integrated stress response (ISR) pathway leading to stress granules (SG) formation to facilitate oxidative stress resolution and promote cell survival. Immunostaining, fluorescence confocal imaging, quantitative analyses, and immunoblotting were performed to test our hypothesis. We report here that acute exposure to CS extract triggers canonical SG formation by activating the ISR pathway via the PERK/eIF2α arm in a reactive oxygen species-dependent manner. SG formation is abolished upon inhibiting PERK or eIF2α function, or by scavenging oxidants prior to smoke exposure. Characterizing SG formation in terms of measuring SG size and abundance and the sequestration of the SG marker G3BP1 reveals that SG formation is maximal at 15% CS extract exposure for 2 h and undergoes gradual disassembly at longer exposure times. This is closely dependent on cytoplasmic p-eIF2α levels. These results demonstrate that acute exposure to CS activates the protective ISR pathway to potentially reduce the detrimental effects of CS and promote stress resolution and cell survival. Full article

Background/Objectives: Pancreatic ductal adenocarcinoma (PDAC) is a lethal disease with limited treatment options. Patients are treated with DNA damaging chemotherapies which act by inducing DNA damage in rapidly dividing tumor cells. Unfortunately, these tumors frequently develop treatment resistance, underscoring the need to understand resistance mechanisms in order to develop better treatment strategies. DNA damage response (DDR) detects and repairs DNA damage, and the DDR pathway has been shown to contribute to chemoresistance. Another factor known to drive chemoresistance in PDAC is the dense stroma, composed of extracellular matrix proteins secreted by cancer-associated fibroblasts (CAFs). Our recent work identified a CAF-induced resistance mechanism involving N-myc downstream regulated gene 1 (NDRG1). CAF-induced signaling resulted in the phosphorylation of NDRG1 and NDRG1-dependent DNA repair and protection from chemotherapies. Loss of NDRG1 resulted in increased chemotherapy-induced DNA damage and decreased replication fork speed and recovery. Methods: To gain insight into the molecular mechanism of NDRG1-mediated DNA repair and replication, we performed a BioID screen to identify binding partners of NDRG1. We further assessed the mechanistic roles of the identified interaction partners on DNA repair using DNA replication and repair assays such as the Comet assay and DNA fiber assays. Results: Our BioID screen identified meiotic recombination 11 (MRE11) protein, a nuclease involved in DDR, as a putative NDRG1 interacting protein. Interaction between MRE11 and NDRG1 was enriched during the late S/early G2 cell cycle phases and under replication stress. However, this interaction is likely indirect as the interaction only occurred in a cellular context and not with in vitro purified proteins. Blocking NDRG1 phosphorylation or blocking MRE11 exonuclease activity both resulted in protection of newly synthesized DNA at stalled replication forks. In NDRG1 knockout cells, blocking MRE11 led to decreased protection of nascent DNA, suggesting that NDRG1 and MRE11 may be acting in the same pathway and that NDRG1 is required for MRE11’s activity at stalled forks. Conclusions: In summary, our work has uncovered a protein complex between NDRG1 and MRE11 that may play a key role in chemoresistance due to its role in the processing of stalled replication forks. Full article

Background: Despite the effectiveness of current SARS-CoV-2 vaccines, the genetic variability of the viral target has led to the emergence of variants capable of evading vaccine-induced protection. To ensure broader and more durable protection, we investigated the efficacy of a novel vaccine strategy. Methods: This vaccine utilizes the highly conserved nucleocapsid (N) protein as its primary antigen, rather than the spike (S) protein. It incorporates the Papaya Mosaic Virus (PapMV) nanoparticle, a Toll-like receptor (TLR) 7/8 agonist with intrinsic adjuvant properties, as a vaccine platform. Results: The vaccine formulations, comprising PapMV nanoparticles and the N antigen covalently or non-covalently attached to the PpaMV nano, generated robust humoral (antibody) and cellular (T-cell) immune responses. Protective efficacy was evaluated in K18-hACE2 transgenic mice challenged with either the ancestral SARS-CoV-2 strain or the Omicron XBB.1.5 variant. In both cases, the vaccine significantly reduced inflammation and viral titers in the lungs of vaccinated animals. Conclusions: These results highlight the potential of this PapMV-N vaccine to induce broad protection against diverse SARS-CoV-2 variants. Full article

Background/Objectives: Our previous study demonstrated that while the third SARS-CoV-2 booster effectively enhanced immunity against the Delta subvariant, its protection declined over time. This study aimed to evaluate and compare the humoral and cellular immune responses, as well as reactogenicity, of the mRNA-1273 vaccine administered as a fourth booster in healthy Thai adults previously vaccinated with two doses of CoronaVac (CV) followed by a third dose of either AZD1222 (AZ) or BNT162b2 (BNT). Methods: Participants received a single 100 µg (0.5 mL) intramuscular dose of mRNA-1273. Blood samples were collected at baseline (D0), D14, D90, and D180 to assess anti-RBD IgG, conduct a surrogate virus neutralization test (sVNT) against the Delta and Omicron variants, and assess IFN-γ levels and reactogenicity. Results: Both 2CV/AZ- and 2CV/BNT-primed groups exhibited comparable local and systemic reactogenicity. The fourth mRNA-1273 dose markedly increased Delta variant inhibition within 14 days in both groups and remained at high levels at Days 90 and 180. sVNT inhibition against Omicron rose similarly in both groups at Day 14; it declined sharply by Days 90 and 180, with the 2CV/AZ-primed group showing significantly lower levels than the 2CV/BNT-primed group. Baseline anti-RBD IgG levels were lower in the 2CV/AZ group (p = 0.003) but surpassed those of the 2CV/BNT group by Day 14, with no significant differences at later time points. IFN-γ responses followed a similar pattern to anti-RBD IgG Conclusions: A heterologous fourth mRNA-1273 booster in both 2CV/AZ- and 2CV/BNT-primed groups effectively enhances B-cell and T-cell responses against SARS-CoV-2. However, emerging variants such as Omicron may still pose challenges. The trial was registered with the Thai Clinical Trials Registry: the name of the registry: “The comparison of immune response to the 4th dose booster with mRNA-1273 COVID-19 vaccine in individuals who had received 2 doses of CoronaVac and booster with ChAdOx-1 or BNT162b2 COVID-19 vaccine”, TCTR20220205002 on 5 February 2022. Full article

Heat stress (HS) affects female reproductive efficiency by disrupting redox homeostasis and activating inflammatory responses in the corpus luteum (CL), a metabolically active tissue essential for pregnancy maintenance. This study reveals the protective effect of resveratrol against HS-induced luteal injury in pregnant mice through the regulation of oxidative stress and cytokine–chemokine-mediated inflammatory and immune responses. The pregnant mice were divided into three groups: control, HS, and resveratrol +HS. Heat stress was applied at 40 ± 0.5 °C for 7 days, with resveratrol (10 mg/kg) given orally 2 h before exposure to HS. The results showed that heat exposure reduced serum total superoxide dismutase activity and increased malondialdehyde level, causing significant disruption of luteal morphology with cellular disorder and vacuolization, which was partially overcome by resveratrol pretreatment. Transcriptomic profiling showed that HS induced a strong immunological and inflammatory response, involving cytokine–cytokine receptor interaction and chemokine signaling. Resveratrol significantly attenuated HS-induced transcriptional changes. The RT-qPCR results showed that HS increased chemokine ligands (Ccl11, Cxcl13, Tslp) and cytokine receptors (Ccr3, Ccr4, Ccr5), which were suppressed by resveratrol. The chemokine-based inflammatory module is one of the most important regulatory properties of the HS response, according to the network analysis. Stable binding of resveratrol with major chemokine receptors was supported by molecular docking and molecular dynamics simulations. Collectively, HS induces oxidative, structural, and inflammatory alterations in luteal tissue, while resveratrol attenuates these changes by being associated with improved antioxidant status and suppression of cytokine–chemokine-mediated responses. Full article

Necrotizing enteritis (NE) is an important intestinal disease threatening the poultry farming industry, and the ban on antibiotic growth promoters has created an urgent demand for safe and effective NE vaccines. Recombinant attenuated Salmonella vectors (RASVs) administered orally can induce mucosal immune responses against delivered antigens, thus showing great potential to elicit protective immunity against NE. The EntB protein is a newly discovered putative enterotoxin of Clostridium perfringens (C. perfringens). Bioinformatic predictions in this study revealed that EntB contains nineteen potential antigenic epitopes, two functional domains (NlpC and YgiM), and interacts with ten proteins, supporting its potential as a target antigen for NE vaccines. To optimize the immunogenicity of EntB-based vaccines, we constructed a novel recombinant attenuated Salmonella Enteritidis (S. Enteritidis) vector rSC0169 harboring a rhamnose-regulated delayed attenuation system, which was then used to deliver EntB to generate the recombinant strain rSC0169(pS-EntB). This system enhanced the immunogenicity of the Salmonella vector rSC0169 and further elicited robust mucosal immune responses against EntB, as well as humoral and cellular immune responses. Compared with the control strain rSC0169(pS0018), rSC0169(pS-EntB) candidate vaccine strain significantly alleviated NE symptoms, increased the intestinal villus height/crypt depth (VH/CD) ratio, upregulated tight junction protein expression, and reduced excessive pro-inflammatory cytokine production. In conclusion, this study provides a promising NE candidate vaccine and offers a valuable strategy for developing vaccines against other intestinal bacterial diseases. Full article

Background/Objectives: Bovine viral diarrhea (BVD) is a major infectious disease of cattle caused by bovine viral diarrhea virus genotypes 1 and 2 (BVDV-1 and BVDV-2). Current inactivated and live attenuated vaccines provide incomplete cross-genotype protection and may exhibit limitations related to durability of immunity or safety. This study evaluated whether co-expression of the BVDV envelope glycoproteins E1 and E2 in a Modified Vaccinia Ankara (MVA) vector could support antigen expression and induce immune responses in a proof-of-concept model. Methods: Recombinant Modified Vaccinia Ankara (MVA) viruses expressing BVDV-1 E1E2 or BVDV-2 E1E2 were generated by homologous recombination. Recombinant viruses were purified and characterized for antigen expression, genetic stability, and growth properties in vitro. Immunogenicity was evaluated in a BALB/c mouse model by measuring E2-specific antibody responses, virus-neutralizing antibodies, and antigen-responsive cellular immune responses. Results: Both recombinant MVA constructs showed detectable E2 expression when E1 and E2 were co-expressed, and exhibited growth characteristics comparable to parental MVA with stable maintenance after serial passage. In contrast, recombinant MVA expressing E2 alone did not yield detectable E2 protein under the same experimental conditions. Immunization induced detectable humoral and cellular immune responses, including E2-specific IgG antibodies, virus-neutralizing antibodies, and increased frequencies of antigen-responsive CD8⁺ T cells with a tendency toward a Th1-biased profile. Conclusions: These findings indicate that co-expression of BVDV E1 and E2 in an MVA vector can support detectable antigen expression and induce measurable immune responses in a mouse proof-of-concept model. Further studies in cattle, including challenge experiments, will be required to determine the protective efficacy and practical applicability of this platform for BVDV vaccine development. Full article

Naked oat (Avena nuda L.) is an important dual-purpose crop for grain and forage in cold regions; however, its high fatty acid content renders seeds prone to deterioration during storage. This study aimed to investigate the protective effects of zinc oxide nanoparticles (ZnO NPs) on artificially aged naked oat seeds and elucidate the underlying molecular mechanisms. Non-aged seeds (Naged) were subjected to artificial aging at 45 °C and 100% relative humidity for 24 h (Aged), followed by priming with 30 mg L⁻¹ ZnO NPs for 6 h (Daged). Antioxidant enzyme activities were determined spectrophotometrically, and transcriptome sequencing was performed on an Illumina platform to identify differentially expressed genes (DEGs) and enriched pathways. We found that ZnO NPs increased catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD) activities by 3–4-fold, restored germination rate from 75% to 98%, and enhanced seed vigor index. A total of 21,403 DEGs were detected, with 15,841 stably expressed in response to nano-priming. Reactive oxygen species (ROS) burst rapidly induced up-regulation of AP2/EREBP transcription factor family members, which subsequently activated antioxidant enzyme genes to maintain cellular redox homeostasis. Metabolic pathway analysis demonstrated that the phenylpropanoid pathway was reprogrammed, characterized by down-regulated lignin biosynthesis and up-regulated flavonoid production, thereby enhancing ROS scavenging capacity. Additionally, the pentose phosphate pathway was activated to provide additional NADPH for antioxidant defense, and up-regulated ADP-glucose pyrophosphorylase (AGPase) facilitated starch accumulation. Notably, the 40S ribosomal protein S13 exhibited the highest connectivity in protein–protein interaction networks, was up-regulated 2.1-fold, and was enriched in post-translational modification processes. These findings suggest that nano-priming with ZnO NPs represents a promising biotechnological strategy for enhancing seed vigor and storability in naked oat, with potential applications in sustainable agriculture and the seed industry. Full article

Ultraviolet (UV) irradiation induces complex skin damage, including hyperpigmentation, oxidative stress, and alterations in proteins related to keratinocyte differentiation and epidermal barrier-associated status. This study investigated the multifunctional protective effects of Padina arborescens ethanolic extract (PAEE) against skin damage in melanocytes, keratinocytes, and zebrafish. In alpha-melanocyte-stimulating hormone (α-MSH)-stimulated B16F10 cells, PAEE effectively suppressed the protein kinase A (PKA)/cyclic adenosine monophosphate (cAMP) response element-binding protein (CREB) signaling pathway, which was associated with reduced expression of microphthalmia-associated transcription factor (MITF) and tyrosinase, leading to decreased melanin synthesis. PAEE also exhibited photoprotective properties by reducing reactive oxygen species (ROS), inhibiting interleukin-1 beta (IL-1β), and attenuating matrix metalloproteinase-1 (MMP-1) upregulation associated with UVB (ultraviolet B)-induced photodamage in HaCaT keratinocytes. Notably, PAEE restored the UVB-reduced expression of filaggrin and involucrin, representative markers of keratinocyte differentiation and epidermal barrier-associated status, in HaCaT keratinocytes. In zebrafish embryos, PAEE suppressed α-MSH-induced melanin accumulation and UVB-induced ROS generation at non-toxic concentrations. Taken together, these results suggest that PAEE exerts anti-melanogenic and photoprotective effects in cellular and zebrasfish models and may serve as a promising marine-derived ingredient for cosmeceutical applications targeting UVB-related skin damage. Full article

Cardiovascular disease continues to impose a substantial global health burden and arises from interconnected pathological processes, including oxidative injury, inflammatory signaling, endothelial dysfunction, metabolic imbalance, and progressive cardiac and vascular structural remodeling. Growing interest has therefore emerged in naturally derived compounds capable of influencing multiple disease pathways simultaneously. Pterostilbene, a dimethoxylated stilbene structurally related to resveratrol, has gained attention due to its enhanced lipophilicity and improved bioavailability. Recent experimental studies have investigated the cardiovascular effects of pterostilbene in both cellular systems and animal models. Evidence from in vitro studies indicates that this compound modulates key regulatory networks involved in cellular energy metabolism, redox homeostasis, endothelial signaling, and stress-associated cardiomyocyte injury. These actions involve pathways linked to 5′ adenosine monophosphate-activated protein kinase and sirtuin-1 signaling, nitric oxide regulation, antioxidant responses, and ferroptosis-related mechanisms. Findings from in vivo investigations further demonstrate protective effects across multiple cardiovascular disease models, including pulmonary hypertension, pressure overload-associated cardiac remodeling, ischemic myocardial injury, toxin-induced cardiotoxicity, and metabolic or atherosclerotic vascular dysfunction. Improvements in functional, structural, and biochemical parameters have been reported in these experimental settings. Overall, current preclinical evidence suggests that pterostilbene may act as a multifunctional modulator of key processes involved in cardiovascular pathology. Although clinical evidence remains limited, the convergence of mechanistic and experimental findings highlights its potential as a multi-target cardiometabolic therapeutic candidate and provides a foundation for future translational and clinical investigation. Full article

The effective treatment of neurodegenerative diseases (NDDs), such as Alzheimer’s disease, Parkinson’s disease, and amyotrophic lateral sclerosis, remains a critical challenge in modern medicine. Given the limitations of current therapies, alternative strategies to slow neurodegeneration are urgently needed. This study presents a critical review of the current evidence regarding low-dose ionizing radiation (IR) as a promising modality for modulating neurodegenerative processes. This study examines current experimental data on the effects of low-dose IR (LDIR) on cellular protective and compensatory mechanisms, including evidence from in vivo models of NDDs. Our analysis demonstrates that LDIR enhances antioxidant activity and DNA repair, stimulates autophagy and neuroplasticity, and modulates neuroinflammatory signaling. Collectively, these findings support the hypothesis of the neuroprotective potential of LDIR, underscoring its translational viability provided that strict dosimetric guidelines are followed and individual biological responses are rigorously monitored. Full article

There is an increasing interest in functional food formulations with added bioactive compounds, such as vitamins, probiotics, polyphenols and bioactive peptides, specifically in dairy and plant-based foods, bakery, and beverages. However, their stability in the food system, release rates and biological activity after consumption/digestion play an important role in the effectiveness of functional foods. There are technical challenges in maintaining the stability and acceptability of added compounds in the formulation design of food items. A novel approach to delivering bioactive compounds in functional foods is their microencapsulation, where stability-sensitive compounds are protected against their degradation during processing and physiological digestion, with targeted release in the gastrointestinal tract (GIT) and elicited cellular responses. Microencapsulation of bioactive compounds has been proven to be beneficial in in vitro models for the stability, antioxidant and immunomodulatory action, and acceptability compared to free (non-capsulated) forms. This technology is worth considering relative to the protection of health benefits of compounds used in food products, with their necessary bioactivity after physiological digestion in GIT. This article reviews important bioactive compounds, challenges, and strategies in the development of functional foods to ensure the required stability for the bioavailability of added compounds. Full article

Oocyte competency is a crucial determinant of fertilisation success and the initial development of embryos in assisted reproductive technologies. The metabolic and biochemical environment of the ovarian follicle is crucial for determining oocyte developmental potential, alongside genetic integrity. The follicular microenvironment includes a complex network of signalling chemicals that regulate mitochondrial activity, steroidogenesis, oxidative balance, and cellular energy metabolism. Recently, metabolic hormones originating from adipose tissue and skeletal muscle, namely, adipokines and myokines, have received considerable focus as crucial regulators of ovarian physiology. Adiponectin, irisin, and the recently identified hormone asprosin have emerged as crucial metabolic regulators influencing granulosa cell activity, mitochondrial bioenergetics, insulin signalling pathways, and redox homeostasis inside the follicular niche. Adiponectin mostly provides metabolic protection by activating AMP-activated protein kinase (AMPK) and improving insulin sensitivity, which in turn enhances mitochondrial efficiency and steroidogenic function in granulosa cells. Irisin, derived from the breakdown of fibronectin type III domain-containing protein 5 (FNDC5), aids the developing oocyte by facilitating mitochondrial biogenesis, augmenting oxidative phosphorylation, and altering cellular defence mechanisms against oxidative stress. Conversely, asprosin has been associated with glucogenic signalling, metabolic stress, and probable mitochondrial malfunction, suggesting a possible relationship between systemic metabolic problems and negative reproductive consequences. Clinical and experimental research indicate that the levels of these metabolic regulators in follicular fluid may correlate with ovarian response, oocyte quality, fertilisation rates, and embryo development during in vitro fertilisation cycles. This review consolidates current molecular, cellular, and clinical information, clarifying the pathways by which adipokines and myokines influence follicular metabolism and impact oocyte competency. Understanding the metabolic connections between systemic endocrine signals and the follicular milieu may provide novel indicators for reproductive prognosis and provide new treatment targets to improve assisted reproduction outcomes. Full article

Background: Effective anti-aging requires dual strategies to stimulate regeneration and counteract damage. While the combination of hydroxypinacolone retinoate (HPR) and carnosine (CA) holds great promise, their effectiveness is hampered by instability and poor skin penetration. Methods: To overcome these challenges, this study developed HPR and CA co-encapsulated nanoliposomes (HC-NLPs) via high-pressure homogenization as an advanced epidermal/dermal delivery system. Results: HC-NLPs markedly improved skin retention of HPR (58.97%) and CA (111.36%) compared to the free combination (Free-HC). In cellular studies, HC-NLPs displayed excellent biocompatibility and demonstrated a 4.7-fold higher cellular uptake. This led to enhanced proliferative (EdU positive rate increased by 78.32%) and migratory (wound closure improved by 31.5%) capacities. Moreover, HC-NLPs effectively reinforced multiple skin-protective processes associated with aging, including enhanced resistance to oxidative and glycation-induced damage, suppressed inflammatory responses, and strengthened cellular barrier integrity. In 3D skin models, HC-NLPs promoted collagen deposition and improved tissue morphology compared to Free-HC. Their superior in vivo antioxidant and anti-aging effects were further validated in Zebrafish assays. HC-NLPs effectively co-deliver HPR and CA, markedly improving their stability, skin penetration, and cellular internalization. Conclusions: The formulation demonstrates comprehensive pro-regenerative, anti-inflammatory, antioxidative, and anti-glycation effects, representing a promising nano-delivery strategy for advanced anti-aging skincare. Full article