Search Results (26)

Background: Pancreatic ductal adenocarcinoma (PDAC) is characterized by its aggressive clinical behavior and intricate microenvironment regulation, leading to dismal prognosis. Elucidating the molecular mechanisms underlying PDAC pathogenesis is crucial for developing improved therapeutic approaches. The functional significance and molecular basis of transcobalamin 1 (TCN1) in PDAC remain largely unexplored. Methods and Results: Through integrated analysis of TCGA and GTEx datasets combined with 80 clinical specimens, we identified significant TCN1 overexpression in PDAC, showing a positive association with tumor stage and negative associations with histological differentiation and overall survival. Functional investigations showed that TCN1 enhanced pancreatic cancer cell proliferation, migration, invasion, and epithelial–mesenchymal transition (EMT) in both in vitro and in vivo models. Mechanistically, TCN1 physically interacts with signal transducer and activator of transcription 4 (STAT4) to enhance its transcriptional activity. Chromatin immunoprecipitation (ChIP) assays showed that STAT4-mediated transcriptional activation of dual oxidase 2 (DUOX2) occurs through direct promoter binding. As a pivotal reactive oxygen species (ROS)-generating enzyme, DUOX2 overexpression elevates intracellular ROS levels, thereby promoting EMT progression and activating proliferation-related signaling cascades. Antioxidant treatment effectively abrogated TCN1-driven oncogenic phenotypes, establishing ROS as the critical downstream mediator. Conclusions: Collectively, our findings reveal a novel TCN1/STAT4/DUOX2 regulatory axis that exacerbates PDAC progression by remodeling redox homeostasis. This signaling cascade may serve as a prognostic biomarker and a potential therapeutic target for ROS-directed precision therapy in PDAC. Full article

Thyroid dyshormonogenesis (THD) is a heterogeneous group of genetic diseases caused by the total or partial defect in the synthesis or secretion of thyroid hormones. Genetic variants in DUOX2 can cause partial to total iodination organification defects and clinical heterogeneity, from transient to permanent congenital hypothyroidism. The aim of this study was to undertake a molecular characterization and genotype–phenotype correlation in patients with THD and candidate variants in DUOX2. A total of 31 (19.38%) patients from the Catalan Neonatal Screening Program presented with variants in DUOX2 that could explain their phenotype. Fifteen (48.39%) patients were compound heterozygous, 10 (32.26%) heterozygous, and 4 (12.90%) homozygous. In addition, 8 (26.67%) of these patients presented variants in other genes. A total of 35 variants were described, 10 (28.57%) of these variants have not been previously reported in literature. The most frequent variant in our cohort was c.2895_2898del/p.(Phe966SerfsTer29), classified as pathogenic according to reported functional studies. The final diagnosis of this cohort was permanent THD in 21 patients and transient THD in 10, according to reevaluation and/or need for treatment with levothyroxine. A clear genotype–phenotype correlation could not be identified; therefore, functional studies are necessary to confirm the pathogenicity of the variants. Full article

Chlorogenic acid (CGA), a polyphenol found mainly in coffee and tea, exerts antioxidant, anti-inflammatory and anti-apoptotic effects at the gastrointestinal level. However, although CGA is known to cross the blood–brain barrier (BBB), its effects on the CNS are still unknown. Oligodendrocytes (OLs), the myelin-forming cells in the CNS, are the main target in demyelinating neuroinflammatory diseases such as multiple sclerosis (MS). We evaluated the antioxidant, anti-inflammatory and anti-apoptotic roles of CGA in M03-13, an immortalized human OL cell line. We found that CGA reduces intracellular superoxide ions, mitochondrial reactive oxygen species (ROS) and NADPH oxidases (NOXs) /dual oxidase 2 (DUOX2) protein levels. The stimulation of M03-13 cells with TNFα activates the nuclear factor kappa-light-chain-enhancer of activated B cell (NF-kB) pathway, leading to an increase in superoxide ion, NOXs/DUOX2 and phosphorylated extracellular regulated protein kinase (pERK) levels. In addition, tumor necrosis factor alpha (TNF-α) stimulation induces caspase 8 activation and the cleavage of poly-ADP-ribose polymerase (PARP). All these TNFα-induced effects are reversed by CGA. Furthermore, CGA induces a blockade of proliferation, driving cells to differentiation, resulting in increased mRNA levels of myelin basic protein (MBP) and proteolipid protein (PLP), which are major markers of mature OLs. Overall, these data suggest that dietary supplementation with this polyphenol could play an important beneficial role in autoimmune neuroinflammatory diseases such as MS. Full article

Recycling and revalorization of waste are currently essential for sustainable growth. Mill scale, a waste product from steel production industries, which contains high levels of iron and minimal impurities, is proposed in this study as the source to synthesize magnetite nanoparticles (Fe₃O₄) for an enhancement of a lactate biosensor range. The synthesized Fe₃O₄ nanoparticles were coated with polydopamine (PDA) to prevent aggregation and degradation, creating a stable platform for immobilizing lactate oxidase enzyme (LOx) on their surfaces. The characterization of the Fe₃O₄@PDA material was carried out using transmission electron microscopy (TEM), dynamic light scattering (DLS), and measurement of the polydispersity index (PdI). The Fe₃O₄@PDA-LOx material was then deposited on a screen-printed carbon electrode modified with Prussian blue (SPCE-PB) for lactate detection. The biosensor exhibited a broad, dual linear concentration-response range, one from 0.1 to 4.62 mM with a limit of detection of 0.32 mM and sensitivity of 1.54 $\mathsf{\mu }\mathrm{A}{\mathrm{m}\mathrm{M}}^{-1}{\mathrm{c}\mathrm{m}}^{-2}$, and another one from 4.62 to 149.21 mM with a limit of detection of 6.31 mM and sensitivity of 0.08 $\mathsf{\mu }\mathrm{A}{\mathrm{m}\mathrm{M}}^{-1}{\mathrm{c}\mathrm{m}}^{-2}$. The dual-range concentration response of the biosensor makes it an ideal tool for lactate determination in various applications, including sports medicine, clinical diagnosis, and industrial bioprocessing. Full article

Reactive oxygen species (ROS) are highly reactive molecules formed from diatomic oxygen. They act as cellular signals, exert antibiotic activity towards invading microorganisms, but can also damage host cells. Dual oxidase 2 (DUOX2) is the main ROS-producing enzyme in the intestine, regulated by cues of the commensal microbiota and functions in pathogen defense. DUOX2 plays multiple roles in different organs and cell types, complicating the functional analysis using systemic deletion models. Here, we interrogate the precise role of epithelial DUOX2 for intestinal homeostasis and host-microbiome interactions. Conditional Duox2^∆IEC mice lacking DUOX2, specifically in intestinal epithelial cells, were generated, and their intestinal mucosal immune phenotype and microbiome were analyzed. Inflammatory susceptibility was evaluated by challenging Duox2^∆IEC mice in the dextran sodium sulfate (DSS) colitis model. DUOX2-microbiome interactions in humans were investigated by paired analyses of mucosal DUOX2 expression and fecal microbiome data in patients with intestinal inflammation. Under unchallenged conditions, we did not observe any obvious phenotype of Duox2^∆IEC mice, although intestinal epithelial ROS production was drastically decreased, and the mucosal microbiome composition was altered. When challenged with DSS, Duox2^∆IEC mice were protected from colitis, possibly by inhibiting ROS-mediated damage and fostering epithelial regenerative responses. Finally, in patients with intestinal inflammation, DUOX2 expression was increased in inflamed tissue, and high DUOX2 levels were linked to a dysbiotic microbiome. Our findings demonstrate that bidirectional DUOX2-microbiome interactions contribute to mucosal homeostasis, and their dysregulation may drive disease development, thus highlighting this axis as a therapeutic target to treat intestinal inflammation. Full article

Intestinal vessels play a critical role in nutrient absorption, whereas the effect and mechanism of low birth weight (LBW) on its formation remain unclear. Here, twenty newborn piglets were assigned to the control (CON) group (1162 ± 98 g) and LBW group (724 ± 31 g) according to their birth weight. Results showed that the villus height and the activity of maltase in the jejunum were lower in the LBW group than in the CON group. LBW group exhibited a higher oxidative stress level and impaired mitochondrial function in the jejunum and was lower than the CON group in the intestinal vascular density. To investigate the role of oxidative stress in intestinal angiogenesis, H₂O₂ was employed to induce oxidative stress in porcine intestinal epithelial cells (IPEC-J2). The results showed that the conditioned media from IPEC-J2 with H₂O₂ treatment decreased the angiogenesis of porcine vascular endothelial cells (PVEC). Transcriptome analysis revealed that a higher expression level of dual oxidase 2 (DUOX2) was found in the intestine of LBW piglets. Knockdown of DUOX2 in IPEC-J2 increased the proliferation and decreased the oxidative stress level. In addition, conditioned media from IPEC-J2 with DUOX2-knockdown was demonstrated to promote the angiogenesis of PVEC. Mechanistically, the knockdown of DUOX2 decreased the reactive oxygen species (ROS) level, thus increasing the angiogenesis in a matrix metalloproteinase 3 (MMP3) dependent manner. Conclusively, our results indicated that DUOX2-induced oxidative stress inhibited intestinal angiogenesis through MMP3 in a LBW piglet model. Full article

We report a 20-year-old, female, adopted Indian patient with over 662 Mb regions of homozy-gosity who presented with intellectual disability, ataxia, schizophrenia, retinal dystrophy, moder-ate-to-severe progressive sensorineural hearing loss (SNHL), congenital hypothyroidism, cleft mi-tral valve with mild mitral valve regurgitation, and dysmorphic features. Exome analysis first on a clinical basis and subsequently on research reanalysis uncovered pathogenic variants in three nu-clear genes following two modes of inheritance that were causal to her complex phenotype. These included (1) compound heterozygous variants in BBS6 potentially causative for Bardet–Biedl syn-drome 6; (2) a homozygous, known pathogenic variant in the stereocilin (STRC) gene associated with nonsyndromic deafness; and (3) a homozygous variant in dual oxidase 2 (DUOX2) gene asso-ciated with congenital hypothyroidism. A variant of uncertain significance was identified in a fourth gene, troponin T2 (TNNT2), associated with cardiomyopathy but not the cleft mitral valve, with mild mitral regurgitation seen in this case. This patient was the product of an apparent first-degree relationship, explaining the multiple independent inherited findings. This case high-lights the need to carefully evaluate multiple independent genetic etiologies for complex pheno-types, particularly in the case of consanguinity, rather than presuming unexplained features are expansions of known gene disorders. Full article

Sulfite determination in foods and alcoholic beverages is a common requirement by food and drug administration organisations in most countries. In this study, the enzyme, sulfite oxidase (SOx), is used to biofunctionalise a platinum-nanoparticle-modified polypyrrole nanowire array (PPyNWA) for the ultrasensitive amperometric detection of sulfite. A dual-step anodisation method was used to prepare the anodic aluminum oxide membrane used as a template for the initial fabrication of the PPyNWA. PtNPs were subsequently deposited on the PPyNWA by potential cycling in a platinum solution. The resulting PPyNWA-PtNP electrode was then biofuntionalised by adsorption of SOx onto the surface. The confirmation of the adsorption of SOx and the presence of PtNPs in the PPyNWA-PtNPs-SOx biosensor was verified by scanning electron microscopy and electron dispersive X-ray spectroscopy. Cyclic voltammetry and amperometric measurements were used to investigate the properties of the nanobiosensor and to optimise its use for sulfite detection. Ultrasensitive detection of sulfite with the PPyNWA-PtNPs-SOx nanobiosensor was accomplished by use of 0.3 M pyrrole, 10 U mL⁻¹ of SOx, adsorption time of 8 h, a polymerisation period of 900 s, and an applied current density of 0.7 mA cm⁻². The response time of the nanobiosensor was 2 s, and its excellent analytical performance was substantiated with a sensitivity of 57.33 μA cm⁻² mM⁻¹, a limit of detection of 12.35 nM, and a linear response range from 0.12 to 1200 μM. Application of the nanobiosensor to sulfite determination in beer and wine samples was achieved with a recovery efficiency of 97–103%. Full article

The cucumber (Cucumis sativa L.) is often subjected to several fungal diseases. Rhizoctonia solani-induced cucumber damping-off and root rot are the most common diseases reported from the commercial greenhouses of the eastern area of Saudi Arabia. The objective of the current study is to explore the antagonistic activity of four Trichoderma species against R. solani in vitro and in vivo. Ten R. solani isolates (eight belonging to AG-4 and two belonging to AG-A and AG2-1) were studied. AG4 isolates were pathogenic to cucumber plants, while AG-A and AG2-1 isolates were non-pathogenic. Seven isolates of Trichoderma spp., named T. hamatum KSATR8, T. harzianum (KSATR9 and KSATR10), T. asperellum (KSATR11, KSATC, and KSAT1E), and T. longibrachiatum KSATS were isolated, and the identities of both R. solani and Trichoderma isolates were confirmed based on the phylogenetic analysis of the DNA sequence of the ITS region. The dual culture findings indicated that T. asperellum KSATC and KSAT1E exhibited the most significant inhibitory activities against R. solani, with values of 79.33 and 70.89%, respectively. Scanning electron microscope (SEM) images showed a considerable degradation in the cell wall and collapsing of R. solani hyphae by all Trichoderma species. Under greenhouse conditions, the application of T. asperellum KSATC and KSAT1E at concentrations of 2 × 10⁸ conidia/mL revealed a reduction in root rot and damping-off incidence percentages with values that did not reveal a significant (p < 0.05) difference from those of Rizolex-T fungicide. Nevertheless, the efficacy of the fungicide attained 86.67%, being higher than that of T. asperellum KSATC, which reached 80%. Trichoderma asperellum KSATC and KSAT1E were the greatest in increasing peroxidase, catalase, and chitinase enzymes activities in cucumber plants. Conversely, a significant (p < 0.05) elevation in polyphenol oxidase enzyme (0.762 and 0.97 U/g FW) and total phenol content (0.55 and 0.62 mg/g FW) was recorded in cucumber plants treated with T. harzianum KSATR9 and KSATR10, respectively. The statistical analysis results displayed no considerable variations among cucumber plants regarding total chlorophyll content as a response to treatments with Trichoderma species and fungicides. Therefore, we endorse using T. asperellum KSATC and KSAT1E as an alternative to fungicides to manage root rot and damping-off in cucumbers. Full article

Metabolic reprogramming is a central hub in tumor development and progression. Therefore, several efforts have been developed to find improved therapeutic approaches targeting cancer cell metabolism. Recently, we identified the 7α-acetoxy-6β-benzoyloxy-12-O-benzoylroyleanone (Roy-Bz) as a PKCδ-selective activator with potent anti-proliferative activity in colon cancer by stimulating a PKCδ-dependent mitochondrial apoptotic pathway. Herein, we investigated whether the antitumor activity of Roy-Bz, in colon cancer, could be related to glucose metabolism interference. The results showed that Roy-Bz decreased the mitochondrial respiration in human colon HCT116 cancer cells, by reducing electron transfer chain complexes I/III. Consistently, this effect was associated with downregulation of the mitochondrial markers cytochrome c oxidase subunit 4 (COX4), voltage-dependent anion channel (VDAC) and mitochondrial import receptor subunit TOM20 homolog (TOM20), and upregulation of synthesis of cytochrome c oxidase 2 (SCO2). Roy-Bz also dropped glycolysis, decreasing the expression of critical glycolytic markers directly implicated in glucose metabolism such as glucose transporter 1 (GLUT1), hexokinase 2 (HK2) and monocarboxylate transporter 4 (MCT4), and increasing TP53-induced glycolysis and apoptosis regulator (TIGAR) protein levels. These results were further corroborated in tumor xenografts of colon cancer. Altogether, using a PKCδ-selective activator, this work evidenced a potential dual role of PKCδ in tumor cell metabolism, resulting from the inhibition of both mitochondrial respiration and glycolysis. Additionally, it reinforces the antitumor therapeutic potential of Roy-Bz in colon cancer by targeting glucose metabolism. Full article

Clostridioides difficile causes the highest number of nosocomial infections. Currently, treatment options for C. difficile infection (CDI) are very limited, resulting in poor treatment outcomes and high recurrence rates. Although the disease caused by CDI is inflammatory in nature, the role of inflammation in the development of CDI symptoms is contradictory and not completely understood. Hence, the use of anti-inflammatory medication is debatable in CDI. In the current study, we evaluated the genetic and microbiome profiles of mice after infection with C. difficile. These mice were categorized based on the severity of CDI and the results were viewed accordingly. Our results indicate that certain genes are upregulated in severe CDI more than in the moderate case. These include oncostatin-M (OSM), matrix metalloprotease 8 (MMP8), triggering receptor expressed on myeloid cells 1 (Trem-1), and dual oxidase 2 (Duox2). We also investigated the microbiome composition of CDI mice before and after infecting with C. difficile. The results show that C. difficile abundance is not indicative of diseases severity. Certain bacterial species (e.g., Citrobacter) were enriched while others (e.g., Turicibacter) were absent in severe CDI. This study identifies novel inflammatory pathways and bacterial species with a potential role in determining the severity of CDI. Full article

High-throughput mass-spectrometry-based quantitative proteomic analysis was performed using formalin-fixed, paraffin-embedded (FFPE) biopsy samples obtained before treatment from 13 patients with locally advanced rectal cancer (LARC), who were treated with concurrent chemoradiation therapy (CCRT) followed by surgery. Patients were divided into complete responder (CR) and non-complete responder (nCR) groups. Immunohistochemical (IHC) staining of 79 independent FFPE tissue samples was performed to validate the predictive ability of proteomic biomarker candidates. A total of 3637 proteins were identified, and the expression of 498 proteins was confirmed at significantly different levels (differentially expressed proteins—DEPs) between two groups. In Gene Ontology enrichment analyses, DEPs enriched in biological processes in the CR group included proteins linked to cytoskeletal organization, immune response processes, and vesicle-associated protein transport processes, whereas DEPs in the nCR group were associated with biosynthesis, transcription, and translation processes. Dual oxidase 2 (DUOX2) was selected as the most predictive biomarker in machine learning algorithm analysis. Further IHC validation ultimately confirmed DUOX2 as a potential biomarker for predicting the response of nCR to CCRT. In conclusion, this study suggests that the treatment response to RT may be affected by the pre-treatment tumor microenvironment. DUOX2 is a potential biomarker for the early prediction of nCR after CCRT. Full article

Nicotinic acid adenine dinucleotide phosphate (NAADP) is a universal Ca²⁺ mobilizing second messenger essential for initiation of Ca²⁺ signaling. Recently, novel molecular mechanisms of both its rapid formation upon receptor stimulation and its mode of action were discovered. Dual NADPH oxidase 2 (DUOX2) and hematological and neurological expressed 1-like protein (HN1L)/Jupiter microtubule-associated homolog 2 (JPT2) were discovered as NAADP-forming enzyme and NAADP receptor/binding protein—the new kids on the block. These novel aspects are reviewed and integrated into the previous view of NAADP signaling. Full article

Parkinson’s disease (PD) is the second most prevalent neurodegenerative disease. It is characterized by a complex network of physiopathological events where oxidative stress plays a central role among other factors such as neuroinflammation and protein homeostasis. Nuclear factor-erythroid 2 p45-related factor 2 (NRF2) has a multitarget profile itself as it controls a plethora of cellular processes involved in the progression of the disease. In this line, we designed a novel family of 2-(1H-indol-3-yl)ethan-1-amine derivatives as NRF2 inducers with complementary activities. Novel compounds are based on melatonin scaffold and include, among other properties, selective monoamine oxidase B (MAO-B) inhibition activity. Novel multitarget compounds exhibited NRF2 induction activity and MAO-B selective inhibition, combined with anti-inflammatory, antioxidant, and blood–brain barrier permeation properties. Furthermore, they exert neuroprotective properties against oxidative stress toxicity in PD-related in vitro. Hit compound 14 reduced oxidative stress markers and exerted neuroprotection in rat striatal slices exposed to 6-hydroxydopamine or rotenone. In conclusion, we developed a promising family of dual NRF2 inducers and selective MAO-B inhibitors that could serve as a novel therapeutic strategy for PD treatment. Full article

Fresh date palm fruits (cv. Barhi) have received much attention due to their sweet taste and popularity in marketing. There is a critical need to prolong their storability, as well as maintain their quality during the postharvest and marketing periods. In this study, the effects of spraying date palm trees with melatonin (Mt) and/or methyl jasmonate (Mj) at 10, 20, and 50 ppm, on their growth and yield were investigated. In addition, impacts on quality and storability of the fruits were also studied. In general, application of Mt was mostly more effective than that of Mj, even at 50 ppm, with regard to all evaluated parameters. However, the dual treatment at 50 ppm recorded the highest relative chlorophyll and nutrient content in date palm leaves, as well as the yield and its components. Regarding the date palm fruits stored at 4 °C for 28 days, this dual treatment recorded the lowest weight loss and fruit decay values (0.14 and 2%, respectively), the highest firmness (6 g·cm⁻²), total soluble solids content (36 °Brix), total sugar content (32.5 g/100 g fresh weight), and the lowest total acidity (0.16 g citric acid/100 mL juice). Moreover, the highest total phenolic content and activity of peroxidase and polyphenol oxidase enzymes in the stored fruits were also recorded for the dual treatment. In contrast to the untreated fruits, scanning electron microscopy observations showed that the sprayed fruits had a very good microstructure, showing intact and thick exocarp tissue with a dense layer of epicuticular wax. The mesocarp tissue showed a normal and clear cellular framework with well organized and arranged cells, after 28 days storage at 4 °C. Based on these results, we can conclude that application of the dual treatment (Mt + Mj) at 50 ppm is a promising way to prolong the storability of date palm fruits and maintain their quality during storage periods. Full article