Search Results (715)

Background/Objectives: High-grade serous carcinoma (HGSC) is the leading cause of ovarian cancer-related mortality. It usually arises from fallopian tube epithelium, with a smaller subset arising in non-tubal sites including the ovary or peritoneum. The origin of HGSCs without evidence of tubal involvement remains unclear. Moreover, in women with genetic predisposition to developing HGSC, the additional protection afforded by prophylactic removal of the ovaries in addition to the fallopian tubes has not yet been established. Methods: We used a well-characterized genetically engineered mouse model (GEMM) of oviductal HGSC based on conditional, somatic inactivation of the Brca1, Trp53, Rb1, and Nf1 tumor suppressor genes (BPRN mice) to compare preventive effects for HGSC via bilateral salpingectomy versus bilateral salpingo-oophorectomy. We also explored the origins of non-tubal HGSCs in ectopic tubal-type epithelium (endosalpingiosis) present in the mouse ovaries and peritoneum. Results: While bilateral salpingectomy significantly reduced the incidence of HGSCs in the GEMM model, bilateral salpingo-oophorectomy completely prevented tumor development. We identified an example of HGSC with apparent origin in endosalpingiosis, implicating endosalpingiosis as a likely precursor for non-tubal HGSC. Conclusions: Our findings confirm the superiority of bilateral salpingo-oophorectomy over salpingectomy alone in reducing HGSC risk and affirm the rationale for surgical strategies to reduce HGSC risk in women carrying pathogenic variants of BRCA1/2 and other genes associated with homologous recombination deficiency. Our findings also illustrate how work with GEMMs can advance new insights into HGSC pathogenesis. Full article

Background: Age-related macular degeneration (AMD) is a major cause of irreversible vision loss in the developed world, and the approved products for geographic atrophy (GA), a late-stage form of dry AMD, have shown limited efficacy and require frequent administration. Therefore, longer-lasting therapies with improved efficacy would be a welcome addition to AMD treatment. One potential therapeutic is ONL1204, a small peptide inhibitor of the Fas receptor that has prevented cell death and inflammation in retinal disease models. This study characterizes the pharmacokinetics (PK) and durability of protection conferred by ONL1204. Methods: Ocular pharmacokinetic profiles were generated over 3 months in rabbit and minipig following a single intravitreal (IVT) injection of ONL1204 at multiple doses. Ocular pharmacodynamics were evaluated in two models: a rabbit model using a single IVT injection of ONL1204 with a delayed sodium iodate challenge coupled with fluorescein angiography to quantify RPE loss, and a chronic mouse model that reflects key features of dry AMD disease pathology to assess the efficacy of repeat IVT administrations of ONL1204. Results: ONL1204 had prolonged residence in the ocular tissues of rabbit and minipig, with a vitreous humor half-life of over 100 days. ONL1204 demonstrated significant protection of the retinal pigment epithelium (RPE) in the rabbit sodium iodate model. In the chronic mouse model, two administrations of ONL1204 preserved RPE morphology, reduced caspase-8 activity, and decreased inflammation. Conclusions: These data represent key characteristics of ONL1204, highlighting its clinical potential as a therapeutic for chronic retinal diseases, including GA. Full article

Background/Objectives: Nutrigenomics explores how dietary components influence genome function, especially via epigenetic mechanisms like DNA methylation. A key challenge is identifying healthy food-derived molecules capable of counteracting epigenetic damage from harmful dietary elements. Pistachio nuts (Pistacia vera L.), particularly the Bronte variety from Sicily, are rich in antioxidant polyphenols. In this study we used a methylomic approach to assess the nutrigenomic potential of a hydrophilic extract from Bronte pistachio (BPHE) in a model of human intestinal epithelium, as well as its capacity to modulate arsenic (As)-induced epigenotoxicity. Methods: BPHE was obtained via ethanol/water Soxhlet extraction. CaCo-2 cells were treated with BPHE, alone and after exposure to sodium arsenite. The methylation pattern of the genomic DNA was assessed by methylation-sensitive arbitrarily primed PCR and the methylomic signature was defined by Next-generation bisulfite sequencing. Results: BPHE alone did not alter DNA methylation pattern but, at the highest dose, modulated the changes induced by As. The identification of differentially methylated gene promoters in cell treatment vs. untreated controls revealed that BPHE and As primarily induced hyper-methylation, with a synergistic effect when combined. In particular, all the treatments increased methylation levels of gene categories such as pseudogenes, key genes of specific pathways, genes for zinc-finger proteins, homeobox proteins, kinases, antisense RNA, and miRNA. Notably, in co-treatment with As, BPHE promoted hypo-methylation of genes involved in tumor suppression, detoxification, mitochondrial function, and cell division. Conclusions: These findings suggest that Bronte pistachio polyphenols may epigenetically steer gene expression toward a protective profile, reducing risks of genomic instability and disease. This supports their potential as nutraceuticals to counter harmful epigenetic effects of toxic food components like arsenic. Full article

Colorectal cancer (CRC) remains the second leading cause of cancer-related mortality worldwide, with aberrant activation of the Wnt/β-catenin signaling pathway constituting a key driver of tumorigenesis. SALL2, a zinc finger transcription factor deregulated in various cancers, has been implicated in Wnt signaling regulation through its Xenopus ortholog; however, its role in human CRC remains unclear. In this study, we investigated the expression and function of SALL2 in CRC. Immunohistochemical analysis revealed that SALL2 is present in the epithelium and stroma of normal colon tissue but is significantly downregulated in adenomas, carcinomas, and CRC cell lines. Reduced SALL2 expression was associated with elevated levels of active β-catenin and poorer overall patient survival. Functional assays demonstrated that SALL2 transcriptionally activates AXIN2, a key negative regulator of the Wnt/β-catenin pathway. Chromatin immunoprecipitation and promoter-reporter assays confirmed SALL2 binding to the AXIN2 proximal promoter and enhanced promoter activity. Furthermore, SALL2 expression potentiated the pro-apoptotic effects of the Wnt pathway inhibitor XAV939 in CRC cells, suggesting a role in sensitizing cells to Wnt-targeted therapies. Collectively, these findings identify SALL2 as a negative regulator of Wnt/β-catenin signaling and support its potential as a prognostic biomarker and therapeutic target in colorectal cancer. Full article

The objective of the present study was to develop injectable solutions of curcumin (CUR) and resveratrol (RES) for intravenous administration as a strategy to increase their solubility and stability, as well as to evaluate their cytotoxic potential, individually and in combination, on human lung non-small adenocarcinoma cells (A549 cells) and non-tumoral cells isolated from normal human bronchial epithelium (BEAS cells) to establish possible synergistic effects and potential therapeutic alternatives for lung cancer. Using factorial experimental designs, the components of the injectable CUR and RES solutions were selected, and their hemolytic potential was evaluated by a static method. In addition, combinations of injectable CUR:RES solutions (25:75, 50:50 and 75:25) were prepared from the individual ones, and their stability under refrigeration conditions and cytotoxic potential on A549 and BEAS cells were evaluated. The stability of the injectable solutions of CUR, RES and their different combinations was maintained for 3 months, except for the 25:75 combination of CUR:RES. Furthermore, the cytotoxic potential of CUR and RES on tumoral cells (A549) and non-tumoral (BEAS) cells was evaluated, indicating a dose-dependent effect; the combination of CUR:RES 50:50 and the combination of CUR:RES 75:25 presented synergistic effects in reducing cell viability. This study suggests that injectable solutions of CUR, RES and their combination for intravenous administration could be potential viable candidates and should be evaluated for their efficacy in animal models of lung cancer to establish new possible treatments. Full article

Background/Objectives: Periodontitis is a chronic infectious inflammatory disorder that affects the supporting structures of the teeth. The gingival epithelium plays a crucial role as a physical and immunological barrier, producing pro-inflammatory cytokines in response to microbial pathogens. Modulation of gingival epithelial function has been proposed as a therapeutic strategy to prevent the progression of periodontal disease. Houttuynia cordata, a perennial herb traditionally used in Asian medicine, is recognized for its anti-inflammatory properties, with documented benefits in the cardiovascular, respiratory, and gastrointestinal systems. However, its potential therapeutic role in oral pathologies, such as periodontitis, remains underexplored. This study aimed to investigate the anti-inflammatory effects of H. cordata extract on interleukin (IL)-1β-stimulated primary gingival keratinocytes (PGKs) subjected to IL-1β-induced inflammatory stress, simulating the conditions encountered during orthodontic treatment. Methods: Inflammation was induced in PGKs using IL-1β, and the impact of H. cordata extract pretreatment was assessed using quantitative real-time reverse transcription polymerase chain reaction, enzyme-linked immunosorbent assay, and immunoblotting. Results: H. cordata extract significantly downregulated the mRNA and protein expression levels of tumor necrosis factor-alpha, IL-8, and intercellular adhesion molecule-1 in IL-1β-stimulated PGKs without inducing cytotoxicity. Conclusions: These findings suggest that H. cordata holds promise as a preventive agent against periodontitis by attenuating inflammatory responses in gingival epithelial tissues. We believe that our findings will inform the development of prophylactic interventions to reduce periodontitis risk in patients undergoing orthodontic therapy. Full article

The retina is highly sensitive to oxygen and blood supply, and hypoxia plays a key role in retinal diseases such as diabetic retinopathy (DR) and age-related macular degeneration (AMD). Müller glial cells, which are essential for retinal homeostasis, respond to injury and hypoxia with reactive gliosis, characterized by the upregulation of the glial fibrillary acidic protein (GFAP) and vimentin, cellular hypertrophy, and extracellular matrix changes, which can impair retinal function and repair. The retinal pigment epithelium (RPE) supports photoreceptors, forms part of the blood–retinal barrier, and protects against oxidative stress; its dysfunction contributes to retinal degenerative diseases such as AMD, retinitis pigmentosa (RP), and Stargardt disease (SD). Extracellular vesicles (EVs) play a crucial role in intercellular communication, protein homeostasis, and immune modulation, and have emerged as promising diagnostic and therapeutic tools. Understanding the role of extracellular vesicles’ (EVs’) signaling machinery of glial cells and the retinal pigment epithelium (RPE) is critical for developing effective treatments for retinal degeneration. In this study, we investigated the bidirectional EV-mediated crosstalk between RPE and Müller cells under hypoxic conditions and its impact on cellular metabolism and retinal cell integrity. Our findings demonstrate that RPE-derived extracellular vesicles (RPE EVs) induce time-dependent metabolic reprogramming in Müller cells. Short-term exposure (24 h) promotes pathways supporting neurotransmitter cycling, calcium and mineral absorption, and glutamate metabolism, while prolonged exposure (72 h) shifts Müller cell metabolism toward enhanced mitochondrial function and ATP production. Conversely, Müller cell-derived EVs under hypoxia influenced RPE metabolic pathways, enhancing fatty acid metabolism, intracellular vesicular trafficking, and the biosynthesis of mitochondrial co-factors such as ubiquinone. Proteomic analysis revealed significant modulation of key regulatory proteins. In Müller cells, hypoxic RPE-EV exposure led to reduced expression of Dyskerin Pseudouridine Synthase 1 (DKc1), Eukaryotic Translation Termination Factor 1 (ETF1), and Protein Ser/Thr phosphatases (PPP2R1B), suggesting alterations in RNA processing, translational fidelity, and signaling. RPE cells exposed to hypoxic Müller cell EVs exhibited elevated Ribosome-binding protein 1 (RRBP1), RAC1/2, and Guanine Nucleotide-Binding Protein G(i) Subunit Alpha-1 (GNAI1), supporting enhanced endoplasmic reticulum (ER) function and cytoskeletal remodeling. Functional assays also revealed the compromised barrier integrity of the outer blood–retinal barrier (oBRB) under hypoxic co-culture conditions. These results underscore the adaptive but time-sensitive nature of retinal cell communication via EVs in response to hypoxia. Targeting this crosstalk may offer novel therapeutic strategies to preserve retinal structure and function in ischemic retinopathies. Full article

Galectin-3 is a multifunctional protein that is associated with diseases of the chorioretinal interface, in which the retinal pigment epithelium (RPE) plays a central role in disease development and progression. Since galectin-3 can function extracellularly as well as intracellularly via different mechanisms, we developed an immortalized human RPE cell line (ARPE-19) with a knockdown for galectin-3 expression (ARPE-19/LGALS3^+/−) using a sgRNA/Cas9 all-in-one expression vector. By Western blot analysis, a reduced galectin-3 expression of approximately 48 to 60% in heterozygous ARPE-19/LGALS3^+/− cells was observed when compared to native controls. Furthermore, ARPE-19/LGALS3^+/− cells displayed a flattened, elongated phenotype with decreased E-cadherin as well as enhanced N-cadherin and α-smooth muscle actin mRNA expression, indicating an epithelial–mesenchymal transition of the cells. Compared to wildtype controls, ARPE-19/LGALS3^+/− cells had significantly reduced metabolic activity to 86% and a substantially decreased proliferation to 73%. Furthermore, an enhanced cell adhesion and a diminished migration of immortalized galectin-3 knockdown RPE cells was observed compared to native ARPE-19 cells. Finally, by Western blot analysis, reduced pAKT, pERK1/2, and β-catenin signaling were detected in ARPE-19/LGALS3^+/− cells when compared to wildtype controls. In summary, in RPE cells, endogenous galectin-3 appears to be essential for maintaining the epithelial phenotype as well as cell biological functions such as metabolism, proliferation, or migration, effects that might be mediated via a decreased activity of the AKT, ERK1/2, and β-catenin signaling pathways. Full article

The deterioration of uterine calcium transport capacity induced by aging is a common problem for late-laying period hens, causing decline in eggshell quality. This study aimed to investigate the effects and possible regulatory mechanisms of dietary puerarin (PU) on calcium transport and eggshell quality in aged hens. Two hundred eighty-eight Hubbard Efficiency Plus broiler breeder hens (50-week-old) were randomly allocated to three dietary treatments containing 0, 40, or 200 mg/kg puerarin (PU), with 8 replicates of 12 birds each, for an 8-week trial. The results demonstrated that dietary PU ameliorated the eggshell thickness and strength, which in turn reduced the broken egg rate (p < 0.05). Histological analysis showed that PU improved uterus morphology and increased epithelium height in the uterus (p < 0.05). Antioxidative capacity was significantly improved via upregulation of Nrf2, HO-1, and GPX1 mRNA expression in the uterus (p < 0.05), along with enhanced total antioxidant capacity (T-AOC) and glutathione peroxidase (GSH-PX) activity, and decreased levels of the oxidative stress marker malondialdehyde (MDA) (p < 0.05). Meanwhile, PU treatment reduced the apoptotic index of the uterus, followed by a significant decrease in expression of pro-apoptotic genes Caspase3 and BAX and the rate of BAX/BCL-2. Additionally, calcium content in serum and uterus, as well as the activity of Ca²⁺-ATPase in the duodenum and uterus, were increased by dietary PU (p < 0.05). The genes involved in calcium transport including ERα, KCNA1, CABP-28K, and OPN in the uterus were upregulated by PU supplementation (p < 0.05). The 16S rRNA gene sequencing revealed that dietary PU supplementation could reverse the age-related decline in the relative abundance of Bacteroidota within the uterus (p < 0.05). Overall, dietary PU can improve eggshell quality and calcium transport through enhanced antioxidative defenses and mitigation of age-related uterine degeneration. Full article

Background/Objectives: During the repair of a wounded epithelium, keratinocytes become invasive via the epithelial-to-mesenchymal transition (EMT) process. Usually temporary and controlled, EMT persists in a chronically inflamed epithelium and is exacerbated in epithelial dysplasia and dysregulated in invasive carcinomas. Here we investigated the effects that IL-1 beta, IL-6, and IL-8, inflammatory cytokines expressed in specimens from OPMDs and OSCCs, have on NOKs and OSCC cells. Methods: AKT activation and EMT induction were assessed along with cellular invasiveness. Results: IL-1 beta, IL-6, and IL-8 induced EMT in NOKs, ex novo conferring them invasive capacity. The same cytokines exacerbated the constitutive EMT and invasiveness of OSCC cells. Since these phenomena were accompanied by AKT activation, we tested whether they could be influenced by RTV, a long-used anti-HIV drug that was previously found to block the activation of human AKT and exert antitumor effects. We observed that therapeutic amounts of RTV counteract all the above-mentioned tumorigenic activities of ILs. Finally, consistent with the key role that AKT and EMT play in OSCC radio-resistance, RTV increased OSCC cells’ sensitivity to therapeutic doses of ionizing radiation. Conclusions: These preliminary in vitro findings encourage the use of RTV to prevent the malignant evolution of OPMDs, reduce the risk of OSCC metastasis, and improve the outcomes of anti-OSCC radiotherapy. Full article

The retinal pigment epithelium (RPE), a monolayer of pigmented cells, is critical for visual function through its interaction with the neural retina. In healthy eyes, RPE cells exhibit a uniform hexagonal arrangement, but under stress or disease, such as age-related macular degeneration (AMD), dysmorphic traits like cell enlargement and apparent multinucleation emerge. Multinucleation has been hypothesized to result from cellular fusion, a compensatory mechanism to maintain cell-to-cell contact and barrier function, as well as conserve resources in unhealthy tissue. However, traditional two-dimensional (2D) imaging using apical border markers alone may misrepresent multinucleation due to the lack of lateral markers. We present high-resolution confocal images enabling three-dimensional (3D) visualization of apical (ZO-1) and lateral (α-catenin) markers alongside nuclei. In two RPE damage models, we find that seemingly multinucleated cells are often single cells with displaced neighboring nuclei and lateral membranes. This emphasizes the need for 3D analyses to avoid misidentifying multinucleation and underlying fusion mechanisms. Lastly, images from the NaIO₃ oxidative damage model reveal variability in RPE damage, with elongated, dysmorphic cells showing increased ZsGreen reporter protein expression driven by EMT-linked CAG promoter activity, while more regular RPE cells displayed somewhat reduced green signal more typical of epithelial phenotypes. Full article

Gingival inflammation compromises the integrity of the gingival epithelium and the underlying tissues, highlighting the need for adjuvant therapies with immunomodulatory and healing properties. Casearia sylvestris, a medicinal plant known as guaçatonga, is traditionally used to treat inflammatory lesions. This study aimed to investigate the effects of C. sylvestris on the synthesis of pro- and anti-inflammatory, proteolytic, and antioxidant molecules and on wound healing in epithelial cells. A human telomerase-immortalized gingival keratinocyte cell line (TIGKs) was used, and cells were exposed to Escherichia coli lipopolysaccharide (LPS) in the presence and absence of C. sylvestris extract, its diterpene-concentrated fraction, and its clerodane diterpene casearin J for 24 h and 48 h. Gene expression and protein synthesis were analyzed by RT-qPCR and ELISA, respectively. Nitric oxide (NO) and NF-κB activation were analyzed by Griess reaction and immunofluorescence, respectively. Additionally, cell viability was evaluated by alamarBlue^® assay, and an automated scratch assay was used for wound healing. LPS significantly increased the expression of cytokines (TNF-α, IL-1β, IL-6, IL-8, IL-10, IL-17), proteases (MMP-1 and MMP-13), iNOS as well as NO synthesis, and triggered NF-κB nuclear translocation. It also reduced IL-4 expression, cell viability, and cellular wound repopulation. Treatment with C. sylvestris derivatives significantly abrogated all aforementioned LPS-induced effects by 80–100%. Furthermore, even at higher concentrations, C. sylvestris did not affect cell viability, thus proving the safety of its derivatives. C. sylvestris exerts anti-inflammatory, antiproteolytic, and antioxidant effects on gingival keratinocytes, highlighting its potential as a valuable adjunct in the prevention and treatment of periodontal diseases. Full article

Background/Objectives: In rhegmatogenous retinal detachment (RRD), vitreous cortex remnants (VCRs) may contribute to the development and progression of proliferative vitreoretinopathy (PVR). This study aimed to evaluate potential toxicity and trauma secondary to VCRs visualization and removal during pars plana vitrectomy (PPV) for RRD. Methods: Prospective study on patients with primary RRD who underwent PPV. Imaging assessment included widefield OCT (WF-OCT), ultra-WF retinography and fundus autofluorescence (FAF). During PPV, a filtered and diluted triamcinolone acetonide (TA) solution (20 mg/mL) was used to evaluate the presence and extension of VCRs, removed using an extendible diamond-dusted sweeper (EDDS). After six months, retinal and retinal pigment epithelium toxicity and retinal trauma due to VCRs removal were investigated. Results: Retinal reattachment was achieved in 21/21 cases included in the study. No signs of retinal or RPE toxicity were detected and WF-OCT performed in the areas of VCRs removal revealed an intact inner retinal architecture in the majority of eyes, with minor and localized inner retinal indentations in 4 cases. Conclusions: VCRs visualization and removal using TA and EDDS appears to be safe, with no retinal toxicity and very limited and circumscribed mechanical trauma. This approach may contribute to reducing the risk of postoperative PVR. Full article

The objective of this study was to evaluate the effect of tyrosine on behavior and ruminal meta-taxonomic profile of Altay sheep. Twelve rams with the lowest behavioral responses to humans and twelve rams with the highest behavioral responses were sorted into calm and nervous groups, respectively. Following the 2 × 2 factorial design, the rams from each group were equally assigned two treatments of a basal diet and a diet with an extra 4 g of tyrosine for 30 d. The temperament trait, growth performance, rumen fermentation parameters, ruminal epithelium antioxidant capacity, and rumen microbial composition were measured at the end of the feeding experiment. The results showed that tyrosine treatment led to a decreased number of crosses in the arena test in nervous sheep (p < 0.05). The tyrosine treatment led to increased antioxidant markers in the rumen epithelium, such as catalase, total antioxidant capacity, and GSH content in rumen epithelial tissues (p < 0.05) in calm and nervous sheep. The total volatile fatty acids and propionic acid content in the rumen were increased by tyrosine treatment in nervous and calm sheep (p < 0.05). The rumen microbial study revealed that the dominant microbial genera were Cryptobacteroides, Prevotella, Limivicinus, Quinella, UBA1711, RUG740, Sachharofermentans, Limomorpha, Soladiphilus, Flexinia, and others in both the tyrosine treatment and temperament groups. A combined effect of treatment and temperament was seen on the abundance of two microbial genera, UBA1711 and RUG740 (p < 0.05). These findings suggest that tyrosine-treated Altay sheep would experience less stress during production, resulting in reduced oxidative stress in the rumen epithelium and improved propionic acid production in the rumen compared with that of basal-diet-treated Altay sheep. Meanwhile, the effect of the dietary tyrosine treatment on ruminal microbial diversity was lower, suggesting lesser degradation of tyrosine by ruminal microbes in Altay sheep. Full article

The extracellular matrix (ECM) is a collagen-based scaffold that provides structural support and regulates nutrient transport and cell signaling. ECM homeostasis depends on a dynamic balance between synthesis and degradation, the latter being primarily mediated by matrix metalloproteinases (MMPs). These enzymes are secreted as pro-forms and require activation to degrade ECM components. Their activity is modulated by tissue inhibitors of metalloproteinases (TIMPs). Aging disrupts this balance, leading to the accumulation of oxidized, cross-linked, and denatured matrix proteins, thereby impairing ECM function. Bruch’s membrane, a penta-laminated ECM structure in the eye, plays a critical role in supporting photoreceptor and retinal pigment epithelium (RPE) health. Its age-related thickening and decreased permeability are associated with impaired nutrient delivery and waste removal, contributing to the pathogenesis of age-related macular degeneration (AMD). In AMD, MMP dysfunction is characterized by the reduced activation and sequestration of MMPs, which further limits matrix turnover. This narrative review explores the structural and functional changes in Bruch’s membrane with aging, the role of MMPs in ECM degradation, and the relevance of these processes to AMD pathophysiology, highlighting emerging regulatory mechanisms and potential therapeutic targets. Full article