Search

UBE2N, a Lys63 ubiquitin-conjugating enzyme, plays critical roles in embryogenesis and immune system development and function. However, its roles in adult epithelial tissue homeostasis and pathogenesis are unclear. We generated conditional mouse models that deleted Ube2n in skin cells in a temporally and spatially controlled manner. We found that Ube2n knockout in the adult skin keratinocytes induced a range of inflammatory skin defects characteristic of psoriatic and actinic keratosis. These included inflammation, epidermal and dermal thickening, parakeratosis, and increased immune cell infiltration as well as signs of edema and blistering. Single-cell transcriptomic analyses and RT-qPCR showed that Ube2n-knockout keratinocytes expressed elevated myeloid cell chemoattractants such as Cxcl1 and Cxcl2 and decreased the homeostatic T lymphocyte chemoattractant Ccl27a. Consistently, the infiltrating immune cells were predominantly myeloid-derived cells, including neutrophils and M1-like macrophages, which expressed high levels of inflammatory cytokines such as Il1β and Il24. Pharmacological blockade of the IL-1 receptor associated kinases (IRAK1/4) alleviated inflammation, epidermal and dermal thickening, and immune infiltration of the Ube2n-mutant skin. Together, these findings highlight a key role of keratinocyte UBE2N in maintenance of epidermal homeostasis and skin immunity and identify IRAK1/4 as potential therapeutic target for inflammatory skin disorders., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: This study focused on the valorization of prickly pear (PP) fruit (Opuntia ficus-indica) into vinegar by spontaneous surface fermentation on different starting matrices (with/without the addition of sucrose and with/without PP peel in the raw material). Different parameters were monitored during the fermentation process in terms of their physicochemical and biological properties., Results: Physicochemical and phytochemical analysis revealed significant differences depending on the starting matrix. An increase in total phenolic content (TPC) was observed for the majority of samples when transformed from PP juice into PP vinegar revealing the role of fermentation in enhancing the bioactive compounds content. Better antioxidant and antibacterial activity were detected for vinegar samples compared with the initial starting matrix. Using whole PP fruit resulted in better TPC and antioxidant activity; in contrast, sugar addition had no significant effect on any studied data. Analysis of variance, taking into account the four factors that were studied (matrix, variety, with/without peel, and with/without sugar), demonstrated that only the factor 'presence or absence of the peel' had a significant influence on the TPC values., Conclusion: This study demonstrated that both whole PP fruit and PP juice could be used as new raw materials for vinegar production. © 2023 Society of Chemical Industry., (© 2023 Society of Chemical Industry.)

Incidence of various cancers including melanoma continues to rise worldwide. While treatment options have expanded in the recent years, the benefit of these treatments suffer from short period of duration for many patients. Hence, new treatment options are highly desired. Here, we propose a method combining a Dextran/reactive-copolymer/AgNPs nanocomposite and a harmless visible light approach to obtain a plasma substitute carbohydrate-based nanoproduct (D@AgNP) that shows strong antitumor activity. Light-driven polysaccharide-based nanocomposite provided essential conditions for extra small (8-12nm) AgNPs capping with subsequent specific self-assembly into spherical-like cloud nanostructures. Obtained biocompatible D@AgNP are stable over six months at room temperature and demonstrated absorbance peak at 406 nm. New formulated nanoproduct revealed efficient anticancer properties against A375 with IC50 0.0035 mg/mL following 24-h incubation; complete cell death is achieved at 0.001 mg/mL and 0.0005 mg/mL by 24- and 48-h time points, respectively. SEM examination shows that D@AgNP altered the shape of the cell structure and damaged the cell membrane. TEM finding shows that D@AgNP are mostly localized at vesicles such as the endosomes, lysosomes and mitochondria. It is anticipated that the introduced new method serves as the cornerstone for improving the generation of biocompatible hydrophilic carbohydrate-based anticancer drugs., Competing Interests: Declaration of competing interest Dr. U.Bunyatova's work has been funded by The Scientific and Technological Research Council of Turkey (TÜBİTAK). She has received the International Post Doctoral Research Fellowship through the BİDEB-2219 Program. Dr.U.B. express gratefulness to the Professor Natalia M. Litchinitser, Department of Electrical and Computer Engineering at Duke University, for providing the facilities and opportunity to execute studies. Prof. NML, Prof. JYZ, and Dr.MBH declare no potential conflict of interest (both financial and non-financial), (Copyright © 2023 Elsevier B.V. All rights reserved.)

To combat emerging antimicrobial-resistant microbes, there is an urgent need to develop new antimicrobials with better therapeutic profiles. For this, a series of 13 new spiropyrrolidine derivatives were designed, synthesized, characterized and evaluated for their in vitro antimicrobial, antioxidant and antidiabetic potential. Antimicrobial results revealed that the designed compounds displayed good activity against clinical isolated strains, with 5d being the most potent (MIC 3.95 mM against Staphylococcus aureus ATCC 25923) compared to tetracycline (MIC 576.01 mM). The antioxidant activity was assessed by trapping DPPH, ABTS and FRAP assays. The results suggest remarkable antioxidant potential of all synthesized compounds, particularly 5c , exhibiting the strongest activity with IC 50 of 3.26 ± 0.32 mM (DPPH), 7.03 ± 0.07 mM (ABTS) and 3.69 ± 0.72 mM (FRAP). Tested for their α -amylase inhibitory effect, the examined analogues display a variable degree of α -amylase activity with IC 50 ranging between 0.55 ± 0.38 mM and 2.19 ± 0.23 mM compared to acarbose (IC 50 1.19 ± 0.02 mM), with the most active compounds being 5d , followed by 5c and 5j , affording IC 50 of 0.55 ± 0.38 mM, 0.92 ± 0.10 mM, and 0.95 ± 0.14 mM, respectively. Preliminary structure-activity relationships revealed the importance of such substituents in enhancing the activity. Furthermore, the ADME screening test was applied to optimize the physicochemical properties and determine their drug-like characteristics. Binding interactions and stability between ligands and active residues of the investigated enzymes were confirmed through molecular docking and dynamic simulation study. These findings provided guidance for further developing leading new spiropyrrolidine scaffolds with improved dual antimicrobial and antidiabetic activities.

The current study is aimed at preparing light-driven novel functional AgNPs- bio-hydrogel and evaluating anticancer potency against human melanoma cells. With an average size of 16-18 nm, the hydrogel nano-silver particle composite (AgNPs@C_MA_O) was synthesized using a soft white LED approach and analyzed by UV-Vis, DLS, FTIR, X-ray, SEM-EDX and TEM techniques. The anticancer activity of the obtained novel functionalized AgNPs@C_MA_O was tested in-vitro in the A375 melanoma cell line. Dose-response analysis showed that AgNPs at 0.01 mg/mL and 0.005 mg/mL doses reduced the viability of A375 cells by 50% at 24 and 48-h time-points, respectively. A375 cells treated with AgNPs@C_MA_O for 24 h at IC50 displayed abnormal morphology such as detachment edges and feet, shrinkage, membrane damage, and the loss of contact with adjacent cells. Our work is the first study showing that non-ionizing radiation mediated biofunctionalized AgNPs have an anti-tumoral effect at such a low concentration of 0.01 mg/mL. Our approach of using harmless wLED increased synergy between soft biopolymer compounds and AgNPs, and enhanced anticancer efficiency of the AgNPs@C_MA_O biohydrogel. Ultimately, the AgNPs accessed through the use of the wLED approach in colloidal syntheses can open new applications and combinatorial advanced cancer treatments and diagnostics.

The c-Jun N-terminal kinases (JNKs), with its members JNK1, JNK2, and JNK3, is a subfamily of (MAPK) mitogen-activated protein kinases. JNK signaling regulates a wide range of cellular processes, including cell proliferation, differentiation, survival, apoptosis, and inflammation. Dysregulation of JNK pathway is associated with a wide range of immune disorders and cancer. Our objective is to provide a review of JNK proteins and their upstream regulators and downstream effector molecules in common skin disorders, including psoriasis, dermal fibrosis, scleroderma, basal cell carcinoma (BCC), squamous cell carcinoma (SCC), and melanoma.

Background: The resistance of melanoma cells to cisplatin restricts its clinical use. Therefore, the search for novel tumor inhibitors and effective combination treatments that sensitize tumor cells to this drug are still needed. We purified macrovipecetin, a novel heterodimeric C-type lectin, from Macrovipera lebetina snake venom and investigated its anti-tumoral effect on its own or combined with cisplatin, in human melanoma cells., Methods: Biochemical characterization, in vitro cells assays such as viability, apoptosis, adhesion, migration, invasion, Western blotting and in silico analysis were used in this study., Results: Macrovipecetin decreased melanoma cell viability 100 times more than cisplatin. Interestingly, when combined with the drug, macrovipecetin enhanced the sensitivity of SK-MEL-28 cells by augmenting their apoptosis through increased expression of the apoptosis inducing factor (AIF) and activation of ERK 1/2 , p38, AKT and NF-κB. Moreover, macrovipecetin alone or combined with cisplatin induced the expression of TRADD, p53, Bax, Bim and Bad and down-regulated the Bcl-2 expression and ROS levels in SK-MEL-28 cells. Interestingly, these treatments impaired SK-MEL-28 cell adhesion, migration and invasion through modulating the function and expression of αvβ3 integrin along with regulating E-cadherin, vimentin, β-catenin, c-Src and RhoA expression. In silico study suggested that only the α chain of macrovipecetin interacts with a region overlapping the RGD motif binding site on this integrin., Conclusions: We validated the antitumor effect of macrovipecetin when combined, or not, with cisplatin on SK-MEL-28 cells., General Significance: The presented work proposes the potential use of macrovipecetin and cisplatin in combination as an effective anti-melanoma treatment., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Melanoma, the most threatening form of skin cancer, has a very poor prognosis and is characterized by its very invasive and chemoresistant properties. Despite the recent promising news from the field of immunotherapy, there is an urgent need for new therapeutic approaches that are free of resistance mechanisms and side effects. Anti-neoplasic properties have been highlighted for different disintegrins from snake venom including Lebein; however, the exact effect of Lebein on melanoma has not yet been defined. In this study, we showed that Lebein blocks melanoma cell proliferation and induces a more differentiated phenotype with inhibition of extracellular signal-regulated kinase (ERK) phosphorylation and microphthalmia-associated transcription factor (MITF) overexpression. Melanoma cells became detached but were less invasive with upregulation of E-cadherin after Lebein exposure. Lebein induced a caspase-independent apoptotic program with apoptosis inducing factor (AIF), BCL-2-associated X protein (BAX) and Bim overexpression together with downregulation of B-cell lymphoma-2 (BCL-2). It generated a distinct response in reactive oxygen species (ROS) generation and p53 levels depending on the p53 cell line status (wild type or mutant). Therefore, we propose Lebein as a new candidate for development of potential therapies for melanoma.

Background: Because oestrogen receptor α (ERα) regulates E2F1 expression to mediate tamoxifen resistance in ERα-positive breast cancer cells, we aimed to define the possible roles of ERα and E2F1 in promoting the resistance of ERα-negative breast cancer cells to 4-hydroxy-tamoxifen (4OHT)., Methods: This study utilised conventional techniques to demonstrate the effects of 4OHT on the expression of ERα and E2F1 and also examined the individual and combined effects of 4OHT with dipyridamole (DIPY) and 3-O-(3,4,5-trimethoxybenzoyl)-(-)-catechin (TMCG) on the oestrogen-negative MDA-MB-231 breast cancer cell line using viability assays, Hoechst staining, MALDI-TOF mass spectroscopy, and confocal microscopy., Results: Despite the ERα-negative status of the MDA-MB-231 cells, we observed that 4OHT efficiently up-regulated ERα in these cells and that this upregulation promoted E2F1-mediated cell growth. Because E2F1 plays a dual role in cell growth/apoptosis, we designed a therapy incorporating TMCG/DIPY to take advantage of the elevated E2F1 expression in these 4OHT-treated cells. 4OHT enhances the toxicity of TMCG/DIPY in these ERα-negative breast cancer cells., Conclusions: Because TMCG/DIPY treatment modulates the methylation status/stability of E2F1, the results demonstrate that therapies targeting the epigenetic machinery of cancer cells in the presence of overexpressed E2F1 may result in efficient E2F1-mediated cell death.

GLU1 encodes the major ferredoxin-dependent glutamate synthase (Fd-GOGAT, EC 1.4.7.1) in Arabidopsis thaliana (ecotype Columbia). With the aim of providing clues on the role of Fd-GOGAT, we analyzed the expression of Fd-GOGAT in tobacco (Nicotiana tabacum L. cv. Xanthi). The 5' flanking element of GLU1 directed the expression of the uidA reporter gene in the palisade and spongy parenchyma of mesophyll, in the phloem cells of vascular tissue and in the roots of tobacco. White light, red light or sucrose induced GUS expression in the dark-grown seedlings in a pattern similar to the GLU1 mRNA accumulation in Arabidopsis. The levels of GLU2 mRNA encoding the second Fd-GOGAT and NADH-glutamate synthase (NADH-GOGAT, EC 1.4.1.14) were not affected by light. Both in the light and in darkness, (15)NH4(+) was incorporated into [5-(15)N]glutamine and [2-(15)N]glutamate by glutamine synthetase (GS, EC 6.3.1.2) and Fd-GOGAT in leaf disks of transgenic tobacco expressing antisense Fd-GOGAT mRNA and in wild-type tobacco. In the light, low level of Fd-glutamate synthase limited the [2-(15)N]glutamate synthesis in transgenic leaf disks. The efficient dark labeling of [2-(15)N]glutamate in the antisense transgenic tobacco leaves indicates that the remaining Fd-GOGAT (15-20% of the wild-type activity) was not the main limiting factor in the dark ammonium assimilation. The antisense tobacco under high CO2 contained glutamine, glutamate, asparagine and aspartate as the bulk of the nitrogen carriers in leaves (62.5%), roots (69.9%) and phloem exudates (53.2%). The levels of glutamate, asparagine and aspartate in the transgenic phloem exudates were similar to the wild-type levels while the glutamine level increased. The proportion of these amino acids remained unchanged in the roots of the transgenic plants. Expression of GLU1 in mesophyll cells implies that Fd-GOGAT assimilates photorespiratory and primary ammonium. GLU1 expression in vascular cells indicates that Fd-GOGAT provides amino acids for nitrogen translocation.