Enniatin B (ENN B) and Beauvericin (BEA) are cyclohexadepsipeptides that can be isolated from Fusarium and Beauveria bassiana, respectively. Both compounds are cytotoxic and ionophoric. In the present study, the mechanism of cell death induced by these compounds was investigated. Epidermal carcinoma-derived cell line KB-3-1 cells were treated with different concentrations of these compounds. The extracellular secretion of cathepsin B increased in a concentration-dependent manner, and the lysosomal staining by lysotracker red was reduced upon the treatment with any of the compounds. However, the extracellular secretion of cathepsin L and cathepsin D were not affected. Inhibition of cathepsin B with specific inhibitor CA074 significantly reduced the cytotoxic effect of both compounds, while inhibition of cathepsin D or cathepsin L did not influence the cytotoxic activities of both compounds. In vitro labelling of lysosomal cysteine cathepsins with Ethyl (2S, 3S)-epoxysuccinate-Leu-Tyr-Acp-Lys (Biotin)-NH2 (DCG04) was not affected in case of cathepsin L upon the treatment with both compounds, while it was significantly reduced in case of cathepsin B. In conclusion, ENN B and BEA increase lysosomal Ph, which inhibits delivery of cathepsin B from Golgi to lysosomes, thereby inducing cathepsin B release in cytosol, which activates caspases and hence the apoptotic pathway.

Introduction: Clinical isolates of Pseudomonas aeruginosa (P. aeruginosa) are among the most recovered bacteria with phenotypic antimicrobial resistance. Bimetallic nanoparticles (BNPs) have received much attention for antimicrobial activity in the last decade. This research aimed to biosynthesize bimetallic copper oxide-selenium nanoparticles (CuO-Se BNPs) and to assess its bioactivity on various P. aeruginosa clinical isolates. Methodology: Based on the possible synergistic effects, CuO-Se BNPs were selected and biosynthesized using leaf extract of Lagenaria siceraria (L. siceraria) for the first time. The obtained BNPs were characterized using UV-vis spectroscopy, X-ray diffraction (XRD), energy dispersive X-ray spectroscopy (EDX), and transmission and scanning electron microscopes. The capability of Cu-Se BNPs to cease the growth of P. aeruginosa isolates and to reduce their virulence characters was evaluated. Also, different cell lines were used to assess its cytotoxicity and anticancer activity. Results: The elemental composition of CuO and Se was revealed by the UV, XRD, and EDX data, indicating the synthesis of CuO-Se core shell BNPs with a size of 50 nm. In well diffusion assay, CuO-Se BNPs P. aeruginosa growth with 10– 21 mm inhibition zone diameter and 38– 95% inhibition. Also, the minimum inhibitory concentration and minimum bactericidal concentration were in a relatively wide range of 7.8– 250 μg/mL and 31.2– 500 μg/mL, respectively, with tolerance level range of 2– 16. Additionally, CuO-Se BNPs shown anti-pyocyanin activity of 4.35– 63.21% inhibition while the anti-proteolytic activity was in a range of 4.96– 12.59% and anti-pyoverdine effect was in a range of 0.24– 83.41%. The IC50 against Wi-38 normal cells was 267.2 μg/mL while the IC50 were 31.1 and 83.4 μg/mL against MCF-7 and Hep-G2, respectively, indicating promising anticancer activity. Conclusion: This research demonstrates the promising antibacterial, anti-virulence, and antitumor properties with safe low concentrations of CuO-Se NPs, synthesized via an eco-friendly green synthesis method without the use of toxic chemicals, offering a sustainable and cost-effective alternative.

Introduction: Skin injuries represent a prevalent form of physical trauma, necessitating effective therapeutic strategies to expedite the wound healing process. Hesperidin, a bioflavonoid naturally occurring in citrus fruits, exhibits a range of pharmacological attributes, including antimicrobial, antioxidant, anti-inflammatory, anticoagulant, and analgesic properties. The main objective of the study was to formulate a hydrogel with the intention of addressing skin conditions, particularly wound healing.Methods: This research introduces a methodology for the fabrication of a membrane composed of a Polyvinyl alcohol - Sodium Alginate (PVA/A) blend, along with the inclusion of an anti-inflammatory agent, Hesperidin (H), which exhibits promising wound healing capabilities. A uniform layer of a homogeneous solution comprising PVA/A was cast. The process of crosslinking and the enhancement of hydrogel characteristics were achieved through the application of gamma irradiation at a dosage of 30 kGy. The membrane was immersed in a Hesperidin (H) solution, facilitating the permeation and absorption of the drug. The resultant system is designed to deliver H in a controlled and sustained manner, which is crucial for promoting efficient wound healing. The obtained PVA/AH hydrogel was evaluated for cytotoxicity, antioxidant and free radical scavenging activities, anti-inflammatory and membrane stability effect. In addition, its action on oxidative stress, and inflammatory markers was evaluated on BJ-1 human normal skin cell line.Results and Discussion: We determined the effect of radical scavenging activity PVA/A (49 %) and PVA/AH (87%), the inhibition of Human red blood cell membrane hemolysis by PVA/AH (81.97 and 84.34 %), hypotonicity (83.68 and 76.48 %) and protein denaturation (83.17 and 85.8 %) as compared to 250 μg/ml diclofenac (Dic.) and aspirin (Asp.), respectively. Furthermore, gene expression analysis revealed an increased expression of genes associated with anti-oxidant and anti-inflammatory properties and downregulated TNFα, NFκB, iNOS, and COX2 by 67, 52, 58 and 60%, respectively, by PVA/AH hydrogel compared to LPS-stimulated BJ-1 cells. The advantages associated with Hesperidin can be ascribed to its antioxidant and anti-inflammatory attributes. The incorporation of Hesperidin into hydrogels offers promise for the development of a novel, secure, and efficient strategy for wound healing. This innovative approach holds potential as a solution for wound healing, capitalizing on the collaborative qualities of PVA/AH and gamma irradiation, which can be combined to establish a drug delivery platform for Hesperidin.

Epithelial sodium channel (ENaC) are integral to maintaining salt and water homeostasis in various biological tissues, including the kidney, lung, and colon. They enable the selective reabsorption of sodium ions, which is a process critical for controlling blood pressure, electrolyte balance, and overall fluid volume. ENaC activity is finely controlled through proteolytic activation, a process wherein specific enzymes, or proteases, cleave ENaC subunits, resulting in channel activation and increased sodium reabsorption. This regulatory mechanism plays a pivotal role in adapting sodium transport to different physiological conditions. In this review article, we provide an in-depth exploration of the role of proteolytic activation in regulating ENaC activity. We elucidate the involvement of various proteases, including furin-like convertases, cysteine, and serine proteases, and detail the precise cleavage sites and regulatory mechanisms underlying ENaC activation by these proteases. We also discuss the physiological implications of proteolytic ENaC activation, focusing on its involvement in blood pressure regulation, pulmonary function, and intestinal sodium absorption. Understanding the mechanisms and consequences of ENaC proteolytic activation provides valuable insights into the pathophysiology of various diseases, including hypertension, pulmonary disorders, and various gastrointestinal conditions. Moreover, we discuss the potential therapeutic avenues that emerge from understanding these mechanisms, offering new possibilities for managing diseases associated with ENaC dysfunction. In summary, this review provides a comprehensive discussion of the intricate interplay between proteases and ENaC, emphasizing the significance of proteolytic activation in maintaining sodium and fluid balance in both health and disease.

Working memory is short-term memory, so temporal improvement does not reflect the consolidation of a memory trace, rather the functionality of the underlying neuronal circuits and molecular signaling cascades. The administration of drugs-either one-time or through daily injectioncan elucidate the underlying mechanisms. The T-maze is especially suitable for studying dopamine-dependent working memory, since it is less stressful than other tests, for example, water maze-based paradigms (Bezu et al., 2016 and 2017). Here, we present a training protocol for evaluating the underlying mechanisms that lead to the development of spatial working memory in rats. Our approach uses a T-maze, and it can be used to get high temporal resolution.