Streptococcus suis is an important zoonotic pathogen that threatens human and pig health. During infection, the host can impose oxidative stress to resist pathogen invasion. Resistance to oxidative toxicity is an important factor for pathogens. Glutathione synthesis contributes to reactive oxygen species (ROS) detoxification in bacterial cells. Little is known about the roles of glutathione synthesis and transport in S. suis. In this study, we demonstrated that glutathione treatment increased oxidative stress tolerance in S. suis. GshAB and GshT were found in S. suis glutathione synthesis and import by bioinformatics. In vitro, inactivation of gshAB and gshT led to increased sensitivity to oxidative stress. Inactivation of gshT led to growth defects in the medium. The intracellular glutathione content of gshAB or gshT deletion mutants was lower than that of wild type (WT) strain. The phagocytic resistance of gshAB and gshT mutants was lower than that of the WT strain. Moreover, the virulence of gshAB and gshT deletion mutants was significantly lower than that of the WT strain in mouse survival and tissue loading experiments. In conclusion, these results revealed the functions of GshAB and GshT in the pathogenesis of S. suis. These findings enhance our understanding of bacterial virulence mechanisms and may provide a new avenue for therapeutic intervention aimed at curbing S. suis infections.

Glutathione (GSH) is a very important antioxidant and also participates in many important physiological processes, accurate determination of GSH in food and blood fluid is crucial for human health. Herein, we introduce a simple colorimetric and paper-based-smartphone dual signal output platform for point-of-care (POC) detection of GSH in garlic and human serum. The experiment revealed that the peroxidase-like activity of Aluminum-doped Prussian blue (AlPB) could be enhanced by the doping of Al element on Prussian blue. AlPB exhibited excellent peroxidase-like activity and efficiently catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2 to generate blue oxidized TMB, resulting in an absorbance and color-based-smartphone dual signal readouts. Enzyme kinetic studies also indicated that the AlPB showed high affinity towards TMB. Hence, a reliable colorimetric and color-based-smartphone dual signal readouts assay was constructed based on AlPB-mediated the peroxidase-like activity, it was used for highly sensitive colorimetric and POC detection of GSH in food and human serum within 7 min with a wide range of linear response from 1 to 20 μM, a low detection limit of 0.1748 μM. This work demonstrates a novel and versatile strategy to develop superior peroxidase mimics and holds great potential for rapid and portable detection of GSH in food, healthcare and clinical diagnosis, and also open promising avenues for more powerful paper-based POC tests.

Encapsulated in a self-produced negatively charged extracellular polymeric substance (EPS) matrix, the wound infected bacterial biofilms exhibit formidable resistance to conventional positively charged antibiotics and host's immune responses, which can undoubtedly lead to persistent infections and lethal complications. Nevertheless, developing efficacious strategies to root out stubborn biofilm and promote tissue regeneration still remains a challenge. To resolve this dilemma, a versatile vacancies-rich Z-scheme MoSSe Van der Waals heterojunction (MoSSe VdW HJ) is rationally fabricated as nanoplatform for hydrogen sulfide (H2S)-sensitized synergistic therapy of wound bacterial biofilm infection. The rich anion vacancies and Z-scheme heterostructure make the fabricated MoSSe VdW HJ can effectively augment H2S, localized hyperthermia, and reactive oxygen species production under the stimulation of biofilm microenvironments (BME) and irradiation of 808 nm near-infrared (NIR) light. Therefore, MoSSe VdW HJ is capable to integrate H2S gas, chemodynamic, photothermal, and photodynamic therapies to effectively destroy eDNA and polysaccharides in the EPS matrix, thereby breaching the biofilm barrier to eradicate bacteria and facilitate wound healing. The synergistic strategy exhibits superior anti-biofilm and wound repair effects both in vivo and in vitro, thus providing guideline for the development of BME and NIR light activated synergistic therapeutics to fight against refractory biofilm infections.

This study aimed to evaluate the effect of thiolated micelles on the cellular uptake of their payload. Reduced and oxidized glutathione were covalently attached to palmitic acid via amide bond formation. Micelles formed with these thiolated surfactants (MP-GSH and MP-GSSG-P) were evaluated regarding critical micellar concentration (CMC) and hemolytic activity. Cytotoxicity was evaluated on HEK 293 and HeLa cells. Diffusion of micelles in these cells was evaluated by fluorescence correlation spectroscopy (FCS). Furthermore, cellular uptake of micelles containing coumarin-6 as model drug was analyzed by flow cytometry and confocal laser scanning microscopy. CMC of MP-GSSG-P, MP-GSH, and palmitic acid micelles (MPA) was determined to be 0.455 mM, 0.166 mM, and 0.046 mM, respectively. At a concentration of 0.5 % MP-GSSG-P, MP-GSH, and MPA caused around 80 % hemolysis. In HEK cells, MP-GSSG-P exhibited toxicity at a concentration of 0.25 %, while MP-GSH showed toxicity at 0.06 % after 4 hours of incubation. In contrast, HeLa cells were more resilient, with only MP-GSH showing toxicity at 0.25 %. Diffusivity of MP-GSH and MP-GSSG-P within the cells was higher than that of MPA. Cellular uptake studies demonstrated a significantly (p < 0.05) enhanced internalization of MP-GSH and MP-GSSG-P, that was 112-fold and 270-fold higher in HEK 293 cells and 12-fold and 60-fold higher in HeLa cells when compared to MPA. These findings suggest that MP-GSH and MP-GSSG-P could serve as promising vehicles for enhancing cellular uptake of drugs.

As a replacement for perfluorooctanoic acid, hexafluoropropylene oxide dimer acid, commercially referred to as "GenX", has attracted significant attention. However, a comprehensive understanding of the reproductive systems of male offspring exposed to GenX is lacking. This study aimed to investigate how embryonic exposure to GenX affects the reproductive development of male offspring and the underlying mechanisms. We administered GenX daily via gavage (2 mg/kg body weight/day) to the mice from day 12.5 of pregnancy until delivery. Our results suggested that embryonic exposure to GenX led to delayed onset of puberty in male offspring, with destruction of the testicular structure, disruption of the blood-testis barrier, decreased serum testosterone levels, decreased sperm count, impaired sperm motility, and increased rates of sperm abnormalities. We investigated the mechanism of blood-testis barrier breakdown in vitro by treating Sertoli cells (TM4) with GenX. GenX exposure caused the accumulation of senescent TM4 cells, decreased their glutathione (GSH) levels, and increased their oxidized glutathione levels. GenX inhibited glutaminase activity in TM4 cells, leading to decreased GSH synthesis, increased intracellular oxidative stress, and subsequent TM4 cell senescence, ultimately compromising the blood-testis barrier. Our findings indicated that embryonic exposure to GenX may cause Sertoli cell senescence by altering glutamine metabolism, disrupting the blood-testis barrier, and resulting in abnormal reproductive development in male offspring.

Ocean acidification (OA) is one of the greatest threats to marine species, with widespread impacts on their physiological functions. However, the adaptive capacities of many marine species to OA and the underlying mechanisms remain unclear. In this study, we investigated the effects of short-term (4 days) and medium-term (30 days) CO2 exposure (pH 8.0, 7.6, and 7.3) on black rockfish (Sebastes schlegelii), focusing on histopathological changes in gill tissues, ion transport biomarkers, oxidative stress indicators, and transcriptomic responses. The results showed that both short-term and medium-term OA induced significant morphological changes in gill tissues, including epithelial lifting, hyperplasia, hypertrophy, and lamellar clubbing, which are likely adaptive mechanisms for maintaining homeostasis. Both Na+/K+-ATPase and carbonic anhydrase (CA) activities increased significantly in both short- and medium-term exposure, while Ca2+-ATPase activity was elevated only in the short-term, suggesting differential enzyme regulation over time to sustain ionic balance. Additionally, oxidative stress indicators (superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), reduced glutathione (GSH) and glutathione peroxidase (GPx)) were significantly elevated after both exposure durations, indicating that the antioxidant defense system was activated. Moreover, the integrated biomarker response (IBR) index further indicated that the stress response was more pronounced during short-term exposure. Transcriptomic analysis reveals significant alterations in pathways related to calcium signaling, cytoskeletal structure, energy metabolism, and oxidative stress following short-term exposure. In contrast, medium-term exposure leads to significant enrichment of pathways associated with cell-environment interactions, highlighting the molecular adaptations of S. schlegelii to OA-induced stress. These findings provide valuable insights into the mechanisms of OA tolerance in S. schlegelii and contribute to understanding the adaptability of marine species in future ocean environments.

In previous studies we reported the S-glutathionylation at Cys residues (C76 and C221) of the α5 subunit of the 20S catalytic unit of the yeast proteasome, later mutated to Ser residues. Notably, the strain with the α5-C76S mutation exhibited a reduced chronological life span when grown in glucose as the carbon source. In the present study, we aimed to explore the interplay between mitochondria and the proteasome, considering the α5-C76S-mutated strain as a model of proteasomal impairment. For this purpose, we focused on the growth of the C76S strain in glycerol/ethanol as the carbon source. C76S strain exhibited poor growth and morphological alterations under these conditions, while the proteasomal activity was significantly decreased. We observed decreased activity of the 30S and 26S complexes in the C76S strain, which were accompanied by increased pool of poly-ubiquitinylated proteins. Regarding mitochondrial function, O2 consumption and the concentration of total cellular ATP were significantly increased in the C76S strain. However, levels of peroxiredoxin-1 (Prx1), an important mitochondrial Cys-based peroxidase, were reduced in the C76S strain. In parallel, H2O2 release by mitochondrial respiration was augmented as well as decreased GSH/GSSG ratios, an important parameter of oxidative stress. These findings suggest that, despite increased O2 consumption and ATP production, the mitochondria from the C76S strain promotes an increased oxidative stress most probably due to decreased Prx1 levels. DNA fragmentation and increased cytoplasmic cytochrome C, two apoptotic markers, were observed in the C76S strain. To assess the role of Prx1 in the survival of the C76S strain, we overexpressed this peroxiredoxin in both wild type and C76S strains, which resulted in the partial recovery of the C76S strain phenotype and proteasome activity. The relationship between decreased Prx1 concentration and proteasome impairment remains under investigation.

Phytic acid (PA) is a common anti-nutritional factor found in plant-based protein sources. Our previous research demonstrated that dietary PA negatively affected the growth of grass carp. Intestinal health plays a vital role in the growth, development, and disease resistance of fish. Therefore, in order to comprehensively reveal the impact of PA on the intestines of fish, we further used the grass carp to investigate the impact of PA on the intestines of fish. The 540 grass carp (120.56 ± 0.51 g) were separated into 6 groups and provided with diets that included varying levels of PA (0, 0.8, 1.6, 2.4, 3.2, and 4.0 %) over 60 days. The findings suggested that a higher level of PA in diet, particularly at 3.2 %-4.0 %, led to a decrease in the goblet cell number as well as a reduced expression of mucin 2 and mucin 3 in the intestine. Concurrently, there was an increase in 8-hydroxy-2'-deoxyguanosine, active oxygen species, protein carbonyl, and malondialdehyde. These changes were accompanied by lower anti-superoxide anion activity, total antioxidant capacity, and anti-hydroxyl radical activity, as well as lower activity and gene expression of antioxidant enzymes. The DNA fragmentation in the intestine increased. Additionally, the bcl-2-associated X protein, Fas-ligand, apoptotic protease activating factor-1, cysteine aspartate protease (caspase) 8, caspase 9, and caspase 3 gene expression was increased, while the expression of B-cell lymphoma 2, myeloid cell leukemia-1b, and inhibitor of apoptosis protein was decreased. The gene expression of tight junction-associated molecules (such as claudin-b, -3c, and -7b and zonula occludens-2b) was decreased, whereas the expression of claudin-15b and myosin light chain kinase was increased. These data suggested that dietary PA may compromise the intestinal mucus barrier and the structural integrity of grass carp by inducing a goblet cell number decrease, causing oxidative damage, promoting apoptosis, and disrupting tight junctions. These results indicated that we must consider the potential threat posed by PA to the intestine of grass carp when utilizing plant-based protein sources.

Nanoparticles (NPs) have actively contributed to nanotechnologies advancement over the last years, due to the unique properties they possess compared to their pristine counterparts. Consequently, NPs found wide applications in various fields such as the medical, biomedical, chemical, agro-food industries, and cosmetology. NP's extensive uses could lead to their release into the environment, especially in the marine ecosystems, considered as NPs sink, resulting in harmful effects on organisms. Concerns regarding NPs' toxicity in aquatic organisms have emerged, however, several points remain unexplored. In the present study, the toxicity of chromium oxide (Cr2O3 = 42 nm) and aluminum oxide (Al2O3 = 38 nm) NPs (1 mg/L, 2.5 mg/L, and 5 mg/L) in the gills of the marine gastropod Stramonita haemastoma was assessed through time (7, 14, and 28 days) by a multi-biomarker, Integrated biomarkers response (IBR), and Histological analysis. Both NPs induced varied changes in the antioxidant system, suggesting the onset of oxidative stress marked by superoxide dismutase (SOD), catalase (CAT), acetylcholinesterase (AChE), metallothionein (MT), and malondialdehyde (MDA) levels imbalance. Varied histological alterations in the gills of S. haemastoma were also observed including inflammation, hypertrophy, and lamellar fusion, IBR proved to be a promising tool for assessing NPs toxicity in gastropods. In this study results indicated the co-response of reduced glutathione (GSH), glutathione S-transferase (GST), glutathione peroxidase (GPx), CAT, SOD, and MT after 28 days of exposure. S. haemastoma showed sensitivity to all exposure concentrations of NPs thus validating this species as a suitable indicator of NPs contamination and toxicity.

Melanoma is a devastating form of skin cancer characterized by a high mutational burden, limited treatment success, and dismal prognosis. Although immunotherapy and targeted therapies have significantly revolutionized melanoma treatment, the majority of patients fail to achieve durable responses, highlighting the urgent need for novel therapeutic strategies. Ferroptosis, an iron-dependent form of regulated cell death driven by the overwhelming accumulation of lipid peroxides, has emerged as a promising therapeutic approach in preclinical melanoma models. A deeper understanding of the ferroptosis landscape in melanoma based on its biology characteristics, including phenotypic plasticity, metabolic state, genomic alterations, and epigenetic changes, as well as the complex role and mechanisms of ferroptosis in immune cells could provide a foundation for developing effective treatments. In this review, we outline the molecular mechanisms of ferroptosis, decipher the role of melanoma biology in ferroptosis regulation, reveal the therapeutic potential of ferroptosis in melanoma, and discuss the pressing questions that should guide future investigations into ferroptosis in melanoma.

The term cardiomyopathy refers to a group of heart diseases that cause severe heart failure over time. Cardiomyopathies have been proven to be associated with ferroptosis, a non-apoptotic form of cell death. It has been shown that some small molecule drugs and active ingredients of herbal medicine can regulate ferroptosis, thereby alleviating the development of cardiomyopathy. This article reviews recent discoveries about ferroptosis, its role in the pathogenesis of cardiomyopathy, and the therapeutic options for treating ferroptosis-associated cardiomyopathy. The article aims to provide insights into the basic mechanisms of ferroptosis and its treatment to prevent cardiomyopathy and related diseases.

Autoimmune diseases (ADs) arise from the breakdown of immune tolerance to self-antigens, leading to pathological tissue damage. Proinflammatory cytokine overproduction disrupts redox homeostasis across diverse cell populations, generating oxidative stress that induces DNA damage through multiple mechanisms. Oxidative stress-induced alterations in membrane permeability and DNA damage can lead to the recognition of double-stranded DNA (dsDNA), mitochondrial DNA (mtDNA) and micronuclei-DNA (MN-DNA) by DNA sensors, thereby initiating activation of the cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway. While previous reviews have characterized cGAS-STING activation in autoimmunity, the reciprocal regulation between redox homeostasis and cGAS-STING activation remains insufficiently defined. This narrative review examines oxidative stress-mediated DNA damage as a critical driver of pathological cGAS-STING signaling and delineates molecular mechanisms linking redox homeostasis to autoimmune pathogenesis. Furthermore, we propose therapeutic strategies that combine redox restoration with the attenuation of aberrant cGAS-STING activation, thereby establishing a mechanistic foundation for precision interventions in autoimmune disorders. METHODS: The manuscript is formatted as a narrative review. We conducted a comprehensive search strategy using electronic databases such as PubMed, Google Scholar and Web of Science. Various keywords were used, such as "cGAS-STING," "Redox homeostasis," "Oxidative stress," "pentose phosphate pathway," "Ferroptosis," "mtDNA," "dsDNA," "DNA damage," "Micronuclei," "Reactive oxygen species," "Reactive nitrogen species," "Nanomaterial," "Autoimmune disease," "Systemic lupus erythematosus," "Type 1 diabetes," "Rheumatoid arthritis," "Multiple sclerosis," "Experimental autoimmune encephalomyelitis," "Psoriasis," etc. The titles and abstracts were reviewed for inclusion into this review. After removing duplicates and irrelevant studies, 174 articles met inclusion criteria (original research, English language).

To explore the characteristics of the gut microbiota and metabolites in infants with perianal abscess compared with healthy infants, thereby providing a reference for treating perianal abscess in infants.

The gut microbiota of 19 infants with perianal abscess and 21 healthy infants were compared using 16S rRNA gene sequencing. Metabolite compositions were compared between a subset of 16 infants with perianal abscess and 8 healthy infants.

Both groups showed significant differences in the abundance of the genera Ruminococcus (P = 0.002) and Parasutterella (P = 0.004). Five metabolic pathways, namely, steroid biosynthesis, one-carbon pool by folate, synthesis, secretion, and action of the parathyroid hormone, cholesterol metabolism, and tuberculosis, were significantly enriched. Three metabolites, namely, calcidiol, dihydrofolic acid, and taurochenodesoxycholic acid, were involved in these enriched pathways.

The study revealed significant differences in the composition of the gut microbiota and metabolites between healthy infants and those with perianal abscess, suggesting a potential association between the gut microbiota and infantile perianal abscess.

Cuproptosis, a novel form of copper (Cu)-dependent programmed cell death, is induced by directly binding Cu species to lipoylated components of the tricarboxylic acid (TCA) cycle. Since its discovery in 2022, cuproptosis has been closely linked to the field of materials science, offering a biological basis and bright prospects for the use of Cu-based nanomaterials in various disease treatments. Owing to the unique physicochemical properties of nanomaterials, Cu delivery nanosystems can specifically increase Cu levels at disease sites, inducing cuproptosis to achieve disease treatment while minimizing the undesirable release of Cu in normal tissues. This innovative nanomaterial-mediated cuproptosis, termed as "nanocuproptosis", positions at the intersection of chemistry, materials science, pharmaceutical science, and clinical medicine. This review aims to comprehensively summarize and discuss recent advancements in cuproptosis across various diseases, with a particular focus on cancer. It delves into the biochemical basis of nanomaterial-mediated cuproptosis, the rational design for cuproptosis inducers, strategies for enhancing therapeutic specificity, and cuproptosis-centric synergistic cancer therapeutics. Beyond oncology, this review also explores the expanded applications of cuproptosis, such as antibacterial, wound healing, and bone tissue engineering, highlighting its great potential to open innovative therapeutic strategies. Furthermore, the clinical potential of cuproptosis is assessed from basic, preclinical to clinical research. Finally, this review addresses current challenges, proposes potential solutions, and discusses the future prospects of this burgeoning field, highlighting cuproptosis nanomedicine as a highly promising alternative to current clinical therapeutics.

Fructose-1, 6-bisphosphatase (FBP1) is a tumor suppressor and frequently deficient in various cancers, including clear cell renal cell carcinoma (ccRCC). VHL inactivation mutations are usually observed in ccRCC, which can lead to abnormal activation of the HIF signaling pathway. FBP1 could enter the nucleus and restrain HIF function in a non-enzymatic manner. However, its regulatory mechanism in ccRCC tumorigenesis remains poorly understood. Here, we report that nuclear FBP1 is degraded through the ubiquitin-proteasome pathway, and CUL4B acts as Cullin-RING E3 ubiquitin ligase (CRL) to promote the degradation of FBP1 in nucleus, while the neddylation inhibitor MLN4924 could inactivate CUL4B E3 ligase, block proteasomal degradation of FBP1 and suppress HIF target gene expression, including GLUT1, LDHA, PDK1 and VEGF, leading to decreased glucose uptake and lactate and NADPH production, thereby repressing tumor growth of ccRCC. Furthermore, MLN4924 sensitizes ccRCC to γ-glutamylcysteine synthetase inhibitor Buthionine sulfoximine (BSO) treatment in vivo. Collectively, these findings proposed that MLN4924 could inhibit the tumor growth of VHL deficiency-driven ccRCC by stabilizing FBP1, providing new target and strategy for clinic treatment of ccRCC.

Lesbicoumestans are a class of bioactive compounds isolated from the roots of Lespedeza bicolor (L. bicolor). These compounds have been reported to exhibit anticancer and antiproliferative activities. In this study, our primary focus was to examine the antioxidant capabilities of lesbicoumestan (7) (LC-7), a newly isolated coumestan from roots of L. bicolor, using lipopolysaccharide (LPS)-stimulated immortalized mouse Kupffer cells (ImKCs) as the experimental model. The investigation revealed that LC-7 played a pivotal role in inhibiting the production of reactive oxygen species (ROS). Additionally, LC-7 effectively restored the imbalanced glutathione(GSH)/glutathione disulfide ratio and enhanced the activity of glutathione peroxidase (GPx) following cellular exposure to LPS. Moreover, our investigation revealed that LC-7 exhibited the capacity to enhance the expression of heme oxygenase-1 (HO-1), leading to inhibition of nitric oxide (NO) and proinflammatory cytokines production induced by LPS. Notably, LC-7 did not significantly impact the nuclear factor kappa B cells (NF-κB) and mitogen-activated protein kinase (MAPK) pathways. Intriguingly, LC-7 modulated the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway by direct interaction with Kelch-like-ECH-associated protein 1 (Keap1). These findings suggest that LC-7 possesses antioxidant and anti-inflammatory properties in LPS-stimulated ImKCs by upregulating Nrf2 expression.

Resveratrol was subjected to a diversity-oriented synthesis using oxidative transformations by various biorelevant, biomimetic, or biomimetic-related chemical reagents. Using a combined strategy of ultrahigh-resolution profiling, bioactivity screening, and bioactivity-guided isolation, 19 metabolites were obtained. The compounds were tested for their in vitro enzyme inhibitory activity on angiotensin-1 converting enzyme (ACE), cyclooxygenase-1 and -2, and 15-lipoxygenase (LOX), and evaluated for their relevant drug-like properties in silico. The compounds demonstrated a generally increased cardiovascular protective and anti-inflammatory potential and better drug-likeness compared to resveratrol. Trans-δ-viniferin (6) was identified as a competitive, C-domain-selective ACE inhibitor that is over 20 times more potent than resveratrol. Further, trans-ε-viniferin (2) acted as an over 40 times stronger LOX inhibitor than resveratrol. While our results cannot be directly translated to the health benefits of dietary resveratrol consumption without further studies, it is demonstrated that biologically relevant oxidative environments transform resveratrol into potent cardiovascular protective and anti-inflammatory metabolites.

The well-being and development of fish are affected to varying degrees under the intensive aquaculture model, and the use of Chinese herbs for aquaculture disease control and feed additives has received increasing attention. This study examined fraxetin supplementation in juvenile grass carp to investigate its effects on growth and intestinal structure. There were 1080 grass carp (11.58 ± 0.01 g) assigned to 6 treatments, fed with fraxetin (0, 3.9, 7.9, 15.8, 31.5, and 63.1 mg/kg) for 60 days in each treatment. In our study, appropriate fraxetin significantly increased final body weight (FBW), percent weight gain (PWG), and specific growth rate (SGR) compared to the unadded group (P < 0.05), but did not affect feed efficiency (FE) (P > 0.05). The administration of 7.9 mg/kg of fraxetin significantly improved fish intestinal development and body composition. Appropriate dietary fraxetin significantly enhanced intestinal digestive enzymes and brush border enzyme activity (P < 0.05), decreased serum diamine oxidase (DAO) levels (P < 0.05), and decreased intestinal cell apoptosis (P < 0.05). Appropriate levels of fraxetin inhibited the RhoA/ROCK signaling pathway while upregulating both mRNA and protein expression of tight junction (TJ) and adherens junction (AJ) (P < 0.05). These changes significantly improved apical junction complex (AJC) integrity. In conclusion, dietary supplementation with appropriate levels of fraxetin added to the diets had a facilitating effect on digestion and absorption, improved intestinal structure, and promoted fish growth performance in juvenile grass carp. In addition, the optimal dietary fraxetin levels were evaluated to be 6.06 and 7.79 mg/kg based on linear regression analysis of PWG and DAO, respectively.

The selective delivery of small molecule compounds such as Gemcitabine to tumor cells is a promising methodology for enhancing therapeutic efficacy and attenuating the side effects of anticancer drugs. Aptamers are useful as target-directed ligands for tumor-selective drug delivery due to their ability to bind specific proteins. However, the drug must be released from the aptamer after the conjugate is taken up by the cell to exert its pharmacological effect. In this study, we designed and synthesized a conjugate in which a linker cleaved by glutathione, which is highly expressed in tumor cells, was inserted between the aptamer (AS1411) and Gemcitabine. Almost all Gemcitabine was released from the conjugate after 30 min in the presence of 6 mM glutathione. AS1411 is known to bind to nucleolin, which is highly expressed on tumor cells. The cytotoxicity of the AS1411 and Gemcitabine conjugate with a disulfide bond on A549 cells was higher than that of the conjugate without a disulfide bond. Furthermore, the cytotoxicity of the disulfide-linked conjugate of AS1411 and Gemcitabine was higher in A549 cells than in MCF10A cells, which were used as the model of normal cells. These results indicate that disulfide conjugation enhanced the tumor cell-selective cytotoxicity of Gemcitabine with AS1411.

Reactive oxygen species (ROS), regulators of cellular behaviors ranging from signaling to cell death, have complex production and control mechanisms to maintain a dynamic redox balance under physiological conditions. Redox imbalance is frequently observed in tumor cells, where ROS within tolerable limits promote oncogenic transformation, while excessive ROS induce a range of regulated cell death (RCD). As such, targeting ROS-mediated regulated cell death as a vulnerability in cancer. However, the precise regulatory networks governing ROS-mediated cancer cell death and their therapeutic applications remain inadequately characterized. In this Review, we first provide a comprehensive overview of the mechanisms underlying ROS production and control within cells, highlighting their dynamic balance. Next, we discuss the paradoxical nature of the redox system in tumor cells, where ROS can promote tumor growth or suppress it, depending on the context. We also systematically explored the role of ROS in tumor signaling pathways and revealed the complex ROS-mediated cross-linking networks in cancer cells. Following this, we focus on the intricate regulation of ROS in RCD and its current applications in cancer therapy. We further summarize the potential of ROS-induced RCD-based therapies, particularly those mediated by drugs targeting specific redox balance mechanisms. Finally, we address the measurement of ROS and oxidative damage in research, discussing existing challenges and future prospects of targeting ROS-mediated RCD in cancer therapy. We hope this review will offer promise for the clinical application of targeting oxidative stress-mediated regulated cell death in cancer therapy.

Excessive reactive oxygen species (ROS) are closely associated with the initiation and progression of cancers. As the most abundant intracellular antioxidant, glutathione (GSH) plays a critical role in regulating cellular ROS levels, modulating physiological processes, and is intricately linked to tumor progression and drug resistance. However, the underlying mechanisms remain not fully elucidated. Non-coding RNAs (ncRNAs), including long non-coding RNAs (lncRNAs) and microRNAs (miRNAs), are key regulators of GSH levels. Different ncRNAs modulate various pathways involved in GSH metabolism, and these regulatory targets have the potential to serve as therapeutic targets for enhancing cancer treatment. In this review, we summarize the functions of GSH metabolism and highlight the significance of ncRNA-mediated regulation of GSH in cancer progression, drug resistance, and clinical applications.

Maternal conditions during pregnancy can influence the long-term health of offspring. In particular, maternal malnutrition (MM), such as protein restriction, affects the development of several organs, including the male reproductive system. This study examined how a low-protein maternal diet impacts the structure and function of the dorsolateral prostate (DLP) in aging male rats. Male offspring were divided into two groups: A control group (CTR), whose mothers received a normal protein diet (17%) during pregnancy and lactation, and a low-protein group (GLLP), whose mothers received a low-protein diet (6%) during the same period. At 540 days of age, the offspring were euthanized, and the DLPs were collected for analysis. The GLLP group showed significant structural changes in the DLP, including increased epithelial and reduced stromal compartments. These rats also had lower levels of probasin (a prostate-specific protein), along with a higher number of mast cells, CD68 + macrophages, and IL-10 protein expression, indicating inflammation. Antioxidant balance was disrupted: Glutathione (GSH) levels increased, while catalase (CAT) and superoxide dismutase (SOD) decreased. The expression of SIRT1, a protein linked to aging and oxidative stress control, was reduced. In silico analysis using human prostate cancer data (PRAD-TCGA) revealed that biological pathways related to oxidative stress, immune response, and tissue remodeling were disrupted in both the rat model and human prostate cancer. In summary, maternal protein restriction leads to long-term changes in the dorsolateral prostate of aging male offspring, including inflammation, oxidative stress, and tissue remodeling. The reduced expression of SIRT1 may play a key role in these effects.

Glutathione (GSH) as a key endogenous antioxidant plays the essential role in many bioprocesses, and the lack of GSH would result in a series of diseases. In order to develop a fluorescent indicator for monitoring the fluctuation of GSH and provide information for clinical diagnosis, the red fluorescence carbon dots containing double bonds (DB-CDs) were developed by one-pot hydrothermal process. Owing to photoinduced electron transfer (PET) process between the surface amine groups and the carbon core, the DB-CDs presented the weak fluorescence. Upon addition of GSH, PET process was inhibited by addition reaction between sulfhydryl group of GSH and double bonds, and the bright red fluorescence was exhibited with an emission maximum (λflmax) of 630 nm. A good linear relationship was exhibited in the range of 0.12-75 μM with the detection limit as low as 34.6 nM. Moreover, the cell imaging and the fast kinetic data all demonstrated that the DB-CDs could detect the endogenous GSH without interferences from metal ions and other amino acids, suggesting that the DB-CDs could be used as fluorescence probe for GSH detection in living systems.

MYC-driven group-3 medulloblastomas (MBs) are malignant pediatric brain cancers without cures. To define actionable metabolic dependencies, we identify upregulation of dihydrolipoyl transacetylase (DLAT), the E2-subunit of pyruvate dehydrogenase complex (PDC) in a subset of group-3 MB with poor prognosis. DLAT is induced by c-MYC and targeting DLAT lowers TCA cycle metabolism and glutathione synthesis. We also note upregulation of isocitrate dehydrogenase 1 (IDH1) gene expression in group-3 MB patient tumors and suppression of IDH1 epigenetically reduces c-MYC and downstream DLAT levels in multiple c-MYC amplified cancers. DLAT is a central regulator of cuproptosis (copper-dependent cell death) induced by the copper ionophore elesclomol. DLAT expression in group-3 MB cells correlates with increased sensitivity to cuproptosis. Elesclomol is brain-penetrant and suppresses tumor growth in vivo in multiple group-3 MB animal models. Our data uncover an IDH1/c-MYC dependent vulnerability that regulates DLAT levels and can be targeted to kill group-3 MB by cuproptosis.

Hepatocellular carcinoma (HCC) has a poor prognosis because it is often diagnosed after clinical deterioration and lacks effective therapies. The advent of tumor-targeting therapeutics provided a promise in the landscape of advanced HCC but it is still ongoing due to the indistinguishable receptor expression and receptor abundance between hepatocytes and HCC cells. Herein, a GSH-responsive prodrug, CA-PEG-ss-PTX, was synthesized with cholic acid (CA), paclitaxel (PTX) and polyethylene glycol (PEG) and further applied to physically encapsulate PTX, forming PTX/CA-PEG-ss-PTX (PTX/CPSP). PTX/CPSP gained enhanced liver accumulation via CA-mediated active targeting. After internalization in HCC cells, PTX/CPSP could rapidly disassociate and release PTX in response to the high-level GSH for tumor killing. However, it could remain intact in hepatocytes. Furthermore, CA-modification significantly increased the biliary excretion of PTX/CPSP and performed a "fast in ∼ fast out" drug delivery pattern in hepatocytes, thereby reducing the toxicity caused by excessive drug accumulation. Finally, PTX/CPSP displayed superior anti-HCC efficacy with tolerable toxicity. It is worth noting that PTX/CPSP achieved satisfied PTX loading efficiency (more than 30 %) by both chemical synthesis and physical encapsulation. In summary, with all parts being clinically available or endogenous, PTX/CPSP is considered a clinical potential HCC treatment strategy.

γ-Glutamylcysteine (γ-EC) can increase intracellular glutathione (GSH) levels, which may prevent and alleviate age-related disorders and chronic diseases caused by oxidative damage. However, the commercial availability of γ-EC remains limited owing to its complex chemical synthesis from glutamate and cysteine. In this study, we have developed the method of the effective conversion of GSH to γ-EC to achieve the optimal reaction conditions for repeated batch production and potential application in industrial γ-EC production using the phytochelatin synthase-like enzyme NsPCS. For repeated batch conversion reactions, the optimal temperature was determined at 25 °C, where γ-EC showed good stability compared with that at 37 °C, leading to higher overall productivity. Cellulose sponges and microcrystalline cellulose (MCC) showed superior mechanical strength as immobilization carriers and greater stability and productivity than other materials. The total amounts of γ-EC obtained by NsPCS immobilized on the cellulose sponge and MCC were 305 mg and 291 mg, respectively, in a 5 mL reaction over five repeated batch reactions. These simple production processes are easily reproduced, and their high volumetric efficiency is promising for the industrial production of stable and low-cost γ-EC.

Oxidative stress (OS) is implicated in dementia. While elevated peripheral OS biomarkers were observed in vascular mild cognitive impairment (vMCI), the role of central antioxidants remains unclear. We assessed levels of the major brain antioxidant glutathione (GSH) in vMCI compared to cognitively normal coronary artery disease (CAD) controls (CN).

In vivo tissue-corrected GSH in the anterior cingulate cortex (ACC) and occipital cortex (OC) were quantified in persons with vMCI and CN using MEscher-GArwood Point RESolved magnetic resonance Spectroscopy.

Among participants (vMCI, n = 22, age [mean ± SD] = 67.4 ± 7.3; CN, n = 21, age = 66.7 ± 7.8), ACC-GSH (i.u. ± SD) was higher in vMCI (4.42 ± 0.59) versus CN (3.72 ± 1.01) (Z = -2.5, p = .01), even after controlling for age and sex (B [SE] = 0.74 [0.26], p = .007). Increased ACC-GSH correlated with poorer executive function (EF) (B [SE] = -0.31 [0.14], p = .04). OC-GSH showed no effect.

Higher ACC-GSH in vMCI may reflect a compensatory response to OS. ACC-GSH was negatively correlated with EF, suggesting a linkage between regional brain antioxidants and disease-relevant cognitive domains.

Brain GSH was measured in vascular MCI and matched controls using MEGA-PRESS. In contrast to GSH deficits in AD, anterior cingulate GSH was elevated in vMCI. Brain GSH was correlated with disease-relevant cognitive domains in vMCI. The GSH antioxidant system may be etiologically implicated in vMCI.

Konjac glucomannan (KGM) is a polysaccharide with potential medical and functional properties. Here, the antioxidant and cytoprotective effects of sulfated and differently sized KGM fractions were investigated using various in vitro assays. The sulfated KGMs (SKGMs) were prepared via a relay strategy. First, Vitamin C (Vc)-H2O2 degradation was employed to obtain three soluble KGM fractions with different molecular weights. Second, nine KGM derivatives with varying sulfate content were obtained by the sulfur trioxide-pyridine method. The scavenging of DPPH, superoxide, and hydroxyl radicals was measured in vitro. The antioxidant activity of SKGM correlated positively with sulfate content. SKGM-I-2 displayed the most potent radical scavenging activity. Its purification by cellulose DEAE-52 column chromatography yielded four homogeneous fractions (SKGM-I-2a, SKGM-I-2b, SKGM-I-2c, and SKGM-I-2d). Pretreatment with SKGM-I-2d increased the viability of RAW264.7 cells exposed to H2O2. Moreover, SKGM-I-2d significantly increased the activity of superoxide dismutase and catalase, as well as the levels of glutathione, while regulating the expression of Keap1, Nrf2, and HO-1 in RAW264.7 cells. The present study suggests that SKGM-I-2d protects RAW264.7 cells against H2O2-induced oxidative injury through the activation of the Nrf2/Keap1 signaling pathway. These results provide a scientific basis for future studies linking the structural and functional features of KGM.

Hypothalamic inflammation plays a pivotal role in appetite dysregulation across various pathological conditions, including cancer cachexia. However, delivering anti-inflammatory agents to microglia, key mediators of hypothalamic inflammation, remains challenging due to the unsurmountable blood-brain barrier (BBB). To overcome this challenge, dual peptide-functionalized polymeric nanocarriers capable of both BBB penetration and microglial targeting are engineered for systemic delivery of IRAK4 inhibitors to treat hypothalamic inflammation. After intravenous administration, the nanocarriers demonstrated efficient brain and hypothalamic accumulation in both acute (lipopolysaccharide-induced) and chronic (pancreatic cancer cachexia) neuroinflammation mouse models. Their microglial targeting capability is confirmed through hypothalamic immunohistochemistry and flow cytometry analysis using a BBB-microglia co-culture model. Systemic administration of IRAK4 inhibitor-loaded nanocarriers effectively attenuated hypothalamic inflammation in both animal models, as evidenced by marked reductions in pro-inflammatory cytokine expression. Treated animals displayed significantly increased food intake and improved body weight compared to the saline-treated group. In the cancer cachexia model, the treatment preserved muscle mass, reducing cachexia-induced gastrocnemius muscle loss by 50% relative to controls. These findings highlight the potential of this nanocarrier system as a promising therapeutic strategy for conditions characterized by hypothalamic dysfunction, particularly cancer cachexia, where neuroinflammation plays a crucial role in disease progression.

Oxidative injury has been implicated in a range of common retinal neurodegenerative disorders. Protecting the retina from such an insult could therefore prove clinically beneficial. We sought to investigate whether glucose, acting via the pentose phosphate pathway (PPP), was able to counteract oxidative cytotoxicity to retinal cells in culture.

Mixed retinal neuron-glial cultures were prepared from Sprague-Dawley rat neonates and used at 7 days in vitro; neuron-only and Müller glial cell-only mono-cultures were subsequently prepared from these cultures. At appropriate stages, cultures were treated with t-butyl hydroperoxide (tbH; 10 nM-1 mM) in glucose/pyruvate-free DMEM to induce oxidative stress. Some cultures were co-treated with glucose. Additional compounds were co-applied to inhibit glycolysis, PPP, cystine uptake, glutathione biosynthesis and glutathione reductase (GR). The effect of glucose on stimulation of reactive oxygen species (ROS), as well as levels of glutathione and NADPH were also investigated.

Oxidative stress resulted in cytotoxicity to both retinal neurons and glial cells. Glucose was able to abrogate the toxicity to glial cells in mono-cultures and mixed cultures, but could only provide protection to neurons in the mixed cultures when glial cells were also present. Glucose was additionally shown to prevent stimulation of ROS and oxidative stress-induced depletions of glutathione and NADPH. Inhibition of PPP, cystine uptake or GR all diminished the protective response of glucose.

Glucose prevented oxidative stress to retinal cells via the PPP. Neurons were not subjected to glucose-induced protection except when glial cells were present, implying the passage of a transmissible mediator or other protective action between the two cell types.

This study assessed the antioxidative and protective effects of peptide extracts from selected Euphorbia species on Aluminum Chloride (AlCl3)-induced toxicity in Drosophila melanogaster. Euphorbia humifusa (EHU), Euphorbia hirta (EHI), and Euphorbia graminae (EHG) were screened for bioactive peptides. The crude peptide extract and partially purified peptide fractions of all the plants were subjected to preliminary antioxidants activities through 2, 2-diphenyl-1-picrylhyhdrazyl (DPPH), and nitric oxide (NO) scavenging activities. The most active peptide fraction was subjected to biochemical studies using Drosophila melanogaster. Flies were treated with AlCl3 (100 mg/kg diet), peptide fraction (5 and 10 mg/kg diet), and cotreatment of AlCl3 and the fraction, respectively. After treatment, flies were homogenized for the determination of total thiol and Glutathione (non-protein thiol) content, catalase and Glutathione-S-transferase activities, and nitric oxide (nitrite/nitrate) and hydroperoxide levels. The antioxidant screening revealed that the peptide fraction from Euphorbia humifusa (PEHU) was the most significant compared to the control (ascorbic acid). The PEHU (5 and 10 mg/kg diet) maintained the redox status of the flies in the biochemical study. The PEHU significantly counteracted AlCl3-induced reduction in antioxidants (catalase, GST, GSH and Total thiol), increased nitric oxide levels, and acetylcholinesterase activity and prevented behavioral deficits flies. Hence, the peptide fraction of Euphorbia humifusa may shield against the life-threatening effects of free radicals associated with aluminum chloride toxicity.

Salt stress poses a significant challenge to the growth and productivity of rice (Oryza sativa L.). Histone deacetylases (HDACs) play a vital role in modulating responses to various abiotic stresses. However, how OsHDAC1 responds to salt stress remains largely unknown. Here, we report that OsHDAC1 decreases salt tolerance in rice through posttranslational modification of metabolic enzymes. Specifically, the rice OsHDAC1 RNAi lines exhibited enhanced resilience to salt stress, while plants overexpressing OsHDAC1 were notably more sensitive. OsHDAC1 interacts with the aldehyde dehydrogenase (ALDH) OsALDH2B1 and deacetylates it at K311 and K531, triggering ubiquitin-proteasome-mediated degradation of OsALDH2B1. OsALDH2B1 can directly target OsGR3, which encodes a type of glutathione reductase critical for reactive oxygen species scavenging. Compared with wild-type plants, OsALDH2B1-overexpressing plants exhibited higher OsGR3 expression levels and increased salt resistance, whereas OsALDH2B1 RNAi lines showed reduced OsGR3 expression and lower salt resistance. Collectively, our data suggest that salt stress downregulates OsHDAC1, resulting in an increase in the acetylation level of OsALDH2B1, which in turn stabilizes OsALDH2B1 and promotes its activity in the regulation of OsGR3 transcription. This OsHDAC1/OsALDH2B1/OsGR3 regulatory module represents an alternative pathway for governing salt stress adaptation in rice.

Nanoparticle-based therapeutics hold promise for the treatment of atherosclerosis, but challenges such as low drug-loading capacity and a lack of scalable, controllable production hinder their clinical translation. Flash nanoprecipitation, a continuous synthesis method, offers a potential solution for scalable and reproducible nanoparticle production. In this study, we employed a custom-designed multi-inlet vortex mixer to perform cross-linking reaction-enabled flash nanoprecipitation, facilitating controlled and scalable synthesis of cross-linked polyamidoamine (PAMAM) dendrimer nanoparticles. Notably, this approach allows simultaneous nanoparticle cross-linking and drug loading in a single step. The mannose moiety enabled specific targeting of macrophages via mannose receptors, enhancing the localization of the nanoparticles to atherosclerotic plaques. Atorvastatin calcium, a widely used clinical drug for atherosclerosis treatment, was selected as the model drug. This approach achieved both high production rates and high drug-loading capacities, with an output flow rate of 9.6 L/h and a nanoparticle concentration of approximately 0.4 g/L. The optimized formulation exhibited a drug-loading capacity of 37% and an encapsulation efficiency of 76%. In vitro and in vivo experiments demonstrated effective macrophage and plaque targeting, leading to significant therapeutic benefits. Treatment with these nanoparticles resulted in approximately 40% inhibition of aortic root plaque progression compared to the free drug-treated group. This scalable and efficient nanoparticle platform is a promising strategy for improving atherosclerosis treatment.

Drug-induced liver injury (DILI) is a leading cause of acute liver failure, which is closely associated with oxidative stress. Glutathione (GSH), a vital sulfhydryl peptide, maintains cellular redox balance and signaling. In this study, we have successfully developed a highly selective near-infrared fluorescent probe, AH-F, which exhibits a 357-fold enhancement in fluorescence upon detection of GSH. With the aid of AH-F, the pertinent physiological parameters in a murine model were characterized by cellular and drug-induced liver injury, concurrently allowing for the assessment of the therapeutic efficacy of relevant pharmaceutical interventions.

Glutathione acts as a natural antioxidant in the human body and the reduction of its content is a sign of oxidative stress. In this study, a sensitive electrochemical sensor was developed using laccase enzyme immobilized onto graphene oxide (GO) for detection of glutathione. The surface of the indium tin oxide (ITO) was modified with GO via a drop casting method. Subsequently, laccase was immobilized onto the modified ITO decorated with GO. The modified electrode was characterized using field-emission scanning electron microscopy (FESEM), X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The FTIR spectra of laccase/GO confirmed the successful immobilization of laccase onto GO sheets. FESEM analysis revealed the transformation from a layered, wrinkled structure to a compact, smooth surface with spherical laccase, confirming successful enzyme integration. Raman analysis confirmed successful laccase immobilization onto GO, as evidenced by structural changes in the D and G bands, highlighting the modification of the material. The cyclic voltammetry measurements revealed that laccase/GO/ITO exhibited better electrocatalytic activity toward oxidation of GSH in acetate buffer solution than the bare ITO electrode. This newly developed electrode exhibited a good response to glutathione with a wide linear range from 1 μM to 100 μM, a limit of detection of 0.89 μM and high sensitivity (6.51 μA μM-1). Furthermore, it exhibited excellent selectivity, repeatability, and long-term stability. The modified electrode was successfully used for the detection of GSH in a real sample, offering satisfactory results.

Sensorineural hearing loss (SNHL), the most commonly-occurring form of hearing loss, is caused mainly by injury to or the loss of hair cells and spiral ganglion neurons in the cochlea. Numerous environmental and physiological factors have been shown to cause acquired SNHL, such as ototoxic drugs, noise exposure, aging, infections, and diseases. Several programmed cell death (PCD) pathways have been reported to be involved in SNHL, especially some novel PCD pathways that have only recently been reported, such as ferroptosis, necroptosis, and pyroptosis. Here we summarize these PCD pathways and their roles and mechanisms in SNHL, aiming to provide new insights and potential therapeutic strategies for SNHL by targeting these PCD pathways.