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Plant glutathione transferases (GSTs) constitute a large and diverse family of enzymes that are involved in plant stress response, metabolism and defence, yet their physiological functions remain largely elusive. Consistent with the traditional view on GSTs across organisms as detoxification enzymes, in vitro most plant GSTs catalyse glutathionylation, conjugation of the tripeptide glutathione (GSH; γ-Glu-Cys-Gly) onto reactive molecules. However, when it comes to elucidating GST functions, it remains a key challenge that the endogenous plant glutathione conjugates (GS-conjugates) that would result from such glutathionylation reactions are rarely reported. Furthermore, GSTs often display high substrate promiscuity, and their proposed substrates are prone to spontaneous chemical reactions with GSH; hence, single-gene knockouts rarely provide clear chemotypes or phenotypes. In a few cases, GS-conjugates are demonstrated to be biosynthetic intermediates that are rapidly further metabolized towards a pathway end product, explaining their low abundance and rare detection. In this review, we summarize the current knowledge of plant GST functions and how and possibly why evolution has resulted in a broad and extensive expansion of the plant GST family. Finally, we demonstrate that endogenous GS-conjugates are more prevalent in plants than assumed and suggest they are overlooked as clues towards the identification of plant GST functions. This article is part of the theme issue 'The evolution of plant metabolism'. [ABSTRACT FROM AUTHOR]

Salinity stress is a major concern in regions where irrigation relies on saline water. This study aimed to investigate the relative water content (RWC), electrolytic leakage (EL), total chlorophyll content, free amino acid content, and total soluble sugar content were analyzed in different groundnut species subjected to various salinity treatments. The results showed that salinity stress significantly reduced the RWC in groundnut leaves, with A. duranensis (wild type) exhibiting higher RWC values compared to the Arachis hypogaea species. RNA sequencing was performed to identify differentially expressed genes (DEGs) during salt stress. A total of 9079 DEGs were identified, with 1372 genes upregulated and 2509 genes downregulated. Genes belonging to transcription factor families, such as WRKY, MYB, bHLH, E2F, and Auxin efflux carrier proteins, were induced under salt stress in the tolerant genotype. Conversely, genes encoding NADH dehydrogenase, glutathione S-transferase, protein kinases, UDP-glycosyltransferase, and peroxidase were downregulated. Gene ontology and pathway analyses revealed several enriched categories and metabolic pathways associated with salt stress response, including catalytic activity, response to salt stress, ATP-dependent activity, and oxidative phosphorylation. The findings of this study provide insights into the physiological and molecular responses of groundnut to salinity stress. A. duranensis exhibited better salinity tolerance than Arachis hypogaea , as indicated by higher RWC values, lower electrolytic leakage, and differential gene expression patterns. These results contribute to our understanding of the mechanisms underlying salt stress tolerance in groundnut and may guide future efforts to develop salinity-tolerant groundnut species, ultimately improving crop yield in saline-affected regions. • Salinity stress decreases crop yield by affecting water content and leaf damage in groundnut plants. • A. duranensis variety shows higher tolerance to salinity stress compared to GG20. • Salinity stress reduces chlorophyll levels in groundnut leaves, with A. duranensis being more resilient. • Salinity stress triggers the accumulation of free amino acids and soluble sugars, especially in A. duranensis. • RNA-seq analysis reveals key genes involved in groundnut's response to salt stress, providing valuable insights. [ABSTRACT FROM AUTHOR]

ABSTRACT As an egg parasitoid, Trichogramma ostriniae (T. ostriniae) exhibits a broad host range and plays a crucial role in controlling various lepidopteran agricultural pests. However, the application of chemical pesticides negatively impacts its development and survival. Therefore, it is essential to assess the toxicity of commonly used insecticides against T. ostriniae and evaluate their compatibility. This study aims to determine the toxic effects of five common insecticides (dinotefuran, abamectin, imidacloprid, beta‐cypermethrin, and chlorantraniliprole) on the development, reproduction, and enzymatic activity of T. ostriniae. The contact, lethal, and developmental toxicities were evaluated. Activities of detoxification enzymes, including cytochrome P450 (CYP450), carboxylesterase (CarE), and glutathione S‐transferase (GST), and protective enzymes, including superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) as well as malondialdehyde (MDA) and reactive oxygen species (ROS), mitochondrial respiratory chain complex I (MRCC I), adenosine triphosphate (ATP) of T. ostriniae were examined after being treated with the five insecticides. The results showed that the order of lethal toxicity in adult T. ostriniae was beta‐cypermethrin > dinotefuran > imidacloprid > abamectin > chlorantraniliprole. The emergence rates of adult T. ostriniae exposed to insecticides during the egg and larval stages were higher than those exposed to insecticides during the prepupal and pupal stages. The activities of CYP450, GST, and SOD were increased, but CarE activity and ROS content were decreased in T. ostriniae treated with the five insecticides compared with the control. Beta‐cypermethrin increased the POD and CAT activities. Chlorantraniliprole decreased CAT activity and increased MDA content. The MRCCI of T. ostriniae was not significantly affected by any of the five insecticides tested. The ATP content of T. ostriniae was not significantly affected by chlorantraniliprole but was significantly decreased by the other four insecticides. In conclusion, the toxicities of the five insecticides to T. ostriniae were different, among which imidacloprid, dinotefuran, abamectin, and beta‐cypermethrin had a high risk of toxicity to T. ostriniae, and chlorantraniliprole had a low risk to T. ostriniae. [ABSTRACT FROM AUTHOR]

The DNA damage response is a highly conserved protective mechanism that enables cells to cope with various lesions in the genome. Extensive studies across different eukaryotic cells have identified the crucial roles played by components required for response to DNA damage. When compared to the essential signal transducers and repair factors in the DNA damage response circuitry, the negative regulators and underlying mechanisms of this circuitry have been relatively under-examined. In this study, we investigated Gst1, a putative glutathione transferase in the fungal pathogen Candida albicans. We found that under stress caused by the DNA damage agent MMS, GST1 expression was significantly upregulated, and this upregulation was further enhanced by the loss of the checkpoint kinases and DNA repair factors. Somewhat counterintuitively, deletion of GST1 conferred increased resistance to MMS, potentially via enhancing the phosphorylation of Rad53. Furthermore, overexpression of RAD53 or deletion of GST1 resulted in upregulated transcription of DNA damage repair genes, including CAS1, RAD7, and RAD30, while repression of RAD7 transcription in the GST1 deletion reversed the strain's heightened resistance to MMS. Finally, Gst1 physically interacted with Rad53, and their interaction weakened in response to MMS-induced stress. Overall, our findings suggest a negative regulatory role for GST1 in DNA damage response in C. albicans, and position Gst1 within the Rad53-mediated signaling pathway. These findings hold significant implications for understanding the mechanisms underlying the DNA damage response in this fungal pathogen and supply new potential targets for therapeutic intervention. [ABSTRACT FROM AUTHOR]

Fasciolosis is a zoonotic infection and is considered a developing deserted tropical illness threatening ruminant productivity and causing financial losses. Herein, we applied immunoinformatics and biophysics studies to develop an epitopes vaccine against Fasciola hepatica using glutathione transferase and Cathepsin L-like proteinase as possible vaccine candidates. Using the selected proteins, B- and Tcell epitopes were predicted. After epitopes prediction, the epitopes were clarified over immunoinformatics screening, and only five epitopes, EFGRWQQEKCTIDLD, RRNIWEKNVKHIQEH, FKAKYLTEMSRASDI, TDMTFEEFKAKYLTE, and YTAVEGQCR were selected for vaccine construction; selected epitopes were linked with the help of a GPGPG linker and attached with an adjuvant through another linker, EAAAK linker. Cholera toxin B subunit was used as an adjuvant. The ExPASy ProtParam tool server predicted 234 amino acids, 25.86257 kDa molecular weight, 8.54 theoretical pI, 36.86 instability index, and -0.424 grand average of hydropathicity. Molecular docking analysis predicted that the vaccine could activate the immune system against F. hepatica. We calculated negative binding energy values. A biophysics study, likely molecular docking molecular dynamic simulation, further validated the docking results. In molecular dynamic simulation analysis, the top hit docked compounds with the lowest binding energy values were subjected to MD simulation; the simulation analysis showed that the vaccine and immune cell receptors are stable and can activate the immune system. MMGBSA of -146.27 net energy (kcal/mol) was calculated for the vaccine-TLR2 complex, while vaccine-TLR4 of -148.11 net energy (kcal/mol) was estimated. Furthermore, the C-ImmSim bioinformatics tool predicted that the vaccine construct can activate the immune system against F. hepatica, eradicate the infection caused by F. hepatica, and reduce financial losses that need to be spent while protecting against infections of F. hepatica. The computational immune simulation unveils that the vaccine model can activate the immune system against F. hepatica; hence, the experimental scientist can validate the finding accomplished through computational approaches. [ABSTRACT FROM AUTHOR]

Yohimbine is a potent bioactive indole alkaloid, isolated from a variety of biological sources and has long been used as a natural stimulant and aphrodisiac, particularly to treat erectile dysfunction. However, some literature also points toward its anticancer effect in different experimental models. The current study aimed to address a clinical concern on the therapeutic utilization of yohimbine as a repurposed drug. We employed 7,12‐dimethylbenz[a]anthracene (DMBA)‐induced hamster buccal pouch carcinogenesis model juxtaposed with biochemical investigation of several detoxification and antioxidant markers, such as Cyt p450, Cyt b5, thiobarbituric acid reactive substance (TBARS), glutathione (GSH), glutathione reductase (GR), glutathione S transferase (GST), DT‐diaphorase, vitamin C, vitamin E, superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx). The immunohistochemical assessment of cyclooxygenase‐2 (COX‐2), interleukin‐6 (IL‐6), proliferating cell nuclear antigen (PCNA), and cyclin D1 expression were also performed to observe the effect of yohimbine on these markers. The hamsters treated with DMBA presented the growth of tumors in the buccal pouches, accompanied by significant changes in the liver and buccal mucosa levels of Phase I & II detoxification enzymes and lipid peroxidation (LPO). A significant rise in the range of 2‐ to 3.5‐fold was observed in Cyt p450, Cyt b5, and LPO in DMBA‐treated animals. However, oral administration of yohimbine significantly restored the LPO, antioxidant, and detoxifying enzyme activities. Additionally, the levels of COX‐2, IL‐6, PCNA, and cyclin D1 were also found to be downregulated by yohimbine treatment. In conclusion, yohimbine improved the biochemical and immunohistochemical markers of DMBA‐induced oral cancer and reverted to near normal values via ameliorating the underlying inflammation and oxidative stress conditions. Our study highlighted the potential of yohimbine as anticancer agent, especially against oral cancer and suggested its possible use as repurposed drug. Summary: In this study, we have used hamster model to evaluate potential of yohimbine against oral carcinogenesis.Yohimbine reinstated DMBA‐induced oral carcinogenesis and facilitated detoxification by modulating the Phase I and Phase II enzyme activities.The upregulated expression of Cox‐2, IL‐6, PCNA, and cyclin D1 was also found to be reverted to near normal by yohimbine treatment. [ABSTRACT FROM AUTHOR]

Decreasing nitrogen (N) supply affected the normal growth of Oryza sativa (O. sativa) seedlings, reducing CO2 assimilation, stomatal conductance (gs), the contents of chlorophylls (Chl) and the ratio of Chl a/Chl b, but increasing the intercellular CO2 concentration. Polyphasic chlorophyll a fluorescence transient and relative fluorescence parameters (JIP test) results indicated that N deficiency increased Fo, but decreased the maximum quantum yield of primary photochemistry (Fv/Fm) and the maximum of the IPphase, implying that N-limiting condition impaired the whole photo electron transport chain from the donor side of photosystem II (PSII) to the end acceptor side of PSI in O. sativa. N deficiency enhanced the activities of the antioxidant enzymes, such as ascorbate peroxidase (APX), guaiacol peroxidase (GuPX), dehydro–ascorbate reductase (DHAR), superoxide dismutase (SOD), glutathione peroxidase (GlPX), glutathione reductase (GR), glutathione S-transferase (GST) and O-acetylserine (thiol) lyase (OASTL), and the contents of antioxidant compounds including reduced glutathione (GSH), total glutathione (GSH+GSSG) and non-protein thiol compounds in O. sativa leaves. In contrast, the enhanced activities of catalase (CAT), DHAR, GR, GST and OASTL, the enhanced ASC–GSH cycle and content of sulfur-containing compounds might provide protective roles against oxidative stress in O. sativa roots under N-limiting conditions. Quantitative real-time PCR (qRT-PCR) analysis indicated that 70% of the enzymes have a consistence between the gene expression pattern and the dynamic of enzyme activity in O. sativa leaves under different N supplies, whereas only 60% of the enzymes have a consistence in O. sativa roots. Our results suggested that the antioxidant system and sulfur metabolism take part in the response of N limiting condition in O. sativa, and this response was different between leaves and roots. Future work should focus on the responsive mechanisms underlying the metabolism of sulfur-containing compounds in O. sativa under nutrient deficient especially N-limiting conditions. [ABSTRACT FROM AUTHOR]

SUMMARY: Maintaining an optimal redox status is essential for plant growth and development, particularly when the plants are under stress. AT‐hook motif nuclear localized (AHL) proteins are evolutionarily conserved transcription factors in plants. Much of our understanding about this gene family has been derived from studies on clade A members. To elucidate the functions of clade B genes, we first analyzed their spatial expression patterns using transgenic plants expressing a nuclear localized GFP under the control of their promoter sequences. AHL1, 2, 6, 7, and 10 were further functionally characterized owing to their high expression in the root apical meristem. Through mutant analyses and transgenic studies, we showed that these genes have the ability to promote root growth. Using yeast one‐hybrid and dual luciferase assays, we demonstrated that AHL1, 2, 6, 7, and 10 are transcription regulators and this activity is required for their roles in root growth. Although mutants for these genes did not showed obvious defects in root growth, transgenic plants expressing their fusion proteins with the SRDX repressor motif exhibited a short‐root phenotype. Through transcriptome analysis, histochemical staining and molecular genetics experiments, we found that AHL10 maintains redox homeostasis via direct regulation of glutathione transferase (GST) genes. When the transcript level of GSTF2, a top‐ranked target of AHL10, was reduced by RNAi, the short‐root phenotype in the AHL10‐SRDX expressing plant was largely rescued. These results together suggest that AHL genes function redundantly in promoting root growth through direct regulation of redox homeostasis. [ABSTRACT FROM AUTHOR]

Wheat seedlings were hydroponically grown in Hoagland solution containing various levels of Se. Tolerance response to Se toxicity was investigated by determining the level of thiobarbituric acid reactive substance (TBARS), proline and chlorophyll content, the growth parameters, and the activity of antioxidant enzymes. The toxic level of Se treatment significantly retarded the seedling growth. A substantial amount of proline accumulation was also observed in response to toxic Se concentration, but it was more pronounced in putative-sensitive cultivars. Chlorophyll content significantly decreased in Se-intoxicated seedlings and increased at the lowest Se dose in both cultivars. Severe and mild chlorosis was observed in putative-sensitive and tolerant cultivars at the highest Se level. Alterations in the activities of glutathione reductase (GR, 1.6.4.2), glutathione S transferase (GST, 2.5.1.18), guaiacol peroxidase (GPX, 1.11.1.7), catalase (CAT, 1.11.1.6), and ascorbate peroxidase (APX, 1.11.1.11) and superoxide dismutase (SOD, EC 1.15.1.1) were determined. TBAR level did not significantly increase in putative tolerant cultivars as an indicator of membrane lipid peroxidation. However, a significant increase was observed in putative-sensitive cultivars in response to higher selenium concentrations. In higher Se treatment groups, CAT and GST activities significantly increased in putative Se tolerant cultivars. However, excluding SOD, the activity of all the studied enzymes was increased considerably in putative-sensitive cultivars in a dose-dependent manner. Higher antioxidant enzyme activities and a substantial amount of proline accumulation did not significantly contribute to overcoming Se phytotoxicity in wheat seedlings grown in media supplemented with toxic selenium levels. [ABSTRACT FROM AUTHOR]

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Plant defence mechanisms, including physical barriers like toughened bark and chemical defences like allelochemicals, are essential for protecting them against pests. Trees allocate non-structural carbohydrates (NSCs) to produce secondary metabolites like monoterpenes, which increase during biotic stress to fend off pests like the Eurasian spruce bark beetle, ESBB (Ips typographus). Despite these defences, the ESBB infests Norway spruce, causing significant ecological damage by exploiting weakened trees and using pheromones for aggregation. However, the mechanism of sensing and resistance towards host allelochemicals in ESBB is poorly understood. We hypothesised that the exposure of ESBB to spruce allelochemicals, especially monoterpenes, leads to an upsurge in the important detoxification genes like P450s, GSTs, UGTs, and transporters, and at the same time, genes responsible for development must be compromised. The current study demonstrates that exposure to monoterpenes like R-limonene and sabiene effectively elevated detoxification enzyme activities. The differential gene expression (DGE) analysis revealed 294 differentially expressed (DE) detoxification genes in response to R-limonene and 426 DE detoxification genes in response to sabiene treatments, with 209 common genes between the treatments. Amongst these, genes from the cytochrome P450 family 4 and 6 genes (CP4 and CP6), esterases, glutathione S-transferases family 1 (GSTT1), UDP-glucuronosyltransferase 2B genes (UDB), and glucose synthesis-related dehydrogenases were highly upregulated. We further validated 19 genes using RT-qPCR. Additionally, we observed similar high expression levels of detoxification genes across different monoterpene treatments, including myrcene and α-pinene, suggesting a conserved detoxification mechanism in ESBB, which demands further investigation. These findings highlight the potential for molecular target-based beetle management strategies targeting these key detoxification genes. [ABSTRACT FROM AUTHOR]

The glyphosate herbicide is a pesticide widely used in the world and can contaminate soil, air, and water. The objective of this work was to evaluate the toxicity of a glyphosate‐based herbicide (GBH) in zebrafish (Danio rerio). Fish were exposed to different concentrations of GBH (0, 50, 250, and 500 µg/L) for 96 hours. Brain, liver, and blood were collected for biochemical and genotoxicity analyses, and behavioral tests were performed. The results showed that there was a reduction in the activity of the antioxidant enzymes of catalase (CAT) and glutathione‐S‐transferase (GST) in the liver at all concentrations and at the highest concentration in the brain. There was also a reduction in lipid peroxidation in the liver at all concentrations of glyphosate. There was an increase in micronuclei in the blood at the 500 µg/L concentration. However, the count of nuclear abnormalities showed no differences from the control. Interleukin‐1beta (IL‐1β) generation was inhibited at all concentrations in the liver and at the highest concentration in the brain. No significant differences were found in the behavioral test compared to the control. The results showed that acute exposure to GBH promoted an inflammatory event, which reduced the efficiency of antioxidants, thus producing a disturbance in tissues, mainly in the liver, causing immunosuppression and generating genotoxicity. [ABSTRACT FROM AUTHOR]

This study was designed to evaluate the acaricidal effect and the mode of action of lemongrass essential oil (LEO) against Aleuroglyphus ovatus. Contact and fumigant mortality indicated that LEO had higher acaricidal activities at a low concentration, and the acaricidal action of LEO was significantly more effective by contact effects than by fumigant bioassay. Repellent rate against A. ovatus at different concentrations of LEO increased with application time. Observation on the toxic symptoms revealed that almost all mites were in immobilized type (IM type) in 30 min, but the symptom of knock-down type (KD type) appeared later. Cuticles of mites killed by LEO exhibited obvious colour change and deformation. Detoxification enzyme activity assay showed that glutathione S-transferase (GST) activity increased in mites treated by LEO compared to the controls. LEO increased catalase (CAT) activity of A. ovatus but inhibited superoxide dismutase (SOD) after 20 h. LEO also increased nitric oxide synthetase (NOS) and inhibited acetylcholine esterase (AChE) activities. Moreover, LEO acted as a fast-acting neurotoxin on A. ovatus. Our results suggest that LEO has an acaricidal potential and is a promising agent in control of A. ovatus. [ABSTRACT FROM AUTHOR]

In breeding programs, stress memory in plants can develop drought stress tolerance. Memory stress, as an approach, can keep stress data by activating tolerance mechanisms. This research was conducted to evaluate some physiologically effective mechanisms in inducing memory drought stress in the seeds that were exposed to water stress three times in four treatments including rainfed, 33%, 66%, and 100% of field capacity (FC). After the production of the seeds, the third-generation seeds were placed under different irrigation treatments, seed and seedling traits, starch to carbohydrate ratio in seed, protein concentration and glutathione reductase were investigatied in a factorial format based on a randomized complete block design with three replications. Results showed that percentage of changes from the lowest to the highest value for traits including seed vigor, seed endosperm weight, seed coat weight, accelerated aging, cold test, seedling biomass and seedling length were 25, 37, 65, 65, 55, 77, 55, 65 and 79, respectively and germination uniformity was 3.9 times higher than the lowest amount. According to the deterioration percentage, seed vigor and the percentage of seed germination in cold test data, it can be reported that seed production by 100% FC was not appropriate for rainfed plots. However, considering the the appropriate results in the percentage of germination for a cold test, germination uniformity percentage, and the lowest accelerated aging seeds, seed production under the rainfed conditions with 33% FC watering can be recommended. In-silico analysis was coducted on Glutathione reductase (GR) enzymes in Gossypium hirsutum. It is clear that GR has a Redox-active site and NADPH binding, and it interacts with Glutathione S transferase (GST). So, memory drought stress through inducing physiological drought tolerance mechanisms such as starch-to-carbohydrate ratio and GR can determine the suitable pattern for seed production for rainfed and low rainfall regions in a breeding program. Our study thus illustrated that seed reprduction under 33% FC equipped cotton with the tolerance against under draught stress from the seedling stage. This process is done through activating glutathione reductase and balancing the ratio of starch to carbohydrates concentration. [ABSTRACT FROM AUTHOR]

Aims This study identifies a unique glutathione S-transferase (GST) in extremophiles using genome, phylogeny, bioinformatics, functional characterization, and RNA sequencing analysis. Methods and Results Five putative GSTs (H0647, H0729, H1478, H3557, and H3594) were identified in Halothece sp. PCC7418. Phylogenetic analysis suggested that H0647, H1478, H0729, H3557, and H3594 are distinct GST classes. Of these, H0729 was classified as an iota-class GST, encoding a high molecular mass GST protein with remarkable features. The protein secondary structure of H0729 revealed the presence of a glutaredoxin (Grx) Cys-Pro-Tyr-Cys (C-P-Y-C) motif that overlaps with the N-terminal domain and harbors a topology similar to the thioredoxin (Trx) fold. Interestingly, recombinant H0729 exhibited a high catalytic efficiency for both glutathione (GSH) and 1-chloro-2, 4-dinitrobenzene (CDNB), with catalytic efficiencies that were 155- and 32-fold higher, respectively, compared to recombinant H3557. Lastly, the Halothece gene expression profiles suggested that antioxidant and phase II detoxification encoding genes are crucial in response to salt stress. Conclusion Iota-class GST was identified in cyanobacteria. This GST exhibited a high catalytic efficiency toward xenobiotic substrates. Our findings shed light on a diversified evolution of GST in cyanobacteria and provide functional dynamics of the genes encoding the enzymatic antioxidant and detoxification systems under abiotic stresses. [ABSTRACT FROM AUTHOR]

The assessment of endothelial dysfunction and free radical homeostasis parameters were performed in 92 women, aged 45 to 69 years, divided into the following groups: women without COVID-19 (unvaccinated, no antibodies, control); women with acute phase of COVID-19 infection (main group, COVID-19+); 12 months post COVID-19+; women with anti-SARS-CoV-2 IgG with no symptoms of COVID-19 in the last 12 months (asymptomatic COVID-19). Compared to the control, patients of the main group had lower glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities, decreased advanced glycation end products (AGEs) level, higher glutathione reductase (GR) activity, and higher glutathione S transferases pi (GSTpi), thiobarbituric acid reactants (TBARs), endothelin (END)-1, and END-2 concentrations (all p ≤ 0.05). The group with asymptomatic COVID-19 had lower 8-OHdG and oxidized glutathione (GSSG) levels, decreased total antioxidant status (TAS), and higher reduced glutathione (GSH) and GSH/GSSG levels (all p ≤ 0.05). In the group COVID-19+, as compared to the group without clinical symptoms, we detected lower GPx and SOD activities, decreased AGEs concentration, a higher TAS, and greater GR activity and GSTpi and TBARs concentrations (all p ≤ 0.05). The high content of lipid peroxidation products 12 months post COVID-19+, despite decrease in ENDs, indicates long-term changes in free radical homeostasis. These data indicate increased levels of lipid peroxidation production contribute, in part, to the development of free radical related pathologies including long-term post COVID syndrome. [ABSTRACT FROM AUTHOR]

Magnolia wufengensis, a newly discovered ornamental species in the Magnoliaceae family, is susceptible to salinity. Moreover, Ca2+ is an essential element for plant growth and is receiving increasing attention for its ability to mitigate the negative effects of environmental stress on plants. In the present study, we investigated the effect of Ca2+ on the growth and transcriptome of M. wufengensis under salt stress. The treatments used here were as follows: control, NaCl (150 mmol/L), CaCl2 (5 mmol/L), and NaCl (150 mmol/L) + CaCl2 (5 mmol/L). After a 60-day treatment period, plant growth indices were determined, and leaves were collected for physiological analysis and transcriptome investigation. The combined application of NaCl and CaCl2 alleviated phenotypic damage and restored seedling growth. Moreover, RNA sequencing data revealed that in the Na vs. control group and the NaCa vs. Na group, there were 968 and 2632 differentially expressed genes, respectively, which were both primarily enriched in secondary metabolism, glutathione metabolism, signaling hormone metabolism, glucose metabolism, and amino acid metabolism. These pathways were analyzed to screen key genes: the adenosine triphosphate (ATP)-binding cassette efflux transporter G1 (ABCG1) genes, which are related to transmembrane transport; the calmodulin genes, which are related to signal transmission; and the glutathione S-transferase (GST), glutathione peroxidase (GPX), and peroxidase (POD) genes related to antioxidant enzymes. Lastly, we constructed a hypothesis model of Ca2+-enhanced salt tolerance in M. wufengensis. This study reveals the potential mechanisms by which Ca2+ enhances the salt tolerance of M. wufengensis and provides a theoretical reference for its cultivation in saline areas. [ABSTRACT FROM AUTHOR]

The proliferation of antibiotic usage has precipitated the emergence of drug-resistant variants of bacteria, thereby augmenting their capacity to withstand pharmaceutical interventions. Among these variants, Cronobacter sakazakii (C. sakazakii), prevalent in powdered infant formula (PIF), poses a grave threat to the well-being of infants. Presently, global contamination by C. sakazakii is being observed. Consequently, research endeavors have been initiated to explore the strain's drug resistance capabilities, alterations in virulence levels, and resistance mechanisms. The primary objective of this study is to investigate the resistance mechanisms and virulence levels of C. sakazakii induced by five distinct antibiotics, while concurrently conducting transcriptomic analyses. Compared to the susceptible strains prior to induction, the drug-resistant strains exhibited differential gene expression, resulting in modifications in the activity of relevant enzymes and biofilm secretion. Transcriptomic studies have shown that the expression of glutathione S-transferase and other genes were significantly upregulated after induction, leading to a notable enhancement in biofilm formation ability, alongside the existence of antibiotic resistance mechanisms associated with efflux pumps, cationic antimicrobial peptides, and biofilm formation pathways. These alterations significantly influence the strain's resistance profile. [ABSTRACT FROM AUTHOR]

Heat stress (HS) poses a great challenge to the poultry industry by inducing oxidative damage to the liver, endangering the health and production of broilers. As an important type of seaweed polyphenols, phlorotannin has been shown to have antioxidant properties. The present study evaluated the protective effects of dietary phlorotannin on HS-induced liver injury in broilers based on oxidative damage parameters. A total of 108 twenty-one days old male Arbor Acres plus (AA+) broilers were randomly divided into three groups: TN group (thermoneutral, 24 ± 1 °C, fed with basal diet), HS group (HS, 33 ± 1 °C for 8 h/day, fed with basal diet), and HS + phlorotannin group (HS + 600 mg/kg phlorotannin). Each group has six replicate cages with six birds per cage. The feeding experiment lasted 21 days. At the termination of the feeding experiment (42 days old), samples were collected for analysis of morphological and biochemical features. The results showed that HS decreased the liver index, serum albumin (ALB) content, hepatic antioxidant enzymes activities of catalase (CAT), total superoxide dismutase (T-SOD), glutathione S-transferase (GST), and glutathione peroxidase (GSH-Px) (p < 0.05), while increasing the hepatic histopathology score, apoptosis rate, and malondialdehyde (MDA) content (p < 0.05) in 42-day-old broilers. Compared with the HS group, dietary phlorotannin improved the activities of antioxidant enzymes (GST and GSH-Px) but decreased the histopathology score and apoptosis rate in the liver (p < 0.05). Moreover, HS down-regulated hepatic mRNA expression of CAT1, NQO1, HO-1, and SLC7A11 (p < 0.05), while up-regulated hepatic mRNA expression of Keap1, MafG, IκBα, NF-κB P65, IFN-γ, TFR1, ACSL4, Bax, and Caspase-9 (p < 0.05). Compared with HS group, dietary phlorotannin up-regulated hepatic mRNA expression of Nrf2, CAT1, MafF, GSTT1, NQO1, HO-1, GCLC, GPX1, TNF-α, Fpn1, and SLC7A11 (p < 0.05), while down-regulated hepatic mRNA expression of IκBα, Bax, Caspase-9, and TFR1 (p < 0.05). In conclusion, dietary supplementation of 600 mg/kg phlorotannin could alleviate HS-induced liver injury via regulating oxidative status, apoptosis, and ferroptosis in broilers; these roles of phlorotannin might be associated with the regulation of the Nrf2 signaling pathway. [ABSTRACT FROM AUTHOR]

Titanium dioxide nanoparticles (TiO2) are released into the natural waters in different ways and cause toxicity to aquatic life. Although various studies have reported the toxicity of nanoparticles for aquatic animals, there is limited information regarding the effects of environmental factors, especially salinity, on their toxicity. Simultaneous effects of TiNPs and water salinity were studied in the Pacific oyster, Crassostrea gigas. Oysters were exposed to sub-lethal concentrations of TiNPs (low concentration: 50 µg/l, middle concentration: 500 µg/l, high concentration: 5000 µg/l) at different water salinities (Extremely low salinity: 5 g/l, low salinity: 15 g/l, middle salinity: 25 g/l, high salinity (normal salinity): 35 g/l) for 96 h. After nano-TiO2 exposure, the immune components in hemolymph (total hemocyte concentration [THC], phagocytosis), antioxidant enzyme activities in digestive gland (superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase), oxidation stress index (malondialdehyde) in gills and the expression of immune-related genes (Vav-3 protein, cofilin, Galectin 4, pro-Cathepsin L, Rho protein, Ficolin, LPS/-1,3-glucan protein, Lysozyme, Defensin) in hemocytes were measured. The THC, phagocytosis, antioxidant enzyme activities, and immune-related gene expressions mostly decreased at salinities of 5 and 15 g/l in response to 500–5000 µg/l nano-TiO2 compared to non-exposed oysters and 50 µg/l nano-TiO2 group. An increase in water salinity up to 35 g/l mitigated the depressing effects of the nano-TiO2 on the immune and antioxidant components of the oysters. Exposure of oysters to 500–5000 µg/l nano-TiO2-induced oxidative stress at salinities of 5 and 15, since malondialdehyde levels were higher in these treatments compared to control and those in 50 µg/l nano-TiO2 group. The increase of salinity up to 35 g/l ameliorated the TiO2-induced oxidative stress. The results of the present study showed that the toxic impacts of nano-TiO2 can be ameliorated in the Pacific oyster with increase in water salinity, as the immune components, antioxidant enzyme activities recovered and oxidative stress mitigated when the salinity increased up to 35 g/l. [ABSTRACT FROM AUTHOR]

Abstract The DNA damage response is a highly conserved protective mechanism that enables cells to cope with various lesions in the genome. Extensive studies across different eukaryotic cells have identified the crucial roles played by components required for response to DNA damage. When compared to the essential signal transducers and repair factors in the DNA damage response circuitry, the negative regulators and underlying mechanisms of this circuitry have been relatively under-examined. In this study, we investigated Gst1, a putative glutathione transferase in the fungal pathogen Candida albicans. We found that under stress caused by the DNA damage agent MMS, GST1 expression was significantly upregulated, and this upregulation was further enhanced by the loss of the checkpoint kinases and DNA repair factors. Somewhat counterintuitively, deletion of GST1 conferred increased resistance to MMS, potentially via enhancing the phosphorylation of Rad53. Furthermore, overexpression of RAD53 or deletion of GST1 resulted in upregulated transcription of DNA damage repair genes, including CAS1, RAD7, and RAD30, while repression of RAD7 transcription in the GST1 deletion reversed the strain’s heightened resistance to MMS. Finally, Gst1 physically interacted with Rad53, and their interaction weakened in response to MMS-induced stress. Overall, our findings suggest a negative regulatory role for GST1 in DNA damage response in C. albicans, and position Gst1 within the Rad53-mediated signaling pathway. These findings hold significant implications for understanding the mechanisms underlying the DNA damage response in this fungal pathogen and supply new potential targets for therapeutic intervention. Graphical Abstract

ARHGAP25, a crucial molecule in immunological processes, serves as a Rac-specific GTPase-activating protein. Its role in cell migration and phagocyte functions, affecting the outcome of complex immunological diseases such as rheumatoid arthritis, renders it a promising target for drug research. Despite its importance, our knowledge of its intracellular interactions is still limited. This study employed proteomic analysis of glutathione S-transferase (GST)-tag pulldowns and co-immunoprecipitation from neutrophilic granulocyte cell lysate, revealing 76 candidates for potential physical interactions that complement ARHGAP25's known profile. Notably, four small GTPases (RAC2, RHOG, ARF4, and RAB27A) exhibited high affinity for ARHGAP25. The ARHGAP25–RAC2 and ARHGAP25–RHOG interactions appeared to be affected by the activation state of the small GTPases, suggesting a GTP–GDP cycle-dependent interaction. In silico dimer prediction pinpointed ARHGAP25's GAP domain as a credible binding interface, suggesting its suitability for GTP hydrolysis. Additionally, a list of Fc receptor-related kinases, phosphatases, and three of the 14-3-3 members were identified as potential partners, with in silico predictions highlighting eight binding sites, presenting novel insight on a potential regulatory mechanism for ARHGAP25. [ABSTRACT FROM AUTHOR]

As rate-limiting enzymes of peroxisomal b-oxidation, acyl-coenzyme A oxidase (ACOXs) play vital roles in maintaining energy homeostasis and regulating reactive oxygen species (ROS) metabolism. However, there are no studies on the functions of ACOXs in crustaceans. In the present study, four full-length cDNA sequences of acoxs, namely the acox-1a (2403 bp), acox-1b (2733 bp), acox-3a (2878 bp) and acox-3b (3445 bp), were successfully isolated from mud crab Scylla paramamosain, which encoded 666, 673, 701 and 658 amino acids, respectively. Sequence analysis showed that the ACOX-1a, ACOX-1b and ACOX-3a possessed conserved structural domains like FAD-binding motif, fatty acyl CoA oxidase domain and peroxisomal targeting signal, while the ACOX-3b lacked peroxisomal targeting signal. Results of phylogenetic tree indicated that the four ACOXs of mud crab grouped gathered with their corresponding orthologues from crustaceans. The acox-1a, acox-3a and acox-3b were highly expressed in hepatopancreas, and the acox-1b was mainly distributed in muscle and hepatopancreas. Compared with feeding groups, the expression levels of acox-1a, acox-3a and acox-3b in hepatopancreas and acox-3a in muscle were markedly up-regulated in fasting groups, suggesting that the acoxs had significant effects in modulating energy balance during fasting. In addition, fasting significantly increased the transcriptional levels of nuclear factor erythroid 2-related factor (nrf2) and its downstream antioxidant genes (catalase (cat), glutathione peroxidase (gpx) and glutathione S-transferase (gst)) to improve antioxidant capacity for removing excessive ROS produced by ACOX-mediated peroxisomal b-oxidation. These results would be conducive to providing new insights into evolutionary characteristics and functions of acoxs in crustaceans. [ABSTRACT FROM AUTHOR]

Objectives: Previous studies have shown that gene expressions can be regulated in the hippocampus of rats after seizures induced by kainic acid (KA). The aim of this study was to examine the potential regulatory impact of KA administration on gene expression levels of enzymes responsible for drug metabolism in rat hippocampal tissue. Materials and Methods: Rats received intraperitoneal injections of KA and saline at a dose of 10 mg/kg. Behavioral changes were observed in experimental animals following the administration of KA. Four hours after receiving treatments, all rats were decapitated, and the brains were removed. Hippocampal tissues were used for total RNA isolation, and cDNA synthesis was performed by reverse transcription polymerase chain reaction (PCR). Gene expression levels of enzymes responsible for drug metabolism were determined by quantitative PCR using the RT2 Profiler PCR Array Rat Drug Metabolism PCR array system containing the relevant primers for a total of 84 genes. The gene expression levels of drug-metabolizing enzymes were quantified using the comparative Ct (2-ΔΔ(delta delta)Ct) method. The Student’s t-test was used for data analysis. Results: Our results indicate that KA treatment caused significant changes in the gene expression levels of metallothionein 3, glucose phosphate isomerase, adenosine triphosphate-binding cassette protein C1, cytochrome P450 enzymes (Cyp2c6v1, Cyp3a23/3a1, Cyp2c7), glutathione peroxidase 1, 4, and 5, glutamic acid decarboxylase 1 and 2, paraoxonase 2, carbohydrate sulfotransferase 1, glutathione S-transferases (Gsta3, Gstm1, Gstm4), microsomal glutathione S-transferase 3, carboxylesterase 2C, fatty acid amide hydrolase, pyruvate kinase-muscle, arachidonate 5-lipoxygenase, apolipoprotein E, cytochrome b5 reductase 5, xanthine dehydrogenase, N-acetyltransferase 1, glucokinase regulator, hexokinase 2, myristoylated alanine rich protein kinase C substrate, and stannin in the hippocampus compared with the control (p < 0.05). Conclusion: As a conclusion, it can be said that the seizure activity triggered by KA has the potential to change the gene expression levels of the enzymes responsible for drug metabolism in the hippocampus of rats. [ABSTRACT FROM AUTHOR]

Successful completion of spermatogenesis is crucial for the perpetuation of the species. In Drosophila, spermatid individualization, a process involving changes in mitochondrial structure and function is critical to produce functional mature sperm. Ant2, encoding a mitochondrial adenine nucleotide translocase, is highly expressed in male testes and plays a role in energy metabolism in the mitochondria. However, its molecular function remains unclear. Here, we identified an important role of Ant2 in spermatid individualization. In Ant2 knockdown testes, spermatid individualization complexes composed of F‐actin cones exhibited a diffuse distribution, and mature sperms were absent in the seminal vesicle, thus leading to male sterility. The most striking effects in Ant2‐knockdown spermatids were decrease in tubulin polyglycylation and disruption of proper mitochondria derivatives function. Excessive apoptotic cells were also observed in Ant2‐knockdown testes. To further investigate the phenotype of Ant2 knockdown in testes at the molecular level, complementary transcriptome and proteome analyses were performed. At the mRNA level, 868 differentially expressed genes were identified, of which 229 genes were upregulated and 639 were downregulated induced via Ant2 knockdown. iTRAQ‐labeling proteome analysis revealed 350 differentially expressed proteins, of which 117 proteins were upregulated and 233 were downregulated. The expression of glutathione transferase (GstD5, GstE5, GstE8, and GstD3), proteins involved in reproduction were significantly regulated at both the mRNA and protein levels. These results indicate that Ant2 is crucial for spermatid maturation by affecting mitochondrial morphogenesis. [ABSTRACT FROM AUTHOR]

Glutathione transferase Pi (GST-P) reportedly overexpressed markedly in cancer cell lines. It was correlated to the resistance towards drugs used in chemotherapy treatment. The study demonstrated the isolation of peptide as potential inhibitor to the enzyme. Glutathione transferase Pi (GST-P) was purified from human colon adenocarcinoma HT-29 cell line using glutathione (GSH)-affinity chromatography. Active components presence in ethanolic extract (50% ethanol) of leaves Leptospermum flavescens Sm. was shown possessing inhibitory property (IC50 of 0.088 mg/mL) towards GST-P in vitro. Further fractionation using polyamide the 50% methanol in 2% acetic acid eluate possessed an inhibitory property at IC50 of 0.191 mg/mL. In the study the IC50 values of doxorubicin are 0.788 and 0.816 μg/mL on HT-29 and MRC-5 cell lines respectively while IC50 values of cisplatin were at 9.49 and 4.07 μg/mL on HT-29 and MRC-5 cell lines respectively. The 50% methanol eluate has significantly non-toxic to both of the cell lines with 100% cell viability at more than 100 μg/mL sample applied. In combination with doxorubicin, 50% methanol eluate enhanced cytotoxicity of the drug towards HT-29 by reduction of IC50 value significantly to 66%. The eluate however only reduced the IC50 to 11% when combined with cisplatin. The study indicates that the eluate can potentiate cytotoxicity of doxorubicin on HT-29 cell line and this effect is correlated to the ability of the eluate to inhibit GST-P in vitro. The purified active molecule was a peptide with molecular weight of 3.5 kDa. [ABSTRACT FROM AUTHOR]

The most abundant tripeptide—glutathione (GSH)—and the major GSH-related enzymes—glutathione peroxidases (GPxs) and glutathione S-transferases (GSTs)—are highly significant in the regulation of tumor cell viability, initiation of tumor development, its progression, and drug resistance. The high level of GSH synthesis in different cancer types depends not only on the increasing expression of the key enzymes of the γ-glutamyl cycle but also on the changes in transport velocity of its precursor amino acids. The ability of GPxs to reduce hydroperoxides is used for cellular viability, and each member of the GPx family has a different mechanism of action and site for maintaining redox balance. GSTs not only catalyze the conjugation of GSH to electrophilic substances and the reduction of organic hydroperoxides but also take part in the regulation of cellular signaling pathways. By catalyzing the S-glutathionylation of key target proteins, GSTs are involved in the regulation of major cellular processes, including metabolism (e.g., glycolysis and the PPP), signal transduction, transcription regulation, and the development of resistance to anticancer drugs. In this review, recent findings in GSH synthesis, the roles and functions of GPxs, and GST isoforms in cancer development are discussed, along with the search for GST and GPx inhibitors for cancer treatment. [ABSTRACT FROM AUTHOR]

Aluminium (Al) is one of the most abundant chemical elements in nature and metal in the earth’s crust. Accumulation of Aluminium ions (Al3+) in target tissues results into formation of oxygen radicals causing oxidative damage through inducing cytotoxicity. The aim of this study is to analyse the antioxidant effect of selected polyphenols (quercetin, rutin and a medicinal plant phagnalon rupestre) on rats intoxicated by aluminium with specific focus on the heart.This experiment was carried out on 6 groups of wistar albino rats,group 1; positive control, group 2; Al male intoxicated group treated with quercetin, group 3; Al male intoxicated group treated with rutin, group 4; Al female intoxicated, group 5; Al intoxicated treated with phagnalon rupestre, group 6; female control. Several Biochemical assays were carried out such as protein test, Thiobarbituric acid reactive substance (TBARS) assay, catalase assay (CAT), glutathione-s-transferase (GST) assay, superoxide dismutase (SOD) assay, Glutathione reduced and glutathione peroxidase (GSH & GPX) assay, lipids (cholesterol & triglycerides) assays. Aluminium intoxicated group showed decrease in the content of protein compared to the control and treated groups. Aluminium intoxicated group showed an increase in the activity of thiobarbituric reactive substances (TBARS) but a significant decrease in the activity of catalase (CAT), glutathione S transferase assays (GST) compared to the control group. Biomarkers of oxidative stress significantly reduced in heart Al-induced oxidative stress by administration of Quercetin. Therefore, Quercetin is an effective antioxidant against oxidative stress caused by free radicals produced because of aluminium exposure. [ABSTRACT FROM AUTHOR]

Endothelial cell injury is a hallmark of IgA vasculitis (IgAV), possibly associated with various factors, including oxidative stress. Certain single nucleotide polymorphisms (SNPs) of glutathione S-transferases (GST) genes have been shown to increase susceptibility to oxidative stress. The objective of our study was to evaluate the gene polymorphisms of GSTM1, GSTT1, GSTP1, and GSTA1 in patients with IgAV. DNA was extracted from the blood of 124 children with IgAV and 168 age-matched healthy controls. A higher frequency of the GSTM1 null genotype was observed in patients with gastrointestinal (GI) system involvement compared to those without GI system involvement (51.5% vs. 28.6%, p = 0.011). Additionally, the GSTM1 null genotype was less prevalent (30.8% vs. 69.2%, p = 0.032), while the GSTP1 Val/Val genotype was significantly more prevalent in patients who developed urogenital complications (scrotal swelling) during the course of the disease (60% vs. 40%, p = 0.039). This study is the first to suggest an association between GSTM1 and GSTP1 polymorphisms and various phenotypes observed during the clinical course of IgAV in the pediatric population. However, it was performed on a national and likely single ethnic cohort, too small for definitive conclusions, so larger studies are needed to confirm this association. [ABSTRACT FROM AUTHOR]

GSTs catalyze detoxification reactions of harmful xenobiotics via conjugation with glutathione (GSH) while glutathione reductase (GR) is the sole enzyme that acts in the recovery reaction of GSH from oxidized glutathione (GSSG). In this study, in vitro inhibitory impacts of 3‐amino‐benzoic acid methyl ester compounds on GR and GST were investigated. For this firstly, GR and GST were obtained from human blood with specific activity of 6.26 EU/mg protein and 8.57 EU/mg protein respectively. Then, inhibition studies were performed. It was found that methyl 3‐amino‐5‐chlorobenzoate had the highest inhibitory effect on hGR with Ki value of 0.524±0.109 μM) and methyl 3‐amino‐4‐nitrobenzoate was the most effective inhibitor on hGST with Ki value of 37.05±4.487 μM. Besides, molecular docking analysis was used to estimate the binding energies of molecules. Methyl 3‐amino‐4‐nitrobenzoate and methyl 3‐amino‐4‐chlorobenzoate were predicted to have the highest binding affinity into GR and GST receptors respectively. [ABSTRACT FROM AUTHOR]

This study investigates the impact of imidacloprid insecticide, on the flesh fly Sarcophaga ruficornis, (Diptera: Sarcophagidae), a medically and veterinary significant species associated with myiasis. Lethal (0.02%) and sub‐lethal (0.004%) doses were administered over 24 and 48 h, focusing on midgut histopathological, ultrastructural and biochemical alterations. Imidacloprid‐induced disruptions in physiological and metabolic functions, leading to adult fly mortality. Histological and cytological analyses revealed significant cellular changes, including cell degeneration, cytoplasmic vacuolization, chromatin condensation, irregular epithelial borders, disrupted peritrophic membrane and the release of cell components into the midgut lumen. Biochemical analyses demonstrated a decrease in total protein, carbohydrate and lipid contents, correlating with insecticide concentration and duration. Antioxidant enzymes, particularly glutathione S‐transferase (GST) and superoxide dismutase (SOD), resulted in a significant increase compared to the control, proportional to time and concentration. This research provides a comprehensive understanding of imidacloprid‐induced toxicity in the midgut of Sarcophaga ruficornis, emphasizing its potential for effective pest population management. [ABSTRACT FROM AUTHOR]

Marine red yeast is a good feed additive for the aquaculture industry that can promote the growth of aquatic animals, and significantly improve their antioxidant capacity, survival rate, and nonspecific immune ability of the body. Our hypothesis is that dietary supplementation with marine red yeast could affect the serum biochemistry, muscle composition, muscle fatty acid composition, lipid metabolism enzyme activity, and expression of antioxidant and inflammatory genes in juvenile genetically improved farmed tilapia (GIFT, Oreochromis niloticus). Five diets with different levels of marine red yeast (0 %, 0.25 %, 0.50 %, 0.75 %, and 1.00 %) were used to feed juvenile GIFT (initial weight: 21.12 ± 0.86 g) for 60 days. The main results are as follows: Compared with the control group (0 % marine red yeast), the contents of total protein (TP), albumin (ALB), high-density lipoprotein (HDL), and free fatty acid (FFA) in the serum of juvenile GIFT tilapia significantly increased (P < 0.05), while the contents of glucose (GLU), triglyceride (TG), total cholesterol (T-CHO), low-density lipoprotein (LDL), alanine aminotransferase (ALT) significantly decreased (P < 0.05). The contents of crude protein, crude fat, and docosahexaenoic acid + eicosapentaenoic acid (DHA+EPA) in the muscle significantly increased (P < 0.05), while the contents of moisture and monounsaturated fatty acids (∑MUFAs) significantly decreased (P < 0.05). The activities of acetyl CoA carboxylase a (ACCα), glucose-6-phosphate dehydrogenase (G6PD), and stearoyl-CoA desaturase (SCD) in the liver significantly increased (P < 0.05). The expression levels of catalase (cat), alkaline phosphatase (alp), nuclear factor erythroid 2-related factor 2 (nrf2), lysozyme (lyz), glutathione S-transferase (gst), glutathione peroxidase (gsh-px), and superoxide dismutase (sod) genes in the liver significantly increased (P < 0.05). The expression levels of tumor necrosis factor α (tnf-α), interferon-γ (inf-γ), interleukin 6 (il-6), interleukin 8 (il-8), interleukin 1β (il-1β), and transforming growth factor-β (tgf-β1) genes in the liver, spleens, and head kidney significantly increased (P < 0.05), while the expression level of interleukin 10 (il-10) gene significantly decreased (P < 0.05). In conclusion, the addition of different levels of marine red yeast could significantly affect the serum biochemistry, muscle composition, muscle fatty acid composition, lipid metabolism enzyme activity, and expression of antioxidant and inflammatory genes in juvenile GIFT tilapia. Based on the results, the optimal dietary marine red yeast level was 0.50 %. [ABSTRACT FROM AUTHOR]

Paraquat (PQT) is among the most toxic herbicides that possesses the potential to exert deleterious effects on animal and human health. Kaempferide (KMF) is a natural flavanol that manifests profound therapeutic potential. Rats (n=24) were separated into 4 groups i.e., Control, PQT 5 mgkg-1, PQT 5 mgkg-1 + KMF 20 mgkg-1 and KMF 20 mgkg-1 administered group. After 30 days of treatment, our results revealed that PQT exposure reduced the activities of antioxidant enzymes i.e., glutathione S-transferase (GST), glutathione (GSH), superoxide dismutase (SOD), glutathione reductase (GSR), catalase (CAT), glutathione peroxidase (GPx) and heme oxygenase-1 (HO-1) while increased the levels of malondialdehyde (MDA) and reactive oxygen species (ROS). Furthermore, PQT intoxication reduced the level of renal biomarkers i.e., creatinine clearance, and increased the level of creatinine, urea, neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 (KIM-1). In addition to this, it increased the levels of nuclear factor-kappa B (NF-kB), tumor necrosis factor-alpha (TNF-a), interleukin-1beta (IL-1β) and interleukin-6 (IL-6) as well as increased the activity of cyclooxygenase-2 (COX-2). Furthermore, PQT exposure increased the levels of caspase-9, Bax and caspase-3 while it decreased the levels of Bcl-2. Additionally, histological analysis showed substantial renal tissue damage in PQT intoxicated rats. However, KMF treatment recovered the renal impairments. Therefore, it is proposed that KMF may be used as a nephroprotective agent against PQT-prompted renal toxicity. [ABSTRACT FROM AUTHOR]

Glutathione transferase (GST) is a superfamily of ubiquitous enzymes, multigenic in numerous organisms and which generally present homodimeric structures. GSTs are involved in numerous biological functions such as chemical detoxification as well as chemoperception in mammals and insects. GSTs catalyze the conjugation of their cofactor, reduced glutathione (GSH), to xenobiotic electrophilic centers. To achieve this catalytic function, GSTs are comprised of a ligand binding site and a GSH binding site per subunit, which is very specific and highly conserved; the hydrophobic substrate binding site enables the binding of diverse substrates. In this work, we focus our interest in a model organism, the fruit fly Drosophila melanogaster (D. mel), which comprises 42 GST sequences distributed in six classes and composing its GSTome. The goal of this study is to describe the complete structural GSTome of D. mel to determine how changes in the amino acid sequence modify the structural characteristics of GST, particularly in the GSH binding sites and in the dimerization interface. First, we predicted the 3D atomic structures of each GST using the AlphaFold (AF) program and compared them with X-ray crystallography structures, when they exist. We also characterized and compared their global and local folds. Second, we used multiple sequence alignment coupled with AF-predicted structures to characterize the relationship between the conservation of amino acids in the sequence and their structural features. Finally, we applied normal mode analysis to estimate thermal B-factors of all GST structures of D. mel. Particularly, we extracted flexibility profiles of GST and identify key residues and motifs that are systematically involved in the ligand binding/dimerization processes and thus playing a crucial role in the catalytic function. This methodology will be extended to guide the in silico design of synthetic GST with new/optimal catalytic properties for detoxification applications. [ABSTRACT FROM AUTHOR]

S-acylation is a reversible post-translational modification catalyzed by protein S-acyltransferases (PATs), and acyl protein thioesterases (APTs) mediate de-S-acylation. Although many proteins are S-acylated, how the S-acylation cycle modulates specific biological functions in plants is poorly understood. In this study, we report that the S-acylation cycle of transcription factor MtNAC80 is involved in the Medicago truncatula cold stress response. Under normal conditions, MtNAC80 localized to membranes through MtPAT9-induced S-acylation. In contrast, under cold stress conditions, MtNAC80 translocated to the nucleus through de-S-acylation mediated by thioesterases such as MtAPT1. MtNAC80 functions in the nucleus by directly binding the promoter of the glutathione S-transferase gene MtGSTU1 and promoting its expression, which enables plants to survive under cold stress by removing excess malondialdehyde and H2O2. Our findings reveal an important function of the S-acylation cycle in plants and provide insight into stress response and tolerance mechanisms. The S-acylation and de-S-acylation of transcription factor MtNAC80 under cold stress form a key S-acylation cycle that contributes to the establishment of cold tolerance in Medicago truncatula. [ABSTRACT FROM AUTHOR]

With the rising prevalence of end-stage kidney disease, the use of expanded criteria donor allografts, seen as essential for meeting organ demand, still proves challenging due to their higher risk of graft loss, delayed function, and rejection. Machine perfusion, a technique in preserving allografts, offers improved allograft outcomes compared to static cold storage while allowing for the noninvasive measurement of kidney injury biomarkers in the perfusate solution. This offers an objective method to assess graft function at various preservation stages. We conducted a narrative review of the databases PubMed and Scopus, including studies written in the English language and published after 2010. In this narrative review, we identified biomarkers, like 4-hydroxyproline, taurine, and glutathione transferase, as predictive markers of delayed graft function. Additionally, biomarkers, like extracellular histone h3, vascular cell adhesion protein, and matrix metalloprotease protein, have shown correlation with decreased graft function, although their predictive ability remains inconclusive. The review outlines various suggestions for potential areas of research focus to enhance future expanded criteria donor allograft utilization. However, limitations exist, including the absence of a singular reliable biomarker and the challenges of validating biomarker effectiveness across diverse outcomes. [ABSTRACT FROM AUTHOR]

Mango (Mangifera indica) is a highly valuable horticultural crop known for its quality and productivity. This study investigates the dynamic changes in physicochemical properties and glutathione and ascorbic acid metabolic pathways in mango pulp across various growth and development stages over two consecutive years (2021–2022 and 2022–2023) by transcriptomic analysis. Overall, the results demonstrate that during different ripening periods, the pulp shows increased levels of total soluble solids, relative conductivity, glutathione, and enzymes, while titratable acidity, malondialdehyde, reactive oxygen species, and ascorbic acid contents decreased. Moreover, transcriptomic analysis identified key differentially expressed genes from the glutathione and ascorbic acid metabolic pathways and validated them with qRT-PCR. In different comparisons, a total of 1776, 2513, and 828 DEGs were identified in 30 vs. 60, 30 vs. 90, and 60 vs. 90 days after flowering, respectively. Among them, seven DEGs were primarily enriched in relevant pathways, which included ascorbate peroxidase, ascorbate oxidase, glutathione peroxidase, gamma-glutamyl transferase, glutathione transferases, and glucose-6-phosphate dehydrogenase. The upregulation of these genes indicates that glutathione and AsA respond well to scavenging reactive oxygen species and maintain normal functioning in plants. This research sheds light on the molecular mechanisms of glutathione and ascorbic acid dynamic changes in mango pulp, providing valuable insights into the regulation of antioxidant and metabolic pathways during fruit growth and development. [ABSTRACT FROM AUTHOR]

Simple Summary: In aquaculture, probiotics are widely used to improve growth, digestion, immunity, and so on. Employing a combination of multiple probiotic strains to enhance host health promotion is commonly acknowledged. However, studies assessing the synergistic effects of mixed probiotic additives on hosts and comparing the properties of each strain in their composition remain unclear. Therefore, this study aims to elucidate the mechanisms underlying the effects of individual and combined endogenous probiotics on hybrid groupers. Our findings revealed that the addition of mixed probiotics (Bacillus cereus and Exiguobacterium acetylicum) effectively enhances the growth performance, immune response, and intestinal health of hybrid groupers. Therefore, we recommend selecting combination strains of endogenous probiotics for optimal growth and immunity in hybrid groupers. This study aims to examine the effects of the mixture of Bacillus cereus G1–11 and Exiguobacterium acetylicum G1–33, isolated from the gut of hybrid groupers (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂), on the host. The hybrid groupers were divided into a control (C, without any probiotics), B. cereus (BC, 1010 cfu/g), E. acetylicum (EA, 108 cfu/g), compound (mix, a 1:1 mixture of B. cereus and E. acetylicum), and positive reference group (P, Lactobacillus acidophilus, 5 × 108 cfu/L). Each group had four replicates, with 30 fish per replicate (53.30 ± 0.50 g), and were fed for 60 days. The results showed that adding probiotics to the feed significantly improved the weight gain, weight growth rate, specific growth rate, and digestive enzyme activities of hybrid groupers compared to the C group. The compound group was the most significant. In addition, composite probiotics added to feed significantly upregulated the expression levels of several growth-related genes in the liver and muscles. The activities of alkaline phosphatase, catalase, glutathione peroxidase, glutathione transferase, lysozyme, and total antioxidant capacity in the serum and liver were significantly influenced through mixed probiotic feeding. Moreover, the expression levels of several immune-related genes in the liver, spleen, and head kidney were significantly enhanced by adding single and mixed probiotics to feed, with the synergy of mixed probiotics being the best. An analysis of the gut microbiota showed that adding composite bacteria enhanced the richness and diversity of the gut microbiota, significantly increasing the relative abundance of potential probiotics (Cetobacterium and Microbacterium) while decreasing the presence of potential pathogens (Mycoplasma). Overall, our findings highlighted the efficacy of mixed probiotics (B. cereus and E. acetylicum) in enhancing growth performance, nutritional value of hybrid grouper feed, antioxidant capacity, immune response, and intestinal health, in finding the best combination of functional feed additives. [ABSTRACT FROM AUTHOR]

The autophagosomal SNARE STX17 (syntaxin 17) promotes lysosomal fusion and degradation, but its autophagosomal recruitment is incompletely understood. Notably, PtdIns4P is generated on autophagosomes and promotes fusion through an unknown mechanism. Here we show that soluble recombinant STX17 is spontaneously recruited to negatively charged liposomes and adding PtdIns4P to liposomes containing neutral lipids is sufficient for its recruitment. Consistently, STX17 colocalizes with PtdIns4P-positive autophagosomes in cells, and specific inhibition of PtdIns4P synthesis on autophagosomes prevents its loading. Molecular dynamics simulations indicate that C-terminal positively charged amino acids establish contact with membrane bilayers containing negatively charged PtdIns4P. Accordingly, Ala substitution of Lys and Arg residues in the C terminus of STX17 abolishes membrane binding and impairs its autophagosomal recruitment. Finally, only wild type but not Ala substituted STX17 expression rescues the autophagosome-lysosome fusion defect of STX17 loss-of-function cells. We thus identify a key step of autophagosome maturation that promotes lysosomal fusion. Abbreviations: Cardiolipin: 1',3'-bis[1-palmitoyl-2-oleoyl-sn-glycero-3-phospho]-glycerol; DMSO: dimethyl sulfoxide; GST: glutathione S-transferase; GUV: giant unilamellar vesicles; LAMP1: lysosomal associated membrane protein 1; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; PA: 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphate; PC/POPC: 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine; PG: 1-palmitoyl-2-linoleoyl-sn-glycero-3-phospho-(1'-rac-glycerol); PI: L-α-phosphatidylinositol; PI4K2A: phosphatidylinositol 4-kinase type 2 alpha; PIK3C3/VPS34: phosphatidylinositol 3-kinase catalytic subunit type 3; POPE/PE: 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine; PS: 1-stearoyl-2-linoleoyl-sn-glycero-3-phospho-L-serine; PtdIns(3,5)P2: 1,2-dioleoyl-sn-glycero-3-phospho-(1"-myo-inositol-3',5'-bisphosphate); PtdIns3P: 1,2- dioleoyl-sn-glycero-3-phospho-(1'-myo-inositol-3'-phosphate); PtdIns4P: 1,2-dioleoyl-sn-glycero-3-phospho-(1"-myo-inositol-4'-phosphate); SDS-PAGE: sodium dodecyl sulfate-polyacrylamide gel electrophoresis; STX17: syntaxin 17. [ABSTRACT FROM AUTHOR]

Chemotherapy-provoked peripheral neuropathy and its associated affective disorders are important adverse effects in cancer patients, and its treatment is not completely resolved. A recent study reveals a positive interaction between molecular hydrogen (H2) and a heme oxygenase (HO-1) enzyme inducer, cobalt protoporphyrin IX (CoPP), in the inhibition of neuropathic pain provoked by nerve injury. Nevertheless, the efficacy of CoPP co-administered with hydrogen-rich water (HRW) on the allodynia and emotional disorders related to paclitaxel (PTX) administration has not yet been assessed. Using male C57BL/6 mice injected with PTX, we examined the effects of the co-administration of low doses of CoPP and HRW on mechanical and thermal allodynia and anxiodepressive-like behaviors triggered by PTX. Moreover, the impact of this combined treatment on the oxidative stress and inflammation caused by PTX in the amygdala (AMG) and dorsal root ganglia (DRG) were studied. Our results indicated that the antiallodynic actions of the co-administration of CoPP plus HRW are more rapid and higher than those given by each of them when independently administered. This combination inhibited anxiodepressive-like behaviors, the up-regulation of the inflammasome NLRP3 and 4-hydroxynonenal, as well as the high mRNA levels of some inflammatory mediators. This combination also increased the expression of NRF2, HO-1, superoxide dismutase 1, glutathione S-transferase mu 1, and/or the glutamate-cysteine ligase modifier subunit and decreased the protein levels of BACH1 in the DRG and/or AMG. Thus, it shows a positive interaction among HO-1 and H2 systems in controlling PTX-induced neuropathy by modulating inflammation and activating the antioxidant system. This study recommends the co-administration of CoPP plus HRW as an effective treatment for PTX-provoked neuropathy and its linked emotive deficits. [ABSTRACT FROM AUTHOR]

Short‐chain dehydrogenases/reductases (SDRs) are ubiquitously distributed across diverse organisms and play pivotal roles in the growth, as well as endogenous and exogenous metabolism of various substances, including drugs. The expression levels of SDR genes are reportedly upregulated in the fenpropathrin (FEN)‐resistant (FeR) strain of Tetranychus cinnabarinus. However, the functions of these SDR genes in acaricide tolerance remain elusive. In this study, the activity of SDRs was found to be significantly higher (2.26‐fold) in the FeR strain compared to the susceptible strain (SS) of T. cinnabarinus. A specific upregulated SDR gene, named SDR112C1, exhibited significant overexpression (3.13‐fold) in the FeR population compared with that in the SS population. Furthermore, the expression of SDR112C1 showed a significant increase in the response to FEN induction. Additionally, knockdown of the SDR112C1 gene resulted in decreased SDR activity and reduced mite viability against FEN. Importantly, heterologous expression and in vitro incubation assays confirmed that recombinant SDR112C1 could effectively deplete FEN. Moreover, the overexpression of the SDR112C1 gene in Drosophila melanogaster significantly decreased the toxicity of FEN to transgenic fruit flies. These findings suggest that the overexpression of SDR SDR112C1 is a crucial factor contributing to FEN tolerance in T. cinnabarinus. This discovery not only enhances our understanding of SDR‐mediated acaricide tolerance but also introduces a new family of detoxification enzymes to consider in practice, beyond cytochrome P450s, carboxyl/choline esterases and glutathione S‐transferases. [ABSTRACT FROM AUTHOR]

So far, the physiological and molecular mechanisms of the impact of arbuscular mycorrhizal fungus (AMF) on Cd absorption, transport and detoxification in Ipomoea aquatica (water spinach) are still unclear. In the present study, a pot experiment was performed to investigate the impact of AMF-Glomus versiforme (Gv) on the photosynthetic characteristics, Cd uptake, antioxidative system and transcriptome in water spinach in the soils supplemented with 5 mg Cd kg−1. Gv inoculation improved significantly the photosynthetic characteristics and growth of water spinach. Furthermore, Gv colonization significantly promoted the activities of catalase (CAT), peroxidase (POD) and glutathione reductase (GR), contents of glutathione (GSH) and ascorbic acid (AsA), and the total antioxidant capacity (TCA), but decreased malondialdehyde (MDA) content in water spinach. In addition, Gv inoculation significantly increased pH in rhizosphere soils and decreased the Cd concentrations and uptakes in water spinach. Importantly, 2670 differentially expressed genes (DEGs) were screened in water spinach root colonized with Gv in 5 mg Cd kg−1 soil, of which 2008 DEGs were upregulated and 662 DEGs were downregulated. Especially, the expression levels of POD, CAT, GR, dehydroascorbate reductase 2 (DHAR2), glutathione S-transferase U8 (GSTU8) and glutathione synthetase (GSHS) and cytochrome P450 (Cyt P450) genes were significantly up-regulated in water spinach inoculated with Gv. Meanwhile, the plant cadmium resistance protein 2 (PCR2), metal tolerance protein 4 (MTP4), ATP-binding cassette transporter C family member (ABCC), ABC-yeast cadmium factor 1 (ABC-YCF1) and metallothionein (MT) genes were also up-regulated in mycorrhizal water spinach. Our results firstly elucidated the mechanism by which AMF reduced the uptake and phytotoxicity of Cd in water spinach through a transcriptome analysis. [ABSTRACT FROM AUTHOR]

Ochratoxin A (OTA) is one of the mycotoxins in the agriculture and livestock sectors. The poultry sector suffered from significant economic losses due to the adverse impacts of OTA on the growth rate, feed conversion ratio, and livability. Thus, the present investigation aimed to determine the impact of chamomile essential oil supplementation against OTA toxicity in growing quails. 360 one‐week‐old growing quails were distributed into six groups (n = 60) with four replicates of 15 birds. The groups were G1 (control negative), G2 (OTA 1 mg/kg diet, control positive), G3 (chamomile oil 0.5 g/kg diet), G4 (chamomile oil 1 g/kg diet), G5 (OTA 1 mg/kg diet + chamomile oil 0.5 g/kg diet), and G6 (OTA 1 mg/kg diet + chamomile oil 1 g/kg diet). Adding OTA significantly (P < 0.05) reduced live body weight and weight gain at 5 weeks. Feed intake at 5 weeks was nonsignificantly reduced in G3 and G4 compared to G1. G4 showed a significant (P < 0.05) increase in weight gain and the lowest feed conversion ratio. The G2 showed the lowest superoxide dismutase (SOD), total antioxidant capacity (TAC), glutathione transferase (GST) activity, and the highest levels of malondialdehyde (MDA). Moreover, they showed a significant improvement in liver enzymes and kidney function tests and a significant (P < 0.05) reduction in the levels of total cholesterol and triglycerides. Chamomile supplementation alone or with OTA significantly (P < 0.05) increased immunoglobulin M, G, A, and complement 3 than OTA alone. Chamomile oil with an OTA diet or alone reduced the negative effects of OTA and improved the performance, antioxidant status, lipid profile, and immunological state of growing Japanese quails. [ABSTRACT FROM AUTHOR]

One of the biggest problems in the treatment of idiopathic Parkinson's disease is the lack of new drugs that slow its progression. L-Dopa remains the star drug in the treatment of this disease, although it induces severe side effects. The failure of clinical studies with new drugs depends on the use of preclinical models based on neurotoxins that do not represent what happens in the disease since they induce rapid and expansive neurodegeneration. We have recently proposed a single-neuron degeneration model for idiopathic Parkinson's disease that requires years to accumulate enough lost neurons for the onset of motor symptoms. This single-neuron degeneration model is based on the excessive formation of aminochrome during neuromelanin synthesis that surpass the neuroprotective action of the enzymes DT-diaphorase and glutathione transferase M2-2, which prevent the neurotoxic effects of aminochrome. Although the neurotoxic effects of aminochrome do not have an expansive effect, a stereotaxic injection of this endogenous neurotoxin cannot be used to generate a preclinical model in an animal. Therefore, the aim of this review is to evaluate the strategies for pharmacologically increasing the expression of DT diaphorase and GSTM2-2 and molecules that induce the expression of vesicular monoamine transporter 2, such as pramipexole. [ABSTRACT FROM AUTHOR]

Glutathione transferases (GSTs) are the primary catalysts protecting from reactive electrophile attack. In this review, the quantitative levels and distribution of glutathione transferases in relation to physiological function are discussed. The catalytic properties (random sequential) tell us that these enzymes have evolved to intercept reactive intermediates. High concentrations of enzymes (up to several hundred micromolar) ensure efficient protection. Individual enzyme molecules, however, turn over only rarely (estimated as low as once daily). The protection of intracellular protein and DNA targets is linearly proportional to enzyme levels. Any lowering of enzyme concentration, or inhibition, would thus result in diminished protection. It is well established that GSTs also function as binding proteins, potentially resulting in enzyme inhibition. Here the relevance of ligand inhibition and catalytic mechanisms, such as negative co-operativity, is discussed. There is a lack of knowledge pertaining to relevant ligand levels in vivo, be they exogenous or endogenous (e.g., bile acids and bilirubin). The stoichiometry of active sites in GSTs is well established, cytosolic enzyme dimers have two sites. It is puzzling that a third of the site's reactivity is observed in trimeric microsomal glutathione transferases (MGSTs). From a physiological point of view, such sub-stoichiometric behavior would appear to be wasteful. Over the years, a substantial amount of detailed knowledge on the structure, distribution, and mechanism of purified GSTs has been gathered. We still lack knowledge on exact cell type distribution and levels in vivo however, especially in relation to ligand levels, which need to be determined. Such knowledge must be gathered in order to allow mathematical modeling to be employed in the future, to generate a holistic understanding of reactive intermediate protection. [ABSTRACT FROM AUTHOR]

Sri Lankan cassava mosaic virus (SLCMV) is a prominent causative agent of cassava mosaic disease in Asia and relies on the whitefly Bemisia tabaci cryptic complex for its transmission. However, the molecular mechanisms involved in SLCMV transmission by B. tabaci have yet to be understood. In this study, we identified an aminopeptidase N‐like protein (BtAPN) in B. tabaci Asia II 1, an efficient vector of SLCMV, which is involved in the SLCMV transmission process. Through the use of glutathione S‐transferase pull‐down assay and LC‐MS/MS analysis, we demonstrated the interaction between BtAPN and the coat protein (CP) of SLCMV. This interaction was further confirmed in vitro, and we observed an induction of BtAPN gene expression following SLCMV infection. By interfering with the function of BtAPN, the quantities of SLCMV were significantly reduced in various parts of B. tabaci Asia II 1, including the whole body, midgut, hemolymph, and primary salivary gland. Furthermore, we discovered that BtAPN is conserved in B. tabaci Middle East‐Asia Minor 1 (MEAM1) and interacts with the CP of tomato yellow leaf curl virus (TYLCV), a begomovirus known to cause severe damage to tomato production. Blocking BtAPN with antibody led to a significant reduction in the quantities of TYLCV in whitefly whole body and organs/tissues. These results demonstrate that BtAPN plays a generic role in interacting with the CP of begomoviruses and positively regulates their acquisition by the whitefly. [ABSTRACT FROM AUTHOR]

Simple Summary: Varroa destructor is a mite that parasitises honeybee hives, weakening colonies and transmitting viruses. Beekeepers apply acaricide treatments to bee colonies to limit the spread of Varroa and the resulting negative effects on bee health. Oxalic acid treatment is the most widely applied, but little is known about its potential effects on the physiology of bees, particularly their immune system. This study investigated the short-term effects of oxalic acid treatment on the immune and antioxidant systems of house bees (i.e., glucose oxidase, phenoloxidase, glutathione S-transferase, catalase activities, and vitellogenin content). Residual concentrations of oxalic acid on the bees' external body and in the haemolymph were also measured. The results showed that the treatment did not affect the concentration of oxalic acid in the haemolymph, in which it was constitutively present. Residues of oxalic acid remained on the outer body of the bees for up to 48 h after treatment. Both glucose oxidase activity and vitellogenin concentration were affected by the treatment, increasing significantly after 48 and 24 h, respectively. No effects were observed on the other parameters investigated (phenoloxidase, glutathione S-transferase, and catalase activities). Honeybees (Apis mellifera L.) have to face many challenges, including Varroa destructor infestation, associated with viral transmission. Oxalic acid is one of the most common treatments against Varroa. Little is known about the physiological effects of oxalic acid, especially those on honeybees' immune systems. In this study, the short-term effects (0–96 h) of oxalic acid treatment on the immune system components (i.e., glucose oxidase, phenoloxidase, glutathione S-transferase, catalase activities, and vitellogenin contents) of house bees were preliminarily investigated. Oxalic acid contents of bee bodies and haemolymphs were also measured. The results confirm that oxalic acid is constitutively present in bee haemolymphs and its concentration is not affected by treatment. At 6 h after the treatment, a maximum peak of oxalic acid content was detected on bees' bodies, which gradually decreased after that until physiological levels were reached at 48 h. In the immune system, the oxalic acid treatment determined a peak in glucose oxidase activity at 48 h, indicating a potential defence response and an increase in vitellogenin content at 24 h. No significant changes were recorded in phenoloxidase, glutathione S-transferase, and catalase activities. These results suggest a time-dependent response to oxalic acid, with potential immune system activation in treated bees. [ABSTRACT FROM AUTHOR]

While research has identified several inhibitors of the main protease (Mpro) of SARS-CoV-2, a significant portion of these compounds exhibit reduced activity in the presence of reducing agents, raising concerns about their effectiveness in vivo. Furthermore, the conventional biosafety level 3 (BSL-3) for cellular assays using viral particles poses a limitation for the widespread evaluation of Mpro inhibitor efficacy in a cell-based assay. Here, we established a BSL-1 compatible cellular assay to evaluate the in vivo potential of Mpro inhibitors. This assay utilizes mammalian cells expressing a tagged Mpro construct containing N-terminal glutathione S-transferase (GST) and C-terminal hemagglutinin (HA) tags and monitors Mpro autodigestion. Using this method, GC376 and boceprevir effectively inhibited Mpro autodigestion, suggesting their potential in vivo activity. Conversely, carmofur and ebselen did not exhibit significant inhibitory effects in this assay. We further investigated the inhibitory potential of selenoneine on Mpro using this approach. Computational analyses of binding energies suggest that noncovalent interactions play a critical role in facilitating the covalent modification of the C145 residue, leading to Mpro inhibition. Our method is straightforward, cost-effective, and readily applicable in standard laboratories, making it accessible to researchers with varying levels of expertise in infectious diseases. [ABSTRACT FROM AUTHOR]

Tetrabromobisphenol A (TBBPA) is a fire-retardant containing bromine, produced in large quantities worldwide and extensively used in several industrial products. This compound was identified as a potential contaminant of the environment, causing toxicity to organisms. However, its toxicity remains poorly understood in marine bivalves. The first objective of this work was to evaluate the impact of TBBPA on mussels (Mytilus galloprovincialis) exposed for 28 days to various concentrations of TBBPA (0, 1, 10, and 100 µg·L−1), by assessing stress biomarkers' responses (Glutathione S-transferase, superoxide dismutase, catalase, lipid peroxidation, total antioxidant capacity, total ubiquitin, caspase-3 and acetylcholinesterase). The results showed that lower concentrations (1 and 10 µg·L−1) were efficiently detoxified, as suggested by GST activities, which were supported by the responses of the other biomarkers. The most pronounced effects were observed in animals exposed to the highest concentration of TBBPA (100 µg·L−1), suggesting oxidative stress. Additionally, significant strong correlations were found between total antioxidant capacity and some biomarkers (superoxide dismutase and lipid peroxidation), showing that processes involved in oxidative stress fighting are working to avoid cell injury. In brief, mussels' defense mechanisms were capable of dealing with exposure to the lower concentrations tested. Despite this, the risk of consuming shellfish or other fishery products contaminated with TBBPA should be a cause for concern. [ABSTRACT FROM AUTHOR]