The emergence of carbapenem resistance in gram-negative bacteria such as Escherichia coli is one of the world's most urgent public health problems. E. coli, which encounter a diverse range of niches in host can rapidly adapt to the changes in surrounding environment by coordinating their behavior via production, release and detection of signal molecules called autoinducers through a cell density dependent communication system known as quorum sensing. Here, in this study we investigated whether imipenem, and acyl homoserine lactone quorum sensing signal molecules influence the transcriptional response within lsr and lsrRK operon which are associated with auto inducer-2 mediated quorum sensing in E. coli. Two E. coli isolates carrying blaNDM were treated with 10% SDS for 20 consecutive days, resulting in the successful elimination of the blaNDM encoding plasmid from one isolate. Plasmid was extracted from the isolate and was transformed into recipient E. coli DH5α by electroporation. The native type, plasmid-cured type, transformant, and E. coli DH5α were allowed to grow under eight different inducing conditions and the transcriptional responses of lsr and lsrRK operons were studied by quantitative real-time PCR method.

Triterpenoids are well-known pharmacological components of Sanghuangporus fungi, such as Sanghuangporus lonicericola. This study investigates the inductive effects of paclobutrazol (PBZ) on triterpenoid biosynthesis in the submerged fermentation of S. lonicericola and explores the induction mechanisms via multi-omics and genetic methods. The addition of 100 mg/L PBZ significantly increases the triterpenoid yield by 151.39%. A total of 29 triterpenoids are tentatively identified, of which 18 are newly presented only under PBZ induction. Moreover, 30 genes involved in the MVA pathway and 31 genes encoding cytochrome P450 monooxygenases assumed to be responsible for decoration are identified. Finally, a MYB transcription factor (SlMYB) is identified and found to be downregulated under paclobutrazol induction. Genetic manipulation of SlMYB demonstrates its negative regulatory effect on four putative target genes, including ACAT, MVD, IDI, and FDPS. Electrophoretic mobility gel shift assays verify the direct interactions with the promoters of MVD, IDI, and FDPS. Taken together, PBZ acts as an effective inducer of triterpenoid biosynthesis in S. lonicericola, and the transcription factor SlMYB is negatively regulated.

Gray mold, caused by Botrytis cinerea, poses an escalating threat to ginseng production in China. Excessive application of chemical fungicides has resulted in severe resistance development. To elucidate the resistance mechanism of B. cinerea to pyrimethanil, transcriptomic comparisons were conducted between resistant (HRG21) and sensitive (FSG43) strains following exposure to pyrimethanil for 2 and 6 h. GO and KEGG analyses identified differential expression of genes associated with ABC and MFS transporters as well as methionine biosynthesis. qRT-PCR validation confirmed a marked upregulation of ABC (BcatrA, BcatrB, BcatrD, BcatrO, and Bmr3) and MFS (Bchex1 and BcmfsM2) transporter genes in HRG21, whereas in FSG43, ABC (BcatrA, BcatrB, BcatrD, and BcatrO) and MFS (Bchex1) transporter genes were downregulated. No significant transcriptional changes were observed in Mrr1, a transcription factor gene, following pyrimethanil exposure. Molecular docking analysis identified potential binding sites and interaction targets for five key transporters (BcatrB, BcatrA, BcatrD, Bmr3, and Bchex1). Furthermore, exogenous methionine supplementation partially mitigated pyrimethanil toxicity in FSG43. However, amino acid sequences of enzymes involved in methionine biosynthesis (BcmetC, BcStr2, Bcmet2, Bcmxr1, and Bcmxr2) exhibited no variation between FSG43 and HRG21. The results indicate that BcatrB plays a central role in pyrimethanil resistance, while methionine contributes only marginally to resistance mechanisms.

Gemcitabine combined with cisplatin is the first-line chemotherapy for advanced cholangiocarcinoma, but drug resistance remains a challenge, leading to unsatisfactory therapeutic effect. Here, we elucidate the possibility of chemotherapy regimens sensitized by inhibiting succinylation in patients with cholangiocarcinoma from the perspective of post-translational modification. Our omics analysis reveals that succinylation of PDHA1 lysine 83, a key enzyme in the tricarboxylic acid cycle, alters PDH enzyme activity, modulates metabolic flux, and leads to alpha-ketoglutaric acid accumulation in the tumor microenvironment. This process activates the OXGR1 receptor on macrophages, triggering MAPK signaling and inhibiting MHC-II antigen presentation, which promotes immune escape and tumor progression. Moreover, we show that inhibiting PDHA1 succinylation with CPI-613 enhances the efficacy of gemcitabine and cisplatin. Targeting PDHA1 succinylation may be a promising strategy to improve treatment outcomes in cholangiocarcinoma and warrants further clinical exploration.

Parkinson's disease (PD) poses a global menace, as the available treatment methods solely aim to mitigate symptoms. An effective strategy to address the pathogenesis of PD involves eliminating the accumulation of aggregated alpha-synuclein, emphasizing the role of epigenetics. Aberrant epigenetic changes significantly influence gene expression, which is pivotal in PD progression, impacting neuronal growth and degeneration. Epigenetic-related genes are regulated by histone modification and DNA methylation processes. Nevertheless, their significance in PD has not been confirmed. This research was carried out using both in vitro and in vivo approaches. In the in vitro investigations, N2A neuronal cell lines were utilized, and the neuroprotective effect of decitabine (DB) was observed at concentrations of 0.1 μM and 0.5 μM. In the in vivo study, PD induction led to significant motor deficits, which were notably ameliorated at the highest treatment dose. This improvement was accompanied by a marked attenuation of inflammatory mediators, including TNF-α, IL-6, IL-1β, and CRP levels. Additionally, there was a significant enhancement in antioxidative defense, evidenced by increased GSH (glutathione) levels and reduced oxidative stress marker NO (nitric oxide). Neurochemical analysis revealed a substantial rise in dopamine levels, a critical PD marker, alongside an elevation in BDNF, indicating neuroprotective effects. Furthermore, gene expression analysis indicated a notable upregulation in the mRNA expression of epigenetic genes and proteins linked to PD pathology. Histological assessments, including IHC, H&E, and CV staining of the substantia nigra, showed enhanced structural integrity following treatment. Collectively, these insights reveal DB's promise as a therapeutic solution for mitigating PD symptoms and pathology exacerbated by 6-OHDA.

Catharanthus roseus has various terpenoid indole alkaloids (TIAs) with adaptive mechanisms to withstand both biotic and abiotic stresses. We investigated the effects of methyl jasmonate (MeJA) on alternative splicing (AS) mechanisms in C. roseus to identify differentially expressed alternatively spliced (DAS) genes following MeJA treatment. We found pairs of co-expressed splicing factors (SFs) and DAS genes and potential roles of co-expressed SFs in the maturation of their respective transcripts. Twenty two clusters encompassing 17 MeJA-responsive DAS genes co-expressed with 10 SF genes. DAS genes, C3H62 , WRK41 , PIL57 , NIP21 , and EDL6 , exhibited co-expression with the SF genes SR34a , DEAD29 , SRC33 , DEAH10 , and DEAD29 , respectively. These gene pairs are implicated in plant developmental processes and/or stress responses. We suggest that MeJA activates the expression of genes encoding SFs that are regulated in tandem with their co-expressed DAS genes and MeJA may enhance the regulatory frameworks that control splicing mechanisms, resulting in the generation of specific mRNA isoforms. This triggers the expression of particular DAS gene variants to allow the plant to effectively respond to environmental stimuli and developmental signals. Our study advances our understanding on how MeJA modulates alternative splicing in C. roseus , potentially influencing various aspects of plant physiology and metabolism. It is recommended that future studies focus on validating the functional relationships between the identified SF/DAS gene pairs and their specific roles in plant development and stress responses, and exploring the potential of manipulating these splicing mechanisms to enhance the production of valuable TIAs in C. roseus .

A significant issue in treating bacterial infections is multidrug resistance (MDR) microbes. Drug efflux pumps that reduce cellular drug accumulation are frequently linked to drug resistance. In this study, we set out to determine the effects of CE-MA truncated peptide derivatives against MDR Mycobacterium tuberculosis. Following the assessment of the minimum inhibitory concentrations (MICs) of these peptides against MDR Mycobacterium tuberculosis, a Real-Time PCR was used to examine the expression of six drug efflux pump genes. Next, an MTT assay was performed to test the cytotoxicity of peptides against the A549 cell line. The outcomes demonstrated that CE-MA significantly upregulated gene expression of mmr, and Rv0876c (⩾ 4-fold) than untreated bacteria. Also, under CMt2 stress, significant overexpression of Rv0876c and drrA was seen. However, the results show that upregulation in CMt2-treated bacteria in comparison CE-MA treated bacteria is significantly less for genes tap (P < 0.05), mmr (P < 0.0001), and Rv0876c (P < 0.001). Meanwhile, CMt1 only upregulated the Rv0876c gene and downregulated gene expression of tap, drrA, and mmr. It was also found that all three peptides have no significant effect (P > 0.05) on changing the expression of genes drrC and pstB. Less than 10% of the A549 cell line was susceptible to the toxicity of CMt1 and CMt2 at their MICs range. Our results emphasize the significance of investigating novel peptide-based approaches to combat MDR Mycobacterium tuberculosis and point to these peptides as prospective candidates for additional research.

TNBC is an aggressive metastatic cancer that poses considerable treatment challenges because of its acquired drug resistance towards the existing targeted and hormonal therapies. The epigenetic modulation including HDACs triggers the EMT in TNBC which produces a more aggressive tumor phenotype. Chemotherapy and radiotherapy cause severe side effects which make treatment complex and challenging. To avoid these serious side effects and boost the effectiveness of current anti-cancer medications, plant flavonoids have been investigated.

Metformin is an antidiabetic drug used in type 2 diabetes as well as indicators in polycystic ovary syndrome (PCOS) and cancer. Due to their increase in popularity, high amounts of metformin are being released into aquatic environments. However, the toxic effect of metformin on embryonic development in aquatic organisms remains limited. Therefore, this study aimed to elucidate the lethal embryotoxicity of metformin and determine the underlying molecular pathways influencing embryonic development using a zebrafish model through multi-omics analysis. Metformin was microinjected into zebrafish embryos at the 1-cell stage with varying concentrations (50 mM, 100 mM, 200 mM, 400 mM, and 800 mM). From the results, hatching rates decreased in a dose dependent manner. Fetal malformation and mortality (LC50 = 339.8 mM) increased in a dose dependent manner. In situ hybridization of whole-embryo assays demonstrated that metformin exerts a significant impact on the initial stages of embryonic development, leading to aberrant differentiation of the germ layers, perturbed organogenesis, and delayed development. Furthermore, transcriptomics, metabolomics, and lipidomics were used to study the molecular mechanisms of embryonic toxicity. The results showed that the cell cycle, dorsoventral axis formation, and collecting duct acid secretion pathways were significantly altered in treated embryos. In brief, these results provide useful information on the lethal toxicity mechanism of metformin overdose and provide clues for further studies in humans.

Endocrine-disrupting chemicals (EDCs) in the aquatic environment are an emerging concern and can lead to adverse health effects on humans and aquatic life. EDCsare ubiquitous in several daily use and personal care products and ubiquitous in aquatic ecosystems. The aquatic ecosystems also serve as major sinks of EDCs and have even been found to accumulate in aquatic organisms. Fish are an important sentinel species in the aquatic system and are a reliable indication of environmental water pollution. In the present study, we have assessed the immunotoxicity effects of three important EDCs, i.e., triclosan (TCS), bisphenol A (BPA), and diethyl phthalate (DEP). There is mounting evidence that EDCs impact several physiological systems, including fish immune systems. Hence, to better understand the immune system's complexity, we have investigated how EDCs alter the immune responses and can aggravate immunotoxicity using Labeo catla as a model fish species. The results showed significant upregulation of immune gene expression; exposure to EDCs differentially modulates immunity across the different organs (liver and brain) of Labeo catla. The present study highlighted that endocrine-disrupting compounds (TCS, BPA, and DEP) have a significant immunotoxicity effect in fish and activate several immunological pathways to control the toxic effect and maintain homeostasis. The results also indicate that immune genes can be used as a biomarker for EDC toxicity. However, further studies need to see how immune-disrupting effects happen at actual exposure levels in the environment to EDCs.

Selenium is an indispensable nutrient for plants, and optimizing selenium levels can enhance plant growth and metabolism, leading to improved yield and quality. In comparison to conventional inorganic or organic selenium fertilizers, nano-selenium demonstrates superior safety and enhanced biological activity, making it more suitable for crop production. Although nano-selenium fertilizer is extensively used in various crops, its application in pak choi remains limited. As a vital source of selenium, previous research on pak choi (Brassica chinensis var. pekinensis cv. 'Suzhouqing') has primarily focused on investigating physiological effects with limited exploration of the molecular mechanism. Therefore, this study aims to investigate the impact of nano-selenium on pak choi through an integrated analysis of transcriptome and metabolome. Specifically, we examined the effects of different concentrations of nano-selenium (0, 5, 10 and 20 mg L-1) on the growth and nutritional quality of Suzhouqing. The findings revealed that a low concentration (5 mg L-1) of nano-selenium significantly increased leaf weight and total selenium content, while modulating primary metabolites such as soluble amino acids, proteins, sugars and ascorbic acid. Additionally, it influenced secondary metabolites including glucosinolates, phenolic acids and flavonoids. Consequently, this enhancement in growth performance and nutritional quality was attributed to the regulation of pathways involved in selenocompound metabolism, phenylpropanoid biosynthesis, and flavonoid biosynthesis by key enzymes such as methionine S-methyltransferase, 5-methyltetrahydrofolate-homocysteine methyltransferase, kynurenine-oxoglutarate transaminase, thioredoxin reductase, phenylalanine ammonian-lyase, 4-coumarate-CoA ligase, flavonoid 3', 5'-hydroxylase, naringenin 3-dioxygenase, flavonol synthase and bifunctional dihydroflavonol 4-reductase. These results provide comprehensive insights into the physiological and molecular mechanisms underlying the influence of nano-selenium on plant growth and nutritional quality. Therefore, they offer a solid theoretical basis and technical support for breeding and cultivation strategies aimed at producing selenium-rich pak choi.

This research aimed to explore the effect of Phyllanthus emblica powder on the growth performance, blood hematology, body composition, serum biochemistry, organ histology, gene expression and resistance of Nile Tilapia (Oreochromis niloticus) against Aspergillus flavus. A total of 240 fish (30.5 ± 2.0 g) were distributed in 12 ponds with 20 fish per pond and fed a basal diet supplemented with 0, 1%, 2% and 3% of P. emblica powder for 60 days. A challenge experiment was performed at the end of the trial. All supplemented groups showed better growth performance (final body weight, weight gain, and specific growth rates) (P < 0.05), while feed intake did not affect (P > 0.05). The feed conversion ratio enhanced by the inclusion of dietary P. emblica (P < 0.05). In terms of body composition, crude lipids decreased, while ash percentages increased after fish were fed with P. emblica (P < 0.05). Liver function decreased significantly, while protein fraction, blood hematology and digestive enzymatic profile (amylase, and lipase) were significantly increased in all treated groups. Fish fed with P. emblica exhibited a higher survival rate and increased resistance to A. flavus compared to the control diet. Diets containing P. emblica improved the histoarchitecture of hepatopancreatic and intestine of Nile Tilapia. The inclusion of P. emblica significantly upregulated the expression of CC chemokine, IL-1β, IL-8, SOD, and GPx genes, with a gradual increase in P. emblica levels in the diet (P < 0.05). In conclusion, dietary supplementation with P. emblica powder may serve as an effective strategy for promoting growth and acting as an immunostimulant in Nile Tilapia aquaculture.

In the leaves of C3 species such as rice (Oryza sativa), mesophyll cells contain the largest compartment of photosynthetically active chloroplasts. In contrast, plants that use the derived and more efficient C4 photosynthetic pathway have a considerable chloroplast compartment in both bundle sheath and mesophyll cells. Accordingly, the evolution of C4 photosynthesis from the ancestral C3 state required an increased chloroplast compartment in the bundle sheath. Here, we investigated the potential to increase chloroplast compartment size in rice bundle sheath cells by manipulating brassinosteroid signaling. Treatment with brassinazole, a brassinosteroid biosynthesis inhibitor, raised leaf chlorophyll content and increased the number but decreased the area of chloroplasts in bundle sheath cells. Ubiquitous overexpression of the transcription factor-encoding BRASSINAZOLE RESISTANT 1 (OsBZR1) increased bundle sheath chloroplast area by up to 45%, but these plants became chlorotic. However, when OsBZR1 expression was driven by a bundle sheath-specific promoter, the negative effects on growth and viability were alleviated while chloroplast area still increased. In summary, we report a role for brassinosteroids in controlling chloroplast area and number in rice and conclude that cell-specific manipulation of brassinosteroid signaling can be used to manipulate the chloroplast compartment in rice bundle sheath cells.

To explore the underlying mechanism behind the fine particulate matter's (PM2.5)-mediated regulation of reproductive function in male rats, and to determine the role of vitamins in this process.

Calcium-dependent protein kinases (CPKs) play crucial roles in plant guard cell signal transduction. Ethylene is known to induce stomatal closure, with the hydrogen peroxide (H2O2)-nitric oxide (NO) signalling module being pivotal to this process. However, the specific roles of CPKs in this process and their interactions with H2O2 and NO remain unclear. In this study, we screened Arabidopsis mutants of nine CPKs and found that in the loss-of-function mutants for CPK3, CPK4, CPK6, CPK11, CPK21, and CPK33, exogenous ethylene failed to induce stomatal closure, indicating that these CPKs act as positive regulators in ethylene-induced stomatal closure. Mutants' stomatal responses to H2O2 and NO treatment and changes of endogenous H2O2 and NO levels in guard cells upon ethylene treatment indicated that CPK3, CPK4, CPK11, and CPK33 function upstream of the H2O2-NO module, while CPK6 and CPK21 act downstream. Furthermore, NADPH oxidases play critical roles in ethylene-induced H2O2 production. We identified the interactions of CPK3, CPK4, and CPK11 with AtRBOHF, and CPK4 and CPK11 with AtRBOHD using four different assays, and exogenous ethylene enhanced these interactions. These results suggest that CPK3, CPK4, and CPK11 may mediate ethylene-induced H2O2 formation in guard cells through their interactions with AtRBOHD/F. Additionally, exogenous ethylene significantly upregulates the expression of CPK3, CPK4, CPK6, CPK11 and CPK21, providing a potential mechanism by which ethylene modulates CPKs. Our findings not only establish the role of CPKs in ethylene guard cell signalling but also offer insights into the mechanism by which ethylene activates NADPH oxidases to initiate H2O2 production.

Seed priming is an effective way to activate defense mechanisms before germination, enhancing seed vigor and stress resistance. Ascorbic acid (AsA) is an important signaling molecule that plays a crucial role in balancing cellular reactive oxygen species. However, whether AsA priming can enhance seed vigor in oat (Avena sativa) and the underlying mechanisms remain unclear. This study primed aged seeds (controlled deterioration at 45°C for 5 days) with 1.5 mM AsA for 24 h. Primed seeds were then sampled after 36 h of imbibition for seed assays. Significant increases in germination percentage, vigor index, shoot and root length, coupled with a significant reduction in mean germination time, demonstrated that AsA priming effectively restored seed vigor. Ultrastructural observations of mitochondria isolated from embryos presented that AsA priming repaired structural damage in aged seeds, with intact double membranes and clear internal cristae observed. Excessive H2O2 accumulation was discovered in mitochondria of aged seeds, while AsA priming reduced H2O2 levels by increasing the activities of CAT, GR, MDHAR and DHAR. AsA priming also increased antioxidant content, particularly DHA, contributing to reduced oxidative stress. Furthermore, transcriptomic analysis highlighted the upregulation of genes associated with antioxidant defense, including APX, CAT, DHAR and MDHAR, indicating enhanced repair and protection pathways in the mitochondrial AsA-GSH cycle. This suggests that AsA priming would increase the activity of antioxidant enzymes, the content of antioxidants, and expression of genes related to AsA-GSH cycle in aged oat seeds, which was conducive to repairing mitochondrial damage and enhancing seed vigor.

Bacteria utilize two-component system (TCS) signal transduction pathways to sense environmental and physiological stimuli and mount appropriate responses. In opportunistic pathogens such as Staphylococcus aureus, TCSs activate virulence programs in response to host defense systems. Due to their critical role in pathogenesis, TCSs are important targets for antivirulence drug discovery campaigns. However, challenges associated with screening TCSs in pathogens and in vitro have limited the output of such efforts to a small number of characterized drug candidates. Here, we functionally express the S. aureus virulence-regulating TCS SaeRS from synthetic gene regulatory elements in the model bacterium Bacillus subtilis to reliably screen this system against a small molecule library under simple culturing conditions. Our approach reveals the compound NSC97920 as a strong inhibitor of SaeRS signaling. We combine in situ, in vivo, in silico, and in vitro characterization to demonstrate that NSC97920 suppresses the critical step of autophosphorylation in the SaeS histidine kinase, resulting in strong antivirulence activity. Our work shows that heterologous expression and screening of TCSs in model bacteria could accelerate the development of therapeutics against antibiotic-resistant pathogens.

Cadmium (Cd) contamination in maize poses a significant threat to global food security due to its persistent accumulation in crops. In this study, the effects of foliar application of glycerol on Cd accumulation in maize seedlings were studied. Our results demonstrated that under Cd treatment, biomass, total chlorophyll content, net photosynthetic rate (Pn), Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) activity, Phosphoenolpyruvate carboxylase (PEPC) activity, sucrose levels, and carbohydrate levels in maize seedlings significantly increased after glycerol application. H2O2 and MDA levels in both the aboveground and belowground portions of the maize plants significantly decreased. Moreover, superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities in the aboveground parts significantly increased. Notably, maize plants used glycerol to chelate Cd, which was fixed within the cell wall and soluble fraction of the roots, reducing Cd transport to the shoots and significantly lowering the Cd transport coefficient (TF). Transcriptomic data suggested that glycerol-mediated alleviation of Cd stress in maize seedlings may be associated with phenylpropanoid biosynthesis, plant-pathogen interactions and photosynthesis pathways. These molecular patterns align with the observed physiological improvements. This study provided a novel approach to effectively alleviate excessive Cd in maize and suggested possible applications of glycerol in cultivating plant resistance to heavy metals.

Esophageal squamous cell carcinoma (ESCC) is a deadly cancer with a poor prognosis and a high recurrence rate after chemotherapy, posing a significant clinical challenge. To elucidate the molecular basis of chemotherapy (chemo)-resistance and to develop methods to effectively eliminate chemo-resistant tumor clones, we established an ESCC organoid (ESCCO) library from 24 ESCC patients of various stages, ages, and treatments. These ESCCOs faithfully recapitulate the oncogenic mutations observed in the original ESCC tissues and manifest tumorigenic properties when xenografted. The ESCCOs respond differently to cisplatin and 5-fluorouracil, chemotherapeutic agents commonly used to treat ESCC patients, with 7 ESCCOs exhibiting potent chemo-resistance. Notably, the chemo-resistant ESCCOs show higher genes involved in antioxidant stress response pathways and more accessible chromatin at their loci than the sensitive ESCCOs. These genes can serve as valuable biomarkers to stratify chemo-resistant ESCCs in histopathological specimens. Through drug screening using the ESCCO library, we reveal that fedratinib effectively induces cell death in chemo-resistant ESCCOs. Collectively, our human ESCCO model offers novel insights into the mechanism of chemo-resistance in ESCCs, which is critical for developing effective therapeutic approaches to eradicate the recurrence of ESCCs.

Protein ubiquitination is a dynamic and reversible process involved in gene transcription, protein metabolism, and cellular apoptosis. Ubiquitin specific proteases (USPs), as the largest family of deubiquitinating enzymes, are able to remove the ubiquitin from target proteins, rescuing them from degradation. Here, we characterized the small molecule antitumor agent YM155 as a broad-spectrum USP inhibitor. By inhibiting the deubiquitinase activity of multiple USPs, YM155 causes the degradation of oncogenic substrate proteins, such as c-Myc and intracellular domain of Notch1. In cancers driven by these proteins, YM155 induces profound cell apoptosis and markedly inhibits tumor growth in xenograft models. Together, these findings demonstrate that YM155 is a broad-spectrum USP inhibitor, and a potential drug candidate for cancers which depend on hyper-active oncogenic proteins that are regulated by the ubiquitin-proteasome pathway.