Two-Dimensional Layered Nano-MoS2 Induces Earthworm Immune Cell Apoptosis by Regulating Lysosomal Maintenance and Function: Toward Unbiased Screening and Validation of Suspicious Pathways.

Molybdenum-based nanosheets (NSMoS2) are increasingly applied in various fields and undergoing relevant risk evaluations on subjectively hypothesized toxicity pathways. However, risk assessment should be unbiased and focus on appropriate end points to avoid biased prescreening. Here, we developed an adverse biological outcome screening strategy based on nontargeted functional protein profiles in earthworm (Eisenia fetida) immune cells exposed to NSMoS2 and their ionic counterpart (Na2MoO4). Through this framework, the apoptosis-related processes with distinct mechanisms were rapidly identified and thoroughly validated phenotypically. Specifically, upon exposure to 50 µg Mo/mL Na2MoO4, cellular signaling and energy homeostasis were disrupted within the transcription-translation biological chain. The autophagic pathway was activated, which, together with energy deprivation, phenotypically induced significant autophagy that ultimately led to apoptosis. In contrast, NSMoS2, tested at the same concentration, caused a reprogramming of apoptotic gene and protein expressions. Transcriptome plasticity facilitated the endocytic-adaptive transcriptional profile characterized by cytoskeleton remodeling and lysosome organization/movement under NSMoS2 exposure. Subcellular dynamics further revealed NSMoS2-induced lysosomal damage with a time-sensitive physiological window, ultimately mediating apoptosis. These findings provide a mechanistic and visual understanding of the distinct risk profile of NSMoS2 compared to molybdate, highlighting the importance of integrating nontargeted screening and phenotypic validation in early risk warning.

A glycosylation signature for predicting the progression and immunotherapeutic response of prostate cancer.

BACKGROUND: Glycosylation has been proposed as a new cancer hallmark. However, focusing on specific glycans or glycoproteins may lose much data relevant to glycosylation alterations. The present study aimed to first comprehensively investigate the expression and mutation profiles of glycosylation-related genes (GRgenes) in prostate cancer (PCa) and then develop a glycosylation signature and explore its role in predicting the progression and immunotherapeutic response of PCa. METHODS: Based on The Cancer Genome Atlas database, we comprehensively screened potential prognostic GRgenes and analyzed their expression and mutation profiles in PCa. Through consensus clustering analysis, the study cohort was classified to investigate the effect of glycosylation patterns on the prognosis of PCa. Next, we developed a glycosylation signature (i.e., the glycosylation score [Gly_score]) using the differentially expressed genes between glycosylation pattern groups and evaluated its role in predicting the progression and immunotherapeutic response of PCa. RESULTS: We identified two distinct glycosylation patterns in PCa and found that GRgene expression patterns rather than mutations are associated with the prognosis of PCa. The high Gly_score group had significantly shorter progression-free survival, lower PD-L1 levels, less infiltration of immune cells and lower immunophenoscores than the low Gly_score group. When the patients were grouped according to both the Gly_score and PD-L1 level, patients with a combination of low Gly_score and low PD-L1 expression had the best survival outcomes. CONCLUSIONS: In the present study, for the first time, we developed a glycosylation signature and demonstrated that the proposed glycosylation signature is a promising tool for predicting the prognosis and immunotherapeutic response of PCa.

Coupled Lipidomics and Digital Pathology as an Effective Strategy to Identify Novel Adverse Outcome Pathways in Eisenia fetida Exposed to MoS2 Nanosheets and Ionic Mo.

Molybdenum disulfide (MoS2) nanosheets are increasingly applied in several fields, but effective and accurate strategies to fully characterize potential risks to soil ecosystems are lacking. We introduce a coelomocyte-based in vivo exposure strategy to identify novel adverse outcome pathways (AOPs) and molecular endpoints from nontransformed (NTMoS2) and ultraviolet-transformed (UTMoS2) MoS2 nanosheets (10 and 100 mg Mo/L) on the earthworm Eisenia fetida using nontargeted lipidomics integrated with transcriptomics. Machine learning-based digital pathology analysis coupled with phenotypic monitoring was further used to establish the correlation between lipid profiling and whole organism effects. As an ionic control, Na2MoO4 exposure significantly reduced (61.2-79.5%) the cellular contents of membrane-associated lipids (glycerophospholipids) in earthworm coelomocytes. Downregulation of the unsaturated fatty acid synthesis pathway and leakage of lactate dehydrogenase (LDH) verified the Na2MoO4-induced membrane stress. Compared to conventional molybdate, NTMoS2 inhibited genes related to transmembrane transport and caused the differential upregulation of phospholipid content. Unlike NTMoS2, UTMoS2 specifically upregulated the glyceride metabolism (10.3-179%) and lipid peroxidation degree (50.4-69.4%). Consequently, lipolytic pathways were activated to compensate for the potential energy deprivation. With pathology image quantification, we report that UTMoS2 caused more severe epithelial damage and intestinal steatosis than NTMoS2, which is attributed to the edge effect and higher Mo release upon UV irradiation. Our results reveal differential AOPs involving soil sentinel organisms exposed to different Mo forms, demonstrating the potential of liposome analysis to identify novel AOPs and furthermore accurate soil risk assessment strategies for emerging contaminants.

Earthworm Coelomocyte Internalization of MoS2 Nanosheets: Multiplexed Imaging, Molecular Profiling, and Computational Modeling.

Fully understanding the cellular uptake and intracellular localization of MoS2 nanosheets (NSMoS2) is a prerequisite for their safe applications. Here, we characterized the uptake profile of NSMoS2 by functional coelomocytes of the earthworm Eisenia fetida. Considering that vacancy engineering is widely applied to enhance the NSMoS2 performance, we assessed the potential role of such atomic vacancies in regulating cellular uptake processes. Coelomocyte internalization and lysosomal accumulation of NSMoS2 were tracked by fluorescent labeling imaging. Cellular uptake inhibitors, proteomics, and transcriptomics helped to mechanistically distinguish vacancy-mediated endocytosis pathways. Specifically, Mo ions activated transmembrane transporter and ion-binding pathways, entering the coelomocyte through assisted diffusion. Unlike molybdate, pristine NSMoS2 (P-NSMoS2) induced protein polymerization and upregulated gene expression related to actin filament binding, which phenotypically initiated actin-mediated endocytosis. Conversely, vacancy-rich NSMoS2 (V-NSMoS2) were internalized by coelomocytes through a vesicle-mediated and energy-dependent pathway. Mechanistically, atomic vacancies inhibited mitochondrial transport gene expression and likely induced membrane stress, significantly enhancing endocytosis (20.3%, p < 0.001). Molecular dynamics modeling revealed structural and conformational damage of cytoskeletal protein caused by P-NSMoS2, as well as the rapid response of transport protein to V-NSMoS2. These findings demonstrate that earthworm functional coelomocytes can accumulate NSMoS2 and directly mediate cytotoxicity and that atomic vacancies can alter the endocytic pathway and enhance cellular uptake by reprogramming protein response and gene expression patterns. This study provides an important mechanistic understanding of the ecological risks of NSMoS2.

Methyl eugenol protects the kidney from oxidative damage in mice by blocking the Nrf2 nuclear export signal through activation of the AMPK/GSK3ß axis.

Disrupted redox homeostasis contributes to renal ischemia-reperfusion (IR) injury. Abundant natural products can activate nuclear factor erythroid-2-related factor 2 (Nrf2), thereby providing therapeutic benefits. Methyl eugenol (ME), an analog of the phenolic compound eugenol, has the ability to induce Nrf2 activity. In this study, we investigated the protective effects of ME against renal oxidative damage in vivo and in vitro. An IR-induced acute kidney injury (AKI) model was established in mice. ME (20 mg·kg-1·d-1, i.p.) was administered to mice on 5 consecutive days before IR surgery. We showed that ME administration significantly attenuated renal destruction, improved the survival rate, reduced excessive oxidative stress and inhibited mitochondrial lesions in AKI mice. We further demonstrated that ME administration significantly enhanced Nrf2 activity and increased the expression of downstream antioxidative molecules. Similar results were observed in vitro in hypoxia/reoxygenation (HR)-exposed proximal tubule epithelial cells following pretreatment with ME (40 µmol·L-1). In both renal oxidative damage models, ME induced Nrf2 nuclear retention in tubular cells. Using specific inhibitors (CC and DIF-3) and molecular docking, we demonstrated that ME bound to the binding pocket of AMPK with high affinity and activated the AMPK/GSK3ß axis, which in turn blocked the Nrf2 nuclear export signal. In addition, ME alleviated the development of renal fibrosis induced by nonfatal IR, which is frequently encountered in the clinic. In conclusion, we demonstrate that ME modulates the AMPK/GSK3ß axis to regulate the cytoplasmic-nuclear translocation of Nrf2, resulting in Nrf2 nuclear retention and thereby enhancing antioxidant target gene transcription that protects the kidney from oxidative damage.

An exploration of the interaction mechanism of Direct Red 80 with α-Amylase at the molecular level.

The use and production of Direct Red 80 (DR80) dye are growing rapidly, and a large amount of dye wastewater is discharged into the soil without treatment. DR80 accumulated in soil or sludge can lead to enzyme poisoning, inhibit microbial activity, and affect the transformation of substances in the soil. In this research, the interaction mechanism between DR80 and α-Amylase (a typical enzyme in soil and sludge) was investigated by multi-spectra, molecular docking, thermodynamics analysis and enzyme activity experiment. The results of UV-visible and resonance light scattering (RLS) spectra showed that the skeleton of α-Amylase became loosened and unfolded under the exposure of Direct Red. The size of α-Amylase was smaller and α-Amylase became dispersed under high concentration of DR80. Molecular docking and thermodynamic analysis showed that DR80 bound to the surface of domain A rather than the active site of α-Amylase in the form of hydrogen bonds, and the binding process was an exothermic reaction. In addition, the inhibition of α-Amylase activity by DR80 was verified by enzyme activity experiment. These results indicate that DR80 has an effect on the structure and function of α-Amylase at molecular level, which means that the toxicity of DR80 should receive more attention.

[Effect of methyl eugenol on hypoxia/reoxygenation injury of human renal tubular epithelial cells and its mechanism].

This study aimed to investigate the effect of methyl eugenol(ME) on hypoxia/reoxygenation(H/R)-induced injury of human renal tubular epithelial HK-2 cells and its mechanism. The viability of HK-2 cells cultured with different concentrations of ME and exposed to H/R was detected by cell counting kit-8(CCK-8) assay. The effect of ME on the morphology of HK-2 cells was observed under an inverted microscope. The content of intracellular reactive oxygen species in different groups was detected after 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA) fluorescence staining. Cell apoptosis was determined by flow cytometry. Changes in mitochondrial membrane potential were monitored by JC-1 dye. The concentrations of nuclear factor erythroid 2 related factor 2(Nrf2), heme oxygenase-1(HO-1), and nicotinamide adenine dinucleotide phosphatase oxidase 4(Nox4) were measured by Western blot, followed by the assay of Nrf2 concentration changes in cytoplasm and nucleus by confocal fluorescence staining. The results showed that when the concentration of ME was 0-40 µmol·L~(-1), the activity of HK-2 cells was not affected. Compared with the model group, ME enhanced the activity of HK-2 cells and the cell morphology was normal. As revealed by further experiments, ME inhibited the release of reactive oxygen species and the decline in mitochondrial membrane potential of HK-2 cells after H/R injury, promoted Nrf2/HO-1 expression and Nrf2 translocation to the nucleus, and down-regulated the expression of Nox4, thereby significantly reducing apoptosis. This protective effect of ME could be reversed by the specific Nrf2 inhibitor ML385. These findings have preliminarily proved that ME effectively protected HK-2 cells against H/R injury, which might be related to its promotion of Nrf2/HO-1 signaling pathway and inhibition of Nox4. Such exploration on the possible mechanism of ME in the treatment of renal ischemia-reperfusion injury(IRI) and protection of organ function from the perspective of antioxidant stress has provided reference for related research on the treatment of acute kidney injury with traditional Chinese medicine.

Toxicity assessment of Fluoranthene, Benz(a)anthracene and its mixed pollution in soil: Studies at the molecular and animal levels.

An increasing amount of Fluoranthene (Fla) and Benz(a)anthracene (BaA) is being produced and used, eventually entering the soil sediments. The accumulation of Fla and BaA will cause poisoning to typical enzymes (α-Amylase) and organisms (Eisenia fetida) in soil. However, the studies about exploring and comparing the different effects of Fla, BaA and their joint effect at different levels are rarely reported. In this paper, the different effects of Fla, BaA and their mixed pollutant on α-Amylase were evaluated and compared at the molecular level, and the effect of Fla-BaA to the antioxidant system of earthworm (Eisenia fetida) was investigated from the aspects of concentration and exposure time at the animal level. The results showed that Fla-BaA had the greatest influence on the skeleton structure and the microenvironment of amino acid residue of α-Amylase compared to Fla and BaA, and in the mixed pollutant system, the joint effect mode was additive mode. The inhibitory effect of Fla-BaA on the activity of α-Amylase was also stronger than that of the system alone. The assays at the animal level showed that low concentrations (below 5 mg/kg) of Fla-BaA increased the activity of GSH-Px and SOD while high concentrations inhibited their activity. The POD that was activated throughout the experiment period suggested its key role in the earthworm antioxidant system. Changes in T-AOC and MDA showed that long-term and high-dose of Fla-BaA exposure inhibited the antioxidant capacity of Eisenia fetida, causing lipid peroxidation and damage to cells.

Molecular interaction mechanisms and cellular response of superoxide dismutase and catalase to fluoranthene.

As an important raw material and intermediate product of the petrochemical industry, fluoranthene (Fla) can be emitted with industrial activities and has become a typical polycyclic aromatic hydrocarbon enriched in the Chinese topsoil layer, posing a significant threat to sensitive soil biota. Here, multispectral tools and molecular simulation techniques were integrated to elucidate the molecular mechanism of Fla interaction with key antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) at the molecular level. Meanwhile, we further revealed the cellular responses of SOD and CAT and the associated redox states in earthworm (Eisenia fetida) coelomocytes based on the molecular-level results. Our results showed that the exposure to Fla affected the backbone structure of SOD and CAT molecules and resulted in the formation of Fla-SOD polymers as well as an overall reduction in the size of the Fla-CAT binding system. Fla altered the microenvironment around Tyr residues in the SOD molecule and quenched the endogenous fluorescence of Tyr within the CAT molecule. In earthworm coelomocytes, Fla at 60 and 80 µM resulted in a significant elevation of CAT and SOD activities by 114% (p = 0.032) and 6.09% (p = 0.013), respectively. Molecular simulation results suggested that Fla-induced changes in the structure and conformation of SOD and CAT may be the key reason for their altered activities. The related redox homeostasis detection in earthworm coelomocytes indicated that high concentrations (80 µM) of Fla led to a significant accumulation of intracellular ROS (p = 0.018) and resulted in the development of lipid peroxidation. Our work contributes to an in-depth understanding of the biological effect of Fla to sensitive soil fauna, thus providing new ideas for Fla ecological risk prevention and control.

Surface Defects Regulate the in Vivo Bioenergetic Response of Earthworm Eisenia fetida Coelomocytes to Molybdenum Disulfide Nanosheets.

Two-dimensional molybdenum disulfide (2D MoS2) nanomaterials are seeing increased use in several areas, and this will lead to their inevitable release into soils. Surface defects can occur on MoS2 nanosheets during synthesis or during environmental aging processes. The mechanisms of MoS2 nanosheet toxicity to soil invertebrates and the role of surface defects in that toxicity have not been fully elucidated. We integrated traditional toxicity end points, targeted energy metabolomics, and transcriptomics to compare the mechanistic differences in the toxicity of defect-free and defect-rich MoS2 nanosheets (DF-MoS2 and DR-MoS2) to Eisenia fetida using a coelomocyte-based in vivo assessment model. After organism-level exposure to DF-MoS2 for 96 h at 10 and 100 mg Mo/L, cellular reactive oxygen species (ROS) levels were elevated by 25.6-96.6% and the activity of mitochondrial respiratory electron transport chain (Mito-RETC) complex III was inhibited by 9.7-19.4%. The tricarboxylic acid cycling and glycolysis were also disrupted. DF-MoS2 preferentially up-regulated subcellular component motility processes related to microtubules and caused mitochondrial fission. Unlike DF-MoS2, DR-MoS2 triggered an increased degree of mitochondrial fusion, as well as more severe oxidative stress. The activities of Mito-RETC complexes (I, III, IV, V) associated with oxidative phosphorylation were significantly inhibited by 22.8-68.6%. Meanwhile, apoptotic pathways were activated upon DR-MoS2 exposure, which together with the depolarization of mitochondrial membrane potential, mediated significant apoptosis. In turn, genes related to cellular homeostasis and energy release were up-regulated to compensate for DR-MoS2-induced energy deprivation. Our study indicates that MoS2 nanosheets have nanospecific effects on E. fetida and also that the role of surface defects from synthesis or that accumulate from environmental impacts needs to be fully considered when evaluating the toxicity of these 2D materials.

Efflux transport proteins of Tetrahymena thermophila play important roles in resistance to perfluorooctane sulfonate exposure.

The biotoxicity of perfluorooctane sulfonate (PFOS) has been a concern. However, the effects of PFOS on Tetrahymena thermophila, a unicellular model organism, remain unclear. This study aimed to investigate the toxicity and detoxification mechanism of PFOS in this protozoan. PFOS did not show prominent toxic effects on T. thermophila. Cell viability of T. thermophila can be concentration-dependently increased by PFOS. PFOS also increased the stability of cell membranes and the activity of lysosomes. However, PFOS inhibited efflux transporter activities. Most of the PFOS amount remained in the culture medium during the culture periods. Only a low amount of PFOS was absorbed by cells, where PFOS molecules were mainly combined with membrane proteins. The expressions of four membrane protein genes involved in transporting xenobiotics were analyzed by real time-PCR. The gene abcg25 was significantly up-regulated. The growth of abcg25 gene knockout protozoans under PFOS treatment was slightly inhibited. However, the amount of PFOS adsorbed by the knockout protozoans showed no significant difference from the Wild-type protozoans. We concluded that the ABCG25 protein might play a key role in preventing PFOS from entering the cell or being exported from the cells to protect T. thermophila against PFOS. However, ABCG25 was not the only membrane protein able to bind with PFOS.

Magnetic resonance imaging based deep-learning model: a rapid, high-performance, automated tool for testicular volume measurements.

Background: Testicular volume (TV) is an essential parameter for monitoring testicular functions and pathologies. Nevertheless, current measurement tools, including orchidometers and ultrasonography, encounter challenges in obtaining accurate and personalized TV measurements. Purpose: Based on magnetic resonance imaging (MRI), this study aimed to establish a deep learning model and evaluate its efficacy in segmenting the testes and measuring TV. Materials and methods: The study cohort consisted of retrospectively collected patient data (N = 200) and a prospectively collected dataset comprising 10 healthy volunteers. The retrospective dataset was divided into training and independent validation sets, with an 8:2 random distribution. Each of the 10 healthy volunteers underwent 5 scans (forming the testing dataset) to evaluate the measurement reproducibility. A ResUNet algorithm was applied to segment the testes. Volume of each testis was calculated by multiplying the voxel volume by the number of voxels. Manually determined masks by experts were used as ground truth to assess the performance of the deep learning model. Results: The deep learning model achieved a mean Dice score of 0.926 ± 0.034 (0.921 ± 0.026 for the left testis and 0.926 ± 0.034 for the right testis) in the validation cohort and a mean Dice score of 0.922 ± 0.02 (0.931 ± 0.019 for the left testis and 0.932 ± 0.022 for the right testis) in the testing cohort. There was strong correlation between the manual and automated TV (R2 ranging from 0.974 to 0.987 in the validation cohort; R2 ranging from 0.936 to 0.973 in the testing cohort). The volume differences between the manual and automated measurements were 0.838 ± 0.991 (0.209 ± 0.665 for LTV and 0.630 ± 0.728 for RTV) in the validation cohort and 0.815 ± 0.824 (0.303 ± 0.664 for LTV and 0.511 ± 0.444 for RTV) in the testing cohort. Additionally, the deep-learning model exhibited excellent reproducibility (intraclass correlation >0.9) in determining TV. Conclusion: The MRI-based deep learning model is an accurate and reliable tool for measuring TV.

XBP1 modulates endoplasmic reticulum and mitochondria crosstalk via regulating NLRP3 in renal ischemia/reperfusion injury.

The functional status of mitochondria and the endoplasmic reticulum are central to renal ischemia/reperfusion injury (IRI). X-box binding protein 1 (XBP1) is an important transcription factor in endoplasmic reticulum stress. NLR family pyrin domain containing-3 (NLRP3) inflammatory bodies are closely related to renal IRI. In vivo and in vitro, we examined the molecular mechanisms and functions of XBP1-NLRP3 signaling in renal IRI, which influences ER-mitochondrial crosstalk. In this study, mice were subjected to 45 min of unilateral renal warm ischemia, the other kidney resected, and reperfusion was performed for 24 h in vivo. In vitro, murine renal tubular epithelial cells (TCMK-1) were exposed to hypoxia for 24 h and reoxygenation for 2 h. Tissue or cell damage was evaluated by measuring blood urea nitrogen and creatinine levels, histological staining, flow cytometry, terminal deoxynucleotidyl transferase-mediated nick-end labeling, diethylene glycol staining, and transmission electron microscopy (TEM). Western blotting, immunofluorescence staining, and ELISA were used to analyze protein expression. Whether XBP1 regulates the NLRP3 promoter was evaluated using a luciferase reporter assay. Kidney damage was reduced with decreasing blood urea nitrogen, creatinine, interleukin-1ß, and interleukin-18 levels. XBP1 deficiency reduced tissue damage and cell apoptosis, protecting the mitochondria. Disruption of XBP1 was associated with reduced NLRP3 and cleaved caspase-1 levels and markedly improved survival. In vitro in TCMK-1 cells, XBP1 interference inhibited caspase-1-dependent mitochondrial damage and reduced the production of mitochondrial reactive oxygen species. The luciferase assay showed that spliced XBP1 isoforms enhanced the activity of the NLRP3 promoter. These findings reveal that XBP1 downregulation suppresses the expression of NLRP3, a potential regulator of endoplasmic reticulum mitochondrial crosstalk in nephritic injury and a potential therapeutic target in XBP1-mediated aseptic nephritis.

Organism and molecular-level responses of superoxide dismutase interaction with 2-pentanone.

2-Pentanone is an excellent organic solvent and extractant, which is widely used in industrial production. 2-Pentanone is harmful to soil organisms when it enters the soil. However, current studies have not clarified the response of the antioxidant enzyme superoxide dismutase (SOD) to 2-Pentanone and its mechanism. In this study, the response of earthworm antioxidant enzyme SOD to 2-Pentanone and its molecular mechanism was investigated at organism molecular levels. The results showed that the SOD activity of earthworms under 2-Pentanone stress was significantly inhibited, and the inability of superoxide anion radicals (·O2-) to be scavenged in time might be one of the reasons for the increase of lipid peroxidation. Under 2-Pentanone exposure conditions, catalase (CAT), an antioxidant enzyme closely related to SOD, and the total antioxidant capacity (T-AOC) of earthworms were activated to resist oxidative damage. On the other hand, the observation of earthworm microstructure provided evidence of a direct risk of 2-Pentanone on earthworm body wall tissues. Molecular-level assays have shown that 2-pentanone altered the secondary structure of SOD, which further led to the loosening of the SOD backbone structure and the extension of the polypeptide chain. On the other hand, 2-pentanone quenched the endogenous fluorescence of SOD in the form of static quenching and formed the 2-pentanone/SOD complex. Molecular simulation results suggested that 2-pentanone tended to bind on the surface of SOD rather than close to the active site, and it is speculated that the alteration of SOD structure is the key reason for the change in its activity. This study enriches the toxicological data of 2-Pentanone on soil organisms, thus responding to the current concerns about its ecological risk.

Prevention of alloimmune rejection using XBP1-deleted bone marrow-derived dendritic cells in heart transplantation.

BACKGROUND: Genetically modified dendritic cells (DCs) modulate the alloimmunity of T lymphocytes by regulating antigen presentation. METHODS: We generated mice with specific deletion of the X-box-binding protein 1 (XBP1) allele in bone marrow cells and cultured bone marrow-derived DCs (Xbp1-/- BMDCs) from these animals. We then tested the phenotype of Xbp1-/- BMDCs, evaluated their capability to activate allogeneic T cells and investigated their mechanistic actions. We developed a mouse model of allogeneic heart transplantation in which recipients received PBS, Xbp1-/- BMDCs, a suboptimal dose of cyclosporine A (CsA), or Xbp1-/- BMDCs combined with a suboptimal dose of CsA to evaluate the effects of Xbp1-/- BMDC transfusion on alloimmunity and on the survival of heart allografts. RESULTS: The deletion of XBP1 in BMDCs exploited the IRE1-dependent decay of TAPBP mRNA to reduce the expression of MHC-I on the cell surface, altered the capability of BMDCs to activate CD8+ T cells, and ultimately suppressed CD8+ T-cell-mediated allogeneic rejection. The adoptive transfer of Xbp1-/- BMDCs inhibited CD8+ T-cell-mediated rejection. In addition, XBP1-deficient BMDCs were weak stimulators of allogeneic CD4+ T cells despite expressing high levels of MHC-II and costimulatory molecules on their cell surface. Moreover, the adoptive transfer of Xbp1-/- BMDCs inhibited the production of circulating donor-specific IgG. The combination of Xbp1-/- BMDCs and CsA treatment significantly prolonged the survival of allografts compared to CsA alone. CONCLUSIONS: The deletion of XBP1 induces immunosuppressive BMDCs, and treatment with these immunosuppressive BMDCs prevents alloimmune rejection and improves the outcomes of heart transplantation. This finding provides a promising therapeutic target in combating transplant rejection and expands knowledge of inducing therapeutic DCs.

Discriminating malignant from benign testicular masses using machine-learning based radiomics signature of appearance diffusion coefficient maps: Comparing with conventional mean and minimum ADC values.

PURPOSE: To develop a machine-learning-based radiomics signature of ADC for discriminating between benign and malignant testicular masses and compare its classification performance with that of minimum and mean ADC. METHODS: A total of ninety-seven patients with 101 histopathologically confirmed testicular masses (70 malignancies, 31 benignities) were evaluated in this retrospective study. Eight hundred fifty-one radiomics features were extracted from the preoperative ADC map of each lesion. The mean and minimum ADC values are part of the radiomics features. Thirty lesions were randomly selected to estimate the reliability of the features. The redundant features were eliminated using univariate analysis (independent t test and Mann-Whitney U test, where appropriate) and Spearman's rank correlation. The least absolute shrinkage and selection operator (LASSO) algorithm was employed for feature selection and radiomics signature generation. The classification performance of the radiomics signature and minimum and mean ADC values were evaluated by receiver operating characteristic (ROC) curve analysis and compared by DeLong's test. RESULTS: The whole lesion-based mean ADC showed no difference between benign and malignant testicular masses (P = 0.070, training cohort; P = 0.418, validation cohort). Compared with the minimum ADC, the ADC-based radiomics signature yielded a higher area under the curve (AUC) in both the training (AUC: 0.904, 95% confidence interval [CI]: 0.832-0.975) and validation cohorts (AUC: 0.868, 95% CI: 0.728-1.00). CONCLUSIONS: Conventional mean ADC values are not always helpful in discriminating between testicular benignities and malignancies. The minimum ADC and radiomics signature might be better alternatives, with the radiomics signature performing better than the minimum ADC.

Toxic effects of acetone, 2-pentanone, and 2-hexanone on physiological indices of wheat (Triticum aestivum L.) germination and seedlings.

Petroleum hydrocarbons are important characteristic pollutants in the process of oil exploitation in the Yellow River Delta (China), and they cause a potential hazard to the surrounding ecological environment. The research on eco-toxicological effects of petroleum-derived products still needs to be studied in depth. This paper describes the physiological indices of wheat (Triticum aestivum L.) seeds and seedlings under independent stresses of acetone, 2-pentanone, and 2-hexanone to determine the toxicological effects of ketones derived from petroleum products on typical crops. The experimental results indicated that ketones with concentrations lower than 0.4 mg·cm-2 and 800 mg·kg-1 the germination of wheat seeds and the growth of seedlings were promoted to 113.32-127.27% and 105.41-126.39%, respectively, thus exhibiting low-dose excitatory effects. However, when the concentration was higher than 0.4 mg·cm-2 and 800 mg·kg-1, germination and seedlings' growth were significantly reduced to 7.14-2.12% and 35.09-13.33%, respectively. At the same time, acetone had a greater impact on the growth of wheat seed roots, the malondialdehyde (MDA), and chlorophyll contents in leaf tissues. The low concentration of acetone had a significant promoting effect on the activity of α-amylase in wheat seeds. 2-Pentanone reduced the electrical conductivity of wheat seed extract, and it significantly promoted the catalase (CAT) activity at low concentrations. 2-Hexanone had a strong inhibitory effect on wheat germination and growth. This study provided new research results to determine the toxic effects of petroleum-derived products and provided a basis for the environmental management of such substances.

A review of human and animals exposure to polycyclic aromatic hydrocarbons: Health risk and adverse effects, photo-induced toxicity and regulating effect of microplastics.

Polycyclic aromatic hydrocarbons (PAHs) are one of the most widely distributed persistent organic pollutants (POPs) in the environmental media. PAHs have been widely concerned due to their significant health risk and adverse effects to human and animals. Currently, the main sources of PAHs in the environment are the incomplete combustion of fossil fuels, as well as municipal waste incineration and agricultural non-surface source emissions. In this work, the scope of our attention includes 16 typical PAHs themselves without involving their metabolites and industrial by-products. Exposure of human and animals to PAHs can lead to a variety of adverse effects, including carcinogenicity and teratogenicity, genotoxicity, reproductive- and endocrine-disrupting effects, immunotoxicity and neurotoxicity, the type and severity of which depend on a variety of factors. On the other hand, the regulatory effect of microplastics (MPs) on the bio-toxicity and bioaccumulation capacity of PAHs has now gradually attracted attention. We critically reviewed the adsorption capacity and mechanisms of MPs on PAHs as well as the effects of MPs on PAHs toxicity, thus highlighting the importance of paying attention to the joint bio-toxicity caused by PAHs-MPs interactions. In addition, due to the extensive nature of the common exposure pathway of PAHs and ultraviolet ray, an accurate understanding of biological processes exposed to both PAHs and UV light is necessary to develop effective protective strategies. Finally, based on the above critical review, we highlighted the research gaps and pointed out the priority of further studies.

Methyl eugenol attenuates liver ischemia reperfusion injury via activating PI3K/Akt signaling.

BACKGROUND: Liver ischemia reperfusion injury (LIRI) often occurs during liver transplantation, resection, and various circulatory shock procedures, leading to severe metabolic disorders, inflammatory immune responses, oxidative stress injury, and cell apoptosis. Methyl eugenol (ME) is structurally similar to eugenol and has anti-inflammatory and apoptotic pharmacological effects. However, whether ME protects the liver from LIRI damage requires further investigation. METHODS: We established a partially warm LIRI model by subjecting C57BL/6J mice to 60 min of ischemia, followed by reperfusion for 6 h. We also established a hypoxia-reoxygenation injury (H/R) cell model by subjecting AML12 (a mouse liver cell line) cells to 24 h hypoxia, followed by 18 h normoxia. The extent of liver injury was assessed by serum transaminase concentrations, hematoxylin and eosin staining, quantitative real-time PCR, myeloperoxidase activity, and TUNEL analysis. Apoptosis was detected using flow cytometry. The protein levels of p-PI3K, PI3K, p-Akt, Akt, p-Bad, Bad, Bcl-2, Bax, and cleaved caspase-3 were detected by western blotting. LY294002, an inhibitor of PI3K/Akt signaling, was used to elucidate the relationship between ME and PI3K/Akt signaling. RESULTS: ME successfully alleviated LIRI-induced liver injury, inflammatory response, and apoptosis induced, as well as liver cell injury induced by hypoxia reoxygenation. ME is known to activate the PI3K/Akt signaling pathway in hepatocyte injury in vivo and in vitro, and when this signaling pathway is inhibited, the protective effect of ME is abrogated. CONCLUSIONS: The use of ME is a potential therapeutic approach for regulating LIRI by activating PI3K/Akt signaling.

Downregulation of XBP1 protects kidney against ischemia-reperfusion injury via suppressing HRD1-mediated NRF2 ubiquitylation.

Ischemia-reperfusion (IR) injury to the renal epithelia is associated with endoplasmic reticulum stress (ERS) and mitochondria dysfunction, which lead to oxidative stress-induced acute kidney injury (AKI). X-box binding protein 1 (XBP1), an ERS response protein, could play a prominent role in IR-induced AKI. In this study, we revealed that XBP1 and its downstream target HRD1 participated in the crosstalk between ERS and mitochondrial dysfunction via regulation of NRF2/HO-1-mediated reactive oxidative stress (ROS) signaling. Mice with reduced expression of XBP1 (heterozygous Xbp1±) were resistant to IR-induced AKI due to the enhanced expression of NRF2/HO-1 and diminished ROS in the kidney. Downregulation of XBP1 in renal epithelial cells resulted in reduced HRD1 expression and increased NRF2/HO-1 function, accompanied with enhanced antioxidant response. Furthermore, HRD1 served as an E3-ligase to facilitate the downregulation of NRF2 through ubiquitination-degradation pathway, and the QSLVPDI motif on NRF2 constituted an active site for its interaction with HRD1. Thus, our findings unveil an important physiological role for XBP1/HRD1 in modulating the antioxidant function of NRF2/HO-1 in the kidney under stress conditions. Molecular therapeutic approaches that target XBP1-HRD1-NRF2 pathway may represent potential effective means to treat renal IR injury.