Iron(I) Complex Bearing an Open-Shell Diazenido Ligand.

Low-valent transition-metal diazenido species are important intermediates in transition-metal-mediated dinitrogen reduction reactions. Isolable complexes of the type unanimously feature closed-shell diazenido ligands. Those bearing open-shell diazenido ligands have remained elusive. Herein, we report the synthesis, characterization, and reactivity of a d7 iron(I) complex featuring an open-shell silyldiazenido ligand, [(ICy)Fe(NNSiiPr3)(η2:η2-dvtms)] (1, ICy = 1,3-dicyclohexylimidazole-2-ylidene, dvtms = divinyltetramethyldisiloxane). Complex 1 is prepared in good yield by silylation of the iron(-I)-N2 complex [K(18-crown-6)][(ICy)Fe(N2)(η2:η2-dvtms)] with iPr3SiOTf and has been fully characterized by various spectroscopic methods. Theoretical studies, in combination with characterization data, established an S = 1/2 ground spin-state for 1 that can best be described as a quartet iron(I) center featuring an antiferromagnetically coupled triplet silyldiazenido ligand. The diazenido and alkene ligands in 1 are labile, as indicated by the facile disproportionation reaction of 1 at ambient temperature to transform into the iron(II) bis(diazenido) species [(ICy)(NNSiiPr3)2Fe(dvtms)Fe(NNSiiPr3)2(ICy)] (2) and the iron(0) species [(ICy)Fe(η2:η2-dvtms)] and also the alkene-exchange reaction of 1 with PhCH═CHBC8H14 to form [(ICy)Fe(NNSiiPr3)(η2-trans-PhCH═CHBC8H14)] (3). Complex 1 is light-sensitive. Upon photolysis, it undergoes a SiiPr3 radical-transfer reaction to yield [(ICy)Fe(σ:η2-MeCHSiMe2OSiMe2CH═CHSiiPr3)] (4) and N2. The reactions of 1 with the trityl radical and organic bromides yield iron(II) complexes, which indicates its reducing nature. Moreover, 1 is a weak hydrogen-atom abstractor, as indicated by its inertness toward HSi(SiMe3)3 and cyclohexa-1,4-diene and the low calculated N-H bond dissociation energy (48 kcal/mol) of its corresponding iron(II) iso-hydrazenido species.

Comparison of Different Fusion Radiomics for Predicting Benign and Malignant Sacral Tumors: A Pilot Study.

Differentiating between benign and malignant sacral tumors is crucial for determining appropriate treatment options. This study aims to develop two benchmark fusion models and a deep learning radiomic nomogram (DLRN) capable of distinguishing between benign and malignant sacral tumors using multiple imaging modalities. We reviewed axial T2-weighted imaging (T2WI) and non-contrast computed tomography (NCCT) of 134 patients pathologically confirmed as sacral tumors. The two benchmark fusion models were developed using fusion deep learning (DL) features and fusion classical machine learning (CML) features from multiple imaging modalities, employing logistic regression, K-nearest neighbor classification, and extremely randomized trees. The two benchmark models exhibiting the most robust predictive performance were merged with clinical data to formulate the DLRN. Performance assessment involved computing the area under the receiver operating characteristic curve (AUC), sensitivity, specificity, accuracy, negative predictive value (NPV), and positive predictive value (PPV). The DL benchmark fusion model demonstrated superior performance compared to the CML fusion model. The DLRN, identified as the optimal model, exhibited the highest predictive performance, achieving an accuracy of 0.889 and an AUC of 0.961 in the test sets. Calibration curves were utilized to evaluate the predictive capability of the models, and decision curve analysis (DCA) was conducted to assess the clinical net benefit of the DLR model. The DLRN could serve as a practical predictive tool, capable of distinguishing between benign and malignant sacral tumors, offering valuable information for risk counseling, and aiding in clinical treatment decisions.

Haemolysins are essential to the pathogenicity of deep-sea Vibrio fluvialis.

Vibrio fluvialis is an emerging foodborne pathogen that produces VFH (Vibrio fluvialis hemolysin) and δVFH (delta-Vibrio fluvialis hemolysin). The function of δVFH is unclear. Currently, no pathogenic V. fluvialis from deep sea has been reported. In this work, a deep-sea V. fluvialis isolate (V13) was examined for pathogenicity. V13 was most closely related to V. fluvialis ATCC 33809, a human isolate, but possessed 262 unique genes. V13 caused lethal infection in fish and induced pyroptosis involving activation of the NLRP3 inflammasome, caspase 1 (Casp1), and gasdermin D (GSDMD). V13 defective in VFH or VFH plus δVFH exhibited significantly weakened cytotoxicity. Recombinant δVFH induced NLRP3-Casp1-GSDMD-mediated pyroptosis in a manner that depended on K+ efflux and intracellular Ca2+ accumulation. δVFH bound several plasma membrane lipids, and these bindings were crucial for δVFH cytotoxicity. Together these results provided new insights into the function of δVFH and the virulence mechanism of V. fluvialis.

A Method for Automatically Predicting the Radiation-Induced Vulnerability of Unit Integrated Circuits.

With the rapid development of semiconductor technology, the reduction in device operating voltage and threshold voltage has made integrated circuits more susceptible to the effects of particle radiation. Moreover, as process sizes decrease, the impact of charge sharing effects becomes increasingly severe, with soft errors caused by single event effects becoming one of the main causes of circuit failures. Therefore, the study of sensitivity evaluation methods for integrated circuits is of great significance for promoting the optimization of integrated circuit design, improving single event effect experimental methods, and enhancing the irradiation reliability of integrated circuits. In this paper, we first established a device model for the charge sharing effect and simulated it under reasonable conditions. Based on the simulation results, we then built a neural network model to predict the charge amounts in primary and secondary devices. We also propose a comprehensive automated method for calculating soft errors in unit circuits and validated it through TCAD simulations, achieving an error margin of 2.8-4.3%. This demonstrated the accuracy and effectiveness of the method we propose.

Association between glyphosate exposure and osteoarthritis in US adults: Especially in people who are obese and inactive in leisure time physical activity.

OBJECTIVE: Little has been known on the effect of chronic glyphosate exposure on osteoarthritis (OA). The aim of this study was to investigate the association between glyphosate exposure and OA and to further investigate the different moderating effects of leisure time physical activity (LTPA) and body mass index (BMI) types on the association between glyphosate exposure and OA. METHODS: Cross-sectional data from 2540 participants in the 2015-2018 National Health and Nutrition Examination Survey (NHANES) were used to explore the association between glyphosate exposure and OA. Multivariate logistic regression models and restricted cubic spline models were used to investigate the association between glyphosate exposure and OA, and further analyses were conducted to determine the association between glyphosate exposure and OA under different LTPA and BMI types. RESULTS: Of the 2540 participants, 346 had OA. Participants with the highest glyphosate concentration (Q4) had a higher incidence of OA compared to participants with the lowest glyphosate concentration (Q1) (OR, 1.88; 95 % confidence interval [CI]: 1.13, 3.13), there was no nonlinear association between glyphosate and OA (non-linear P = 0.343). In the no LTPA group, glyphosate concentration in the Q4 group was correlated with OA (OR, 2.65; 95%CI: 1.27, 5.51). In the obese group, glyphosate concentration in the Q4 group was correlated with OA (OR, 2.74; 95 % CI: 1.48, 5.07). Among people with high BMI and inactive in LTPA, glyphosate concentrations in Q4 were associated with OA (OR, 2.19; 95 % CI: 1.07, 4.48). CONCLUSIONS: Glyphosate is associated with OA odd, and physical activity and moderate weight loss can mitigate this association to some degree. This study provides a scientific basis for rational prevention of OA by regulation of LTPA and BMI under glyphosate exposure.

Bacillus cereus cereolysin O induces pyroptosis in an undecapeptide-dependent manner.

Bacillus cereus is a clinically significant foodborne pathogen that causes severe gastrointestinal and non-gastrointestinal disease. Cereolysin O (CLO) is a putative virulence factor of B. cereus, and its function remains to be investigated. In this study, we examined the biological activity of CLO from a deep sea B. cereus isolate. CLO was highly toxic to mammalian cells and triggered pyroptosis through NLRP3 inflammasome-mediated caspase 1 and gasdermin D activation. CLO-induced cell death involved ROS accumulation and K+ efflux, and was blocked by serum lipids. CLO bound specifically to cholesterol, and this binding was essential to CLO cytotoxicity. The structural integrity of the three tryptophan residues in the C-terminal undecapeptide was vital for CLO to interact with membrane lipids and cause membrane perforation. Taken together, these results provided new insights into the molecular mechanism of B. cereus CLO-mediated cytotoxicity.

MRI-Based Machine Learning Fusion Models to Distinguish Encephalitis and Gliomas.

This paper aims to compare the performance of the classical machine learning (CML) model and the deep learning (DL) model, and to assess the effectiveness of utilizing fusion radiomics from both CML and DL in distinguishing encephalitis from glioma in atypical cases. We analysed the axial FLAIR images of preoperative MRI in 116 patients pathologically confirmed as gliomas and clinically diagnosed with encephalitis. The 3 CML models (logistic regression (LR), support vector machine (SVM) and multi-layer perceptron (MLP)), 3 DL models (DenseNet 121, ResNet 50 and ResNet 18) and a deep learning radiomic (DLR) model were established, respectively. The area under the receiver operating curve (AUC) and sensitivity, specificity, accuracy, negative predictive value (NPV) and positive predictive value (PPV) were calculated for the training and validation sets. In addition, a deep learning radiomic nomogram (DLRN) and a web calculator were designed as a tool to aid clinical decision-making. The best DL model (ResNet50) consistently outperformed the best CML model (LR). The DLR model had the best predictive performance, with AUC, sensitivity, specificity, accuracy, NPV and PPV of 0.879, 0.929, 0.800, 0.875, 0.867 and 0.889 in the validation sets, respectively. Calibration curve of DLR model shows good agreement between prediction and observation, and the decision curve analysis (DCA) indicated that the DLR model had higher overall net benefit than the other two models (ResNet50 and LR). Meanwhile, the DLRN and web calculator can provide dynamic assessments. Machine learning (ML) models have the potential to non-invasively differentiate between encephalitis and glioma in atypical cases. Furthermore, combining DL and CML techniques could enhance the performance of the ML models.

MCM2 is involved in subtyping and tamoxifen resistance of ERα-positive breast cancer by acting as the downstream factor of ERα.

Tamoxifen (TAM) resistance is finally developed in over 40% of patients with estrogen receptor α-positive breast cancer (ERα+ -BC), documenting that discovering new molecular subtype is needed to confer perception to the heterogeneity of ERα+ -BC. We obtained representative gene sets subtyping ERα+ -BC using gene set variation analysis (GSVA), non-negative matrix factorization (NMF), and COX regression methods on the basis of METABRIC, TCGA, and GEO databases. Furthermore, the risk score of ERα+ -BC subtyping was established using least absolute shrinkage and selection operator (LASSO) regression on the basis of genes in the representative gene sets, thereby generating the two subtypes of ERα+ -BC. We further found that minichromosome maintenance complex component 2 (MCM2) functioned as the hub gene subtyping ERα+ -BC using GO, KEGG, and MCODE. MCM2 expression was capable for specifically predicting 1-year overall survival (OS) of ERα+ -BC and correlated with T stage, AJCC stage, and tamoxifen (TAM) sensitivity of ERα+ -BC. The downregulation of MCM2 expression inhibited proliferation, migration, and invasion of TAM-resistant cells and promoted G0/G1 arrest. Altogether, tamoxifen resistance entails that MCM2 is a hub gene subtyping ERα+ -BC, providing a novel dimension for discovering a potential target of TAM-resistant BC.

Infection status and molecular detection of pathogens carried by ectoparasites of Miniopterus fuliginosus bats in Yunnan, China.

Bats serve as natural hosts for various infectious agents that can affect both humans and animals, and they are geographically widespread. In recent years, the prevalence of bat-associated pathogens has surged on a global scale, consequently generating significant interest in bats and their ectoparasites. In this study, we specifically selected the Miniopterus fuliginosus as the host and conducted bat captures in Nanjian Yi Autonomous County, Dali Bai Autonomous Prefecture, and the other in Mouding Township, Chuxiong Yi Autonomous Prefecture, located in Yunnan Province, China. Ectoparasites were meticulously collected from the bat body surface, alongside blood samples for subsequent analyses. Following collection, the ectoparasites were methodically identified and subjected to comprehensive ecological analysis. Additionally, DNA was extracted from both the bat blood and bat flies, with conventional PCR techniques utilized for molecular screening of four pathogens: Anaplasma sp., Babesia sp., Hepatozoon sp., and Bartonella sp. The capture efforts yielded a total of 37 M. fuliginosus, from which 388 ectoparasites were recovered, including 197 gamasid mites (Cr = 50.77%, PM = 94.59%, MA = 5.32, MI = 5.63) and 191 bat flies (Cr = 49.23%, PM = 75.68%, MA = 5.16, MI = 6.82). Notably, Steatonyssus nyctali (Y = 0.28, m*/m = 2.44) and Nycteribia allotopa (Y = 0.23,m*/m = 1.54) predominated among different individuals of M. fuliginosus, exhibiting an aggregated distribution pattern. The infection rates of Bartonella sp. were identified to be 18.92% (7/37) among bats and 37.17% (71/191) among bat flies, based on the testing of 37 bats and 191 bat flies. Phylogenetic analysis demonstrated that the Bartonella sequences exhibited similarity to those found in bats and bat flies within China and South Korea. This study not only contributes to our comprehension of ectoparasite infection in M. fuliginosus but also establishes a foundation for potential exploration of their role as vectors.

Fusion Radiomics-Based Prediction of Response to Neoadjuvant Chemotherapy for Osteosarcoma.

RATIONALE AND OBJECTIVES: Neoadjuvant chemotherapy (NAC) is the most crucial prognostic factor for osteosarcoma (OS), it significantly prolongs progression-free survival and improves the quality of life. This study aims to develop a deep learning radiomics (DLR) model to accurately predict the response to NAC in patients diagnosed with OS using preoperative MR images. METHODS: We reviewed axial T2-weighted imaging (T2WI) and contrast-enhanced T1-weighted (T1CE) of 106 patients pathologically confirmed as OS. First, the Auto3DSeg framework was utilized for automated OS segmentation. Second, using three feature extraction methods, nine risk classification models were constructed based on three classifiers. The area under the receiver operating curve (AUC), sensitivity, specificity, accuracy, negative predictive value and positive predictive value were calculated for performance evaluation. Additionally, we developed a deep learning radiomics nomogram with clinical indicators. RESULTS: The model for OS automatic segmentation achieved a Dice coefficient of 0.868 across datasets. To predict the response to NAC, the DLR model achieved the highest prediction performance with an accuracy of 93.8% and an AUC of 0.961 in the test sets. We used calibration curves to assess the predictive ability of the models and performed decision curve analysis to evaluate the clinical net benefit of the DLR model. CONCLUSION: The DLR model can serve as a pragmatic prediction tool, capable of identifying patients with poor response to NAC, providing information for risk counseling, and assisting in making clinical treatment decisions. Poor responders are better advised to undergo immunotherapy and receive the best supportive care.

Multiscale levels CO2 decouple reinforcement in China.

Decoupling economic growth from CO2 emissions is imperative for China. Meanwhile, establishing a consistent and comprehensive decoupling inventory that includes national (N), regional and provincial (RP), and city and county (CC) levels is essential for further policy formulation. This research aims to investigate the decoupling status using the "N-RP-CC" approach while considering changes in decoupling trends at the different levels. A combination of the Tapio decoupling model and cluster analysis is employed to study the decoupling's spatiotemporal characteristics and trends. The study first calculates the decoupling value for "national, 7; regions, 30; provinces, 1501 CCs" in China, 2006-2017. The results show that there continues to be an improvement in the decoupling trend at the national level. Conversely, the regional scale exhibits a more vulnerable decoupling trend compared to the national level, with weak and extended negative decoupling observed in northeastern and northern China. Moreover, provincial heterogeneities are increasingly evident, with poor decoupling statuses appearing in Jilin, Heilongjiang, Liaoning, and Xinjiang, as well as many central provinces. Additionally, although more than half of CCs exhibit weak decoupling during most years, seven different states of decoupling were also identified during the time frame. These findings further indicate that spatiotemporal heterogeneities extend beyond RP scales within CCs. Taking the Yangtze River as a boundary line reveals a severe situation in northern areas along with rapid development trends observed in southern regions. Finally, we clustered 1414 CCs based on their industrial proportions for 2017 which further highlights increasingly prominent heterogeneities that should be carefully considered. Based on these findings, policy recommendations such as spatial organization and optimization and technique investment are proposed to achieve CO2 emission decoupling under the N-RP-CC levels.

Complete mitogenome of Nycteribia allotopa Speiser, 1901 (Diptera, Hippoboscoidea, Nycteribiidae) and comparative analysis of mitochondrial genomes of Nycteribiidae.

In recent years, the global pandemic of bat-associated pathogens has led to increasing attention on bat ectoparasites. Numerous studies have identified human-associated pathogens in Nycteribiidae, indicating their potential as vectors. In this study, the first complete sequencing of the mitochondrial genome of Nycteribia allotopa Speiser, 1901 was sequenced and analyzed. We also compared the mitochondrial sequences of N. allotopa with those available in the database for other Nycteribiidae species. The complete mitochondrial genome of N. allotopa was found to be 15,161 bp in size with an A + T content of 82.49%. Nucleotide polymorphism analysis of 13 protein-coding genes from five species of Nycteribiidae showed that nad6 exhibited the most significant variation, while cox1 was the most conserved. Furthermore, selection pressure analysis revealed cox1 to exhibit the strongest purifying selection, while atp8, nad2, nad4L, and nad5 showed slightly looser purifying selection. Pairwise genetic distances indicated that cox1 and cox2 were evolving comparatively slowly, whereas atp8, nad2, and nad6 were evolving comparatively quickly. Phylogenetic trees constructed using Bayesian inference and maximum likelihood methods demonstrated that all four families within the superfamily Hippoboscoidea clustered into one branch each, indicating their monophyly. N. allotopa was found to be most closely related to the same genus N. parvula. This study significantly enriches the molecular database for Nycteribiidae and provides invaluable reference data for future species identification, phylogenetic analysis, and exploration of their potential as vectors for human-associated pathogens.

Induction of clastogenesis and gene mutations by carbamazepine (at its therapeutically effective serum levels) in mammalian cells and the dependence on human CYP2B6 enzyme activity.

Carbamazepine (CBZ, an antiepileptic) is metabolized by multiple CYP enzymes to its epoxide and hydroxides; however, whether it is genotoxic remains unclear. In this study, molecular docking (CBZ to CYPs) and cytogenotoxic toxicity assays were employed to investigate the activation of CBZ for mutagenic effects, in various mammalian cell models. Docking results indicated that CBZ was valid as a substrate of human CYP2B6 and 2E1, while not for CYP1A1, 1A2, 1B1 or 3A4. In the Chinese hamster (V79) cell line and its derivatives genetically engineered for the expression of human CYP1A1, 1A2, 1B1, 2E1 or 3A4 CBZ (2.5 ~ 40 µM) did not induce micronucleus, while in human CYP2B6-expressing cells CBZ significantly induced micronucleus formation. In a human hepatoma C3A cell line, which endogenously expressed CYP2B6 twofold higher than in HepG2 cells, CBZ induced micronucleus potently, which was blocked by 1-aminobenzotriazole (inhibitor of CYPs) and ticlopidine (specific CYP2B6 inhibitor). In HepG2 cells CBZ did not induce micronucleus; however, pretreatment of the cells with CICTO (CYP2B6 inducer) led to micronucleus formation by CBZ, while rifampicin (CYP3A4 inducer) or PCB126 (CYP1A inducer) did not change the negative results. Immunofluorescent assay showed that CBZ selectively induced centromere-free micronucleus. Moreover, CBZ induced double-strand DNA breaks (γ-H2AX elevation, by Western blot) and PIG-A gene mutations (by flowcytometry) in C3A (threshold being 5 µM, lower than its therapeutic serum concentrations, 17 ~ 51 µM), with no effects in HepG2 cells. Clearly, CBZ may induce clastogenesis and gene mutations at its therapeutic concentrations, human CYP2B6 being a major activating enzyme.

Capacity-enhanced and kinetic-expedited zinc-ion storage ability in a Zn3V3O8/VO2 cathode enabled by heterostructural design.

Heterostructured double-phase composites are promising electrode candidates for high-performance secondary metal batteries due to their superior capacity and ion transfer kinetics compared with the pristine phase. Herein, a Zn3V3O8/VO2 (ZVO/VO) heterostructure with abundant phase boundaries was designed as the cathode for aqueous zinc-ion batteries (ZIBs). The preparation method is based on a solid pre-intercalation approach, and the Zn content in the ZVO/VO heterostructure can be precisely controlled. The electrochemical performance of ZVO/VO containing different amounts of Zn, pristine ZVO, and VO phases was compared. ZVO/VO showed superior capacity and cycling stability compared to pristine ZVO and VO. The ZVO/VO heterostructure showed a capacity of 328.4 mA h g-1 at 0.3 A g-1 after 200 cycles. The long-term cycling performance of ZVO/VO was evaluated at 3 A g-1, and it delivered a capacity retention of 90.5% after 1000 cycles. The ion storage mechanism of the ZVO/VO electrode was analyzed by ex situ X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM). This work provides a simple strategy for designing vanadium-based heterostructure composites as advanced cathodes for ZIBs.

Treatment of Scoliosis with One-Stage Posterior Pedicle Screw System by Paraspinal Intermuscular Approach: A Minimum of Two Years of Follow-Up.

OBJECTIVE: To evaluate the clinical efficacy of the treatment of scoliosis with a pedicle screw system through paraspinal intermuscular approach (PIA). METHODS: This is a retrospective case series study. A total of 10 patients diagnosed with scoliosis had surgical indications and treated with a pedicle screw system in one-stage posterior surgery by PIA from March 2013 to April 2015 at the First Hospital of Jilin University were enrolled in this study. The average age of the patients was 14.9 years, including one male and nine females. The operative information and surgical results, including Cobb angle correction, correction loss, global balance (including Frontal Plane Balance [FPB] and Sagittal Plane Balance [SPB]), and fusion rate were reviewed. Functional outcomes including visual analog scale (VAS) back pain score, leg pain score, and Scoliosis Research Society-22 questionnaire (SRS-22) were used to evaluate the quality of life of patients preoperatively and at last follow-up. RESULTS: Each patient was followed up at least six times. The average follow-up time was 43.2 months. Mean scoliosis and kyphosis improved from 68.5° ± 18.1°to 18.7° ± 11.8° and from 34.4° ± 17.9°to 24.0° ± 6.7°, respectively (p < 0.05); at last follow-up, it was 20.1° and 24.7°, respectively (p > 0.05). During the follow-up, mean coronal and sagittal correction loss was 1.4° ± 1.2°and 0.7° ± 0.8°, respectively (p > 0.05). Mean FPB improved from 32.7 to 11.7 mm (p < 0.05); Mean SPB changed from 0.3 to -0.7 mm (p > 0.05). No dural tears were observed during the corrective surgery or wound infection or implant-related complications. No pseudoarthrosis was identified according to the last follow-up three-dimensional (3D) CT scan. All the domains in SRS-22 questionnaire show statistically significant improvement at the last follow-up (p < 0.05). The VAS back pain scores improved from a mean preoperative score of 1.7 to a mean postoperative score of 0.2 (p < 0.05). CONCLUSION: This original one-stage posterior PIA is safe and effective in the treatment of scoliosis, which is characterized with less blood loss, shorter operation time, and satisfactory bony fusion.

Performance evaluation of differential splicing analysis methods and splicing analytics platform construction.

A proportion of previously defined benign variants or variants of uncertain significance in humans, which are challenging to identify, may induce an abnormal splicing process. An increasing number of methods have been developed to predict splicing variants, but their performance has not been completely evaluated using independent benchmarks. Here, we manually sourced ∼50 000 positive/negative splicing variants from > 8000 studies and selected the independent splicing variants to evaluate the performance of prediction methods. These methods showed different performances in recognizing splicing variants in donor and acceptor regions, reminiscent of different weight coefficient applications to predict novel splicing variants. Of these methods, 66.67% exhibited higher specificities than sensitivities, suggesting that more moderate cut-off values are necessary to distinguish splicing variants. Moreover, the high correlation and consistent prediction ratio validated the feasibility of integration of the splicing prediction method in identifying splicing variants. We developed a splicing analytics platform called SPCards, which curates splicing variants from publications and predicts splicing scores of variants in genomes. SPCards also offers variant-level and gene-level annotation information, including allele frequency, non-synonymous prediction and comprehensive functional information. SPCards is suitable for high-throughput genetic identification of splicing variants, particularly those located in non-canonical splicing regions.

Transcriptional and Metabolic Responses of Maize Shoots to Long-Term Potassium Deficiency.

Potassium is important for plant growth and crop yield. However, the effects of potassium (K+) deficiency on silage maize biomass yield and how maize shoot feedback mechanisms of K+ deficiency regulate whole plant growth remains largely unknown. Here, the study aims to explore the maize growth, transcriptional and metabolic responses of shoots to long-term potassium deficiency. Under the K+ insufficiency condition, the biomass yield of silage maize decreased. The transcriptome data showed that there were 922 and 1,107 differential expression genes in DH605 and Z58, respectively. In the two varieties, 390 differently expressed overlapping genes were similarly regulated. These genes were considered the fundamental responses to K+ deficiency in maize shoots. Many stress-induced genes are involved in transport, primary and secondary metabolism, regulation, and other processes, which are involved in K+ acquisition and homeostasis. Metabolic profiles indicated that most amino acids, phenolic acids, organic acids, and alkaloids were accumulated in shoots under K+ deficiency conditions and part of the sugars and sugar alcohols also increased. It revealed that putrescine and putrescine derivatives were specifically accumulated under the K+ deficiency condition, which may play a role in the feedback regulation of shoot growth. These results confirmed the importance of K+ on silage maize production and provided a deeper insight into the responses to K+ deficiency in maize shoots.

Characterization of the pathogenicity of a Bacillus cereus isolate from the Mariana Trench.

Bacillus cereus is an important opportunistic pathogen widely distributed in the environment. In this study, we reported the isolation and characterization of a B. cereus isolate, MB1, from the Challenger Deep of the Mariana Trench. MB1 is aerobic, motile, and able to form endospores. It possesses 5966 genes distributed on a circular chromosome and two plasmids. The MB1 genome contains 14 sets of 23S, 5S, and 16S ribosomal RNA operons, 106 tRNA genes, 4 sRNA genes, 12 genomic islands, 13 prophages, and 302 putative virulence genes, including enterotoxins and cytolysins. Infection studies showed that MB1 was able to cause acute and lethal infection in fish and mice, and was highly toxic to mammalian cells. MB1 induced, in a dose-dependent manner, pyroptotic cell death, characterized by activation of caspase-1, cleavage of gasdermin D, and release of IL-1ß and IL-18. MB1 spores exhibited swimming and haemolytic capacity, but were severely attenuated in pathogenicity, which, however, was regained to the full extent when the spores germinated under suitable conditions. Taken together, these results provide new insights into the biological and pathogenic mechanism of deep sea B. cereus.

A Type Ib Crustin from Deep-Sea Shrimp Possesses Antimicrobial and Immunomodulatory Activity.

Crustins are small antimicrobial proteins produced by crustaceans. Of the many reported crustins, very few are from deep sea environments. Crustins are categorized into several types. Recently, the Type I crustin has been further classified into three subtypes, one of which is Type Ib, whose function is unknown. Here, we studied the function of a Type Ib crustin (designated Crus2) identified from a deep-sea crustacean. Crus2 has a whey acidic protein (WAP) domain and a long C-terminal region (named P58). Recombinant Crus2 bound to peptidoglycan (PGN), lipoteichoic acid (LTA), and lipopolysaccharide (LPS), and killed Gram-positive and Gram-negative bacteria by permeabilizing the bacterial cytomembrane. Consistently, Crus2 dramatically attenuated the inflammatory response induced by LPS and LTA. Disruption of the disulfide bonds in the WAP domain abolished the bactericidal ability of Crus2, but had no effect on the bacterial binding ability of Crus2. Deletion of the C-terminal P58 region moderately affected the antimicrobial activity of Crus2 against some bacteria. P58 as a synthesized peptide could bind bacteria and inhibit the bactericidal activity of Crus2. Taken together, these results revealed different roles played by the WAP domain and the P58 region in Type Ib crustin, and provided new insights into the antimicrobial and immunomodulatory functions of crustins.

Determination of Cyazofamid and Its Metabolite in Oily Agricultural Products with HPLC-MS/MS.

A high-performance liquid chromatography-tandem mass spectrometry method was developed for the determination of cyazofamid and its major metabolite 4-Chloro-5-(4-tolyl)-1H-imidazole-2-carbonitrile (CCIM) in oily samples. Samples were extracted with acetonitrile, which contained 1% acetic acid and cleaned-up with C18 and Florisil absorbents. Recoveries ranged from 75.91% to 109.85% with coefficients of variation from 5.14% to 10.69%. The limits of detection (LODs) and the limits of quantification (LOQs) were >0.0020 mg kg-1 and ≤0.0040 mg·kg-1, respectively, which were smaller than maximum residue levels established by Australia for oily samples. The proposed fragmentation pathway of cyazofamid and CCIM were discussed.