Characterization, Mechanism, and Application of Dipeptidyl Peptidase III: An Aflatoxin B1-Degrading Enzyme from Aspergillus terreus HNGD-TM15.

Aflatoxin B1 is a notorious mycotoxin with mutagenicity and carcinogenicity, posing a serious hazard to human and animal health. In this study, an AFB1-degrading dipeptidyl-peptidase III mining from Aspergillus terreus HNGD-TM15 (ADPP III) with a molecular weight of 79 kDa was identified. ADPP III exhibited optimal activity toward AFB1 at 40 °C and pH 7.0, maintaining over 80% relative activity at 80 °C. The key amino acid residues that affected enzyme activity were identified as H450, E451, H455, and E509 via bioinformatic analysis and site-directed mutagenesis. The degradation product of ADPP III toward AFB1 was verified to be AFD1. The zebrafish hepatotoxicity assay verified the toxicity of the AFB1 degradation product was significantly weaker than that of AFB1. The result of this study proved that ADPP III presented a promising prospect for industrial application in food and feed detoxification.

Seismic resolution improving by a sequential convolutional neural network.

Thin-bed soft rock is one of the main factors causing large deformations of tunnels. In addition to relying on some innovative construction techniques, detecting thin beds early during surface geological exploration and advanced geological prediction can provide a basis for planning and implementing effective coping measures. The commonly used seismic methods cannot meet the requirement for thin beds detection accuracy. A high-resolution (HR) seismic signal processing method is proposed by introducing a sequential convolutional neural network (SCNN). The deep learning dataset including low-resolution (LR) and HR seismic is firstly prepared through forward modeling. Then, a one-dimension (1D) SCNN architecture is proposed to establish the mapping relationship between LR and HR sequences. Training on the prepared dataset, the HR seismic processing model with high accuracy is achieved and applied to some practical seismic data. The applications on both poststack and prestack seismic data demonstrate that the trained HR processing model can effectively improve the seismic resolution and restore the high-frequency seismic energy so that to recognize the thin-bed rocks.

Thermo-Alkali-Tolerant Recombinant Laccase from Bacillus swezeyi and Its Degradation Potential against Zearalenone and Aflatoxin B1.

The enzymatic biodegradation of mycotoxins in food and feed has attracted the most interest in recent years. In this paper, the laccase gene from Bacillus swezeyi was cloned and expressed in Escherichia coli BL 21(D3). The sequence analysis indicated that the gene consisted of 1533 bp. The purified B. swezeyi laccase was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis -12% with an estimated molecular weight of 56.7 kDa. The enzyme is thermo-alkali-tolerant, displaying the optimal degradation of zearalenone (ZEN) and aflatoxin B1 (AFB1) at pH 8 and 9, with incubation temperatures of 55 and 50 °C, respectively, within 24 h. The degradation potentials of the 50 µg of the enzyme against ZEN (5.0 µg/mL) and AFB1 (2.5 µg/mL) were 99.60 and 96.73%, respectively, within 24 h. To the best of our knowledge, this is the first study revealing the recombinant production of laccase from B. swezeyi, its biochemical properties, and potential use in ZEN and AFB1 degradation in vitro and in vivo.

The phosphorylation landscape of infection-related development by the rice blast fungus.

Many of the world's most devastating crop diseases are caused by fungal pathogens that elaborate specialized infection structures to invade plant tissue. Here, we present a quantitative mass-spectrometry-based phosphoproteomic analysis of infection-related development by the rice blast fungus Magnaporthe oryzae, which threatens global food security. We mapped 8,005 phosphosites on 2,062 fungal proteins following germination on a hydrophobic surface, revealing major re-wiring of phosphorylation-based signaling cascades during appressorium development. Comparing phosphosite conservation across 41 fungal species reveals phosphorylation signatures specifically associated with biotrophic and hemibiotrophic fungal infection. We then used parallel reaction monitoring (PRM) to identify phosphoproteins regulated by the fungal Pmk1 MAPK that controls plant infection by M. oryzae. We define 32 substrates of Pmk1 and show that Pmk1-dependent phosphorylation of regulator Vts1 is required for rice blast disease. Defining the phosphorylation landscape of infection therefore identifies potential therapeutic interventions for the control of plant diseases.

Characterization, Structural Analysis, and Thermal Stability Mutation of a New Zearalenone-Degrading Enzyme Mined from Bacillus subtilis.

Zearalenone (ZEN) is a widespread mycotoxin that causes serious damage to animal husbandry and poses a threat to human health. A screen of ZEN-degrading soil bacteria yielded Bacillus subtilis YT-4, which yielded 80% ZEN degradation after 6 h and 95% after 36 h. The gene sequence encoding the degradative enzyme ZENY was mined from the genome of YT-4 and expressed in yeast. ZENY is an α/ß-hydrolase with an optimal enzyme activity at 37 °C and pH 8. By breaking the lactone ring of ZEN, it produces ZENY-C18H24O5 with a molecular weight of 320.16 g/mol. Sequence comparison and molecular docking analyses identified the catalytic ZENY triad 99S-245H-123E and the primary ZEN-binding mode within the hydrophobic pocket of the enzyme. To improve the thermal stability of the enzyme for industrial applications, we introduced a mutation at the N-terminus, specifically replacing the fifth residue N with V, and achieved a 25% improvement in stability at 45 °C. These findings aim to achieve ZEN biodegradation and provide insight into the structure and function of ZEN hydrolases.

Mechanism of inactivation of Aspergillus flavus spores by dielectric barrier discharge plasma.

Dielectric barrier discharge plasma (DBDP) displays strong against fungal spores, while its precise mechanism of spore inactivation remains inadequately understood. In this study, we applied morphological, in vivo and in vitro experiments, transcriptomics, and physicochemical detection to unveil the potential molecular pathways underlying the inactivation of Aspergillus flavus spores by DBDP. Our findings suggested that mycelium growth was inhibited as observed by SEM after 30 s treatment at 70 kV, meanwhile spore germination ceased and clustering occurred. It led to the release of cellular contents and subsequent spore demise by disrupting the integrity of spore membrane. Additionally, based on the transcriptomic data, we hypothesized that the induction of spore inactivation by DBDP might be associated with downregulation of genes related to cell membranes, organelles (mitochondria), oxidative phosphorylation, and the tricarboxylic acid cycle. Subsequently, we validated our transcriptomic findings by measuring the levels of relevant enzymes in metabolic pathways, such as superoxide dismutase, acetyl-CoA, total dehydrogenase, and ATP. These physicochemical indicators revealed that DBDP treatment resulted in mitochondrial dysfunction, redox imbalance, and inhibited energy metabolism pathways. These findings were consistent with the transcriptomic results. Hence, we concluded that DBDP accelerated spore rupture and death via ROS-mediated mitochondrial dysfunction, which does not depend on cell membranes.

Thiamethoxam dynamics in pepper plants: Deciphering deposition and dissipation pattern across diverse planting modes and regions.

To reduce the application dosage of thiamethoxam (TMX), we investigated the deposition and dissipation patterns in a pepper-planted ecosystem under different planting modes across four regions in China, namely Hainan (HN), Zhejiang (ZJ), Anhui (AH) and Hebei (HB). This study focused on the deposition and dissipation of TMX at concentrations of 63.00, 47.25, 31.50, 23.63 and 15.75 g a.i.hm-2. As the application dose increased, the deposition amount of TMX initially increased in the plants and cultivated soil, showing obvious geographic differences in four cultivation areas. Surprisingly, the initial amount of TMX deposited the pepper-cultivated greenhouse of ZJ and AH was 1.1-2.1-fold and 1.0-3.6-fold higher than that in the open field system at the same application dose, respectively. In pepper leaves, stems, fruits and soil, the dissipation exhibited rapid growth and then slowed. However, the residual concentration showed an increasing trend, followed by a subsequent decrease in the pepper roots. In different planting regions, the dissipation rate of TMX followed the order HN > ZJ > AH > HB in pepper plants and cultivated soil. In comparison to the open field, the total TMX retention rate in greenhouse was higher, indicating overall greater persistence in the greenhouse conditions. These findings reveal the deposition and dissipation characteristics of TMX within the pepper-field ecosystem, offering a significant contribution to the risk assessment of pesticides.

Mining Lactonase Gene from Aflatoxin B1-Degrading Strain Bacillus megaterium and Degrading Properties of the Recombinant Enzyme.

Mycotoxins are toxic secondary metabolites mainly produced by filamentous fungal species that commonly contaminate food and feed. Aflatoxin B1 (AFB1) is extremely toxic and seriously threatens the health of humans and animals. In this work, the Bacillus megaterium HNGD-A6 was obtained and showed a 94.66% removal ability of AFB1 by employing extracellular enzymes as the degrading active substance. The degradation products were P1 (AFD1, C16H14O5) and P2 (C14H16N2O2), and their toxicity was greatly reduced compared to that of AFB1. The AttM gene was mined by BlastP comparison and successfully expressed in Escherichia coli BL21. AttM could degrade 86.78% of AFB1 at pH 8.5 and 80 °C, as well as 81.32% of ochratoxin A and 67.82% of zearalenone. The ability of AttM to degrade a wide range of toxins and its resistance to high temperatures offer the possibility of its use in food or feed applications.

A molecular mechanosensor for real-time visualization of appressorium membrane tension in Magnaporthe oryzae.

The rice blast fungus Magnaporthe oryzae uses a pressurized infection cell called an appressorium to drive a rigid penetration peg through the leaf cuticle. The vast internal pressure of an appressorium is very challenging to investigate, leaving our understanding of the cellular mechanics of plant infection incomplete. Here, using fluorescence lifetime imaging of a membrane-targeting molecular mechanoprobe, we quantify changes in membrane tension in M. oryzae. We show that extreme pressure in the appressorium leads to large-scale spatial heterogeneities in membrane mechanics, much greater than those observed in any cell type previously. By contrast, non-pathogenic melanin-deficient mutants, exhibit low spatially homogeneous membrane tension. The sensor kinase ∆sln1 mutant displays significantly higher membrane tension during inflation of the appressorium, providing evidence that Sln1 controls turgor throughout plant infection. This non-invasive, live cell imaging technique therefore provides new insight into the enormous invasive forces deployed by pathogenic fungi to invade their hosts, offering the potential for new disease intervention strategies.

Development and validation of a nomogram for postoperative sleep disturbance in adults: a prospective survey of 640 patients undergoing spinal surgery.

BACKGROUND: Postoperative sleep disturbance (PSD) is a prevalent clinical complication that may arise due to various factors. The purpose of this investigation is to identify the risk factors for PSD in spinal surgery and establish a risk prediction nomogram. METHODS: The clinical records of individuals who underwent spinal surgery from January 2020 to January 2021 were gathered prospectively. The least absolute shrinkage and selection operator (LASSO) regression, along with multivariate logistic regression analysis, was employed to establish independent risk factors. A nomogram prediction model was devised based on these factors. The nomogram's effectiveness was evaluated and verified via the receiver operating characteristic (ROC) curve, calibration plot, and decision curve analysis (DCA). RESULTS: A total of 640 patients who underwent spinal surgery were analyzed in this investigation, among which 393 patients experienced PSD with an incidence rate of 61.4%. After conducting LASSO regression and logistic regression analyses using R software on the variables in training set, 8 independent risk factors associated to PSD were identified, including female, preoperative sleep disorder, high preoperative anxiety score, high intraoperative bleeding volume, high postoperative pain score, dissatisfaction with ward sleep environment, non-use of dexmedetomidine and non-use of erector spinae plane block (ESPB). The nomogram and online dynamic nomogram were constructed after incorporating these variables. In the training and validation sets, the area under the curve (AUC) in the receiver operating characteristic (ROC) curves were 0.806 (0.768-0.844) and 0.755 (0.667-0.844), respectively. The calibration plots indicated that the mean absolute error (MAE) values in both sets were respectively 1.2% and 1.7%. The decision curve analysis demonstrated the model had a substantial net benefit within the range of threshold probabilities between 20% and 90%. CONCLUSIONS: The nomogram model proposed in this study included eight frequently observed clinical factors and exhibited favorable accuracy and calibration. TRIAL REGISTRATION: The study was retrospectively registered with the Chinese Clinical Trial Registry (ChiCTR2200061257, 18/06/2022).

Genomic surveillance uncovers a pandemic clonal lineage of the wheat blast fungus.

Wheat, one of the most important food crops, is threatened by a blast disease pandemic. Here, we show that a clonal lineage of the wheat blast fungus recently spread to Asia and Africa following two independent introductions from South America. Through a combination of genome analyses and laboratory experiments, we show that the decade-old blast pandemic lineage can be controlled by the Rmg8 disease resistance gene and is sensitive to strobilurin fungicides. However, we also highlight the potential of the pandemic clone to evolve fungicide-insensitive variants and sexually recombine with African lineages. This underscores the urgent need for genomic surveillance to track and mitigate the spread of wheat blast outside of South America and to guide preemptive wheat breeding for blast resistance.

Successful mechanical thrombectomy in acute bilateral M1 middle cerebral artery occlusion: a case report and literature review.

BACKGROUND: Acute bilateral occlusion of the middle cerebral artery (MCA) is a very rare condition, and most cases are accompanied by a poor prognosis. However, mechanical thrombectomy (MT) for bilateral MCA is challenging. Here, we report a case of acute unilateral MCA occlusion with sequential acute occlusion of the bilateral MCA during intravenous thrombolysis (IVT). We urgently performed bilateral MT of the MCA and effective recanalization. CASE PRESENTATION: The patient is a 73-year-old man who complained of a sudden adverse influence on speech and an inability to move his left limb for 2 h. He had a history of paroxysmal atrial fibrillation, but had never used any anticoagulants before. Head and neck computed tomography angiography (CTA) showed embolism in the right M1 MCA. During intravenous alteplase thrombolytic therapy, the patient suddenly became unconscious. Cerebral angiography showed occlusion of the M1 segment of the bilateral MCA in the patients. MT of the bilateral MCA was performed using a combination of a stent retriever and an aspiration catheter with mTici 3 revascularization. On the second day, the patient became conscious, although he had remaining symptoms of speech insufficiency and weakness of the left limb. The mRS score was 2 90 days after the operation. CONCLUSIONS: Acute bilateral occlusion of the M1 segment of the MCA is extremely rare and is accompanied by high morbidity and high mortality. Intravenous alteplase thrombolysis can increase the risk of atrial thrombus shedding in patients with atrial fibrillation, so patients with acute bilateral MCA occlusion in the M1 segment chose direct MT or bridging therapy, which remains controversial, and the sequence of MT remains to be discussed. Nevertheless, early endovascular treatment can decrease the morbidity and mortality of such patients.

The transcriptional landscape of plant infection by the rice blast fungus Magnaporthe oryzae reveals distinct families of temporally co-regulated and structurally conserved effectors.

The rice blast fungus Magnaporthe oryzae causes a devastating disease that threatens global rice (Oryza sativa) production. Despite intense study, the biology of plant tissue invasion during blast disease remains poorly understood. Here we report a high-resolution transcriptional profiling study of the entire plant-associated development of the blast fungus. Our analysis revealed major temporal changes in fungal gene expression during plant infection. Pathogen gene expression could be classified into 10 modules of temporally co-expressed genes, providing evidence for the induction of pronounced shifts in primary and secondary metabolism, cell signaling, and transcriptional regulation. A set of 863 genes encoding secreted proteins are differentially expressed at specific stages of infection, and 546 genes named MEP (Magnaportheeffector protein) genes were predicted to encode effectors. Computational prediction of structurally related MEPs, including the MAX effector family, revealed their temporal co-regulation in the same co-expression modules. We characterized 32 MEP genes and demonstrate that Mep effectors are predominantly targeted to the cytoplasm of rice cells via the biotrophic interfacial complex and use a common unconventional secretory pathway. Taken together, our study reveals major changes in gene expression associated with blast disease and identifies a diverse repertoire of effectors critical for successful infection.

Using drug-coated balloons for symptomatic vertebral artery origin stenosis: A systematic review and meta-Analysis.

BACKGROUND: Endovascular stenting has emerged as an effective treatment for patients with symptomatic vertebral artery origin stenosis (VAOS), but the incidence of severe restenosis is concerning. Angioplasty alone with a drug-coated balloon (DCB) is a potential treatment for VAOS. The purpose of this systematic review and meta-analysis was to assess the utility of DCB angioplasty for VAOS. METHODS: A systematic search of the Medline (PubMed), Embase, CNKI, and Cochrane databases for studies on the treatment of VAOS by DCB angioplasty published in English and Chinese before June 15, 2022 was conducted. Data were extracted using standardized methods. The incidence rates of restenosis, technical success, and perioperative complication in the follow-up period were pooled using Freeman-Tukey double arcsine transformation with random or fixed-effect models. Tests for heterogeneity and publication bias were performed. RESULTS: A total of seven studies containing 159 patients were included in this review and meta-analysis. The pre-treatment stenosis rate of the vertebral artery in the DCB group ranged from 70.0 % to 86.3 %, and the median follow-up time ranged from 6.0 to 14.1 months. The pooled restenosis incidence was 11.9 % (95 % CI: 3.4 %-23.4 %; I2 = 59 %, p = 0.02) during the follow-up period. The pooled technical success rate was 96.6 % (95 % CI: 91.4 %-99.7 %; I2 = 37 %, p = 0.14). The overall perioperative complication rate was 2.9 % (95 % CI: 0.3 %-6.9 %; I2 = 0 %, p = 0.64). According to the funnel diagram and Egger's test, there was no evidence of publication bias. CONCLUSION: It is suggested in this review and meta-analysis that angioplasty with DCB may be a potential treatment for VAOS. However, randomized studies including a large representative sample of VAOS patients are needed to validate our findings.

Research progress of ochratoxin a bio-detoxification.

Ochratoxins (OTs) is an extremely toxic mycotoxin in which Ochratoxin A (OTA) is the most toxic and prevalent in the ochratoxin family. OTA is among the five most critical mycotoxins that are subject to legal regulations. Animals and humans may be exposed to OTA through dietary intake, inhalation, and dermal contact. OTA is considered nephrotoxic, genotoxic, cytotoxic, teratogenic, carcinogenic, mutagenic, immunotoxic, and myelotoxic. So, intake of OTA contaminated foods and feeds can impact the productivity of animals and health of people. According to this review, several studies have reported on the approaches that have been established for OTA removal. This review focused on the control approaches to mitigate OTA contamination, OTA bio-detoxification materials and their applicable techniques, recombinant strains for OTA bio-detoxification, and their detoxification effects, recombinant OTA-degrading enzymes and their sources, recombinant fusion enzymes for OTA, ZEN and AFB1 mycotoxins detoxification, as well as the current application and commercialized OTA bio-detoxification products. However, there is no single technique that has been approved to detoxify OTA by 100% to date. Some preferred current strategies for OTA bio-detoxification have been recombinant degrading enzymes and genetic engineering technology due to their efficiency and safety. Therefore, prospective studies should focus on standardizing pure enzymes from genetically engineered microbial strains that have great potential for OTA detoxification.

Tracing and quantifying the source of heavy metals in agricultural soils in a coal gangue stacking area: Insights from isotope fingerprints and receptor models.

Historic coal gangue stacking probably brings heavy metals (HMs) into the surrounding agricultural soil, posing potential harm to human and environmental health. For better controlling and preventing agricultural soil HMs pollution, the screening of priority pollutants and identification of their pollution pathways are urgent in coal gangue stacking areas. Thus, this study selected a coal gangue stacking area in Chongqing, China as the research object and conducted the pollution evaluation, spatial distribution and source apportionment of the HMs (Cd, Cr, Ni, Cu, Zn, As, Pb and Hg) in surrounding agricultural soil. Results showed that the soil was moderately to heavily contaminated by Cd with average concentrations of 1.23 mg/kg, which were 4.1 times higher than the Environmental Quality Standards for Soils of China. Cd was considered as the soil precedent-controlled pollutant in this study area and subsequent soil δ114/110Cd values indicated that Cd in surface soils primarily originated from the leachate of coal gangue stacking, which contributed about 89.9 % and 85.47 % to the total soil Cd according to the absolute principal component scores-multiple linear regression model (APCS-MLR) and positive matrix factorization model (PMF), respectively. In addition, other HMs mainly resulted from the leachate of coal gangue, natural and agricultural mixed pollution as well as traffic pollution. Therefore, this study provided basic information for pollution control of the HMs in agricultural soil in the coal gangue stacking area.

Formation, structure and stability of high internal phase Pickering emulsions stabilized by BSPI-C3G covalent complexes.

Food-grade high internal phase Pickering emulsions (HIPPEs) are stabilized by protein-based particles, which have attracted extensive attention due to their good gel-like structure. The black soybean isolate protein/cyanidin-3-O-glucoside (BSPI-C3G) covalent particles were used as a particulate emulsifier to form stable HIPPEs with oil phase fractions (74 % v/v) and low particle concentrations (0.5 %-3 % w/v) The particle size distribution and microstructure demonstrated that the BSPI-C3G covalent particles acted as an interfacial layer and surrounded the oil droplets. As the concentration of BSPI-C3G particles increased from 0.5 % to 3 %, the droplet size, elasticity, antioxidant capacity of the heated or stored HIPPEs more stable. So, the HIPPEs had the best stability with the BSPI-C3G particle at 3 % (w/v) concentration. These findings may extend the application of BSPI and C3G in foods and provide the guidelines for the rational design of food-grade HIPPEs stabilized by protein/anthocyanin complexes.

Distribution of Mercury and Methylmercury in Farmland Soils Affected by Manganese Mining and Smelting Activities.

Manganese (Mn)-related activities would affect the mercury (Hg) cycling in farmlands, whereas this was not well understood. Here, one of the largest Mn ores in China was selected to study the effects of Mn-related activities on the accumulation and distribution of total Hg (THg) and methylmercury (MeHg) in farmland soils. The soil THg concentrations in the mining area were 0.56 ± 0.45, 0.56 ± 0.45, 0.53 ± 0.44, and 0.50 ± 0.46 mg kg−1 in the 0−10, 10−20, 20−30, and 30−40 cm layers, respectively, while they were increased to 0.75 ± 0.75, 0.72 ± 0.60, 0.62 ± 0.46, and 0.52 ± 0.38 mg kg−1 in the smelting area. Similarly, the soil MeHg concentrations in the smelting area were also elevated by 1.04−1.34 times as compared to those in the mining area. Concentrations of THg (0.59 ± 0.50 mg kg−1) and MeHg (0.64 ± 0.82 µg kg−1) in soils were higher than the regional background value but lower than in vicinal Hg-mining areas, while they were largely elevated at the intersection of two rivers in the smelting area. Significant positive Mn-THg relationship (p < 0.01) and negative Mn-MeHg relationship (p < 0.01) favored the conclusion that soil Mn could promote Hg accumulation while inhibiting MeHg production. Approximately 70% of soil Hg was distributed in the residual phase, and the environmental hazard was not elevated according to a geochemical model. Overall, mining and smelting activities of Mn ores have resulted in obvious and distinct effects on the accumulation and methylation of Hg in farmland soils, but the environmental hazards are currently manageable.

Classification of the loosening zones and estimation of the loosening pressures of tunnels in layered jointed rock strata.

Due to the complexity of tunnels, accurate prediction of their loosening pressures in layered jointed rock strata is a very difficult engineering task. To recognize loosening patterns and estimate loosening pressures, numerical tests were employed in areas where tunnels were excavated in layered jointed rock strata. A total of 12 influential factors, including joints, tunnel depth, and strata, were considered in each of the numerical models. Three loosening patterns were found in the numerical testing: a ringent trumpet-shaped boundary, an arch-shaped boundary, and a closing-trend trumpet-shaped loosening zone. Empirical expressions for the loosening zone boundaries were further established and, in the form of the 12 influential factors, considered in the simulation. Given the boundary function, the loosening pressures were further deduced, which were categorized according to loosening pattern, i.e., ringent trumpet shape or arch shape, and the excavation condition of whether or not the embedded depth was deeper than the soft layer. Two case studies were used to test this method. The newly-proposed method was found to perform better than existing methods, with loosening pressure values that were slightly larger than, but very close to, actual measured field data.