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Fire incidents in energy storage stations are frequent, posing significant firefighting safety risks. To simulate the fire characteristics and inhibition performances by fine water mist for lithium-ion battery packs in an energy-storage cabin, the PyroSim software is used to build a 1:1 experimental geometry model of a containerized lithium-ion energy storage cabin. The simulation reveals five stages and their characteristic parameter variations during a fire incident: initial temperature rise, flame generation, flame spread, stable combustion, and flame extinguishment. By adjusting various parameters of the fine water mist, the design of the fine water mist firefighting system for containerized energy storage units is optimized. The simulation results indicate that the optimal inhibition effect for the energy storage cabin's fine water mist firefighting system is achieved when the spray intensity is ≥24 l/min, the fog cone angle is 76°, nozzle velocity is 10 m/s, and the optimal particle size of the fine water mist is 50 μm. The research findings not only provide a rational method and theoretical guidance for the numerical simulation of thermal runaway in lithium batteries but also offer theoretical data support for the safety design and protection of future energy storage cabins. [ABSTRACT FROM AUTHOR]

AI-driven mineral prospectivity mapping (MPM) is a valid and increasingly accepted tool for delineating the targets of mineral exploration, but it suffers from noisy and unrepresentative input features. In this study, a set of fractal and multifractal methods, including box-counting calculation, concentration–area fractal modeling, and multifractal analyses, were employed to excavate the underlying nonlinear mineralization-related information from geological features. Based on these methods, multiple feature selection criteria, namely prediction–area plot, K-means clustering, information gain, chi-square, and the Pearson correlation coefficient, were jointly applied to rank the relative importance of ore-related features and their fractal representations, so as to choose the optimal input feature dataset readily used for training predictive AI models. The results indicate that fault density, the multifractal spectrum width (∆α) of the Yanshanian intrusions, information dimension (D1) of magnetic anomalies, correlation dimension (D2) of iron-oxide alteration, and the D2 of argillic alteration serve as the most effective predictor features representative of the corresponding ore-controlling elements. The comparative results of the model assessment suggest that all the AI models trained by the fractal datasets outperform their counterparts trained by raw datasets, demonstrating a significant improvement in the predictive capability of fractal-trained AI models in terms of both classification accuracy and predictive efficiency. A Shapley additive explanation was employed to trace the contributions of these features and to explain the modeling results, which imply that fractal representations provide more discriminative and definitive feature values that enhance the cognitive capability of AI models trained by these data, thereby improving their predictive performance, especially for those indirect predictor features that show subtle correlations with mineralization in the raw dataset. In addition, fractal-trained models can benefit practical mineral exploration by outputting low-risk exploration targets that achieve higher capturing efficiency and by providing new mineralization clues extracted from remote sensing data. This study demonstrates that the fractal representations of geological features filtered by multi-criteria feature selection can provide a feasible and promising means of improving the predictive capability of AI-driven MPM. [ABSTRACT FROM AUTHOR]

High-strength steel reinforcement offers promising application prospects in buildings with large spans and loads, as it can reduce steel congestion and simplify construction process. To investigate the mechanical behavior of flexural members reinforced with high-strength steel and measures to control crack width, in this study, static loading tests were conducted on 17 full-scale reinforced concrete (RC) beams with HRB635 high-strength steel reinforcement, six of which were reinforced with steel fiber. The failure mode, flexural strength, ductility, crack width, and deflection of the specimens were investigated, and the effects of different key parameters, including the reinforcement ratio and diameter, concrete strength, and steel fiber volume fraction were also analyzed. Test results indicated that the tensile strength of HRB635 reinforcement could be fully utilized, while the ductility ratios of high-strength steel reinforced concrete (HSS-RC) beams were less than those of beams with normal strength reinforcement. The maximum crack widths of most specimens without steel fibers exceeded the threshold defined in GB 50010-2010, indicating that the design of HSS-RC beams may be controlled by the requirements of serviceability limit state. Furthermore, the crack widths predicting methods defined in GB 50010-2010 and EN 1992-1-1 are more accurate for HSS-RC beams, while the deflection predicting methods defined in GB 50010-2010 and ACI 318-19 both underestimated the deflection of HSS-RC beams. This study could provide a theoretical basis for the engineering application of HRB635 high-strength steel reinforcement. [ABSTRACT FROM AUTHOR]

The development of mineral prospectivity mapping (MPM), which aims to outline and prioritize mineral exploration targets, has been spurred by advances in data-driven machine learning algorithms. Supervised data-driven MPM is a typical few-shot task, suffering from a scarcity of labeled data, the over-fitting of models and an uncertainty of predictions. The main objective of this contribution is to propose a robust framework of few-shot learning (FSL), combining data augmentation and transfer learning to enable the generation of prospectivity models with excellent predictive efficiency and low uncertainty. The mineral systems approach was used to transfer a conceptual mineral system into mappable exploration criteria. Synthetic minority over-sampling technique (SMOTE) was employed to augment and balance the labeled dataset, allowing for model pre-training with the large synthetic training dataset of a source domain. The knowledge derived from pre-trained models was then transferred to the target domain by fine-tuning, and the prospectivity model was generated in light of over-fitting and uncertainty assessments. The proposed FSL framework was applied to tungsten prospectivity mapping in southern Jiangxi Province. The results indicated that the SMOTE-ed balanced dataset boosted the classification accuracy in the training process. The FSL models yielded an arch-shaped prediction point pattern which was favorable for focusing potential targets with high probability and low uncertainty. The FSL models achieved a high predictive performance (test AUC = 0.9172) and the lowest quantitative over-fitting value compared to the models derived from the benchmark algorithms of random forest and support vector machine. Four levels of potential targeting zones, considering both predictive efficiency and uncertainty, were extracted from the resulting FSL prospectivity map. The final high-potential and low-risk exploration targets only cover 4.27% of the area, but capture 41.53% of known tungsten deposits, thus achieving a superior predictive performance. This study highlights the capability of FSL framework to control over-fitting and generate high-confidence exploration targets with low levels of uncertainty. [ABSTRACT FROM AUTHOR]

Objective: Survival and reconstruction in osteosarcoma is quite challenging. The study aimed to investigate the prognosis in patients treated with neoadjuvant chemotherapy and determine the clinical outcomes of expandable endoprosthesis reconstruction in children. Methods: From January 2009 to December 2014, we retrospectively analyzed 29 skeletally immature children (mean age, 10.5 years; range, 6–15 years) with osteosarcoma around the knee. Of the 29 patients who underwent neoadjuvant chemotherapy and limb salvage surgery, an expandable prosthesis was implanted for reconstruction. No patients were missed during follow‐up. The evaluation index involved follow‐up time, complication, functional results, and lengthening procedures. The survivorship and recurrence were assessed by GraphPad Software, and the function was evaluated by the Musculoskeletal Tumor Society (MSTS) scoring system. Results: A mean follow‐up time was 8.9 years (range, 6–12 years), and the overall 5‐year survival was 89.1% based on Kaplan–Meier analysis. Three patients suffered a relapse and one underwent amputation. Lung metastasis developed in one patient. At 6 months after the operation, patients had a mean MSTS score of 27 points (range, 24–29). Two patients underwent revision surgery, one for implant infection and one for aseptic loosening. Prognosis is correlated with alkaline phosphatase change after treatment. Conclusions: Chemotherapy scheme and limb salvage can achieve high survival rates. This expandable prosthesis was associated with good function and low complication rates. The character of expandability could be a method to overcome discrepancies in the growth period. [ABSTRACT FROM AUTHOR]

A hybrid structure constructed by replacing external steel beams and columns in the steel frame (SF) to precast concrete shear walls with insulation (PCSWs) and precast concrete cladding panels (PCCPs, SFPCSW) is proposed in this paper to avoid the problem of columns protruding from walls and PCCPs directly connected with SF. Numerical models of the SF and SFPCSW structures are established in ETABS software based on an 18‐story assembled steel residential building to investigate the seismic performance and collapse resistance capacity of SFPCSW. Time history analysis results show that the SFPCSW exhibits bending‐shearing lateral deformation and has more uniform interstory drift ratios than the SF. Under major earthquakes (i.e., probability of exceedance of 2% in 50 years), the damage of SF is concentrated in stories 1–6; for SFPCSW, the plastic hinges mainly appear in the concrete beams between PCSWs at each story, and the damage in PCSWs and the internal frame is small. Using pinned connections between the steel beam and PCSW reduces the structural lateral stiffness and increases the earthquake load carried by the internal frame. The incremental dynamic analysis results show that the SF and SFPCSW have similar collapse resistance capacities. [ABSTRACT FROM AUTHOR]

Anthracyclines, such as doxorubicin, represent one group of chemotherapy drugs with the most cardiotoxicity. Despite that anthracyclines are capable of treating assorted solid tumors and hematological malignancies, the side effect of inducing cardiac dysfunction has hampered their clinical use. Currently, the mechanism underlying anthracycline cardiotoxicity remains obscure. Increasing evidence points to mitochondria, the energy factory of cardiomyocytes, as a major target of anthracyclines. In this review, we will summarize recent findings about mitochondrial mechanism during anthracycline cardiotoxicity. In particular, we will focus on the following aspects: 1) the traditional view about anthracycline-induced reactive oxygen species (ROS), which is produced by mitochondria, but in turn causes mitochondrial injury. 2) Mitochondrial iron-overload and ferroptosis during anthracycline cardiotoxicity. 3) Autophagy, mitophagy and mitochondrial dynamics during anthracycline cardiotoxicity. 4) Anthracycline-induced disruption of cardiac metabolism. [ABSTRACT FROM AUTHOR]

Purpose: Hepatocellular carcinoma (HCC), a worldwide leading cause of morbidity and mortality, is the most frequent primary liver tumor. Most HCC patients are diagnosed with advanced liver cancer, resulting in a very low 5-year survival rate. Thus, there is an urgent need for the development of targeted therapies. In this study, we aimed to investigate the effect and mechanism of the miR-20a/EZH1 axis on the proliferation and metastasis of HCC and the inhibitory effect of the EZH1/EZH2 inhibitor UNC1999 on HCC. Materials and Methods: The expression of miR-20a in human HCC tissues and cell lines was detected using quantitative real-time PCR (qRT-PCR). The expressions of proteins were analyzed with immunohistochemistry and Western blotting. Luciferase assay was used to verify whether miR-20a targets EZH1 or EZH2. The effect of miR-20a on HCC progression was studied in vivo and in vitro. The tumor inhibitory effect of UNC1999 was confirmed in vivo. CCK8 assay, wound healing assay, cell migration and invasion assay were used to evaluate the synergistic effect of UNC1999 with sorafenib. RNA sequencing (RNA-seq) was performed to screen the differentially expressed genes in the Huh7 and SMMC7721 cell lines after UNC1999, sorafenib, and combination treatments. Results: In this study, miR-20a showed a lower expression in both HCC tissues and cell lines. MiR-20a inhibited the proliferation and migration of SMMC7721 and Huh7 cells. The results of the luciferase assay and Western blot analysis revealed that miR-20a directly targeted EZH1, a histone methyltransferase. We demonstrated that miR-20a negatively regulated the expression of EZH1 and inhibited the proliferation and metastasis of HCC by reducing H3K27 methylation. We found UNC1999 inhibited tumor cells proliferation and enhanced the inhibitory effect of sorafenib. Conclusion: We demonstrated that miR-20a suppresses the tumor proliferation and metastasis in HCC by directly targeting EZH1. UNC1999 can inhibit tumor proliferation in vivo and increase the sensitivity of hepatoma cell lines to sorafenib. [ABSTRACT FROM AUTHOR]

Detecting pavement cracks from images is a complex computer vision task due to their varying shapes, backgrounds, and sizes. We propose CrackF-Net, an end-to-end convolutional neural network for automatic crack detection in road images. We construct the CrackF-Net network using an encoder–decoder architecture to extract image features in convolutional blocks with residuals and fuse the multiscale convolutional features produced by the decoder. Convolutional blocks with residuals are used to capture the strong semantic features of cracks, and an adaptive filter fusion module is proposed to assist the network make a selection of filter fusion features on the channels. CrackF-Net fuses the multiscale features in decoder to improve crack detection performance. The proposed CrackF-Net is compared to other advanced crack detection methods using three public datasets. The experimental results show that CrackF-Net achieves state-of-the-art performance, which obtains F-measures of 0.866, 0.737, and 0.852 on the three datasets. [ABSTRACT FROM AUTHOR]

The combustion of lithium-ion batteries is characterized by fast ignition, prolonged duration, high combustion temperature, release of significant energy, and generation of a large number of toxic gases. Fine water mist has characteristics such as a high fire extinguishing efficiency and environmental friendliness. In order to thoroughly investigate the temperature control effect of fine water mist on lithium-ion battery fires. This study employs numerical simulation methods, utilizing PyroSim software to simulate the fire process in lithium-ion battery energy storage compartments. First, we focus on the variation patterns of flame, changes in combustion temperature, and heat release rate over time at environmental temperatures of 10, 25, and 35 °C. Subsequently, the suppression of flame, reduction in temperature, and changes in heat release rate are simulated for water mist in lithium-ion battery fires. The simulation results indicate that the environmental temperature has a considerable impact on the flame but a lesser effect on the heat release rate. Fine water mist effectively impedes the spread of thermal runaway between internal battery core cells, leading to a reduction in the flame size and a significant decrease in the maximum temperature and heat release rate. The numerical simulation results can provide scientific guidance for the prevention and control of fires in lithium-ion battery energy storage compartments., Competing Interests: The authors declare no competing financial interest., (© 2024 The Authors. Published by American Chemical Society.)

Background: The emergence of ceftazidime-avibactam (CZA) resistance among carbapenem-resistant Klebsiella pneumoniae (CRKP) is of major concern due to limited therapeutic options., Methods: In this study, 10 CRKP strains were isolated from different samples of a patient with CRKP infection receiving CZA treatment. Whole-genome sequencing (WGS) and conjugation experiments were performed to determine the transferability of the carbapenem resistance gene., Results: This infection began with a KPC-2-producing K. pneumoniae (CZA MIC = 2 μg/mL, imipenem MIC ≥ 16 μg/mL). After 20 days of CZA treatment, the strains switched to the amino acid substitution of T263A caused by a novel KPC - producing gene, bla KPC-145 , which restored carbapenem susceptibility but showed CZA resistance (CZA MIC ≥ 256 μg/mL, imipenem MIC = 1 μg/mL). The bla KPC-145 gene was located on a 148,185-bp untransformable IncFII-type plasmid. The subsequent use of carbapenem against KPC-145-producing K. pneumoniae infection led to a reversion of KPC-2 production (CZA MIC = 2 μg/mL, imipenem MIC ≥ 16 μg/mL). WGS analysis showed that all isolates belonged to ST11-KL47, and the number of SNPs was 14. This implied that these bla KPC -positive K. pneumoniae isolates might originate from a single clone and have been colonized for a long time during the 120-day treatment period., Conclusion: This is the first report of CZA resistance caused by bla KPC-145 , which emerged during the treatment with CZA against bla KPC-2 -positive K. pneumoniae -associated infection in China. These findings indicated that routine testing for antibiotic susceptibility and carbapenemase genotype is essential during CZA treatment., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Chen, Yang, Guo, Liu, Deng, Wu, Wu, Huang and Liao.)

Objective: To investigate the effectiveness of complete resection of bone tumor in pelvic zone Ⅱ and reconstruction with allogeneic pelvis, modular prosthesis, and three-dimensional (3D) printing prosthesis., Methods: The clinical data of 13 patients with primary bone tumor in pelvic zone Ⅱ who underwent tumor resection and acetabular reconstruction between March 2011 and March 2022 were retrospectively analyzed. There were 4 males and 9 females with an average age of 39.0 years ranging from 16 to 59 years. There were 4 cases of giant cell tumor, 5 cases of chondrosarcoma, 2 cases of osteosarcoma, and 2 cases of Ewing sarcoma. The Enneking classification of pelvic tumors showed that 4 cases involved zone Ⅱ, 4 cases involved zone Ⅰ and zone Ⅱ, and 5 cases involved zone Ⅱ and zone Ⅲ. The disease duration ranged from 1 to 24 months, with an average of 9.5 months. The patients were followed up to observe the recurrence and metastasis of the tumor, and the imaging examination was performed to observe the status of implant in place, fracture, bone resorption, bone nonunion, and so on. The improvement of hip pain was evaluated by visual analogue scale (VAS) score before operation and at 1 week after operation, and the recovery of hip function was evaluated according to the Musculoskeletal Tumor Society (MSTS) scoring system after operation., Results: The operation time was 4-7 hours, with an average of 4.6 hours; the intraoperative blood loss ranged from 800 to 1 600 mL, with an average of 1 200.0 mL. There was no reoperation or death after operation. All patients were followed up 9-60 months (mean, 33.5 months). No tumor metastasis was found in 4 patients receiving chemotherapy during follow-up. Postoperative wound infection occurred in 1 case, and prosthesis dislocation occurred in 1 case at 1 month after prosthesis replacement. One case of giant cell tumor recurred at 12 months after operation, and the puncture biopsy showed malignant transformation of giant cell tumor, and hemipelvic amputation was performed. The postoperative hip pain significantly relieved, and the VAS score was 6.1±0.9 at 1 week after operation, which was significantly different from the preoperative score (8.2±1.3) ( t =9.699, P <0.001). At 12 months after operation, the MSTS score was 23.0±2.1, including 22.8±2.1 for patients with allogenic pelvis reconstruction and 23.3±2.3 for patients with prosthsis reconstruction. There was no significant difference in the MSTS score between the two reconstruction methods ( t =0.450, P =0.516). At last follow-up, 5 patients could walk with cane assistance and 7 patients could walk without cane assistance., Conclusion: The resection and reconstruction of primary bone tumor in pelvic zone Ⅱ can obtain satisfactory hip function, and the interface of allogeneic pelvis and 3D printing prosthesis have better bone ingrowth, which is more in line with the requirements of biomechanics and biological reconstruction. However, pelvis reconstruction is difficult, the patient's condition should be evaluated comprehensively before operation, and the long-term effectiveness needs further follow-up.

Ferroptosis is a form of regulated cell death that relies on iron and is characterized by the accumulation of lipid peroxides, resulting in oncotic cell swelling and eventual disruption of cellular membranes. Lipid peroxidation, a hallmark of ferroptosis, refers to the oxidative deterioration of lipids that contain carbon-carbon double bonds, particularly polyunsaturated fatty acids (PUFAs). Understanding the molecular mechanisms underlying the interplay between ferroptosis and lipid peroxidation and identifying reliable techniques for assessing lipid peroxidation levels are crucial for further advancements in this field of research. Various methods have been developed to detect lipid peroxidation levels, including C11-BODIPY (BODIPY™ 581/591 C11), liperfluo, 4-hydroxynonenal (4-HNE), malondialdehyde (MDA), Click-iT LAA (linoleamide alkyne), and liquid chromatography-mass spectrometry (LC-MS)-based epilipidomics (redox lipidomics). Currently, one of the most commonly used and effective methods is the C11-BODIPY assay, which utilizes a fluorescent probe that selectively sensitizes lipid peroxidation in cell membranes. Incorporating advanced techniques such as flow cytometry and fluorescence microscopy with C11-BODIPY dye is essential for accurate assessment of lipid peroxidation levels in ferroptosis. This chapter aims to provide comprehensive experimental protocols for detecting lipid peroxidation levels indicative of ferroptosis using C11-BODIPY staining and subsequent detection via flow cytometry and fluorescence microscopy., (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Introduction: Immune checkpoint blockade inhibitor (ICI) therapy offers significant survival benefits for malignant melanoma. However, some patients were observed to be in disease progression after the first few treatment cycles. As such, it is urgent to find convenient and accessible indicators that assess whether patients can benefit from ICI therapy., Methods: In the training cohort, flow cytometry was used to determine the absolute values of 66 immune cell subsets in the peripheral blood of melanoma patients (n=29) before treatment with anti-PD-1 inhibitors. The least absolute shrinkage and selection operator (LASSO) Cox regression model was followed for the efficacy of each subset in predicting progression-free survival. Then we validated the performance of the selected model in validation cohorts (n=20), and developed a nomogram for clinical use., Results: A prognostic immune risk score composed of CD1c + dendritic cells and three subsets of T cells (CD8 + CD28 + , CD3 + TCRab + HLA-DR + , CD3 + TCRgd + HLA-DR + ) with a higher prognostic power than individual features (AUC = 0.825). Using this model, patients in the training cohort were divided into high- and low-risk groups with significant differences in mean progression-free survival (3.6 vs. 12.3 months), including disease control rate (41.2% vs. 91.7%), and objective response rate (17.6% vs. 41.6%). Integrating four-immune cell-subset based classifiers and three clinicopathologic risk factors can help to predict which patients might benefit from anti-PD-1 antibody inhibitors and remind potential non-responders to pursue effective treatment options in a timely way., Conclusions: The prognostic immune risk score including the innate immune and adaptive immune cell populations could provide an accurate prediction efficacy in malignant melanoma patients with ICI therapy., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Chi, Jiang, Li, Li, Wen, Ding, Chen, Zhang, Huang and Ding.)

Actomyosin contractile force produced by myosin II molecules that bind and pull actin filaments is harnessed for diverse functions, from cell division by the cytokinetic contractile ring to morphogenesis driven by supracellular actomyosin networks during development. However, actomyosin contractility is intrinsically unstable to self-reinforcing spatial variations that may destroy the actomyosin architecture if unopposed. How cells control this threat is not established, and while large myosin fluctuations and punctateness are widely reported, the full course of the instability in cells has not been observed. Here, we observed the instability run its full course in isolated cytokinetic contractile rings in cell ghosts where component turnover processes are absent. Unprotected by turnover, myosin II merged hierarchically into aggregates with increasing amounts of myosin and increasing separation, up to a maximum separation. Molecularly explicit simulations reproduced the hierarchical aggregation which precipitated tension loss and ring fracture and identified the maximum separation as the length of actin filaments mediating mechanical communication between aggregates. In the final simulated dead-end state, aggregates were morphologically quiescent, including asters with polarity-sorted actin, similar to the dead-end state observed in actomyosin systems in vitro. Our results suggest the myosin II turnover time controls actomyosin contractile instability in normal cells, long enough for aggregation to build robust aggregates but sufficiently short to intercept catastrophic hierarchical aggregation and fracture.

Purpose: To clarify the underlying regulatory mechanisms of progression from liver cirrhosis to hepatocellular carcinoma (HCC), we analyzed the microbiomics, metabolomics, and proteomics in plasma and tissues from patients with HCC or decompensated liver cirrhosis (DC)., Patients and Methods: Tissues and plasma from 44 HCC patients and 28 patients with DC were collected for metabolomic analysis. 16S rRNA sequencing was performed in nine HCC tissues (HCCT), four distal noncancerous tissues (HCCN), and 11 DC tissues (DCT). Five HCC tissues had liver cirrhosis (HCCT-LC). Five hepatocellular carcinoma tissues without liver cirrhosis (HCCT-NLC) and five DCT were selected for proteomic sequencing. After combining proteomic and metabolomic analysis, we constructed a mouse model of chronic liver injury using carbon tetrachloride (CCl4) and treated them with vitamin B6 (VB6)., Results: 16s rRNA sequence results showed that HCC tissues had higher alpha diversity. The highest LDA scores were detected for Elizabethkingia in HCCT, Subsaxibacter in DCT, and Stenotrophomon in HCCN. Metabolomics results demonstrated some metabolites, including capric acid, L-threonate, choline, alpha-D-Glucose, D-ribose, betaine, 2E-eicosenoic acid, linoleic acid, L-palmitoylcarnitine, taurodeoxycholic acid, L-pyroglutamic acid, androsterone sulfate, and phthalic acid mono-2-ethylhexyl ester (MEHP), had better diagnostic efficacy than AFP (AUC: 0.852; 95% CI: 0.749, 0.954). In a combined analysis of metabolomics and proteomics, we found that HCCT-LC had more obvious disorders of VB6 metabolism and pentose and glucuronate interconversions than DCT, and kynurenine metabolism disorder was more significant in HCCT-LC than in HCCT-NLC. In the CCl4-induced chronic liver injury model, after VB6 supplementation, inflammatory cell infiltration, hepatocyte edema, and degeneration were significantly improved., Conclusion: We found significant differences in the flora distribution between HCCT and DC; MEHP was a new diagnostic biomarker of HCC, and VB6 ameliorated the inflammatory cell infiltration, hepatocyte edema, and degeneration in chronic liver injury., Competing Interests: The authors report no conflicts of interest in this work., (© 2022 Mei et al.)

The fungal plant pathogen, Fusarium graminearum , contains two genes, FgCPK1 and FgCPK2 , encoding the catalytic subunits of cAMP-dependent protein kinase A. FgCPK1 and FgCPK2 are responsible for most of the PKA activities and have overlapping functions in various cellular processes in F. graminearum . The cpk1 cpk2 double mutant was significantly reduced in growth, rarely produced conidia, and was non-pathogenic. In this study, we found that the cpk1 cpk2 double mutant was unstable and produced fast-growing spontaneous sectors that were defective in plant infection. All spontaneous suppressor strains had mutations in FgSFL1 , a transcription factor gene orthologous to SFL1 in yeast. Thirteen suppressor strains had non-sense mutations at Q501, three suppressor strains had frameshift mutations at W198, and five suppressor strains had mutations in the HSF binding domain of FgSfl1. Only one suppressor strain had both a non-synonymous mutation at H225 and a non-sense mutation at R490. We generated the SFL1 deletion mutant and found that it produced less than 2% of conidia than that of the wild-type strain PH-1. The sfl1 mutant was significantly reduced in the number of perithecia on carrot agar plates at 7 days post-fertilization (dpf). When incubated for more than 12 days, ascospore cirrhi were observed on the sfl1 mutant perithecia. The infection ability of the sfl1 deletion mutant was also obviously defective. Furthermore, we found that in addition to the S223 and S559 phosphorylation sites, FgSFL1 had another predicted phosphorylation site: T452. Interestingly, the S223 phosphorylation site was responsible for sexual reproduction, and the T452 phosphorylation site was responsible for growth and sexual reproduction. Only the S559 phosphorylation site was found to play an important role in conidiation, sexual reproduction, and infection. Overall, our results indicate that FgSFL1 and its conserved PKA phosphorylation sites are important for vegetative growth, conidiation, sexual reproduction, and pathogenesis in F. graminearum .