The role of NAD-dependent deacetylase sirtuin-2 in liver metabolic stress through regulating pyruvate kinase M2 ubiquitination.

NAD-dependent deacetylase Sirt2 is involved in mammalian metabolic activities, matching energy demand with energy production and expenditure, and is relevant to a variety of metabolic diseases. Here, we constructed Sirt2 knockout and adeno-associated virus overexpression mice and found that deletion of hepatic Sirt2 accelerated primary obesity and insulin resistance in mice with concomitant hepatic metabolic dysfunction. However, the key targets of Sirt2 are unknown. We identified the M2 isoform of pyruvate kinase (PKM2) as a key Sirt2 target involved in glycolysis in metabolic stress. Through yeast two-hybrid and mass spectrometry combined with multi-omics analysis, we identified candidate acetylation modification targets of Sirt2 on PKM2 lysine 135 (K135). The Sirt2-mediated deacetylation-ubiquitination switch of PKM2 regulated the development of glycolysis. Here, we found that Sirt2 deficiency led to impaired glucose tolerance and insulin resistance and induced primary obesity. Sirt2 severely disrupted liver function in mice under metabolic stress, exacerbated the metabolic burden on the liver, and affected glucose metabolism. Sirt2 underwent acetylation modification of lysine 135 of PKM2 through a histidine 187 enzyme active site-dependent effect and reduced ubiquitination of the K48 ubiquitin chain of PKM2. Our findings reveal that the hepatic glucose metabolism links nutrient state to whole-body energetics through the rhythmic regulation of Sirt2.

Predicting the evolution of clinical skin aging in a multi-ethnic population: Developing causal Bayesian networks using dermatological expertise.

INTRODUCTION: Software to predict the impact of aging on physical appearance is increasingly popular. But it does not consider the complex interplay of factors that contribute to skin aging. OBJECTIVES: To predict the +15-year progression of clinical signs of skin aging by developing Causal Bayesian Belief Networks (CBBNs) using expert knowledge from dermatologists. MATERIAL AND METHODS: Structures and conditional probability distributions were elicited worldwide from dermatologists with experience of at least 15 years in aesthetics. CBBN models were built for all phototypes and for ages ranging from 18 to 65 years, focusing on wrinkles, pigmentary heterogeneity and facial ptosis. Models were also evaluated by a group of independent dermatologists ensuring the quality of prediction of the cumulative effects of extrinsic and intrinsic skin aging factors, especially the distribution of scores for clinical signs 15 years after the initial assessment. RESULTS: For easiness, only models on African skins are presented in this paper. The forehead wrinkle evolution model has been detailed. Specific atlas and extrinsic factors of facial aging were used for this skin type. But the prediction method has been validated for all phototypes, and for all clinical signs of facial aging. CONCLUSION: This method proposes a skin aging model that predicts the aging process for each clinical sign, considering endogenous and exogenous factors. It simulates aging curves according to lifestyle. It can be used as a preventive tool and could be coupled with a generative AI algorithm to visualize aging and, potentially, other skin conditions, using appropriate images.

Fault Diagnosis Method of Special Vehicle Bearing Based on Multi-Scale Feature Fusion and Transfer Adversarial Learning.

To address the issues of inadequate feature extraction for rolling bearings, inaccurate fault diagnosis, and overfitting in complex operating conditions, this paper proposes a rolling bearing diagnosis method based on multi-scale feature fusion and transfer adversarial learning. Firstly, a multi-scale convolutional fusion layer is designed to effectively extract fault features from the original vibration signals at multiple time scales. Through a feature encoding fusion module based on the multi-head attention mechanism, feature fusion extraction is performed, which can model long-distance contextual information and significantly improve diagnostic accuracy and anti-noise capability. Secondly, based on the domain adaptation (DA) cross-domain feature adversarial learning strategy of transfer learning methods, the extraction of optimal domain-invariant features is achieved by reducing the gap in data distribution between the target domain and the source domain, addressing the call for research on fault diagnosis across operating conditions, equipment, and virtual-real migrations. Finally, experiments were conducted to verify and optimize the effectiveness of the feature extraction and fusion network. A public bearing dataset was used as the source domain data, and special vehicle bearing data were selected as the target domain data for comparative experiments on the effect of network transfer learning. The experimental results demonstrate that the proposed method exhibits an exceptional performance in cross-domain and variable load environments. In multiple bearing cross-domain transfer learning tasks, the method achieves an average migration fault diagnosis accuracy rate of up to 98.65%. When compared with existing methods, the proposed method significantly enhances the ability of data feature extraction, thereby achieving a more robust diagnostic performance.

Surface-capping engineering for electrically neutral surface of perovskite films and stable solar cells.

Metal halide perovskite solar cells (PSCs) have developed rapidly in recent years, due to their high performance and low-cost solution-based fabrication process. These excellent properties are mainly attributed to the high defect tolerance of polycrystalline perovskite films. Meanwhile, these defects can also facilitate ion migration and carrier recombination, which cause the device performance and the long-term stability of PSCs to deteriorate heavily. Therefore, it is critical to passivate the defects, especially at the surfaces of perovskite grains where the defects are most concentrated due to the dangling bonds. Here we propose a surface-capping engineering (SCE) method to construct 'dangling-bond-free' surfaces for perovskite grains. Diamine iodide (methylenediammonium diiodide, MDAI2) was used to construct an electroneutral PbX6-MDA-PbX6(X = Cl, Br or I) layer at the perovskite surfaces. Compared to the monovalent FA+which can only coordinate one [PbX6]4-slab, the bivalent MDA2+can coordinate two [PbX6]4-slabs on both sides, thus realizing a dangling-bond-free surface. Solar cells based on SCE-perovskite films exhibited a higher power conversion efficiency (PCE) of 21.6%, compared with 19.9% of the control group; and maintained over 96% of its initial PCE after 13 h during the maximum power point tracking test under continuous AM1.5G illumination, whereas the control group only lasted 1.5 h. Constructing a dangling-bond-free capping layer on the grain boundary opens new avenues for the fabrication of ultralow-defect polycrystalline semiconductors, paving the way to further improve the PCE and lifetime of PSCs.

Three-Dimensional Arterial Distribution Over the Midline of the Nasal Bone.

BACKGROUND: A comprehensive understanding of arterial variations around the midline of the nose is of great importance for the safety of filler injection. OBJECTIVES: The aim of the study was to clearly define the 3-dimensional location of the arteries along the midline of the nasal bone. METHODS: The arterial structures overlapping the nasal bone along the midline were observed in 79 cadavers. RESULTS: The present study found that 0 to 3 named arteries per nose segment could be identified. All the arterial structures were located in or above the superficial musculoaponeurotic system layer overlapping the nasal bone. The probability of encountering named arteries at 5 defined points, P1 to P5, was 5/79 (6.3%), 4/79 (5.1%), 1/79 (1.3%), 6/79 (7.6%), and 9/79 (11.4%), respectively. The depth of the main arterial trunk was 1.2 ± 0.4 mm, 1.6 ± 0.6 mm, 1.8 ± 0 mm, 1.0 ± 0.4 mm, and 0.9 ± 0.5 mm below the skin at P1 to P5, respectively. CONCLUSIONS: The authors confirmed that sub-superficial musculoaponeurotic system injection along the midline through a needle is anatomically reliable and that a technique with 1 entry point through the rhinion via a cannula can easily keep the needle sufficiently deep for safe nasal filler injection.

The Arf-GAP AoGlo3 regulates conidiation, endocytosis, and pathogenicity in the nematode-trapping fungus Arthrobotrys oligospora.

Small GTPases of the ADP-ribosylation factor (Arf) family and their activating proteins (Arf-GAPs) regulate mycelial development and pathogenicity in yeast and filamentous fungi; however, little is known about their roles in nematode-trapping (NT) fungi. In this study, an ortholog of Arf-GAP Glo3 (AoGlo3) in Saccharomyces cerevisiae was characterized in the NT fungus Arthrobotrys oligospora. Deletion of the Aoglo3 gene resulted in growth defects and an increase in hyphal septum. Meanwhile, the sporulation capacity of the ΔAoglo3 mutant was decreased by 98%, and 67.1-71.2% spores became gourd or claviform in shape (from obovoid), which was accompanied by a significant decrease in the spore germination rate. This reduced sporulation capacity correlated with the transcriptional repression of several sporulation-related genes including fluG, rodA, abaA, medA, and lreA. The ΔAoglo3 mutant was also sensitive to several chemical stressors such as Congo red, NaCl, and sorbitol. Additionally, AoGlo3 was found to be involved in endocytosis, and more myelin figures were observed in the ΔAoglo3 mutant than in the wild-type strain, which was consistent with the presence of more autophagosomes observed in the mutant. Importantly, AoGlo3 affected the production of mycelial traps and serine proteases for nematode predation. In summary, AoGlo3 is involved in the regulation of multiple cellular processes such as mycelial growth, conidiation, environmental adaption, endocytosis, and pathogenicity in A. oligospora.

Improvement of Surgical Scars by Early Intervention With Carbon Dioxide Fractional Laser.

BACKGROUND AND OBJECTIVES: Laser therapy during the early stage of wound healing can reduce surgical scar formation. Previous clinical trials showed that ablative fractional laser (AFL) therapy might have achieved the best outcomes; however, there was no consensus on the laser therapy protocol. The current study investigated the efficacy of early intervention with carbon dioxide FL (CO2 FL) in reducing the surgical scars in patients of all age groups to understand the effects of specific intervention timing, light dose, number of treatments, treatment interval on the therapeutic outcome, and side effects. STUDY DESIGN/MATERIALS AND METHODS: Within 1 week after suture removal, 18 patients received low-energy CO2 FL therapy on their incisional wounds from facial or neck surgery. The treatments were performed once every 3 months for 1-3 sessions. After the surgical treatment, we applied a topical gel that contained recombinant human epidermal growth factor (EGF) on the incisional wounds to facilitate wound healing. The outcomes were evaluated by the Vancouver Scar Scale (VSS) and a satisfaction survey (the quartile grading scale) before and 3 months after completing the therapy was conducted. During the same period, 15 patients who had not received any treatment for about one year after face and neck surgery were randomly selected as the untreated control group. RESULTS: In the treatment group, 18 patients had no obvious scar marks after the laser therapy. The satisfaction survey showed excellent outcomes in 16 (88.9%) patients and good outcomes in two (11.1%). After treatment, the average VSS scale and satisfaction score of the treatment group were 1.11 and 3.89, respectively; while the average VSS scale of the control group was 3.07 and the satisfaction score was 1.93. There were significant differences in VSS scale and satisfaction score between the treatment group and the control group (P < 0.01). No adverse effects were observed. CONCLUSIONS: Early intervention with CO2 FL on surgical wounds achieved satisfactory outcomes in improving the surgical scars. Lasers Surg. Med. © 2019 Wiley Periodicals, Inc.

Effect of Pb Filling and Synthesis Pressure Regulation on the Thermoelectric Properties of CoSb3.

To take the advantage of the synergy of atom filling and pressure regulation, atom Pb was assumed to play the role of fillers to occupy the Sb-icosahedron voids. In this paper, skutterudite Pb xCo4Sb11.5Te0.5 materials have been synthesized by high-pressure and high-temperature (HPHT) method with 0.5 h processing time. Although the increase of Pb filling rate increased the electrical resistivity of samples, it reduced the thermal conductivity of samples significantly. When the synthesis pressure increased, the Seebeck coefficients of Pb0.2Co4Sb11.5Te0.5 increased, and the thermal conductivity decreased. The crystal morphology and structure of samples, which can effectively affect the thermoelectric properties of materials, were investigated with XRD, Raman spectra, SEM mapping, and HR-TEM analysis.

The effect of ice-cold water spray following the model for symptom management on postoperative thirst in patients admitted to intensive care unit: A randomized controlled study.

BACKGROUND: Postoperative thirst is common in patients admitted to the intensive care unit. Existing methods like wet cotton swabs or oral care prove ineffectual or operationally intricate. Currently, an efficacious postoperative thirst alleviation method remains elusive. Exploring a prompt, safe, and efficacious solution is of paramount importance. OBJECTIVE: To assess the effect of ice-cold water spray applied following a symptom management model on postoperative thirst and to establish a framework for mitigating thirst in intensive care unit patients. RESEARCH DESIGN: Single-center randomized controlled study. SETTING: Surgical intensive care unit in a university-affiliated hospital. MAIN OUTCOME MEASURES: 56 intensive care unit patients were selected and equally randomized. The experimental group received ice-cold water spray in conjunction with eight symptom management strategies, while the control group underwent standard care involving wet cotton swabs. Thirst intervention was initiated 0.5 hours after postoperative extubation, followed by subsequent interventions at 2-hour, 4-hour, and 6-hour intervals post-extubation. Thirst intensity, oral comfort, and the duration of relief from thirst were assessed and compared between groups before and 0.5 hours after each thirst intervention. RESULTS: Across different interventions, the experimental group exhibited superior scores in thirst intensity and oral comfort compared to the control group. Additionally, the nursing time required to alleviate thirst in the experimental group was significantly shorter than that in the control group (P < 0.01). CONCLUSION: Ice-cold water spray following the model for symptom management can effectively mitigate the postoperative thirst intensity in intensive care unit patients, improve oral comfort, and reduce the nursing time for relieving thirst. IMPLICATIONS FOR CLINICAL PRACTICE: Clinical nurses can employ ice-cold water spray following the model for symptom management to ameliorate postoperative thirst intensity in ICU patients while enhancing oral comfort. Furthermore, the utilization of ice-cold water spray can reduce the nursing time required for relieving postoperative thirst in intensive care unit patients.

Bridging-Solvent Strategy for Quasi-Single-Crystal Perovskite Films and Stable Solar Cells.

Controllable fabrication of formamidinium (FA)-based perovskite solar cells (PSCs) with both high efficiency and long-term stability is the key to their further commercialization. However, the diversity of PbI2 complexes and perovskite compositions usually leads to light sensitive PbI2 residues and phase impurities in the film, which can accelerate the device degradation. Here, the crystallization kinetics of FA-based perovskite films are studied and a bridging-solvent strategy is proposed to modulate the reaction kinetics between PbI2 and ammonium salts by prohibiting the formation of undesired intermediates. N-methylpyrrolidone (NMP) solvent is introduced into the PbI2 precursor solution to obtain stable and homogeneous PbI2-NMP complex films. The strong interaction between NMP and formamidinium iodide (FAI) molecules promotes the conversion from PbI2-NMP into (001)-oriented quasi-single-crystal perovskite films with negligible impurities, long carrier lifetime of 1.5 µs and a large grain size of 3 µm. The optimized PSCs exhibit a high power conversion efficiency of 24.1%, as well as superior shelf stability which maintains 95% initial efficiency after storage in air for 1200 h (T95 = 1200 h), and operating stability with T96 = 300 h under continuous working at the maximum power point. This work offers a simple and reproducible method for fabricating phase-pure and uniaxially oriented perovskite films.

Uniaxial-Oriented Perovskite Films with Controllable Orientation.

Perovskite films with large crystal size, preferred orientation, and facile fabrication process, combining advantages of single-crystal and polycrystalline films, have gained considerable attention recently. However, there is little research on the facet properties of perovskite films. Here, (111)- and (001)-oriented perovskite films with bandgaps ranging from 1.53 to 1.77 eV, and systematically investigated their orientation-dependent properties are achieved. The (111)-oriented films show electron-dominated traps and the (001)-oriented films show hole-dominated traps, which are related to their atomic arrangement at the surface. Compared with the (001)-oriented films, the (111)-oriented films exhibit lower work function and superior water/oxygen robustness. For the wide-bandgap films, the lattice of the (001)-oriented film provides an unobstructed passage for ion migration. Comparably, the (111)-oriented films exhibit suppressed ion migration and excellent phase stability. The optimized unencapsulated solar cells based on both (001) and (111) orientations show a similar high efficiency of ≈23%. The (111)-oriented solar cell exhibits excellent stability, maintaining 95% of its initial efficiency after 1500 h maximum power point (MPP) tracking test, and 97% initial efficiency after 3000 h aging in ambient conditions. This work paves the way for the rational design, controllable synthesis, and targeted optimization of uniaxial-oriented perovskite films for various electronic applications.

Peroxin Pex14/17 Is Required for Trap Formation, and Plays Pleiotropic Roles in Mycelial Development, Stress Response, and Secondary Metabolism in Arthrobotrys oligospora.

The peroxins encoded by PEX genes involved in peroxisome biogenesis play a crucial role in cellular metabolism and pathogenicity in fungi. Herein, we characterized a filamentous fungus-specific peroxin Pex14/17 in the Arthrobotrys oligospora, a representative species of nematode-trapping fungi. The deletion of AoPEX14/17 resulted in a remarkable reduction in mycelial growth, conidia yield, trap formation, and pathogenicity. Compared with the wild-type strain, the ΔAopex14/17 mutant exhibited more lipid droplet and reactive oxygen species accumulation accompanied with a significant decrease in fatty acid utilization and tolerance to oxidative stress. Transcriptomic analysis indicated that AoPEX14/17 was involved in the regulation of metabolism, genetic information processing, environmental information processing, and cellular processes. In subcellular morphology, the deletion of AoPEX14/17 resulted in a decrease in the number of cell nuclei, autophagosomes, and Woronin bodies. Metabolic profile analysis showed that AoPex14/17 affects the biosynthesis of secondary metabolites. Yeast two-hybrid assay revealed that AoPex14/17 interacted with AoPex14 but not with AoPex13. Taken together, our results suggest that Pex14/17 is the main factor for modulating growth, development, and pathogenicity in A. oligospora. IMPORTANCE Peroxisome biogenesis genes (PEX) play an important role in growth, development, and pathogenicity in pathogenic fungi. However, the roles of PEX genes remain largely unknown in nematode-trapping (NT) fungi. Here, we provide direct evidence that AoPex14/17 regulates mycelial growth, conidiation, trap formation, autophagy, endocytosis, catalase activity, stress response to oxidants, lipid metabolism, and reactive oxygen species production. Transcriptome analysis and metabolic profile suggested that AoPex14/17 is involved in multiple cellular processes and the regulation of secondary metabolism. Therefore, our study extends the functions of PEX genes, which helps to elucidate the mechanism of organelle development and trap formation in NT fungi and lays the foundation for the development of efficient nematode biocontrol agents.

Morin exhibits a neuroprotective effect in MPTP-induced Parkinson's disease model via TFEB/AMPK-mediated mitophagy.

BACKGROUND: Parkinson's disease (PD) is one of the most common neurodegenerative diseases in the world. Mitophagy has been implicated in PD etiology for decades and its pharmacological activation is recognized as a promising treatment strategy for PD. For mitophagy initiation, low mitochondrial membrane potential (ΔΨm) is essential. We identified a natural compound morin that could induce mitophagy without affecting ΔΨm. Morin is a flavonoid that can be isolated from fruits like mulberry. PURPOSE: To reveal the effect of morin on the PD mice model and their potential underlying molecular mechanism. METHODS: Mitophagy process induced by morin in N2a cells meditation were measured using flow cytometry and immunofluorescence. JC-1 fluorescence dye used to detect the mitochondrial membrane potential (ΔΨm). The TFEB nuclear translocation were examined by immunofluorescence staining and western blot assay. The PD mice model was induced by MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) intraperitoneal administration. RESULTS: We found that morin also promoted nuclear translocation of the mitophagy regulator TFEB and activated the AMPK-ULK1 pathway. In MPTP-induced PD in vivo models, morin protected DA neurons from MPTP neurotoxicity and ameliorated behavioral deficit. CONCLUSION: Although morin was previously reported to be neuroprotective in PD, the detailed molecular mechanisms remain to be elucidated. For the first time, we report morin served as a novel and safe mitophagy enhancer underlying AMPK-ULK1 pathway and exhibited anti-Parkinsonian effects indicating its potential as a clinical drug for PD treatment.

A reinterpretation of the gap fraction of tree crowns from the perspectives of computer graphics and porous media theory.

The gap fraction (GF) of vegetative canopies is an important property related to the contained bulk of reproductive elements and woody facets within the tree crown volume. This work was developed from the perspectives of porous media theory and computer graphics techniques, considering the vegetative elements in the canopy as a solid matrix and treating the gaps between them as pores to guide volume-based GFvol calculations. Woody components and individual leaves were extracted from terrestrial laser scanning data. The concept of equivalent leaf thickness describing the degrees of leaf curling and drooping was proposed to construct hexagonal prisms properly enclosing the scanned points of each leaf, and cylinder models were adopted to fit each branch segment, enabling the calculation of the equivalent leaf and branch volumes within the crown. Finally, the volume-based GFvol of the tree crown following the definition of the void fraction in porous media theory was calculated as one minus the ratio of the total plant leaf and branch volume to the canopy volume. This approach was tested on five tree species and a forest plot with variable canopy architecture, yielding an estimated maximum volume-based GFvol of 0.985 for a small crepe myrtle and a minimal volume-based GFvol of 0.953 for a sakura tree. The 3D morphology of each compositional element in the tree canopy was geometrically defined and the canopy was considered a porous structure to conduct GFvol calculations based on multidisciplinary theory.

Biological Activities of p-Hydroxycinnamic Acids in Maintaining Gut Barrier Integrity and Function.

It is well established that p-Hydroxycinnamic acids (HCAs), including ferulic, caffeic, sinapic, and p-coumaric acids, possess a characteristic phenylpropanoid C6-C3 backbone and account for about one-third of the phenolic compounds in our diet. HCAs are typically associated with various plant cell wall components, including mono-, di-, and polysaccharides, sterols, polyamines, glycoproteins, and lignins. Interestingly, enzymes produced by intestinal microbes liberate HCAs from these associations. HCAs are completely absorbed in their free form upon ingestion and undergo specific reactions upon absorption in the small intestine or liver. The gut epithelium, composed of intestinal epithelial cells (IECs), acts as a physical barrier against harmful bacteria and a site for regulated interactions between bacteria and the gut lumen. Thus, maintaining the integrity of the epithelial barrier is essential for establishing a physiochemical environment conducive to homeostasis. This review summarizes the protective effects of HCAs on the intestinal barrier, achieved through four mechanisms: preserving tight junction proteins (TJPs), modulating pro-inflammatory cytokines, exerting antioxidant activity, and regulating the intestinal microbiota.

(111)-Dominated Perovskite Films by Antisolvent Engineering.

Fabricating perovskite films with a dominant crystal orientation is an effective path to realizing quasi-single-crystal perovskite film, which can eliminate the fluctuation of the electrical properties in films arising from grain-to-grain variations, and improve the performance of perovskite solar cells (PSCs). Perovskite (FAPbI3 ) films based on one-step antisolvent methods usually suffer from chaotic orientations due to the inevitable intermediate phase conversion from intermediates of PbI2 •DMSO, FA2 Pb3 I8 •4DMSO, and δ-FAPbI3 to α-FAPbI3 . Here, a high-quality perovskite film with (111) preferred orientation ((111)-α-FAPbI3 ) using a short-chain isomeric alcohol antisolvent, isopropanol (IPA) or isobutanol (IBA), is reported. The interaction between IPA and PbI2 leads to a corner-sharing structure instead of an edge-sharing PbI2 octahedron, sidestepping the formation of these intermediates. With the volatilization of IPA, FA+ can replace IPA in situ to form α-FAPbI3 along the (111) direction. Compared to randomly orientated perovskites, the dominantly (111) orientated perovskite ((111)-perovskite) exhibits improved carrier mobility, uniform surface potential, suppressed film defects and enhanced photostability. PSCs based on the (111)-perovskite films show 22% power conversion efficiency and excellent stability, which remains unchanged after 600 h continuous working at maximum power point, and 95% after 2000 h of storage in atmosphere environment.

(Z)-N-{(E)-10-[(2,6-Diisopropyl-phen-yl)-imino]-9,10-dihydro-phenanthren-9-yl-idene}-2,6-dimethyl-aniline.

The title compound, C(34)H(34)N(2), adopts a Z,E configuration with respect to the N=C-C=N backbone, with an N-C-C-N torsion angle of 41.1 (4)° The dihedral angle between the benzene rings in the 9,10-dihydro-phenanthrene moiety is 18.0 (1)°.

AoPEX1 and AoPEX6 Are Required for Mycelial Growth, Conidiation, Stress Response, Fatty Acid Utilization, and Trap Formation in Arthrobotrys oligospora.

Arthrobotrys oligospora (A. oligospora) is a typical nematode-trapping (NT) fungus that can capture nematodes by producing adhesive networks. Peroxisomes are single membrane-bound organelles that perform multiple physiological functions in filamentous fungi. Peroxisome biogenesis proteins are encoded by PEX genes, and the functions of PEX genes in A. oligospora and other NT fungi remain largely unknown. Here, our results demonstrated that two PEX genes (AoPEX1 and AoPEX6) are essential for mycelial growth, conidiation, fatty acid utilization, stress tolerance, and pathogenicity in A. oligospora. AoPEX1 and AoPEX6 knockout resulted in a failure to produce traps, conidia, peroxisomes, and Woronin bodies and damaged cell walls, reduced autophagosome levels, and increased lipid droplet size. Transcriptome data analysis showed that AoPEX1 and AoPEX6 deletion resulted in the upregulation of the proteasome, membranes, ribosomes, DNA replication, and cell cycle functions, and the downregulation of MAPK signaling and nitrogen metabolism. In summary, our results provide novel insights into the functions of PEX genes in the growth, development, and pathogenicity of A. oligospora and contribute to the elucidation of the regulatory mechanism of peroxisomes in trap formation and lifestyle switching in NT fungi. IMPORTANCE Nematode-trapping (NT) fungi are important resources for the biological control of plant-parasitic nematodes. They are widely distributed in various ecological environments and capture nematodes by producing unique predatory organs (traps). However, the molecular mechanisms of trap formation and lifestyle switching in NT fungi are still unclear. Here, we provided experimental evidence that the AoPEX1 and AoPEX6 genes could regulate mycelial growth and development, trap formation, and nematode predation of A. oligospora. We further analyzed the global transcription level changes of wild-type and mutant strains using RNA-seq. This study highlights the important role of peroxisome biogenesis genes in vegetative growth, conidiation, trap formation, and pathogenicity, which contribute to probing the mechanism of organelle development and trap formation of NT fungi and lays a foundation for developing high-efficiency nematode biocontrol agents.

Functional Analysis of Two Affinity cAMP Phosphodiesterases in the Nematode-Trapping Fungus Arthrobotrys oligospora.

Phosphodiesterases are essential regulators of cyclic nucleotide signaling with diverse physiological functions. Two phosphodiesterases, PdeH and PdeL, have been identified from yeast and filamentous fungi. Here, the orthologs of PdeH and PdeL were characterized in a typical nematode-trapping fungus Arthrobotrys oligospora by gene disruption and phenotypic comparison. Deletion of AopdeH caused serious defects in mycelial growth, conidiation, stress response, trap formation, and nematicidal efficiency compared to the wild-type strain. In contrast, these phenotypes have no significant difference in the absence of AopdeL. In addition, deletion of AopdeH and AopdeL resulted in a remarkable increase in cAMP level during vegetative growth and trap formation, and the number of autophagosomes was decreased in ΔAopdeH and ΔAopdeL mutants, whereas their volumes considerably increased. Moreover, metabolomic analyses revealed that many metabolites were downregulated in ΔAopdeH mutant compared to their expression in the wild-type strain. Our results indicate that AoPdeH plays a crucial role in mycelial growth, conidiation, stress response, secondary metabolism, and trap formation. In contrast, AoPdeL only plays a minor role in hyphal and conidial morphology, autophagy, and trap formation in A. oligospora. This work expands the roles of phosphodiesterases and deepens the understanding of the regulation of trap formation in nematode-trapping fungi.

Functional analysis of seven regulators of G protein signaling (RGSs) in the nematode-trapping fungus Arthrobotrys oligospora.

Regulators of G protein signaling (RGSs) are proteins that negatively regulate G protein signal transduction. In this study, seven putative RGSs were characterized in the nematode-trapping (NT) fungus, Arthrobotrys oligospora. Deleting Rgs genes significantly increased intracellular cAMP levels, and caused defects in mycelia growth, stress resistance, conidiation, trap formation, and nematocidal activity. In particular, the ΔAoFlbA mutant was unable to produce conidia and traps. Transcriptomic analysis showed that amino acid metabolic and biosynthetic processes were significantly enriched in the ΔAoFlbA mutant compared to WT. Interestingly, Gas1 family genes are significantly expanded in A. oligospora and other NT fungi that produce adhesive traps, and are differentially expressed during trap formation in A. oligospora. Disruption of two Gas1 genes resulted in defective conidiation, trap formation, and pathogenicity. Our results indicate that RGSs play pleiotropic roles in regulating A. oligospora mycelial growth, development, and pathogenicity. Further, AoFlbA is a prominent member and required for conidiation and trap formation, possibly by regulating amino acid metabolism and biosynthesis. Our results provide a basis for elucidating the signaling mechanism of vegetative growth, lifestyle transition, and pathogenicity in NT fungi.