Molecular mechanisms of metabolic dysregulation in diabetic cardiomyopathy.

Diabetic cardiomyopathy (DCM), one of the most serious complications of diabetes mellitus, has become recognized as a cardiometabolic disease. In normoxic conditions, the majority of the ATP production (>95%) required for heart beating comes from mitochondrial oxidative phosphorylation of fatty acids (FAs) and glucose, with the remaining portion coming from a variety of sources, including fructose, lactate, ketone bodies (KB) and branched chain amino acids (BCAA). Increased FA intake and decreased utilization of glucose and lactic acid were observed in the diabetic hearts of animal models and diabetic patients. Moreover, the polyol pathway is activated, and fructose metabolism is enhanced. The use of ketones as energy sources in human diabetic hearts also increases significantly. Furthermore, elevated BCAA levels and impaired BCAA metabolism were observed in the hearts of diabetic mice and patients. The shift in energy substrate preference in diabetic hearts results in increased oxygen consumption and impaired oxidative phosphorylation, leading to diabetic cardiomyopathy. However, the precise mechanisms by which impaired myocardial metabolic alterations result in diabetes mellitus cardiac disease are not fully understood. Therefore, this review focuses on the molecular mechanisms involved in alterations of myocardial energy metabolism. It not only adds more molecular targets for the diagnosis and treatment, but also provides an experimental foundation for screening novel therapeutic agents for diabetic cardiomyopathy.

Toxic effects of nanoplastics on biological nitrogen removal in constructed wetlands: Evidence from iron utilization and metabolism.

Nanoplastics (NPs) in wastewaters may present a potential threat to biological nitrogen removal in constructed wetlands (CWs). Iron ions are pivotal in microbially mediated nitrogen metabolism, however, explicit evidence demonstrating the impact of NPs on nitrogen removal regulated by iron utilization and metabolism remains unclear. Here, we investigated how NPs disturb intracellular iron homeostasis, consequently interfering with the coupling mechanism between iron utilization and nitrogen metabolism in CWs. Results indicated that microorganisms affected by NPs developed a siderophore-mediated iron acquisition mechanism to compensate for iron loss. This deficiency resulted from NPs internalization limited the activity of the electron transport system and key enzymes involved in nitrogen metabolism. Microbial network analysis further suggested that NPs exposure could potentially trigger destabilization in microbial networks and impair effective microbial communication, and ultimately inhibit nitrogen metabolism. These adverse effects, accompanied by the dominance of Fe3+ over certain electron acceptors engaged in nitrogen metabolism under NPs exposure, were potentially responsible for the observed significant deterioration in nitrogen removal (decreased by 30 %). This study sheds light on the potential impact of NPs on intracellular iron utilization and offers a substantial understanding of the iron-nitrogen coupling mechanisms in CWs.

Remediation of soda-saline-alkali soil through soil amendments: Microbially mediated carbon and nitrogen cycles and remediation mechanisms.

Due to the high salt content and pH value, the structure of saline-sodic soil was deteriorated, resulting in decreased soil fertility and inhibited soil element cycling. This, in turn, caused significant negative impacts on crop growth, posing a major challenge to global agriculture and food security. Despite numerous studies aimed at reducing the loss of plant productivity in saline-sodic soils, the knowledge regarding shifts in soil microbial communities and carbon/nitrogen cycling during saline-sodic soil improvement remains incomplete. Consequently, we developed a composite soil amendment to explore its potential to alleviate salt stress and enhance soil quality. Our findings demonstrated that the application of this composite soil amendment effectively enhanced microbial salinity resistance, promotes soil carbon fixation and nitrogen cycling, thereby reducing HCO3- concentration and greenhouse gas emissions while improving physicochemical properties and enzyme activity in the soil. Additionally, the presence of CaSO4 contributed to a decrease in water-soluble Na+ content, resulting in reduced soil ESP and pH by 14.64 % and 7.42, respectively. Our research presents an innovative approach to rehabilitate saline-sodic soil and promote ecological restoration through the perspective of elements cycles.

Investigation of non-classical secretion of oxalate decarboxylase in Bacillus mojavensis XH1 mediated by exopeptide YydF: Mechanism and application.

Non-classical secretory proteins are widely found in bacteria and have been extensively studied due to their important physiological roles. However, the relevant non-classical secretory mechanisms remain unclear. In this study, we found that oxalate decarboxylase (Bacm OxDC) from Bacillus mojavensis XH1 belongs to non-classical secretory proteins. Its N-terminus showed high hydrophilicity, which was different from the conventional signal peptide. The truncation test revealed that the deletion of the N-terminus affects the structure resulting in its inability to cross the cell membrane. Further studies verified that the exported peptide YydF played an important role in the secretion process of Bacm OxDC. Experimental results on the secretion mechanism indicated that Bacm OxDC bound to the exported peptide YydF and they are translocated to the cell membrane together, after which Bacm OxDC caused cell membrane relaxation for transmembrane secretion. Thereafter, three recombinant proteins were successfully secreted with certain enzymatic activity by fusing Bacm OxDC as a guide protein with various target proteins. To the best of our knowledge, this was the first time that non-classical secretion mechanism in bacteria has been analyzed. The novel discovery may provide a reference and broaden the horizons of the secretion pathway and expression regulation of proteins.

Raloxifene-driven benzothiophene derivatives: Discovery, structural refinement, and biological evaluation as potent PPARγ modulators based on drug repurposing.

By virtue of the drug repurposing strategy, the anti-osteoporosis drug raloxifene was identified as a novel PPARγ ligand through structure-based virtual high throughput screening (SB-VHTS) of FDA-approved drugs and TR-FRET competitive binding assay. Subsequent structural refinement of raloxifene led to the synthesis of a benzothiophene derivative, YGL-12. This compound exhibited potent PPARγ modulation with partial agonism, uniquely promoting adiponectin expression and inhibiting PPARγ Ser273 phosphorylation by CDK5 without inducing the expression of adipongenesis associated genes, including PPARγ, aP2, CD36, FASN and C/EBPα. This specific activity profile resulted in effective hypoglycemic properties, avoiding major TZD-related adverse effects like weight gain and hepatomegaly, which were demonstrated in db/db mice. Molecular docking studies showed that YGL-12 established additional hydrogen bonds with Ile281 and enhanced hydrogen-bond interaction with Ser289 as well as PPARγ Ser273 phosphorylation-related residues Ser342 and Glu343. These findings suggested YGL-12 as a promising T2DM therapeutic candidate, thereby providing a molecular framework for the development of novel PPARγ modulators with an enhanced therapeutic index.

Gold nanoparticles synthesis mediated by fungus isolated from aerobic granular sludge: Process and mechanisms.

Due to the low toxicity, biocompatibility and eco-friendliness, microorganisms have received a lot of attention for gold nanoparticles (AuNPs) synthesis. This work isolated a fungal strain capable of efficiently generating AuNPs from aerobic granular sludge, named XY3. Comparison of 18S rDNA sequence results showed that fungus XY3 belongs to Candida rugopelliculosa. AuNPs were synthesized by initiating an Au3+-induced stress response that prompted the reduction of Au3+ to Au0 by the fungus XY3. It is worth noting that the addition of nutritional substrates weakens the stress response induced by Au3+, resulting in a decrease in the yield of AuNPs. As evidenced by nystatin inhibition studies, the synthesis of AuNPs is based on biochemical reactions rather than purely physical changes. The XRD results suggested that XY3-secreted biomolecules were involved in the reduction of Au3+ and AuNPs synthesis. The results of the three variation patterns of reducing power, biomolecules, and AuNPs absorbance revealed that Au3+ reduction was mostly dependent on the reducing polysaccharides. In addition, extracellular proteins were shown to be involved in the synthesis of AuNPs, which is responsible for the uniform distribution of AuNPs. This work provided a wide and cost-effective seed source for AuNPs synthesis, and also offered a resourceful solution for residual sludge treatment of fungal type aerobic granular sludge.

Chemotaxis-mediated degradation of PAHs and heterocyclic PAHs under low-temperature stress by Pseudomonas fluorescens S01: Insights into the mechanisms of biodegradation and cold adaptation.

As wellknown persistent contaminants, polycyclic aromatic hydrocarbons (PAHs) and heterocyclic polyaromatic hydrocarbons (Heterocyclic PAHs)'s fates in cryogenic environments are remains uncertain. Herein, strain S01 was identified as Pseudomonas fluorescens, a novel bacterium tolerant to low temperature and capable of degrading PAHs and heterocyclic PAHs. Strain S01 exhibited growth at 5-40 â„ƒ and degradation rate of mixed PAHs and heterocyclic PAHs reached 52% under low-temperature. Through comprehensive metabolomic, genomic, and transcriptomic analyses, we reconstructed the biodegradation pathway for PAHs and heterocyclic PAHs in S01 while investigating its response to low temperature. Further experiments involving deletion and replacement of methyl-accepting chemotaxis protein (MCP) confirmed its crucial role in enabling strain S01's adaptation to dual stress of low temperature and pollutants. Additionally, our analysis revealed that MCP was upregulated under cold stress which enhanced strain S01's motility capabilities leading to increased biofilm formation. The establishment of biofilm promoted preservation of distinct cellular membrane stability, thereby enhancing energy metabolism. Consequently, this led to heightened efficiency in pollutant degradation and improved cold resistance capabilities. Our findings provide a comprehensive understanding of the environmental fate of both PAHs and heterocyclic PAHs under low-temperature conditions while also shedding light on cold adaptation mechanism employed by strain S01.

Virtual screening of natural products targeting ubiquitin-specific protease 7.

Ubiquitin-specific protease 7 (USP7) is a promising prognostic and druggable target for cancer therapy. Inhibition of USP7 can activate the MDM2-P53 signaling pathway, thereby promoting cancer cell apoptosis. This study based on watvina molecular docking of virtual screening method and biological evaluation found the new USP7 inhibitors targeting catalytic active site. Three hits were screened from 3760 natural products and validated as USP7 inhibitors by enzymatic and kinetic assays. The IC50 values of scutellarein (Scu), semethylzeylastera (DML) and salvianolic acid C (SAC) were 3.017, 6.865 and 8.495 µM, respectively. Further, we reported that the hits could downregulate MDM2 and activate p53 signal pathway in HCT116 cells. Molecular dynamics simulation was used to investigate the binding mechanism of USP7 to Scu, the compound with the best performance, which formed stable contact with Val296, Gln297, Phe409, Tyr465 and Tyr514. These interactions are essential for maintaining the biological activity of Scu. Three natural products are suitable as lead compounds for the development of novel USP7 inhibitors, especially anti-colon cancer drugs.Communicated by Ramaswamy H. Sarma.

Energy utilization of agricultural waste: Machine learning prediction and pyrolysis transformation.

The rapid screening of agricultural waste materials for capacitor preparation holds significant importance in comprehending the relationship between material properties and enhancing experimental efficiency. In this study, we developed two machine learning models to predict electrode material characteristics using 2997 data points extracted from 235 articles. The identification and influence of key features on prediction indices provide a theoretical foundation for subsequent practical preparation. Through regression analysis and index evaluation, corn straw emerged as the optimal material for capacitor preparation, leading us to propose a one-step activation and two-step modification approach to convert corn straw into porous biochar. By modifying biochar with Co(NO3)2·6H2O, the maximum electrode capacitance of porous carbon reached 732.6 F/g. Furthermore, the electrode exhibited exceptional cycle stability with a remaining capacitance of 96 % after 5000 cycles. The prepared symmetric capacitor demonstrated pseudocapacitance behavior with a capacitance of 183.15 F/g at a current density of 1.0 A/g, power density of 22 kW/kg, and energy density of 9.03 Wh/kg. Considering the increasing annual output of corn straw and its superior industrial application prospects compared to acid-, base-, or precious metal-based alternatives due to their cost-effectiveness and environmental friendliness, these findings highlight the potential practical value in utilizing modified corn straw biochar as an efficient energy storage electrode material.

Automatic Diagnosis of Major Depressive Disorder Using a High- and Low-Frequency Feature Fusion Framework.

Major Depressive Disorder (MDD) is a common mental illness resulting in immune disorders and even thoughts of suicidal behavior. Neuroimaging techniques serve as a quantitative tool for the assessment of MDD diagnosis. In the domain of computer-aided magnetic resonance imaging diagnosis, current research predominantly focuses on isolated local or global information, often neglecting the synergistic integration of multiple data sources, thus potentially overlooking valuable details. To address this issue, we proposed a diagnostic model for MDD that integrates high-frequency and low-frequency information using data from diffusion tensor imaging (DTI), structural magnetic resonance imaging (sMRI), and functional magnetic resonance imaging (fMRI). First, we designed a meta-low-frequency encoder (MLFE) and a meta-high-frequency encoder (MHFE) to extract the low-frequency and high-frequency feature information from DTI and sMRI, respectively. Then, we utilized a multilayer perceptron (MLP) to extract features from fMRI data. Following the feature cross-fusion, we designed the ensemble learning threshold voting method to determine the ultimate diagnosis for MDD. The model achieved accuracy, precision, specificity, F1-score, MCC, and AUC values of 0.724, 0.750, 0.882, 0.600, 0.421, and 0.667, respectively. This approach provides new research ideas for the diagnosis of MDD.

CcpA-Knockout Staphylococcus aureus Induces Abnormal Metabolic Phenotype via the Activation of Hepatic STAT5/PDK4 Signaling in Diabetic Mice.

Catabolite control protein A (CcpA), an important global regulatory protein, is extensively found in S. aureus. Many studies have reported that CcpA plays a pivotal role in regulating the tricarboxylic acid cycle and pathogenicity. Moreover, the CcpA-knockout Staphylococcus aureus (S. aureus) in diabetic mice, compared with the wild-type, showed a reduced colonization rate in the tissues and organs and decreased inflammatory factor expression. However, the effect of CcpA-knockout S. aureus on the host's energy metabolism in a high-glucose environment and its mechanism of action remain unclear. S. aureus, a common and major human pathogen, is increasingly found in patients with obesity and diabetes, as recent clinical data reveal. To address this issue, we generated CcpA-knockout S. aureus strains with different genetic backgrounds to conduct in-depth investigations. In vitro experiments with high-glucose-treated cells and an in vivo model study with type 1 diabetic mice were used to evaluate the unknown effect of CcpA-knockout strains on both the glucose and lipid metabolism phenotypes of the host. We found that the strains caused an abnormal metabolic phenotype in type 1 diabetic mice, particularly in reducing random and fasting blood glucose and increasing triglyceride and fatty acid contents in the serum. In a high-glucose environment, CcpA-knockout S. aureus may activate the hepatic STAT5/PDK4 pathway and affect pyruvate utilization. An abnormal metabolic phenotype was thus observed in diabetic mice. Our findings provide a better understanding of the molecular mechanism of glucose and lipid metabolism disorders in diabetic patients infected with S. aureus.

ROP-GAN: an image synthesis method for retinopathy of prematurity based on generative adversarial network.

Objective. Training data with annotations are scarce in the intelligent diagnosis of retinopathy of prematurity (ROP), and existing typical data augmentation methods cannot generate data with a high degree of diversity. In order to increase the sample size and the generalization ability of the classification model, we propose a method called ROP-GAN for image synthesis of ROP based on a generative adversarial network.Approach. To generate a binary vascular network from color fundus images, we first design an image segmentation model based on U2-Net that can extract multi-scale features without reducing the resolution of the feature map. The vascular network is then fed into an adversarial autoencoder for reconstruction, which increases the diversity of the vascular network diagram. Then, we design an ROP image synthesis algorithm based on a generative adversarial network, in which paired color fundus images and binarized vascular networks are input into the image generation model to train the generator and discriminator, and attention mechanism modules are added to the generator to improve its detail synthesis ability.Main results. Qualitative and quantitative evaluation indicators are applied to evaluate the proposed method, and experiments demonstrate that the proposed method is superior to the existing ROP image synthesis methods, as it can synthesize realistic ROP fundus images.Significance. Our method effectively alleviates the problem of data imbalance in ROP intelligent diagnosis, contributes to the implementation of ROP staging tasks, and lays the foundation for further research. In addition to classification tasks, our synthesized images can facilitate tasks that require large amounts of medical data, such as detecting lesions and segmenting medical images.

Off-label use of antimicrobials among hospitalized children: a retrospective study of 3,406 patients.

Introduction: Off-label drug use is a global problem for which many countries and regions have issued legal provisions or reached an expert consensus. Off-label use is sometimes a necessity, especially since antibacterial drugs have become one of the most widely used drugs in pediatric settings and the issue of causing antimicrobial resistance has increasingly become unavoidable. It also poses additional risks, such as adverse drug reactions. Methods: Our study analyzed the antimicrobial prescriptions of pediatric inpatients in a large Chinese hospital in the first half of 2021. This retrospective investigation included 6,829 prescriptions, including 2,294 off-label prescriptions. We performed descriptive analyses of prescription antimicrobial agents among pediatric populations and reported the percentages and frequencies. Results: It was found that off-label use of antibiotics was present in many children (n = 1,665, 48.9%) and was most common in newborns (n = 328, 82.8%). Among the commonly used antibiotics in pediatric patients, cephalosporins (n = 2,778, 40.7%) accounted for a relatively low proportion of offlabel use (n = 360, 15.7%), while macrolides (n = 628, 27.4%) and penicillins (n = 610, 26.6%) accounted for a higher proportion. The off-label type mainly referred to the appropriate population (46.5%) and dosage (dose, 10.0%; frequency of administration, 48.3%). Discussion: Off-label use was due to imperfect labels, improper medications, or medication errors. Only a few consensuses could apply to pediatric patients. More clinical trials are required to update the consensus, and drug labels must be continuously improved. The prescription behavior of doctors is also needed to be regulated. Rational use of drugs, especially antimicrobials, is the responsibility of all people, including the states, medical institutions, and individuals.

Application of low-intensity pulsed ultrasound on tissue resident stem cells: Potential for ophthalmic diseases.

Introduction: Tissue-resident stem cells (TRSCs) have the ability to self-renew and differentiate throughout an individual's lifespan, and they utilize both mechanisms to maintain homeostasis and regenerate damaged tissues. Several studies suggest that these stem cells can serve as a potential source for cell-replacement-based therapy by promoting differentiation or expansion. In recent years, low-intensity pulsed ultrasound (LIPUS) has been demonstrated to effectively stimulate stem cell proliferation and differentiation, promote tissue regeneration, and inhibit inflammatory responses. Aims: To present a comprehensive overview of current application and mechanism of LIPUS on tissue resident stem cells. Methods: We searched PubMed, Web of Science for articles on the effects of LIPUS on tissue resident stem cells and its application. Results: The LIPUS could modulate cellular activities such as cell viability, proliferation and differentiation of tissue resident stem cells and related cells through various cellular signaling pathways. Currently, LIPUS, as the main therapeutic ultrasound, is being widely used in the treatment of preclinical and clinical diseases. Conclusion: The stem cell research is the hot topic in the biological science, while in recent years, increasing evidence has shown that TRSCs are good targets for LIPUS-regulated regenerative medicine. LIPUS may be a novel and valuable therapeutic approach for the treatment of ophthalmic diseases. How to further improve its efficiency and accuracy, as well as the biological mechanism therein, will be the focus of future research.

Insight into the soil aggregate-mediated restoration mechanism of degraded black soil via biochar addition: Emphasizing the driving role of core microbial communities and nutrient cycling.

Soil microbial communities are responsive to biochar application. However, few studies have investigated the synergistic effects of biochar application in the restoration of degraded black soil, especially soil aggregate-mediated microbial community changes that improve soil quality. From the perspective of soil aggregates, this study explored the potential microbial driving mechanism of biochar (derived from soybean straw) addition in black soil restoration in Northeast China. The results showed that biochar significantly improved the soil organic carbon, cation exchange capacity and water content, which play crucial roles in aggregate stability. The addition of biochar also significantly increased the concentration of the bacterial community in mega-aggregates (ME; 0.25-2 mm) compared with micro-aggregates (MI; <0.25 mm). Microbial co-occurrence networks analysis showed that biochar enhanced microbial interactions in terms of the number of links and modularity, particularly in ME. 16 S rRNA sequencing predicted that the expression of genes related to carbon (rbcL, acsA, gltS, aclB, and mcrA) and nitrogen (nifH and amoA) transformation increased after the addition of biochar. Furthermore, the functional microbes involved in carbon fixation (Firmicutes and Bacteroidetes) and nitrification (Proteobacteria) were significantly enriched and are the key regulators of carbon and nitrogen kinetics. Structural equation model (SEM) analysis further showed that the application of biochar promoted soil aggregates to positively regulate the abundance of soil nutrient conversion-related microorganisms, thereby increasing soil nutrient content and enzyme activities. These results provide new insights into the mechanisms of soil restoration through biochar addition.

Microalgae-based constructed wetland system enhances nitrogen removal and reduce carbon emissions: Performance and mechanisms.

Combination of constructed wetlands (CWs) and microalgae-based technologies has been proved as effective wastewater treatment option; however, little attention was paid to investigate the optimal combination ways. This study showed that the integrated system (IS) connecting microalgal pond with CWs exhibited improved pollutant-removal efficiencies and preferred carbon reduction effects compared to other alternatives such as coupled system or independent CWs. Microbial analysis demonstrated that core microorganisms (e.g., Acinetobacter and Thermomonas) of the IS were mostly associated with carbon, nitrogen, and energy metabolism. Based on co-occurrence networks, microbial quantity with denitrification function in the IS accounted for 71.01 % of the microorganism related to nitrogen metabolism, which was higher than that of 48.84 % in the independent CWs, indicating that the presence of microalgae in IS played important role in promoting biological denitrification. These findings provide insights into the microbial mechanism and highlights the complementary effects between microalgae and CWs.

Advances in targeted retinal photocoagulation in the treatment of diabetic retinopathy.

Aim: Targeted retinal photocoagulation (TRP) is an emerging laser technology for retinal targeted therapy. TRP can specifically act on unperfused retinal capillaries and retinal intermediate ischemic areas, reduce damage to tissue perfusion areas and panretinal photocoagulation (PRP) complications or adverse events. In this regard, this review discusses the treatment options, efficacy, and latest progress of TRP for diabetic retinopathy (DR) based on randomized controlled trial (RCT), meta-analysis, case review, and other existing studies. Methods: In-depth research was conducted on articles about the proposal and development of TRP, its simple application in DR, and combined therapy. In order to review the new progress, application methods, effects, and prospects of TRP in the treatment of DR, the articles related to TRP in the databases of PubMed and Web Of Science since this century were comprehensively analyzed. Results: TRP is effective in treating DR and may become a substitute for PRP in the future. In addition, the treatment regimen of TRP combined with intravitreal injection of anti-vascular endothelial growth factor (anti-VEGF) drugs can also be used as a new therapeutic approach to expand the treatment regimen for the treatment of DR, and this combination therapy also has effects on other retinal vascular diseases. Conclusions: With the advancement of technology, TRP has been continuously applied in clinical practice, and its potential benefits have opened up broad prospects for the treatment of DR. The combination therapy of TRP and anti-VEGF is expected to become a new option for patients with DR an retinal diseases.

[Characteristics of Soil Carbon, Nitrogen, Phosphorus, and Ecological Stoichiometry in Vegetable Fields and Orchard in the Coastal Area of Fuzhou].

The soil ecological stoichiometric characteristics of different agricultural land use types have a certain indicator function for characterizing the level of soil nutrient supply and are of great significance to the management of nutrient resources in farmland ecosystems. In order to reveal the soil carbon (C), nitrogen (N), and phosphorus (P) contents and their ecological stoichiometric characteristics in different vegetable fields and orchard agricultural land use types, this study took vegetable fields (taro field and jicama field) and orchards (citrus tree orchard, watermelon field, and pear tree orchard) as the research objects in the coastal area of Fuzhou City. The contents of soil C, N, and P and their ecological stoichiometric characteristics in different vegetable fields and orchard agricultural land uses were measured and analyzed. The soil C and N contents were in the order of orchard>vegetable field (P<0.05). The C content in the citrus tree orchard was the highest (4.44 g·kg-1), and the N content in the watermelon field was the highest (1.46 g·kg-1). The soil P content was vegetable field>orchard (P<0.05), and the jicama field had the highest P content (0.19 g·kg-1). The soil carbon and nitrogen ratio (C/N), carbon and phosphorus ratio (C/P), and nitrogen and phosphorus ratio (N/P) were orchard>vegetable field (P<0.05). Among them, the citrus tree orchard had the highest C/N (7.40) and C/P (61.43), and the watermelon field had the highest N/P (10.27). Soil N content was significantly and negatively correlated with bulk density and conductivity (r=-0.49, r=-0.28, P<0.05), and there was a significant and positive correlation with soil water content (r=0.61, P<0.05). C/P and C/N were significantly and positively correlated with SOM (r=0.71, r=0.64, P<0.01). In the process of crop planting and management in the coastal area of Fuzhou City, it is necessary to reasonably add nitrogen fertilizer to compensate for the N limitation, and slow-release nitrogen fertilizer is better for promoting the sustainable supply of nitrogen nutrients in the growth and development of crops.

A Novel Estrone Degradation Gene Cluster and Catabolic Mechanism in Microbacterium oxydans ML-6.

Global-scale estrone (E1) contamination of soil and aquatic environments results from the widespread use of animal manure as fertilizer, threatening both human health and environmental security. A detailed understanding of the degradation of E1 by microorganisms and the associated catabolic mechanism remains a key challenge for the bioremediation of E1-contaminated soil. Here, Microbacterium oxydans ML-6, isolated from estrogen-contaminated soil, was shown to efficiently degrade E1. A complete catabolic pathway for E1 was proposed via liquid chromatography-tandem mass spectrometry (LC-MS/MS), genome sequencing, transcriptomic analysis, and quantitative reverse transcription-PCR (qRT-PCR). In particular, a novel gene cluster (moc) associated with E1 catabolism was predicted. The combination of heterologous expression, gene knockout, and complementation experiments demonstrated that the 3-hydroxybenzoate 4-monooxygenase (MocA; a single-component flavoprotein monooxygenase) encoded by the mocA gene was responsible for the initial hydroxylation of E1. Furthermore, to demonstrate the detoxification of E1 by strain ML-6, phytotoxicity tests were performed. Overall, our findings provide new insight into the molecular mechanism underlying the diversity of E1 catabolism in microorganisms and suggest that M. oxydans ML-6 and its enzymes have potential applications in E1 bioremediation to reduce or eliminate E1-related environmental pollution. IMPORTANCE Steroidal estrogens (SEs) are mainly produced by animals, while bacteria are major consumers of SEs in the biosphere. However, the understanding of the gene clusters that participate in E1 degradation is still limited, and the enzymes involved in the biodegradation of E1 have not been well characterized. The present study reports that M. oxydans ML-6 has effective SE degradation capacity, which facilitates the development of strain ML-6 as a broad-spectrum biocatalyst for the production of certain desired compounds. A novel gene cluster (moc) associated with E1 catabolism was predicted. The 3-hydroxybenzoate 4-monooxygenase (MocA; a single-component flavoprotein monooxygenase) identified in the moc cluster was found to be necessary and specific for the initial hydroxylation of E1 to generate 4-OHE1, providing new insight into the biological role of flavoprotein monooxygenase.