Taming Two-Dimensional Polymerization by a Machine-Learning Discovered Crystallization Model.

Rapidly synthesizing high-quality two-dimensional covalent organic frameworks (2D COFs) is crucial to their practical applications. Here, we use a machine-learning approach that overcomes the challenges associated with bottom-up model derivation for the non-classical 2D COF crystallization processes. The resulting model, referred to as NEgen1, establishes correlations among the induction time, nucleation rate, growth rate, bond-forming rate constants, and common solution synthesis conditions for 2D COFs that grow by a nucleation-elongation mechanism. The results elucidate the detailed competition between the nucleation and growth dynamics in solution, which has been inappropriately described previously by classical, empirical models with assumptions invalid for 2D COF polymerization. By understanding the dynamic processes at play, the NEgen1 model reveals a simple strategy of gradually increasing monomer addition speed for growing large 2D COF crystals. This insight enables us to rapidly synthesize large COF-5 colloids, which could only be achieved previously by prolonged reaction times or by introducing chemical modulators. These results highlight the potential for systematically improving the crystal quality of 2D COFs, which has wide-reaching relevance for many of the applications for which 2D COFs are speculated to be valuable.

Exploring explainable AI features in the vocal biomarkers of lung disease.

This review delves into the burgeoning field of explainable artificial intelligence (XAI) in the detection and analysis of lung diseases through vocal biomarkers. Lung diseases, often elusive in their early stages, pose a significant public health challenge. Recent advancements in AI have ushered in innovative methods for early detection, yet the black-box nature of many AI models limits their clinical applicability. XAI emerges as a pivotal tool, enhancing transparency and interpretability in AI-driven diagnostics. This review synthesizes current research on the application of XAI in analyzing vocal biomarkers for lung diseases, highlighting how these techniques elucidate the connections between specific vocal features and lung pathology. We critically examine the methodologies employed, the types of lung diseases studied, and the performance of various XAI models. The potential for XAI to aid in early detection, monitor disease progression, and personalize treatment strategies in pulmonary medicine is emphasized. Furthermore, this review identifies current challenges, including data heterogeneity and model generalizability, and proposes future directions for research. By offering a comprehensive analysis of explainable AI features in the context of lung disease detection, this review aims to bridge the gap between advanced computational approaches and clinical practice, paving the way for more transparent, reliable, and effective diagnostic tools.

Genome-Wide Identification of the WRKY Gene Family in Four Cotton Varieties and the Positive Role of GhWRKY31 in Response to Salt and Drought Stress.

The WRKY gene family is ubiquitously distributed in plants, serving crucial functions in stress responses. Nevertheless, the structural organization and evolutionary dynamics of WRKY genes in cotton have not been fully elucidated. In this study, a total of 112, 119, 217, and 222 WRKY genes were identified in Gossypium arboreum, Gossypium raimondii, Gossypium hirsutum, and Gossypium barbadense, respectively. These 670 WRKY genes were categorized into seven distinct subgroups and unequally distributed across chromosomes. Examination of conserved motifs, domains, cis-acting elements, and gene architecture collectively highlighted the evolutionary conservation and divergence within the WRKY gene family in cotton. Analysis of synteny and collinearity further confirmed instances of expansion, duplication, and loss events among WRKY genes during cotton evolution. Furthermore, GhWRKY31 transgenic Arabidopsis exhibited heightened germination rates and longer root lengths under drought and salt stress. Silencing GhWRKY31 in cotton led to reduced levels of ABA, proline, POD, and SOD, along with downregulated expression of stress-responsive genes. Yeast one-hybrid and molecular docking assays confirmed the binding capacity of GhWRKY31 to the W box of GhABF1, GhDREB2, and GhRD29. The findings collectively offer a systematic and comprehensive insight into the evolutionary patterns of cotton WRKYs, proposing a suitable regulatory framework for developing cotton cultivars with enhanced resilience to drought and salinity stress.

Sulfur-Vacancy Engineering Accelerates Rapid Surface Reconstruction in Ni-Co Bimetal Sulfide Nanosheet for Urea Oxidation Electrocatalysis.

Developing a highly efficient catalyst for electrocatalytic urea oxidation reaction (UOR) is not only beneficial for the degradation of urea pollutants in wastewater but also provides a benign route for hydrogen production. Herein, a sulfur-vacancy (Sv) engineering is proposed to accelerate the formation of metal (oxy)hydroxide on the surface of Ni-Co bimetal sulfide nanosheet arrays on nickel foam (Sv-CoNiS@NF) for boosting the urea oxidation electrocatalysis. As a result, the obtained Sv-CoNiS@NF demonstrates an outstanding electrocatalytic UOR performance, which requires a low potential of only 1.397 V versus the reversible hydrogen electrode to achieve the current density of 100 mA cm-2. The ex situ Raman spectra and density functional theory calculations reveal the key roles of the Sv site and Co9S8 in promoting the electrocatalytic UOR performance. This work provides a new strategy for accelerating the transformation of electrocatalysts to active metallic (oxy)hydroxide for urea electrolysis via engineering the surface vacancies.

DIAS: A dataset and benchmark for intracranial artery segmentation in DSA sequences.

The automated segmentation of Intracranial Arteries (IA) in Digital Subtraction Angiography (DSA) plays a crucial role in the quantification of vascular morphology, significantly contributing to computer-assisted stroke research and clinical practice. Current research primarily focuses on the segmentation of single-frame DSA using proprietary datasets. However, these methods face challenges due to the inherent limitation of single-frame DSA, which only partially displays vascular contrast, thereby hindering accurate vascular structure representation. In this work, we introduce DIAS, a dataset specifically developed for IA segmentation in DSA sequences. We establish a comprehensive benchmark for evaluating DIAS, covering full, weak, and semi-supervised segmentation methods. Specifically, we propose the vessel sequence segmentation network, in which the sequence feature extraction module effectively captures spatiotemporal representations of intravascular contrast, achieving intracranial artery segmentation in 2D+Time DSA sequences. For weakly-supervised IA segmentation, we propose a novel scribble learning-based image segmentation framework, which, under the guidance of scribble labels, employs cross pseudo-supervision and consistency regularization to improve the performance of the segmentation network. Furthermore, we introduce the random patch-based self-training framework, aimed at alleviating the performance constraints encountered in IA segmentation due to the limited availability of annotated DSA data. Our extensive experiments on the DIAS dataset demonstrate the effectiveness of these methods as potential baselines for future research and clinical applications. The dataset and code are publicly available at https://doi.org/10.5281/zenodo.11401368 and https://github.com/lseventeen/DIAS.

Intramolecular Dehydrogenative Photocyclization of N-Phenyl-1-naphthamides.

Here, we explore a dehydrogenative 6π photocyclization method for N-substituted naphthalene carboxamides, which can be conducted in air. This method employs DMSO as both the reaction solvent and oxidant while also stabilizing the excited state of the substrate. Furthermore, the addition of photosensitizer enables the reaction to proceed under a 440-445 nm LED source via energy transfer. The proposed mechanism is initially validated through DFT calculations.

Role of polyferric sulphate in hydration regulation of phosphogypsum-based excess-sulphate slag cement: A multiscale investigation.

Current demand for waste recycling, phosphogypsum-based excess-sulphate slag cement (PESSC) as a sustainable cement prepared by solid wastes, urges enhancing its performance development based on microstructure optimisation. For the purpose of improving the performance and durability of PESSC used in normal or corrosive environments, it is deemed an efficient technique to produce iron-doped compounds with high thermodynamic stability. This paper presents a systematic study of the effect of iron modification on PESSC binders by introducing 0%-2% polyferric sulphate (PFS) from a multiscale viewpoint. XPS, 29Si and 27Al NMR, and TEM were used to characterise the nanostructure of solid particles firstly at Level I. Then, the chemical composition and phase assemblage of PESSC binders were revealed at Level II in terms of ICC, ICP, DTG-DSC, FTIR, BSE-EDS and XRD. Finally, setting time and strength development were determined at Level III. Results indicated that the soluble FeOH4- supplied by the hydrolysis of PFS promotes the generation of iron-doped ettringite with a greater length-to-diameter ratio and thermodynamic stability. Seeding effect of iron doping also promotes the production of spherical and retiform gels with a slight influence on the chemical components and polymerisation. Despite the fact that iron doping weakens the early strength of PESSC mortars, it promotes the persistent hydration rate by retarding precipitation and encapsulation of hydrates on the surface of the slag, showing excellent strength in the later stages. In view of microstructure evolution and performance development during each stage, PFS supplementation within 1.0% is considered a feasible modification of PESSC relying on the formation control of iron-doped hydrates.

Role of mitochondria-associated membranes in the hippocampus in the pathogenesis of depression.

BACKGROUND: Pathological changes, such as microglia activation in the hippocampus frequently occur in individuals with animal models of depression; however, they may share a common cellular mechanism, such as endoplasmic reticulum (ER) stress and mitochondrial dysfunction. Mitochondria associated membranes (MAMs) are communication platforms between ER and mitochondria. This study aimed to investigate the role of intracellular stress responses, especially structural and functional changes of MAMs in depression. METHODS: We used chronic social defeat stress (CSDS) to mimic depression in C57 mice to investigate the pathophysiological changes in the hippocampus associated with depression and assess the antidepressant effect of electroacupuncture (EA). Molecular, histological, and electron microscopic techniques were utilized to study intracellular stress responses, including the ER stress pathway reaction, mitochondrial damage, and structural and functional changes in MAMs in the hippocampus after CSDS. Proteomics technology was employed to explore protein-level changes in MAMs caused by CSDS. RESULTS: CSDS caused mitochondrial dysfunction, ER stress, closer contact between ER and mitochondria, and enrichment of functional protein clusters at MAMs in hippocampus along with depressive-like behaviors. Also, EA showed beneficial effects on intracellular stress responses and depressive-like behaviors in CSDS mice. LIMITATION: The cellular specificity of MAMs related protein changes in CSDS mice was not explored. CONCLUSIONS: In the hippocampus, ER stress and mitochondrial damage occur, along with enriched mitochondria-ER interactions and MAM-related protein enrichment, which may contribute to depression's pathophysiology. EA may improve depression by regulating intracellular stress responses.

Mechanisms of acid generation from ionic photoacid generators for extreme ultraviolet and electron beam lithography.

Photoacid generators (PAGs) are important components of chemically amplified resists. The properties of PAGs directly affect the sensitivity of photoresists, line edge roughness, and resolution. Understanding the photoacid generation process in extreme ultraviolet (EUV) and electron beam (EB) lithography is helpful for photoresist design. However, the microscopic mechanisms remain largely unclear and the large variety in the molecular structure of PAGs presents a challenge to overcome. In this work, we investigate the microscopic processes of photoacid production of ionic PAGs for EUV and EB lithography. The PAG dissociation pathway is found to depend on the molecular structure and conformations. The processes of photoacid production and by-product generation are also revealed. The results contribute to a better understanding of the photochemical reactions in EUV and EB lithography, providing insights into the molecular design of novel PAGs and photoresists.

Strain-Controlled Formation Energy and Migration of Nitrogen Vacancy in Al1-xScxN: A First-Principles Study.

The impact of strain on the formation energy and migration behavior of nitrogen vacancies (VNs) in Al1-xScxN has been investigated by first-principles calculations. The formation energy of VNs is obtained by total energy calculations. The migration barrier calculation utilizes the climbing nudged elastic band method. It is found that the formation energy of VNs is highly tunable with respect to the strain. The formation energy of VNs increases with the tensile strain increasing to +4% and decreases with the increasing compressive strain to -4%. A minimum formation energy of 4.11 eV is obtained when -4% strain is applied. Furthermore, the migration behavior of VNs is studied by calculating the migration barriers. Calculation results show that the migration barrier is strongly affected by strain. When the strain is -4%, the barrier is 2.46 eV while the barrier is increased to 2.71 eV under +4% strain. Therefore, a tensile strain can prevent the formation and migration of VNs. These findings suggest that strain engineering may serve as a tool for regulating VNs behavior in Al1-xScxN, potentially alleviating the ferroelectric degradations associated with VNs.

Accelerating diabetic wound healing by ROS-scavenging lipid nanoparticle-mRNA formulation.

Current treatment options for diabetic wounds face challenges due to low efficacy, as well as potential side effects and the necessity for repetitive treatments. To address these issues, we report a formulation utilizing trisulfide-derived lipid nanoparticle (TS LNP)-mRNA therapy to accelerate diabetic wound healing by repairing and reprogramming the microenvironment of the wounds. A library of reactive oxygen species (ROS)-responsive TS LNPs was designed and developed to encapsulate interleukin-4 (IL4) mRNA. TS2-IL4 LNP-mRNA effectively scavenges excess ROS at the wound site and induces the expression of IL4 in macrophages, promoting the polarization from the proinflammatory M1 to the anti-inflammatory M2 phenotype at the wound site. In a diabetic wound model of db/db mice, treatment with this formulation significantly accelerates wound healing by enhancing the formation of an intact epidermis, angiogenesis, and myofibroblasts. Overall, this TS LNP-mRNA platform not only provides a safe, effective, and convenient therapeutic strategy for diabetic wound healing but also holds great potential for clinical translation in both acute and chronic wound care.

Multifunctional carbon aerogels loaded with pea-pod-like carbon nanotubes for outstanding electromagnetic wave absorption performance.

Although ordered porous carbon materials (PCMs) have shown promising potential in the field of electromagnetic wave absorption (EWA), creating multifunctional PCMs with outstanding microwave absorption performance remains a significant challenge. Herein, ordered porous carbon aerogels loaded with pea-pod-like nitrogen-doped carbon nanotubes (CNTs) were fabricated via orientation freeze-drying followed by high-temperature pyrolysis. The optimized aerogel exhibits extraordinary EWA performance with a broad effective absorption bandwidth of 7.68 GHz and exceptionally strong absorption of -91.58 dB at a low filling ratio of only 3 wt%, which is the largest absorption strength among all known aerogels to date. The exceptional EWA performance is attributed to the synergistic effect of abundant loss mechanisms resulting from a unique pod-like structure in ordered porous carbon aerogel, where nitrogen-doped CNTs encapsulate magnetic alloy nanoparticles. Optimized aerogel exhibits superior compressive elasticity, thermal insulation, and light weight, laying the groundwork for designing practical next-generation EWA materials.

Mitochondrial tRNA pseudouridylation governs erythropoiesis.

Pseudouridine is the most prevalent RNA modification, and its aberrant function is implicated in various human diseases. However, the specific impact of pseudouridylation on hematopoiesis remains poorly understood. In this study, we investigated the role of tRNA pseudouridylation in erythropoiesis and its association with mitochondrial myopathy, lactic acidosis, and sideroblastic anemia syndrome (MLASA) pathogenesis. By utilizing patient-specific induced pluripotent stem cells (iPSCs) carrying a genetic PUS1 mutation and a corresponding mutant mouse model, we demonstrated impaired erythropoiesis in MLASA iPSCs and anemia in the MLASA mouse model. Both MLASA iPSCs and mouse erythroblasts exhibited compromised mitochondrial function and impaired protein synthesis. Mechanistically, we revealed that PUS1 deficiency resulted in reduced mitochondrial tRNA levels due to pseudouridylation loss, leading to aberrant mitochondrial translation. Screening of mitochondrial supplements aimed at enhancing respiration or heme synthesis showed limited effect in promoting erythroid differentiation. Interestingly, the mTOR inhibitor rapamycin facilitated erythroid differentiation in MLASA-iPSCs by suppressing mTOR signaling and protein synthesis, and consistent results were observed in the MLASA mouse model. Importantly, rapamycin treatment effectively ameliorated anemia phenotypes in the MLASA patient. Our findings provide novel insights into the crucial role of mitochondrial tRNA pseudouridylation in governing erythropoiesis and present potential therapeutic strategies for anemia patients facing challenges related to protein translation.

Study of CHF3/CH2F2 Adsorption Separation in TIFSIX-2-Cu-i.

Hydrofluorocarbons (HFCs) have important applications in different industries; however, they are environmentally unfriendly due to their high global warming potential (GWP). Hence, reclamation of used hydrofluorocarbons via energy-efficient adsorption-based separation will greatly contribute to reducing their impact on the environment. In particular, the separation of azeotropic refrigerants remains challenging, such as typical mixtures of CH2F2 (HFC-23) and CHF3 (HFC-32), due to a lack of adsorptive mechanisms. Metal-organic frameworks (MOFs) can provide a promising solution for the separation of CHF3-CH2F2 mixtures. In this study, the adsorption mechanism of CHF3-CH2F2 mixtures in TIFSIX-2-Cu-i was revealed at the microscopic level by combining static pure-component adsorption experiments, molecular simulations, and density-functional theory (DFT) calculations. The adsorption separation selectivity of CH2F2/CHF3 in TIFSIX-2-Cu-i is 3.17 at 3 bar under 308 K. The existence of similar TiF62- binding sites for CH2F2 or CHF3 was revealed in TIFSIX-2-Cu-i. Interactions between the fluorine atom of the framework and the hydrogen atom of the guest molecule were found to be responsible for determining the high adsorption separation selectivity of CH2F2/CHF3. This exploration is important for the design of highly selective adsorbents for the separation of azeotropic refrigerants.

Engineering LNPs with polysarcosine lipids for mRNA delivery.

Since the approval of the lipid nanoparticles (LNP)-mRNA vaccines against the SARS-CoV-2 virus, there has been an increased interest in the delivery of mRNA through LNPs. However, current LNP formulations contain PEG lipids, which can stimulate the generation of anti-PEG antibodies. The presence of these antibodies can potentially cause adverse reactions and reduce therapeutic efficacy after administration. Given the widespread deployment of the COVID-19 vaccines, the increased exposure to PEG may necessitate the evaluation of alternative LNP formulations without PEG components. In this study, we investigated a series of polysarcosine (pSar) lipids as alternatives to the PEG lipids to determine whether pSar lipids could still provide the functionality of the PEG lipids in the ALC-0315 and SM-102 LNP systems. We found that complete replacement of the PEG lipid with a pSar lipid can increase or maintain mRNA delivery efficiency and exhibit similar safety profiles in vivo.

Pseudouridine synthase 1 regulates erythropoiesis via transfer RNAs pseudouridylation and cytoplasmic translation.

Pseudouridylation plays a regulatory role in various physiological and pathological processes. A prime example is the mitochondrial myopathy, lactic acidosis, and sideroblastic anemia syndrome (MLASA), characterized by defective pseudouridylation resulting from genetic mutations in pseudouridine synthase 1 (PUS1). However, the roles and mechanisms of pseudouridylation in normal erythropoiesis and MLASA-related anemia remain elusive. We established a mouse model carrying a point mutation (R110W) in the enzymatic domain of PUS1, mimicking the common mutation in human MLASA. Pus1-mutant mice exhibited anemia at 4 weeks old. Impaired mitochondrial oxidative phosphorylation was also observed in mutant erythroblasts. Mechanistically, mutant erythroblasts showed defective pseudouridylation of targeted tRNAs, altered tRNA profiles, decreased translation efficiency of ribosomal protein genes, and reduced globin synthesis, culminating in ineffective erythropoiesis. Our study thus provided direct evidence that pseudouridylation participates in erythropoiesis in vivo. We demonstrated the critical role of pseudouridylation in regulating tRNA homeostasis, cytoplasmic translation, and erythropoiesis.

Exercise habits and health behaviors on adolescent obesity.

BACKGROUND AND PURPOSE: Obesity among children and adolescents continues to increase globally, and it is important to determine the factors associated with obesity among adolescents for the prevention and reduction of obesity. The purpose of this study is to understand the factors associated with the increase in the obesity rate among adolescents, providing a reference basis for the development of projects aimed at promoting adolescent health. METHODS: Using the raw data of 2021 adolescent health behavior online survey, this study analyzed demographic sociological factors, mental health, exercise habits, health behaviors and other categorical variables, and conducted the frequency, χ2 test for the difference in the proportion of obese and non-obese. According to the hierarchy model of obesity-related variables, binary logistics regression is used for multivariate analysis. This study used the original data of the 2021 Youth Health Behavior Online Survey, and performed frequency, χ2 tests on the differences in the proportion of obese and non-obese for categorical variables such as demographic sociological factors, mental health, exercise habits, and health behaviors. Multivariate analysis was performed using binary logistic regression based on hierarchical models of obesity-related variables. RESULTS: The obesity rate among Korean adolescents was 18.25 %. The obesity risk for females was reduced by 0.344 times compared to males (95 % CI = 0.327-0.361, p < 0.001); high school students had a 1.4 times higher obesity risk than middle school students (95 % CI = 1.379-1.511, p < 0.001); students with "Subjective household economic status" rated as "Medium" and 'Low' had their obesity risk increased by 1.07 times (95 % CI = 1.020-1.124, p < 0.01) and 1.254 times (95 % CI = 1.165-1.350, p < 0.001), respectively, compared to students with 'Subjective household economic status' rated as 'High'; students with 'Moderate' and 'Low' levels of 'Perceived stress' had their obesity risk reduced by 0.78 times (95 % CI = 0.74-0.823, P < 0.001) and 0.75 times (95 % CI = 0.70-0.803, P < 0.001), respectively, compared to students with 'High' levels of 'Perceived stress'; students engaging in 'Muscle strengthening exercise' '1-2 times/week' and "≥ 3 times/week" had their obesity risk reduced by 0.844 times (95% CI = 0.797-0.895, P < 0.001) and 0.575 times (95% CI = 0.537-0.616, P < 0.001), respectively, compared to students not participating in "Muscle strengthening exercise". CONCLUSION: The obesity rate of boys is higher than that of girls and high school students is higher than that of middle school students, and obesity is inversely proportional to family economic status. Mental health factors, exercise habits and eating habits are all important factors affecting adolescent obesity. It is suggested that gender differences, psychological factors, health habits, obesity education and healthy eating habits suitable for different age groups should be considered in the formulation of adolescent obesity policy.

Revision of the macropterous subgenus Curtonotus from east China, with the description of a new species (Carabidae, Zabrini, Amara).

Species from east China belonging to the subgenus Curtonotus were studied, resulting in the description of a new species, Amara (Curtonotus) beijingensissp. nov. The type locality is Xiaolongmen Forest Park in Beijing. All the known macropterous Curtonotus species from eastern China are reviewed and for each species taxonomical notes, illustrations, and new provincial records are noted. An improved key for their identification is provided as well.

IL-1α is required for T cell-driven weight loss after respiratory viral infection.

Respiratory viral infections remain a major cause of hospitalization and death worldwide. Patients with respiratory infections often lose weight. While acute weight loss is speculated to be a tolerance mechanism to limit pathogen growth, severe weight loss following infection can cause quality of life deterioration. Despite the clinical relevance of respiratory infection-induced weight loss, its mechanism is not yet completely understood. We utilized a model of CD 8+ T cell-driven weight loss during respiratory syncytial virus (RSV) infection to dissect the immune regulation of post-infection weight loss. Supporting previous data, bulk RNA sequencing indicated significant enrichment of the interleukin (IL)-1 signaling pathway after RSV infection. Despite increased viral load, infection-associated weight loss was significantly reduced after IL-1α (but not IL-1ß) blockade. IL-1α depletion resulted in a reversal of the gut microbiota changes observed following RSV infection. Direct nasal instillation of IL-1α also caused weight loss. Of note, we detected IL-1α in the brain after either infection or nasal delivery. This was associated with changes in genes controlling appetite after RSV infection and corresponding changes in signaling molecules such as leptin and growth/differentiation factor 15. Together, these findings indicate a lung-brain-gut signaling axis for IL-1α in regulating weight loss after RSV infection.

Largemouth bass galectin, MsGal-9: Mediating various functions as a pattern recognition receptor and a potential damage-associated molecular pattern.

Galectins are lectins that bind to ß-galactose and are widely expressed in immune system tissues, playing pivotal roles in innate immunity through their conserved carbohydrate-recognition domains (CRDs). In this present investigation, a tandem-repeat galectin was discovered in the largemouth bass, Micropterus salmoides (designated as MsGal-9). The open reading frame of MsGal-9 encodes two CRDs, each containing two consensus motifs that are essential for ligand binding. MsGal-9 is expressed in various tissues of the largemouth bass, with particularly high expression levels in the liver and spleen. The full-length form of MsGal-9, as well as the N-terminal (MsGal-9-N) and C-terminal (MsGal-9-C) CRDs, were individually recombined. Their ability for nonself recognition was studied. The three recombinant proteins were able to bind to glucan (GLU), peptidoglycan (PGN), and lipopolysaccharide (LPS), with MsGal-9 displaying the highest binding activity. Furthermore, rMsGal-9-N exhibited higher binding activity towards GLU in comparison to rMsGal-9-C. Further investigations revealed that the full-length rMsGal-9 could significantly bind to Gram-positive bacteria, Gram-negative bacteria, and fungi, while rMsGal-9-C specifically bound to Escherichia coli. However, rMsGal-9-N did not exhibit significant binding activity towards any microbes. These findings indicate that MsGal-9 requires both CRDs to cooperate in order to fulfill its nonself recognition function. All three recombinant proteins demonstrated agglutination activity towards various microbes, with MsGal-9 and MsGal-9-N displaying a similar broad binding spectrum, while MsGal-9-C agglutinated three types of bacteria. Moreover, both MsGal-9 and MsGal-9-N were capable of coagulating largemouth bass red blood cells, whereas MsGal-9-C lacked this ability. However, MsGal-9-C played a significant role in enhancing the encapsulation of leukocytes in comparison to MsGal-9-N. All three proteins acted as potential damage-associated molecular patterns (DAMPs), inducing apoptosis in leukocytes.