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Abstract T helper cells, particularly T follicular helper (TFH) cells, are essential for the neutralizing antibody production elicited by pathogens or vaccines. However, in immunocompromised individuals, the inefficient support from TFH cells could lead to limited protection after vaccine inoculation. Here we showed that the conjugation of inducible T cell costimulatory (ICOS) onto the nanoparticle, together with immunogen, significantly enhanced the immune response of the vaccines specific for SARS-CoV-2 or human immunodeficiency virus type-1 (HIV-1) in TFH-deficient mice. Further studies indicated that ICOSL on B cells was triggered by ICOS binding, subsequently activated the PKCβ signaling pathway, and enhanced the survival and proliferation of B cells. Our findings revealed that the stimulation of ICOS-ICOSL interaction by adding ICOS on the nanoparticle vaccine significantly substitutes the function of TFH cells to support B cell response, which is significant for the immunocompromised people, such as the elderly or HIV-1-infected individuals.

Abstract Background The pathogenesis of osteoarthritis (OA) involves the progressive degradation of articular cartilage. Exosomes derived from mesenchymal stem cells (MSC-EXOs) have been shown to mitigate joint pathological injury by attenuating cartilage destruction. Optimization the yield and therapeutic efficacy of exosomes derived from MSCs is crucial for promoting their clinical translation. The preconditioning of MSCs enhances the therapeutic potential of engineered exosomes, offering promising prospects for application by enabling controlled and quantifiable external stimulation. This study aims to address these issues by employing pro-inflammatory preconditioning of MSCs to enhance exosome production and augment their therapeutic efficacy for OA. Methods The exosomes were isolated from the supernatant of infrapatellar fat pad (IPFP)-MSCs preconditioned with a pro-inflammatory factor, TNF-α, and their production was subsequently quantified. The exosome secretion-related pathways in IPFP-MSCs were evaluated through high-throughput transcriptome sequencing analysis, q-PCR and western blot analysis before and after TNF-α preconditioning. Furthermore, exosomes derived from TNF-α preconditioned IPFP-MSCs (IPFP-MSC-EXOsTNF−α) were administered intra-articularly in an OA mouse model, and subsequent evaluations were conducted to assess joint pathology and gait alterations. The expression of proteins involved in the maintenance of cartilage homeostasis within the exosomes was determined through proteomic analysis. Results The preconditioning with TNF-α significantly enhanced the exosome secretion of IPFP-MSCs compared to unpreconditioned MSCs. The potential mechanism involved the activation of the PI3K/AKT signaling pathway in IPFP-MSCs by TNF-α precondition, leading to an up-regulation of autophagy-related protein 16 like 1(ATG16L1) levels, which subsequently facilitated exosome secretion. The intra-articular administration of IPFP-MSC-EXOsTNF−α demonstrated superior efficacy in ameliorating pathological changes in the joints of OA mice. The preconditioning of TNF-α enhanced the up-regulation of low-density lipoprotein receptor-related protein 1 (LRP1) levels in IPFP-MSC-EXOsTNF−α, thereby exerting chondroprotective effects. Conclusion TNF-α preconditioning constitutes an effective and promising method for optimizing the therapeutic effects of IPFP-MSCs derived exosomes in the treatment of OA. Graphical Abstract

The precise correction of atmospheric zenith tropospheric delay (ZTD) is significant for the Global Navigation Satellite System (GNSS) performance regarding positioning accuracy and convergence time. In the past decades, many empirical ZTD models based on whether the gridded or scattered ZTD products have been proposed and widely used in the GNSS positioning applications. But there is no comprehensive evaluation of these models for the whole China region, which features complicated topography and climate. In this study, we completely assess the typical empirical models, the IGGtropSH model (gridded, non-meteorology), the SHAtropE model (scattered, non-meteorology), and the GPT3 model (gridded, meteorology) using the Crustal Movement Observation Network of China (CMONOC) network. In general, the results show that the three models share consistent performance with RMSE/bias of 37.45/1.63, 37.13/2.20, and 38.27/1.34 mm for the GPT3, SHAtropE and IGGtropSH model, respectively. However, the models had a distinct performance regarding geographical distribution, elevation, seasonal variations, and daily variation. In the southeastern region of China, RMSE values are around 50 mm, which are much higher than that in the western region, approximately 20 mm. The SHAtropE model exhibits better performance for areas with large variations in elevation. The GPT3 model and the IGGtropSH model are more stable across different months, and the SHAtropE model based on the GNSS data exhibits superior performance across various UTC epochs.

Joint diseases greatly impact the daily lives and occupational functioning of patients globally. However, conventional treatments for joint diseases have several limitations, such as unsatisfatory efficacy and side effects, necessitating the exploration of more efficacious therapeutic strategies. Mesenchymal stem cell (MSC)-derived EVs (MSC-EVs) have demonstrated high therapeutic efficacyin tissue repair and regeneration, with low immunogenicity and tumorigenicity. Recent studies have reported that EVs-based therapy has considerable therapeutic effects against joint diseases, including osteoarthritis, tendon and ligament injuries, femoral head osteonecrosis, and rheumatoid arthritis. Herein, we review the therapeutic potential of various types of MSC-EVs in the aforementioned joint diseases, summarise the mechanisms underlying specific biological effects of MSC-EVs, and discuss future prospects for basic research on MSC-EV-based therapeutic modalities and their clinical translation. In general, this review provides an in-depth understanding of the therapeutic effects of MSC-EVs in joint diseases, as well as the underlying mechanisms, which may be beneficial to the clinical translation of MSC-EV-based treatment.The translational potential of this article: MSC-EV-based cell-free therapy can effectively promote regeneration and tissue repair. When used to treat joint diseases, MSC-EVs have demonstrated desirable therapeutic effects in preclinical research. This review may supplement further research on MSC-EV-based treatment of joint diseases and its clinical translation.

Immunotherapy has become the cornerstone of current malignant tumor treatment. However, the response of different patients to immunotherapy is highly heterogeneous, and not all patients can benefit from it. There is an urgent need to find biomarkers that can effectively predict the efficacy of immunotherapy. C-C chemokine ligand 4 (CCL4) is a cytokine, belonging to the inflammatory CCL subfamily. It is mainly secreted by immune cells and tumor cells and shows low or no expression in normal tissues but abnormally high expression in various malignant tumor tissues. After binding to CCL4 and its receptor C-C chemokine receptor type 5 (CCR5), it can recruit and mediate immune cell migration, destroy the stability of the tumor microenvironment (TME), participate in carcinogenesis and promote the development of tumors. In the tumor immune microenvironment, CCL4 can mediate and recruit the directed migration of key immune cells such as monocytes, macrophages, natural killer (NK) cells, and T cells, which makes it a potentially important element affecting the efficacy of immunotherapy and has specific value. This paper reviews the research progresses of CCL4's effects on immune escape in TME, in order to provide clues and references for basic research and clinical diagnosis and treatment.

Abstract The filamentous ‘Pf’ bacteriophages of Pseudomonas aeruginosa play roles in biofilm formation and virulence, but mechanisms governing Pf prophage activation in biofilms are unclear. Here, we identify a prophage regulatory module, KKP (kinase-kinase-phosphatase), that controls virion production of co-resident Pf prophages and mediates host defense against diverse lytic phages. KKP consists of Ser/Thr kinases PfkA and PfkB, and phosphatase PfpC. The kinases have multiple host targets, one of which is MvaU, a host nucleoid-binding protein and known prophage-silencing factor. Characterization of KKP deletion and overexpression strains with transcriptional, protein-level and prophage-based approaches indicates that shifts in the balance between kinase and phosphatase activities regulate phage production by controlling MvaU phosphorylation. In addition, KKP acts as a tripartite toxin-antitoxin system that provides defense against some lytic phages. A conserved lytic phage replication protein inhibits the KKP phosphatase PfpC, stimulating toxic kinase activity and blocking lytic phage production. Thus, KKP represents a phosphorylation-based mechanism for prophage regulation and antiphage defense. The conservation of KKP gene clusters in >1000 diverse temperate prophages suggests that integrated control of temperate and lytic phage infection by KKP-like regulatory modules may play a widespread role in shaping host cell physiology.

Abstract Owing to the instability of gelatine in hygrothermal environments, gelatine-based cultural heritage undergo various deterioration processes, such as cracking, peeling, warping, curling and fracture, posing significant threats to its long-term preservation. Building on previous research, this study investigates the stability of polyol glycidyl ether–gelatine composite films under high-humidity and high-temperature conditions using ethylene glycol diglycidyl ether (EGDE) as a model compound. The hygrothermal ageing properties of EGDE–gelatine composite films are evaluated in terms of macrosize, mesoscopic structure, surface properties and mechanical properties. Results indicate that EGDE enhances the dimensional stability and swelling ratios of the composite films, stabilizes the pore structure and distribution and maintains the hydrophilicity and molecular structural stability under hygrothermal ageing conditions. Furthermore, the incorporation of EGDE leads to superior stress–strain properties of the composite films in such challenging environments. This study provides valuable experimental data for the preparation and conservation applications of gelatine-based cultural heritage materials.

Purpose: This study examined the impact of school leadership on teacher professional collaboration, with collective teacher innovativeness and teacher self-efficacy (TSE) playing the mediating role. Two most commonly used leadership styles, instructional leadership (IL) and distributed leadership (DL), were analyzed using a multilevel design, i.e. teachers are nested within schools. Design/methodology/approach: The proposed model was validated using data of Taiwan TALIS 2018 collected from both teachers and principals and analyzed using hierarchical linear modeling. Findings: Results showed that IL and DL influence teacher professional collaboration through different paths. IL had a significant direct impact on teacher professional collaboration alone, while DL had a significant direct impact on both teachers' collective innovativeness and their professional collaboration. While TSE had a direct effect on collective teacher innovativeness, TSE and collective teacher innovativeness had a direct effect on teacher professional collaboration. Originality/value: This study highlights the significant impact of principal leadership as both principals and teachers work in the same environment and culture co-shaped through the interaction and collaboration. Research evidence regarding the effects of IL and DL on teacher professional collaboration is limited; this is even less evidential when the indirect effects of variables mediating between school leadership and teacher outcomes, including teacher collective innovativeness and TSE, are added to the total effects. The present findings provide useful references for principals and teachers when promoting professional collaboration to achieve desired outcomes in school and student improvement.

Objective: To reveal the distribution characteristics and demographic factors of traditional Chinese medicine (TCM) constitution among elderly individuals in China. Methods: Elderly individuals from seven regions in China were selected as samples in this study using a multistage cluster random sampling method. The basic information questionnaire and Constitution in Chinese Medicine Questionnaire (Elderly Edition) were used. Descriptive statistical analysis, chi-squared tests, and binary logistic regression analysis were used. Results: The single balanced constitution (BC) accounted for 23.9%. The results of the major TCM constitution types showed that BC (43.2%) accounted for the largest proportion and unbalanced constitutions ranged from 0.9% to 15.7%. East China region (odds ratio [OR] = 2.097; 95% confidence interval [CI], 1.912 to 2.301), married status (OR = 1.341; 95% CI, 1.235 to 1.457), and managers (OR = 1.254; 95% CI, 1.044 to 1.505) were significantly associated with BC. Age >70 years was associated with qi-deficiency constitution and blood stasis constitution (BSC). Female sex was significantly associated with yang-deficiency constitution (OR = 1.646; 95% CI, 1.52 to 1.782). Southwest region was significantly associated with phlegm-dampness constitution (OR = 1.809; 95% CI, 1.569 to 2.086). North China region was significantly associated with inherited special constitution (OR = 2.521; 95% CI, 1.569 to 4.05). South China region (OR = 2.741; 95% CI, 1.997 to 1.3.763), Central China region (OR = 8.889; 95% CI, 6.676 to 11.835), senior middle school education (OR = 2.442; 95% CI, 1.932 to 3.088), and managers (OR = 1.804; 95% CI, 1.21 to 2.69) were significantly associated with BSC. Conclusions: This study defined the distribution characteristics and demographic factors of TCM constitution in the elderly population. Adjusting and improving unbalanced constitutions, which are correlated with diseases, can help promote healthy aging through the scientific management of these demographic factors.

本研究旨在瞭解在COVID-19線上教學期間教師情緒勞務中的淺層展現與深層展現對自我效能感的影響，並檢視教學正念於淺層展現與深層展現對自我效能感影響關係的調節效果。研究方法為問卷調查法，研究對象為於新冠疫情期間具線上教學經驗之臺灣國小教師，有效樣本為644份。研究使用多元迴歸分析進行假設檢驗。研究結果發現，因COVID-19疫情採線上教學期間，教師淺層展現程度愈高，則自我效能感愈低；教師深層展現程度愈高，則自我效能感愈高，本研究進一步發現教學正念對深層展現影響自我效能感的加乘效果不顯著、正念程度高的教師在進行淺層展現時對自我效能感的降低程度與正念程度低的教師無顯著差異。本研究建議學校聘請專家進行教師情緒調控之培訓課或鼓勵教師參與相關研習課程，多採深層展現進行教學，提升教師自我效能感。 Introduction Following its outbreak in 2019, the COVID-19 pandemic caused a considerable shift in work patterns across multiple industries. Taiwan continued education through online platforms, maintaining a “no suspension of classes” approach to ensure uninterrupted teaching and student learning. However, this transition imposed challenges on teachers, many of whom faced substantial stress and anxiety due to the demands of online teaching. Accordingly, this study focused on emotional labor, specifically surface acting and deep acting, as antecedent variables. During online teaching, teachers must maintain a strong belief in and enthusiasm for teaching to effectively foster student engagement and learning outcomes. The present study selected teacher self-efficacy as its outcome variable and explored the effects of surface acting and deep acting on teacher self-efficacy in the context of online teaching. The relevant literature suggests that mindfulness in teaching can enhance the effectiveness of teaching strategies and mitigate emotional exhaustion in teachers, thereby warranting its inclusion as a moderating variable in this study. Relevant studies have indicated that surface acting in emotional labor diminishes individual well-being and lowers self-efficacy, whereas deep acting enhances individual well-being and increases self-efficacy. Furthermore, deep acting positively predicts classroom management self-efficacy, whereas surface acting negatively affects such self-efficacy. However, these findings are based on traditional face-to-face teaching methods and lack support from empirical research regarding the relationships among teachers’ surface acting, deep acting, and self-efficacy during the online teaching period prompted by the COVID-19 pandemic in Taiwan. Additionally, understanding of the moderating role of teaching mindfulness in the relationships among surface acting, deep acting, and teacher self-efficacy remains limited. Accordingly, this study aimed to determine (1) how teacher surface acting and deep acting influence teacher self-efficacy in the context of online teaching during the COVID-19 pandemic and (2) the moderating role of teaching mindfulness in the relationships among teacher surface acting, deep acting, and self-efficacy. Research Purpose This study examined how teacher emotional labor affected teacher self-efficacy during the online teaching period prompted by the COVID-19 pandemic in Taiwan and investigated the moderating role of teaching mindfulness in the relationship between teacher emotional labor and self-efficacy. The aim of this study was to clarify the differential effects of surface acting and deep acting on teacher self-efficacy in the context of online teaching. The results of this study are expected to contribute to practical implications for teaching and to suggest timely support and assistance for teachers engaged in online teaching in order to enhance overall teaching effectiveness within school organizations. Data Collection The present research methodology conducted a questionnaire survey targeting primary school teachers in Taiwan with experience of online teaching during the COVID-19 pandemic period. A total of 644 valid responses were collected. The study utilized an English scale from an internationally published journal. To ensure conceptual validity and to avoid any loss of meaning due to translation, a back-translation process was employed. In addition, adjustments were made to some wordings to suit the context of online teaching during the COVID-19 pandemic, and relevant explanations were added accordingly. This study employed the emotional labor scale developed by Diefendorff et al. (2005) to assess teacher surface acting and deep acting during online teaching in the pandemic period. The measurement tool for surface acting consisted of seven items, whereas the measurement tool for deep acting consisted of four items. The Teacher Self-Efficacy Scale developed by Tschannen-Moran and Hoy (2001) was employed to assess self-efficacy among teachers engaged in online teaching. This scale contains three dimensions— namely “Teaching Strategies” (four items), “Classroom Management” (four items), and “Student Engagement” (four items)— yielding a total of 12 items. The Mindfulness in Teaching Scale developed by Frank et al. (2016) was employed to assess teacher mindfulness. This scale comprises eight items across two dimensions: “Intrapersonal Mindfulness” (six items) and “Interpersonal Mindfulness” (two items). Data Analysis IBM SPSS Statistics version 20.0 was used to perform quantitative statistical analysis of the survey data. The data analysis methods utilized in this study included descriptive statistics, Pearson correlation, and multiple regression analysis. Multiple regression analysis was employed to examine how surface acting and deep acting influenced teacher self-efficacy and to investigate the moderating effects of teaching mindfulness. Results The study results revealed that during the period of online teaching prompted by the COVID-19 pandemic, higher levels of surface acting among teachers were associated with lower self-efficacy, whereas higher levels of deep acting were associated with higher self-efficacy. Furthermore, the moderating effect of teaching mindfulness on the relationship between deep acting and self-efficacy was not significant. Finally, no significant difference was found in the reduction of self-efficacy between teachers with high levels of mindfulness who engaged in surface acting and those with low levels of mindfulness. Recommendations At the school level, organizational culture can substantially affect teachers’ emotional labor. Organizations can enhance the substance of a teacher’s job structure while fostering a school culture based on mutual trust and support. These actions are particularly crucial in the context of online teaching, where support for teachers’ instructional efforts is even more essential. Hiring experts to conduct training or relevant workshops related to emotional regulation for teachers can empower educators to apply their professional knowledge to develop personalized emotional expression skills. Such skill development can, in turn, improve teachers’ foundational ability to control their emotions and thus align with external expectations regarding their educational goals. In the context of online teaching, supporting teachers in managing their emotions appropriately and effectively by using emotional labor strategies is crucial for enhancing the effectiveness of online teaching. Suggestions for Future Research Future research should incorporate genuine emotional expression into strategies for emotional labor. Although this study analyzed only teachers from primary schools in Taiwan, it did not specifically analyze numbers of teachers in different regions. Accordingly, future research could investigate the distribution of teachers on the basis of the geographical locations of the primary schools where they teach. Because teachers are embedded within schools and because each school may have unique resources and methods of supporting online teaching, teachers from a single school often share certain common traits. Future research could utilize hierarchical linear modeling to explore relationships among variables at different levels. Finally, this study employed a quantitative research approach, and the self-reported data obtained limited the in-depth understanding of the participants’ performance in emotional labor and self-efficacy. Accordingly, future research is recommended to employ qualitative methods, such as interviews, to collect data and verify the authenticity of self-reported information.

Abstract Host immune responses are tightly controlled by various immune factors during infection, and protozoan parasites also manipulate the immune system to evade surveillance, leading to an evolutionary arms race in host‒pathogen interactions; however, the underlying mechanisms are not fully understood. We observed that the level of superoxide dismutase 3 (SOD3) was significantly elevated in both Plasmodium falciparum malaria patients and mice infected with four parasite species. SOD3-deficient mice had a substantially longer survival time and lower parasitemia than control mice after infection, whereas SOD3-overexpressing mice were much more vulnerable to parasite infection. We revealed that SOD3, secreted from activated neutrophils, bound to T cells, suppressed the interleukin-2 expression and concomitant interferon-gamma responses crucial for parasite clearance. Overall, our findings expose active fronts in the arms race between the parasites and host immune system and provide insights into the roles of SOD3 in shaping host innate immune responses to parasite infection.

Abstract Despite recent developments on the design of dynamic catalysts, none of them have been exploited for the in-situ control of multiple stereogenic centers in a single molecular scaffold. We report herein that it is possible to obtain in majority any amongst the four possible stereoisomers of an amino alcohol by means of a switchable asymmetric catalyst built on supramolecular helices. Hydrogen-bonded assemblies between a benzene-1,3,5-tricarboxamide (BTA) achiral phosphine ligand coordinated to copper and a chiral BTA comonomer are engaged in a copper-hydride catalyzed hydrosilylation and hydroamination cascade process. The nature of the product stereoisomer is related to the handedness of the helices and can thus be directed in a predictable way by changing the nature of the major enantiomer of the BTA comonomer present in the assemblies. The strategy allows all stereoisomers to be obtained one-pot with similar selectivities by conducting the cascade reaction in a concomitant manner, i.e. without inverting the handedness of the helices, or sequentially, i.e. by switching the handedness of the supramolecular helices between the hydrosilylation and hydroamination steps. Supramolecular helical catalysts appear as a unique and versatile platform to control the configuration of molecules or polymers embedding several stereogenic centers.

Apple replant disease (ARD) negatively affects plant growth and reduces yields in replanted orchards. In this study, biochar was applied to apple replant soil with Fusarium oxysporum. Our aim was to investigate whether biochar could promote plant growth and alleviate apple replant disease by reducing the growth of harmful soil microorganisms, changing soil microbial community structure and improving the soil environment. This experiment included five treatments: apple replant soil (CK), methyl bromide fumigation apple replant soil (FM), replant soil with biochar addition (2%), replant soil with F. oxysporum spore solution (8 × 107 spores · mL−1), and replant soil with biochar and F. oxysporum spore solution addition. Seedling biomass, the activity of antioxidant enzymes in the leaves and roots, and soil environmental variables were measured. Microbial community composition and community structure were analyzed using 16S rDNA and ITS2 gene sequencing. Biochar significantly reduced the abundance of F. oxysporum and increased soil microbial diversity and richness. Biochar also increased the soil enzyme activities (urease, invertase, neutral phosphatase, and catalase), the biomass (plant height, fresh weight, dry weight) and the activity of antioxidant enzymes (superoxide dismutase, peroxidase, and catalase). The root indexes of apple seedlings was also increased in replant soil by biochar. In sum, biochar promoted the growth of plants, improved the replant soil environment, and alleviated apple replant disease.

Abstract Background Teeth identification has a pivotal role in the dental curriculum and provides one of the important foundations of clinical practice. Accurately identifying teeth is a vital aspect of dental education and clinical practice, but can be challenging due to the anatomical similarities between categories. In this study, we aim to explore the possibility of using a deep learning model to classify isolated tooth by a set of photographs. Methods A collection of 5,100 photographs from 850 isolated human tooth specimens were assembled to serve as the dataset for this study. Each tooth was carefully labeled during the data collection phase through direct observation. We developed a deep learning model that incorporates the state-of-the-art feature extractor and attention mechanism to classify each tooth based on a set of 6 photographs captured from multiple angles. To increase the validity of model evaluation, a voting-based strategy was applied to refine the test set to generate a more reliable label, and the model was evaluated under different types of classification granularities. Results This deep learning model achieved top-3 accuracies of over 90% in all classification types, with an average AUC of 0.95. The Cohen’s Kappa demonstrated good agreement between model prediction and the test set. Conclusions This deep learning model can achieve performance comparable to that of human experts and has the potential to become a valuable tool for dental education and various applications in accurately identifying isolated tooth.

Nickel oxide (NiOx) serves as one of the most promising hole transport materials for perovskite light‐emitting diodes (PeLEDs). However, only moderate PeLED performances have been reported on the pristine NiOx layer due to insufficient hole injection, interfacial exciton quenching, and poor perovskite quality. Herein, a multifunctional molecule of 3‐mercapto‐1‐propanesulfonate (MPS) is demonstrated to successfully tailor the NiOx–perovskite heterogenous interface by addressing the above issues. In detail, the large binding energy between mercapto sulfur and nickel induces preferential self‐assembly of the mercapto group on the NiOx surface, which simultaneously enlarges the NiOx work function by the formation of interfacial dipole and suppresses the trap‐assisted exciton quenching by the passivation of the oxygen vacancies. Meanwhile, the self‐assembled MPS on NiOx also favors high‐quality perovskite films with good morphology, high crystallinity, and reduced defects for efficient carrier radiative recombination. As a result, blue PeLEDs show a remarkable efficiency of 10.4%, representing one of the highest efficiencies for NiOx‐based blue PeLEDs, as well as a very low turn‐on voltage of 2.8 V. Consequently, this work contributes to an efficient approach to tailor the NiOx–perovskite interface for highly efficient blue PeLEDs.

ABSTRACT Toxoplasma gondii, the causative parasite of toxoplasmosis, is an apicomplexan parasite that infects warm-blooded mammals. The ability of the calcium-binding proteins (CBPs) to transport large amounts of Ca2+ appears to be critical for the biological activity of T. gondii. However, the functions of some members of the CBP family have not yet been deciphered. Here, we characterized a putative CBP of T. gondii, TgpCaBP (TGME49_229480), which is composed of four EF-hand motifs with Ca2+-binding capability. TgpCaBP was localized in the cytosol and ER of T. gondii, and parasites lacking the TgpCaBP gene exhibited diminished abilities in cell invasion, intracellular growth, egress, and motility. These phenomena were due to the abnormalities in intracellular Ca2+ efflux and ER Ca2+ storage, and the reduction in motility was associated with a decrease in the discharge of secretory proteins. Therefore, we propose that TgpCaBP is a Ca2+ transporter and signaling molecule involved in Ca2+ regulation and parasitization in the hosts.IMPORTANCECa2+ signaling is essential in the development of T. gondii. In this study, we identified a calcium-binding protein in T. gondii, named TgpCaBP, which actively regulates intracellular Ca2+ levels in the parasite. Deletion of the gene coding for TgpCaBP caused serious deficits in the parasite’s ability to maintain a stable intracellular calcium environment, which also impaired the secretory protein discharged from the parasite, and its capacity of gliding motility, cell invasion, intracellular growth, and egress from host cells. In summary, we have identified a novel calcium-binding protein, TgpCaBP, in the zoonotic parasite T. gondii, which is a potential therapeutic target for toxoplasmosis.

Planning for the expansion of generation and transmission infrastructure, with a focus on integrating wind farms is presented in this study according to the goals of economic efficiency, operational performance, security, and reliability of the transmission system operator. The approach incorporates a bi-level optimization framework, with the upper level outlining the formulation of power system expansion planning to reduce construction and operational costs while adhering to the investment budget limits. In the lower-level model, which economic reliable-secure functioning is formulated for the transmission network. The objective function of the problem aims to minimize operational costs, energy losses, and anticipated energy not supplied, and the voltage security index. The constraints of this problem include AC optimal power flow model, security, and reliability limits. Scheme extracts a convex-linear model for the formulation using the conventional linearization technique. Pareto optimization driven by the aggregation of weighted functions results in a single-objective framework for the lower-level problem, and the Karush-Kuhn-Tucker technique presents a single-level model for the scheme. The approach utilizes stochastic optimization has been adopted to model load, wind farms, and network equipment availability uncertainties. Finally, the study cases yielded numerical findings that showcase the efficacy of the proposed scheme in achieving the desired economic, operational, security, and reliability status within the transmission network. This is particularly true in situations where there is appropriate generation and transmission expansion.

Apple replant disease (ARD) negatively affects plant growth and reduces yields in replanted orchards. In this study, biochar and humic acid were applied to apple replant soil. We aimed to investigate whether biochar and humic acid could promote plant growth and alleviate apple replant disease by reducing the growth of harmful soil microorganisms, changing soil microbial community structure, and improving the soil environment. This experiment included five treatments: apple replant soil (CK), apple replant soil with methyl bromide fumigation (FM), replant soil with biochar addition (2 %), replant soil with humic acid addition (1.5 ‰), and replant soil with biochar combined with humic acid. Seedling biomass, the activity of antioxidant enzymes in the leaves and roots, and soil environmental variables were measured. Microbial community composition and structure were analyzed using ITS gene sequencing. Biochar and humic acid significantly reduced the abundance of Fusarium and promoted the recovery of replant soil microbial communities. Biochar and humic acid also increased the soil enzymes activity (urease, invertase, neutral phosphatase, and catalase), the plant height, fresh weight, dry weight, the activity of antioxidant enzymes (superoxide dismutase, peroxidase, and catalase), and root indexes of apple seedlings increased in replant soil. In sum, We can use biochar combined with humic acid to alleviate apple replant disease.

As a supplement to medical services, telemedicine is of great significance to alleviate the shortage of health resources in China. Based on the traditional consumer behavior measurement model the Technology Acceptance Mode/Theory of Planned Behavior (TAM/TPB), this paper divides online patient trust into six dimensions: perceived risk, personal trust tendency, doctors’ credibility, hospitals’ credibility, websites’ credibility, and system guarantee. On this basis, a structural equation model (SEM) was used to explore the influence of each dimension of online patient trust on online patient intention, behavior choice, and pre-factors. A total of 582 valid questionnaires were distributed to selected patients with experience in using mobile healthcare services in the vicinity of hospitals and communities, as well as to users who shared their experiences in the discussion forums of mobile healthcare websites. The results show that online patient trust has a significant positive impact on telemedicine behavior intention selection, with a standardized path coefficient being as high as 0.866. Doctors’ credibility, system guarantee, and website credibility have significant positive effects on online patient trust, with standardized path coefficients of 0.401, 0.260, and 0.226, respectively. Hospital trustworthiness and personal trust propensity have no significant effect on online patient trust. Perceived risk has a significant negative effect on online patient trust, with a standardized path coefficient of −0.118. The research findings suggest that health departments and mobile healthcare providers can enhance mobile healthcare services by considering the patients’ perspectives, elevate their online trust levels, and foster a deeper understanding, safety consciousness, and confidence in telehealth services. On this basis, it can be concluded that only the participation of government, medical subjects, and online patients can effectively reduce perceived risks, improve perceived characteristics of online patients, enhance online patient trust, and promote the real willingness and behavior choice for online medical services, effectively improving the positive role of telemedicine in increasing health benefits to people.

Coarctation of the aorta (CoA) ranks among the most prevalent congenital heart defects and poses a life-threatening risk if left undiagnosed. Herein, we utilized fetal HQ technology to improve the prenatal prediction of CoA. A retrospective analysis was conducted on 64 fetal cases with suspected aortic arch constriction, identified through prenatal ultrasound findings between November 2020 and March 2022 at the Department of Ultrasound, Sir Run Run Shaw Hospital, Zhejiang University. According to the follow-up results, these cases were divided into two groups: 35 cases confirmed as CoA by postpartum surgery or induction, and 29 cases initially suspected of CoA prenatally but subsequently ruled outpostnatally. Additionally, 88 cases of normal fetuses were randomly selected as the control group. Both conventional M-mode ultrasound techniques and Fetal HQ software were utilized for fetal analysis across all groups. Parameters related to the heart were measured, including fetal 4-CV length, width, Global Spherical Index (GSI), Mitral Annular Plane Systolic Excursion (MAPSE), areas and ratios of the left and right ventricles, as well as lengths and ratios of the left and right ventricles. Functional measurements of the left and right ventricles included ejection fraction (EF), fractional area change (FAC), global longitudinal strain (GLS), fractional shortening (FS), end-diastolic diameter (ED), and sphericity index (SI). Left ventricular (LV)-GLS, LV-FAC, LV-EF, and LV-EF Z-score could potentially differentiate between true CoA and false CoA or normal groups and serve as potential indicators for the clinical diagnosis of CoA. The receiver operating characteristic (ROC) curves indicated that LV-GLS and LV-EF Z-score have the greatest predictive power for CoA diagnosis. The segments 6-12 of FS in the confirmed CoA group were significantly lower than those in the false CoA and normal groups. Fetal HQ technology, by assessing changes in the size and shape of the heart, can provide relatively reliable parameter support for the prenatal diagnosis of fetal aortic coarctation.

Attention deficit hyperactivity disorder (ADHD) is one of the most common neurodevelopmental disorders in childhood. Numerous resting-state functional magnetic resonance imaging (rs-fMRI) studies in ADHD have been performed using traditional low-frequency bands (0.01–0.08 Hz). However, the neural activity patterns of frequency subbands in ADHD still require further investigation. The purpose of this study is to explore the frequency-dependent characteristics and neural activity patterns of ADHD subtypes. We selected the ADHD combined type (ADHD-C, N = 25), ADHD inattentive type (ADHD-I, N = 26) and typically developing (TD, N = 28) children from the ADHD-200 Consortium. Based on the slow-5 band (0.01–0.027 Hz) and slow-4 band (0.027–0.073 Hz), we generated static and dynamic fractional amplitude of low-frequency fluctuation (fALFF) and regional homogeneity (ReHo) maps for each participant. A flexible-factorial analysis of variance model was performed on static and temporal dynamic rs-fMRI measurements within two subbands. Results revealed that the orbital-frontal gyrus, precuneus, superior temporal gyrus and angular gyrus were found to have obvious frequency band and group interaction effects. The intrinsic neural activity differences among three groups were more prominent in the slow-5 frequency band compared to the slow-4 band. In addition, the indices of significant interaction regions showed correlations with the progression of the disease and the features in slow-5 showed an advantageous diagnostic performance compared with those in slow-4. The results suggested the intrinsic neural activities of ADHD subtypes were frequency-dependent. The frequency-specific analysis of static and dynamic brain activity may provide a deeper understanding of neurophysiological dysfunction patterns in ADHD subtypes and provide supplementary information for assessing ADHD subtypes.

Background: The impact of dietary carbohydrate intake on bone health remains a subject of controversy, potentially influenced by individuals with diabetic osteoporosis who exhibit normal or elevated bone mineral density (BMD). The cross-sectional study was conducted to explore the association between carbohydrate intake and BMD, osteoporosis and fractures among adults without diabetes, based on the National health and nutrition examination survey (NHANES). Methods: Participants were from the NHANES 2005–2010, excluding individuals with diabetes and those with incomplete data. The association between carbohydrate intake and BMD was analyzed using Spearman correlation, linear regression analysis and subgroup analysis, respectively. The association between carbohydrate intake and osteoporosis/fractures was analyzed using weighted logistic regression analysis. Results: A total of 7275 adult participants were included and their dietary carbohydrate intake was inversely associated with BMD in the total femur [β = −0.20 95%CI (−0.30, −0.10); p

In the past few decades, great progress has been made in prenatal diagnosis of congenital heart disease (CHD). Fetal echocardiography is recognized as the main prenatal screening and diagnostic tool that can accurately detect approximately 85 % of fetal cardiac abnormalities. Evaluation of the fetal heart remains a major challenge in prenatal ultrasound screening and diagnosis due to fetal position, involuntary movement, small and complex fetal cardiac anatomy, maternal abdominal wall conditions, and lack of expertise in fetal echocardiography by some physicians engaged in obstetric ultrasound. Artificial intelligence (AI) can automate and standardize the display of each diagnostic section of the fetal heart and thus contribute to accurate diagnosis, which significantly optimizes the clinical application of fetal echocardiography. In this review, we not only clarify the role of AI but also highlight its significance and future solutions in the field of fetal echocardiography.

Abstract Background Quinic acid (QA) and its derivatives have good lipid-lowering and hepatoprotective functions, but their role in atherosclerosis remains unknown. This study attempted to investigate the mechanism of QA on atherogenesis in Apoe−/− mice induced by HFD. Methods HE staining and oil red O staining were used to observe the pathology. The PCSK9, Mac-3 and SM22a expressions were detected by IHC. Cholesterol, HMGB1, TIMP-1 and CXCL13 levels were measured by biochemical and ELISA. Lipid metabolism and the HMGB1-SREBP2-SR-BI pathway were detected by PCR and WB. 16 S and metabolomics were used to detect gut microbiota and serum metabolites. Results QA or low-frequency ABX inhibited weight gain and aortic tissue atherogenesis in HFD-induced Apoe−/− mice. QA inhibited the increase of cholesterol, TMA, TMAO, CXCL13, TIMP-1 and HMGB1 levels in peripheral blood of Apoe−/− mice induced by HFD. Meanwhile, QA or low-frequency ABX treatment inhibited the expression of CAV-1, ABCA1, Mac-3 and SM22α, and promoted the expression of SREBP-1 and LXR in the vascular tissues of HFD-induced Apoe−/− mice. QA reduced Streptococcus_danieliae abundance, and promoted Lactobacillus_intestinalis and Ileibacterium_valens abundance in HFD-induced Apoe−/− mice. QA altered serum galactose metabolism, promoted SREBP-2 and LDLR, inhibited IDOL, FMO3 and PCSK9 expression in liver of HFD-induced Apoe−/− mice. The combined treatment of QA and low-frequency ABX regulated microbe-related Glycoursodeoxycholic acid and GLYCOCHENODEOXYCHOLATE metabolism in HFD-induced Apoe−/− mice. QA inhibited TMAO or LDL-induced HCAECs damage and HMGB1/SREBP2 axis dysfunction, which was reversed by HMGB1 overexpression. Conclusions QA regulated the gut-liver lipid metabolism and chronic vascular inflammation of TMA/TMAO through gut microbiota to inhibit the atherogenesis in Apoe−/− mice, and the mechanism may be related to the HMGB1/SREBP2 pathway.

Abstract Background Whole-genome duplication and long terminal repeat retrotransposons (LTR-RTs) amplification in organisms are essential factors that affect speciation, local adaptation, and diversification of organisms. Understanding the karyotype projection and LTR-RTs amplification could contribute to untangling evolutionary history. This study compared the karyotype and LTR-RTs evolution in the genomes of eight oaks, a dominant lineage in Northern Hemisphere forests. Results Karyotype projections showed that chromosomal evolution was relatively conservative in oaks, especially on chromosomes 1 and 7. Modern oak chromosomes formed through multiple fusions, fissions, and rearrangements after an ancestral triplication event. Species-specific chromosomal rearrangements revealed fragments preserved through natural selection and adaptive evolution. A total of 441,449 full-length LTR-RTs were identified from eight oak genomes, and the number of LTR-RTs for oaks from section Cyclobalanopsis was larger than in other sections. Recent amplification of the species-specific LTR-RTs lineages resulted in significant variation in the abundance and composition of LTR-RTs among oaks. The LTR-RTs insertion suppresses gene expression, and the suppressed intensity in gene regions was larger than in promoter regions. Some centromere and rearrangement regions indicated high-density peaks of LTR/Copia and LTR/Gypsy. Different centromeric regional repeat units (32, 78, 79 bp) were detected on different Q. glauca chromosomes. Conclusion Chromosome fusions and arm exchanges contribute to the formation of oak karyotypes. The composition and abundance of LTR-RTs are affected by its recent amplification. LTR-RTs random retrotransposition suppresses gene expression and is enriched in centromere and chromosomal rearrangement regions. This study provides novel insights into the evolutionary history of oak karyotypes and the organization, amplification, and function of LTR-RTs.

The extraction and decolorization conditions for Spirulina platensis polysaccharides using hot alkali and hydrogen peroxide were investigated, and the process was optimized. Based on the results of a single-factor test, the yield and decolorization rate of polysaccharides were taken as indices to optimize the technological conditions for extracting polysaccharides from Spirulina platensis using the hot alkali method and decolorizing them with hydrogen peroxide using the response surface methodology. The optimal conditions for hot alkali extraction of polysaccharides from Spirulina platensis were determined as follows: NaOH concentration of 1.5%, extraction temperature of 90 ℃, extraction time of 4 hours, and material-to-liquid ratio of 1:20 g/mL. Under these conditions, the yield of polysaccharides from Spirulina platensis was 5.18%±0.23%. The optimal conditions for decolorization using hydrogen peroxide were found to be: 17% hydrogen peroxide solution, decolorization time of 55 minutes, and decolorization temperature of 50 ℃. Under these conditions, the actual decolorization rate was 82.54%±0.03%, and the retention rate of polysaccharides was 83.45%±0.13%. The findings of this study provide valuable insights for the development and utilization of Spirulina platensis polysaccharides.

Lakes and reservoirs are an important sources of drinking water for human life. However, numerous water quality indexes show seasonal fluctuation under different climatic conditions. We conducted a one-year investigation of Taiping Lake in year 2022 to construct relationship between water quality parameters and water quality. High water temperature promotes the growth of algae, and the existence of N species could provide the essential nutrient substance for the growing of algae. Under the irradiation of sunlight, algae will consume carbon dioxide through photosynthesis and release oxygen which can raise oxygen levels in the water. However, the rapid growth of algae could influence the acid-base balance in water through the consumption of carbon dioxide; thus, resulting in the increasing of pH values. These pH changes may pose a threat to drinking water.

Abstract Background Carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-hvKP) co-producing bla KPC and bla NDM poses a serious threat to public health. This study aimed to investigate the mechanisms underlying the resistance and virulence of CR-hvKP isolates collected from a Chinese hospital, with a focus on bla KPC and bla NDM dual-positive hvKP strains. Methods Five CR-hvKP strains were isolated from a teaching hospital in China. Antimicrobial susceptibility and plasmid stability testing, plasmid conjugation, pulsed-field gel electrophoresis, and whole-genome sequencing (WGS) were performed to examine the mechanisms of resistance and virulence. The virulence of CR-hvKP was evaluated through serum-killing assay and Galleria mellonella lethality experiments. Phylogenetic analysis based on 16 highly homologous carbapenem-resistant K. pneumoniae (CRKP) producing KPC-2 isolates from the same hospital was conducted to elucidate the potential evolutionary pathway of CRKP co-producing NDM and KPC. Results WGS revealed that five isolates individually carried three unique plasmids: an IncFIB/IncHI1B-type virulence plasmid, IncFII/IncR-type plasmid harboring KPC-2 and IncC-type plasmid harboring NDM-1. The conjugation test results indicated that the transference of KPC-2 harboring IncFII/IncR-type plasmid was unsuccessful on their own, but could be transferred by forming a hybrid plasmid with the IncC plasmid harboring NDM. Further genetic analysis confirmed that the pJNKPN26-KPC plasmid was entirely integrated into the IncC-type plasmid via the copy-in route, which was mediated by TnAs1 and IS26. Conclusion KPC-NDM-CR-hvKP likely evolved from a KPC-2-CRKP ancestor and later acquired a highly transferable bla NDM-1 plasmid. ST11-KL64 CRKP exhibited enhanced plasticity. The identification of KPC-2-NDM-1-CR-hvKP highlights the urgent need for effective preventive strategies against aggravated accumulation of resistance genes.

Homologous recombination (HR) repair for DNA double‐strand breaks (DSBs) is critical for maintaining genome stability and conferring the resistance of tumor cells to chemotherapy. Nuclear PTEN which contains both phosphatidylinositol 3,4,5‐trisphosphate 3‐phosphatase and protein phosphatase plays a key role in HR repair, but the underlying mechanism remains largely elusive. We find that SUMOylated PTEN promotes HR repair but represses nonhomologous end joining (NHEJ) repair by directly dephosphorylating TP53‐binding protein 1 (53BP1). During DNA damage responses (DDR), tumor suppressor ARF (p14ARF) was phosphorylated and then interacted efficiently with PTEN, thus promoting PTEN SUMOylation as an atypical SUMO E3 ligase. Interestingly, SUMOylated PTEN was subsequently recruited to the chromatin at DSB sites. This was because SUMO1 that was conjugated to PTEN was recognized and bound by the SUMO‐interacting motif (SIM) of breast cancer type 1 susceptibility protein (BRCA1), which has been located to the core of 53BP1 foci on chromatin during S/G2 stage. Furthermore, these chromatin‐loaded PTEN directly and specifically dephosphorylated phosphothreonine‐543 (pT543) of 53BP1, resulting in the dissociation of the 53BP1 complex, which facilitated DNA end resection and ongoing HR repair. SUMOylation‐site‐mutated PTENK254R mice also showed decreased DNA damage repair in vivo. Blocking the PTEN SUMOylation pathway with either a SUMOylation inhibitor or a p14ARF(2‐13) peptide sensitized tumor cells to chemotherapy. Our study therefore provides a new mechanistic understanding of PTEN in HR repair and clinical intervention of chemoresistant tumors.

Abstract Background The human sortilin protein is an important drug target and detection marker for cancer research. The sortilin from Toxoplasma gondii transports proteins associated with the apical organelles of the parasite. In this study, we aimed to determine the intracellular localization and structural domains of T. gondii sortilin, which may mediate protein transportation. Approaches to the functional inhibition of sortilin to establish novel treatments for T. gondii infections were explored. Methods A gene encoding the sortilin protein was identified in the T. gondii genome. Immunoprecipitation and mass spectrometry were performed to identify the protein species transported by T. gondii sortilin. The interaction of each structural domain of sortilin with the transported proteins was investigated using bio-layer interferometry. The binding regions of the transported proteins in sortilin were identified. The effect of the sortilin inhibitor AF38469 on the infectivity of T. gondii was investigated. The binding site of AF38469 on sortilin was determined. Results The subdomains Vps10, sortilin-C, and sortilin-M of the sortilin were identified as the binding regions for intracellular transportation of the target proteins. The sortilin inhibitor AF38469 bound to the Vps10 structural domain of T. gondii sortilin, which inhibited parasite invasion, replication, and intracellular growth in vitro and was therapeutic in mice infected with T. gondii. Conclusion The Vps10, sortilin-C, and sortilin-M subdomains of T. gondii sortilin were identified as functional regions for intracellular protein transport. The binding region for the sortilin inhibitor AF38469 was also identified as the Vps10 subdomain. This study establishes sortilin as a promising drug target against T. gondii and provides a valuable reference for the development of anti-T. gondii drug-target studies. Graphical Abstract

Abstract Background Forests are essential for maintaining species diversity, stabilizing local and global climate, and providing ecosystem services. Exploring the impact of paleogeographic events and climate change on the genetic structure and distribution dynamics of forest keystone species could help predict responses to future climate change. In this study, we combined an ensemble species distribution model (eSDM) and multilocus phylogeography to investigate the spatial genetic patterns and distribution change of Quercus glauca Thunb, a keystone of East Asian subtropical evergreen broad-leaved forest. Results A total of 781 samples were collected from 77 populations, largely covering the natural distribution of Q. glauca. The eSDM showed that the suitable habitat experienced a significant expansion after the last glacial maximum (LGM) but will recede in the future under a general climate warming scenario. The distribution centroid will migrate toward the northeast as the climate warms. Using nuclear SSR data, two distinct lineages split between east and west were detected. Within-group genetic differentiation was higher in the West than in the East. Based on the identified 58 haplotypes, no clear phylogeographic structure was found. Populations in the Nanling Mountains, Wuyi Mountains, and the southwest region were found to have high genetic diversity. Conclusions A significant negative correlation between habitat stability and heterozygosity might be explained by the mixing of different lineages in the expansion region after LGM and/or hybridization between Q. glauca and closely related species. The Nanling Mountains may be important for organisms as a dispersal corridor in the west-east direction and as a refugium during the glacial period. This study provided new insights into spatial genetic patterns and distribution dynamics of Q. glauca.

Abstract Purpose This study aimed to characterise the whole-genome structure of two clinical Klebsiella pneumoniae strains co-harbouring mcr-8.1 and tmexCD1-toprJ1, both resistant to colistin and tigecycline. Methods K. pneumoniae strains TGC-02 (ST656) and TGC-05 (ST273) were isolated from urine samples of different patients hospitalised at separate times in 2021. Characterisation involved antimicrobial susceptibility testing (AST), conjugation assays, whole-genome sequencing (WGS), and bioinformatics analysis. Comparative genomic analysis was conducted on mcr-8.1-carrying and tmexCD1-toprJ1-carrying plasmids. Results Both K. pneumoniae isolates displayed a multidrug-resistant phenotype, exhibiting resistance or reduced susceptibility to ampicillin, ampicillin/sulbactam, cefazolin, aztreonam, amikacin, gentamicin, tobramycin, ciprofloxacin, levofloxacin, nitrofurantoin, trimethoprim/sulfamethoxazole, apramycin, tigecycline and colistin. WGS analysis revealed that clinical strain TGC-02 carried the TmexCD1-toprJ1 gene on a 200-Kb IncFII/IncFIB-type plasmid, while mcr-8 was situated on a 146-Kb IncFII-type plasmid. In clinical strain TGC-05, TmexCD1-toprJ1 was found on a 300-Kb IncFIB/IncHI1B/IncR-type plasmid, and mcr-8 was identified on a 137-Kb IncFII/IncFIA-type plasmid. Conjugation experiments assessed the transferability of these plasmids. While transconjugants were not obtained for TGC-05 despite multiple screening with tigecycline or colistin, pTGC-02-tmex and pTGC-02-mcr8 from clinical K. pneumoniae TGC-02 demonstrated self-transferability through conjugation. Notably, the rearrangement of pTGC-02-tmex and pTGC-02-mcr8 via IS26-based homologous recombination was observed. Moreover, the conjugative and fusion plasmids of the transconjugant co-harboured the tmexCD1-toprJ1 gene cluster and mcr-8.1, potentially resulting from IS26-based homologous recombination. Conclusion The emergence of colistin- and tigecycline-resistant K. pneumoniae strains is concerning, and effective surveillance measures should be implemented to prevent further dissemination.

Abstract Bacterial cystitis, a commonly occurring urinary tract infection (UTI), is renowned for its extensive prevalence and tendency to recur. Despite the extensive utilization of levofloxacin as a conventional therapeutic approach for bacterial cystitis, its effectiveness is impeded by adverse toxic effects, drug resistance concerns, and its influence on the gut microbiota. This study introduces Lev@PADM, a hydrogel with antibacterial properties that demonstrates efficacy in the treatment of bacterial cystitis. Lev@PADM is produced by combining levofloxacin with decellularized porcine acellular dermal matrix hydrogel and exhibits remarkable biocompatibility. Lev@PADM demonstrates excellent stability as a hydrogel at body temperature, enabling direct administration to the site of infection through intravesical injection. This localized delivery route circumvents the systemic circulation of levofloxacin, resulting in a swift and substantial elevation of the antimicrobial agent’s concentration specifically at the site of infection. The in vivo experimental findings provide evidence that Lev@PADM effectively prolongs the duration of levofloxacin’s action, impedes the retention and invasion of E.coli in the urinary tract, diminishes the infiltration of innate immune cells into infected tissues, and simultaneously preserves the composition of the intestinal microbiota. These results indicate that, in comparison to the exclusive administration of levofloxacin, Lev@PADM offers notable benefits in terms of preserving the integrity of the bladder epithelial barrier and suppressing the recurrence of urinary tract infections. Graphical Abstract

The membrane-proximal external region (MPER) represents a highly conserved region of the Human Immunodeficiency Virus (HIV) envelope glycoprotein (env) targeted by several broadly neutralizing antibodies (bnAbs). In this study, we employed single genome amplification to amplify 34 full-length env sequences from the 2005 plasma sample of CBJC504, a chronic HIV-1 clade B infected individual. We identified three amino acid changes (N671S, D674N, and K677R) in the MPER. A longitudinal analysis revealed that the proportion of env sequences with MPER mutations increased from 26.5 % in 2005 to 56.0 % in 2009, and the sequences with the same mutation clustered together. Nine functional pseudoviruses were generated from the 34 env sequences to examine the effect of these mutations on neutralizing activity. Pseudoviruses carrying N674 or R677 mutations demonstrate increased sensitivity to autologous plasma and monoclonal antibodies 2F5, 4E10, and 10E8. Reverse mutations were performed in env including N674, R677, D659, and S671/N677 mutations, to validate the impact of the mutations on neutralizing sensitivity. Neutralization assays indicated that the N671S mutation increased neutralization sensitivity to 2F5 and 10E8. The amino acid R at position 677 increased viral resistance to 10E8, whereas N enhanced viral resistance to 4E10 and 10E8. It has been proposed that critical amino acids in the extra-MPER and the number of potential N-like glycosylation sites (PNGSs) in the V1 loop may have an impact on neutralizing activity. Understanding the mutations and evolution of MPER in chronically infected patients with HIV-1 is crucial for the design and development of vaccines that trigger bnAbs against MPER.

Abstract The coral reef microbiome plays a vital role in the health and resilience of reefs. Previous studies have examined phage therapy for coral pathogens and for modifying the coral reef microbiome, but defence systems against coral‐associated bacteria have received limited attention. Phage defence systems play a crucial role in helping bacteria fight phage infections. In this study, we characterized a new defence system, Hma (HmaA‐HmaB‐HmaC), in the coral‐associated Halomonas meridiana derived from the scleractinian coral Galaxea fascicularis. The Swi2/Snf2 helicase HmaA with a C‐terminal nuclease domain exhibits antiviral activity against Escherichia phage T4. Mutation analysis revealed the nickase activity of the nuclease domain (belonging to PDD/EXK superfamily) of HmaA is essential in phage defence. Additionally, HmaA homologues are present in ~1000 bacterial and archaeal genomes. The high frequency of HmaA helicase in Halomonas strains indicates the widespread presence of these phage defence systems, while the insertion of defence genes in the hma region confirms the existence of a defence gene insertion hotspot. These findings offer insights into the diversity of phage defence systems in coral‐associated bacteria and these diverse defence systems can be further applied into designing probiotics with high‐phage resistance.