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The Classroom Appraisal of Resources and Demands (CARD) was designed to evaluate teacher stress based on subjective evaluations of classroom demands and resources. However, the CARD has been mostly utilized in western countries. The aim of the current study was to provide aspects of the validity of responses to a Chinese version of the CARD that considers Chinese teachers' unique vocational conditions in the classroom. A sample of 580 Chinese elementary school teachers (510 female teachers and 70 male teachers) were asked to respond to the Chinese version of the CARD. Confirmatory factor analyses showed that the data fit the theoretical model very well (e.g., CFI: .982; NFI: .977; GFI: .968; SRMR: .028; RMSEA: .075; where CFI is comparative fit index, NFI is normed fit index, GFI is goodness of fit, SRMR is standardized root mean square residual, RMSEA is root mean square error of approximation), thus providing evidence of construct validity. Latent constructs of the Chinese version of the CARD were also found to be significantly associated with other measures that are related to teacher stress such as self-efficacy, job satisfaction, personal habits to deal with stress, and intention to leave their current job. [ABSTRACT FROM AUTHOR]

Highlights: [•] We reviewed adverse events of live attenuated Japanese encephalitis vaccine. [•] The adverse event rate was 61.24 per million doses. [•] Most adverse events are relatively mild, with relatively rare neurologic events. [Copyright &y& Elsevier]

Summary: Objective: To evaluate the impact of the universal infant hepatitis B vaccination program on hepatitis B virus (HBV) infection in Guangdong Province, China. Methods: In 2006, a serosurvey was conducted in Guangdong Province among children aged <15 years, 14 years after the introduction of universal infant hepatitis B vaccination. The participants were selected by stratified, multi-stage random sampling. Demographic characteristics and hepatitis B vaccination history were collected by a questionnaire and a review of the vaccination records, and serum specimens were tested for hepatitis B surface antigen (HBsAg), antibody to hepatitis B core antigen (anti-HBc), and antibody to hepatitis B surface antigen (anti-HBs) by ELISA. The prevalence rate of HBV serological markers and the rate of immunization coverage in this survey were compared with those of the 1992 and 2002 surveys. Results: A total of 1967 children aged <15 years participated in 2006. The prevalence rate of HBsAg decreased from 19.86% in the 1992 survey to 4.91% in the 2006 survey. The rates of three-dose and timely birth dose coverage of hepatitis B vaccine were 92.40% and 70.84%, respectively, among children born during the period 2002–2005. The prevalence of HBsAg was significantly lower among fully immunized children (1.99%) than among unvaccinated children (5.56%). Conclusions: Guangdong Province has successfully integrated the hepatitis B vaccine into routine immunization programs and this has had a very significant impact on decreasing the HBsAg carrier rate among children. [ABSTRACT FROM AUTHOR]

Fusobacterium nucleatum is an oral commensal bacterium that can act as an opportunistic pathogen, and is implicated in diseases such as periodontitis, adverse pregnancy outcomes, colorectal cancer, and Alzheimer's disease. F. nucleatum synthesizes lanthionine for its peptidoglycan, rather than meso-2,6-diaminopimelic acid (DAP) used by most Gram-negative bacteria. Despite lacking the biosynthetic pathway for DAP, the genome of F. nucleatum ATCC 25586 encodes a predicted DAP epimerase. A recent study hypothesized that this enzyme may act as a lanthionine epimerase, but the authors found a very low turnover rate, suggesting that this enzyme likely has another more favored substrate. Here, we characterize this enzyme as a histidine racemase (HisR), and found that catalytic turnover is ∼10,000× faster with L-histidine than with L,L-lanthionine. Kinetic experiments suggest that HisR functions as a cofactor-independent racemase and that turnover is specific for histidine, while crystal structures of catalytic cysteine to serine mutants (C67S or C209S) reveal this enzyme in its substrate-unbound, open conformation. Currently, the only other reported cofactor-independent histidine racemase is CntK from Staphylococcus aureus, which is used in the biosynthesis of staphylopine, a broad-spectrum metallophore that increases virulence of S. aureus. However, CntK shares only 28% sequence identity with HisR, and their genes exist in different genomic contexts. Knock-out of hisR in F. nucleatum results in a small but reproducible lag in growth compared to wild-type during exponential phase, suggesting that HisR may play a role in growth of this periodontal pathogen., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Fusobacterium nucleatum , prevalent in the oral cavity, is significantly linked to overall human health. Our molecular comprehension of its role in oral biofilm formation and its interactions with the host under various pathological circumstances has seen considerable advancements in recent years, primarily due to the development of various genetic tools for DNA manipulation in this bacterium. Of these, counterselection-based unmarked in-frame mutation methods have proved notably effective. Under suitable growth conditions, cells carrying a counterselectable gene die, enabling efficient selection of rare defined allelic exchange mutants. The sacB gene from Bacillus subtilis , encoding levansucrase, is a widely used counterselective marker partly due to the easy availability of sucrose. Yet, its potential application in F. nucleatum genetic study remains untested. We demonstrated that F. nucleatum cells expressing sacB in either a shuttle or suicide plasmid exhibit a lethal sensitivity to supplemental sucrose. Utilizing sucrose counterselection, we created an in-frame deletion of the F. nucleatum tonB gene, a critical gene for energy-dependent transport processes in Gram-negative bacteria, and a precise knockin of the luciferase gene immediately following the stop codon of the hslO gene, the last gene of a five-gene operon possible related to the natural competence of F. nucleatum . Post counterselection with 5% sucrose, chromosomal plasmid loss occurred in all colonies, leading to gene alternations in half of the screened isolates. This sacB -based counterselection technique provides a reliable method for isolating unmarked gene mutations in wild-type F. nucleatum , enriching the toolkit for fusobacterial research., Importance: Investigations into Fusobacterium nucleatum 's role in related diseases significantly benefit from the strategies of creating unmarked gene mutations, which hinge on using a counterselective marker. Previously, the galk -based allelic exchange method, while effective, faced an inherent limitation - the need for a modified host. This study aims to surmount this limitation by substituting galK with sacB for gene modification in F. nucleatum . Our application of the sacB -based methodology successfully yielded a tonB in-frame deletion mutant and a luciferase gene knockin at the precise chromosomal location in the wild-type background. The new method augments the existing toolkit for F. nucleatum research and has far-reaching implications due to the easy accessibility to the counterselection compound sucrose. We anticipate its broader adoption in further exploration, thereby reinforcing its critical role in propelling our understanding of F. nucleatum .

Introduction: Health records serve not only as a database of a patient's health history and treatment process but also as a crucial tool for doctors to diagnose and treat patients. However, the storage and sharing of these records are sensitive issues as they involve maintaining patient privacy and ensuring data transparency, security, and interoperability between different parties. Challenges to achieving these goals in the current surgical process can impact the allocation of medical resources and surgical outcomes., Methods: This article proposes a healthcare 5.0 framework for medical surgery that deploys a secure and distributed network using Blockchain to demonstrate transactions between different parties in the orthopedic surgery process. The proposed network uses the Hyperledger Composer platform for deployment, and a patient-doctor-supplier orthopedic surgery network is designed and implemented to enable the safe sharing of medical records., Results: A benchmarking tool was implemented for analyzing different scenarios of applying blockchain technology to orthopedic surgery. The application of blockchain technology to orthopedic surgery presents a promising solution for data sharing and supply chain management in the field. The integration of blockchain with cloud storage and hybrid encryption ensures secure and efficient storage of Electronic Health Record (EHR) and Personal Health Record (PHR) data. By leveraging the tamper-proof nature of blockchain and addressing concerns regarding centralized data storage, this scenario demonstrates enhanced security, improved access efficiency, and privacy protection in medical data sharing., Conclusions: The article demonstrates the feasibility of using an IoT-based blockchain network in orthopedic surgery, which can reduce medical errors and improve data interoperability among different parties. This unique application of blockchain enables secure sharing of medical records, ensuring transparency, security, and interoperability. The network design may also be applicable to other surgeries and medical applications in the future., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Fusobacterium nucleatum , a pathobiont inhabiting the oral cavity, contributes to opportunistic diseases, such as periodontal diseases and gastrointestinal cancers, which involve microbiota imbalance. Broad-spectrum antimicrobial agents, while effective against F. nucleatum infections, can exacerbate dysbiosis. This necessitates the discovery of more targeted narrow-spectrum antimicrobial agents. We therefore investigated the potential for the fusobacterial enoyl-ACP reductase II (ENR II) isoenzyme Fn FabK (C4N14&#95; 04250) as a narrow-spectrum drug target. ENRs catalyze the rate-limiting step in the bacterial fatty acid synthesis pathway. Bioinformatics revealed that of the four distinct bacterial ENR isoforms, F. nucleatum specifically encodes Fn FabK. Genetic studies revealed that fabK was indispensable for F. nucleatum growth, as the gene could not be deleted, and silencing of its mRNA inhibited growth under the test conditions. Remarkably, exogenous fatty acids failed to rescue growth inhibition caused by the silencing of fabK . Screening of synthetic phenylimidazole analogues of a known FabK inhibitor identified an inhibitor (i.e., 681) of Fn FabK enzymatic activity and F. nucleatum growth, with an IC 50 of 2.1 μM (1.0 μg/mL) and a MIC of 0.4 μg/mL, respectively. Exogenous fatty acids did not attenuate the activity of 681 against F. nucleatum . Furthermore, Fn FabK was confirmed as the intracellular target of 681 based on the overexpression of Fn FabK shifting MICs and 681-resistant mutants having amino acid substitutions in Fn FabK or mutations in other genetic loci affecting fatty acid biosynthesis. 681 had minimal activity against a range of commensal flora, and it was less active against streptococci in physiologic fatty acids. Taken together, Fn FabK is an essential enzyme that is amenable to drug targeting for the discovery and development of narrow-spectrum antimicrobial agents.

Krieger et al.'s study in this issue of Cell Host & Microbe reveals that Fusobacterium nucleatum subsp. animalis strains, previously underestimated, are significant in disease-affected oral areas. This challenges the long-held notion of the dominance of Fusobacterium nucleatum subsp. nucleatum, reshaping our understanding of Fusobacterium distribution in the oral microbiome., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The study of the relationship between key psychological attributes of learners and their engagement in second language (L2) learning helps to understand the critical personality mechanisms influencing language learning. The present study examined the L2 learning engagement from the perspectives of grit (i.e., consistent efforts and interests devoted to a long-term goal) and affect balance (a notion that takes into account both positive and negative emotions concurrently, assessing and evaluating which side holds more significance or influence). A cohort of English L2 learners ( N = 394) participated in an online survey aimed at gauging their levels of grit, affect balance, and engagement in L2 learning. The results indicated that grit and affect balance were significantly correlated with behavioral engagement and affective engagement in L2 learning. However, among the two components of grit, namely consistency of interest, showed no significant relationship with L2 learning engagement, while perseverance of effort was significantly positively correlated with L2 learning engagement. Affect balance played a partially mediating and full mediating role between perseverance of effort and behavioral engagement as well as affective engagement respectively. These findings confirm the crucial role of perseverance of effort in second language learning and reveal the unique role of affect balance in their relationship.

Gene inactivation by creating in-frame deletion mutations in Fusobacterium nucleatum is time consuming, and most fusobacterial strains are genetically intractable. Addressing these problems, we introduced a riboswitch-based inducible CRISPR interference (CRISPRi) system. This system employs the nuclease-inactive Streptococcus pyogenes Cas9 protein (dCas9), specifically guided to the gene of interest by a constantly expressed single-guide RNA (sgRNA). Mechanistically, this dCas9-sgRNA complex serves as an insurmountable roadblock for RNA polymerase, thus repressing the target gene transcription. Leveraging this system, we first examined two non-essential genes, ftsX and radD , which are pivotal for fusobacterial cytokinesis and coaggregation. Upon adding the inducer, theophylline, ftsX suppression caused filamentous cell formation akin to chromosomal ftsX deletion, while targeting radD significantly reduced RadD protein levels, abolishing RadD-mediated coaggregation. The system was then extended to probe essential genes bamA and ftsZ , which are vital for outer membrane biogenesis and cell division. Impressively, bamA suppression disrupted membrane integrity and bacterial separation, stalling growth, while ftsZ targeting yielded elongated cells in broth with compromised agar growth. Further studies on F. nucleatum clinical strain CTI-2 and Fusobacterium periodonticum revealed reduced indole synthesis when targeting tnaA . Moreover, silencing clpB in F. periodonticum decreased ClpB, increasing thermal sensitivity. In summary, our CRISPRi system streamlines gene inactivation across various fusobacterial strains.IMPORTANCEHow can we effectively investigate the gene functions in Fusobacterium nucleatum , given the dual challenges of gene inactivation and the inherent genetic resistance of many strains? Traditional methods have been cumbersome and often inadequate. Addressing this, our work introduces a novel inducible CRISPR interference (CRISPRi) system in which dCas9 expression is controlled at the translation level by a theophylline-responsive riboswitch unit, and single-guide RNA expression is driven by the robust, constitutive rpsJ promoter. This approach simplifies gene inactivation in the model organism (ATCC 23726) and extends its application to previously considered genetically intractable strains like CTI-2 and Fusobacterium periodonticum . With CRISPRi's potential, it is a pivotal tool for in-depth genetic studies into fusobacterial pathogenesis, potentially unlocking targeted therapeutic strategies., Competing Interests: The authors declare no conflict of interest.

Importance: This paper illuminates the significant question of how the oral commensal Fusobacterium nucleatum adapts to the metabolically changing environments of several extra-oral sites such as placenta and colon to promote various diseases as an opportunistic pathogen. We demonstrate here that the highly conserved R hodobacter n itrogen- f ixation complex, commonly known as Rnf complex, is key to fusobacterial metabolic adaptation and virulence. Genetic disruption of this Rnf complex causes global defects in polymicrobial interaction, biofilm formation, cell growth and morphology, hydrogen sulfide production, and ATP synthesis. Targeted metabolomic profiling demonstrates that the loss of this respiratory enzyme significantly diminishes catabolism of numerous amino acids, which negatively impacts fusobacterial virulence as tested in a preterm birth model in mice., Competing Interests: The authors declare no conflict of interest.

Low human papillomavirus (HPV) vaccine uptake is a key barrier to cervical cancer elimination. We aimed to evaluate the health impact and cost-effectiveness of introducing different HPV vaccines into immunization programs and scaling up the screening program in Guangdong. We used a dynamic compartmental model to estimate the impact of intervention strategies during 2023-2100. We implemented the incremental cost-effectiveness ratio (ICER) in costs per averted disability-adjusted life year (DALY) as an indicator to assess the effectiveness of the intervention. We used an age-standardized incidence of 4 cases per 100,000 women as the threshold for the elimination of cervical cancer. Compared with the status quo, scaling up cervical cancer screening coverage alone would prevent 215,000 (95% CI: 205,000 to 227,000) cervical cancer cases and 49,000 (95% CI: 48,000 to 52,000) deaths during 2023-2100. If the coverage of vaccination reached 90%, domestic two-dose 2vHPV vaccination would be more cost-effective than single-dose and two-dose 9vHPV vaccination. If Guangdong introduced domestic two-dose 2vHPV vaccination at 90% coverage for schoolgirls from 2023 and increased the screening coverage, cervical cancer would be eliminated by 2049 (95% CI 2047 to 2051). Introducing two doses of domestic 2vHPV vaccination for schoolgirls and expanding cervical cancer screening is estimated to be highly cost-effective to accelerate the elimination of cervical cancer in Guangdong.

Controlled septal peptidoglycan hydrolysis is vital for bacterial cell division, preserving cellular integrity and facilitating proper daughter cell separation. In Escherichia coli, the FtsEX ABC system governs cell wall hydrolase activation by regulating the EnvC activator. However, the processes underlying cell division in the Gram-negative oral bacterium Fusobacterium nucleatum are poorly understood, mainly due to its well-known genetic intractability. Herein, we provide a step-by-step procedure for a new gene deletion method in F. nucleatum, focusing on the ftsX gene as a target. This novel approach exploits the HicAB toxin-antitoxin system, using HicA as a counter-selective marker to enable efficient and precise gene deletion. By implementing this technique, we successfully demonstrated its applicability in F. nucleatum, providing new insights into this important microorganism's cell division process. Furthermore, this advanced gene deletion technique offers a valuable resource for future investigation into the functional characterization of genes involved in cell wall biogenesis in F. nucleatum and other genetically intractable microorganisms., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

The present study offered the emotion prototypicality (EmoPro) ratings for 1,083 Chinese emotion words. EmoPro measures the extent to which an emotion word refers to an emotion. Emotion words with high EmoPro are representative emotion-label words, so EmoPro provides an objective evaluation of defining an emotion-label word. The EmoPro rating results had adequate reliability and validity. The correlation results showed that EmoPro was related to valence, arousal, age of acquisition (AoA), and word frequency, but was not associated with concreteness, familiarity, and imageability. The EmoPro was also predicted by valence, arousal, and AoA. However, EmoPro failed to predict lexical decision performance after considering the contribution of valence, arousal, AoA, and concreteness. The present normative study is of high value for selecting the most typical emotion-label words as stimuli in future affective science and psycholinguistic studies., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Morality has been an integral part of social cognition and our daily life, and different languages may exert distinct impacts on human moral judgment. However, it remains unclear how moral concept is encoded in the bilingual brain. This study, therefore, aimed to explore the emotional and cognitive involvement of bilingual morality judgement by using combined event-related potential (ERP) and psychophysiological (including skin, heart, and pulse) measures. In the experiment, thirty-one Chinese-English bilingual participants were asked to make moral judgments in Chinese and English, respectively. Our results revealed increased early frontal N400 and decreased LPC in L1 moral concept encoding as compared to L2, suggesting that L1 was more reliant on automatic processes and emotions yet less on elaboration. In contrast, L2 moral and immoral concepts elicited enhanced LPC, decreased N400, and greater automatic psychophysiological electrocardiograph responses, which might reflect more elaborate processing despite blunted emotional responses and increased anxiety. Additionally, both behavioral and P200 data revealed a reliable immorality bias across languages. Our results were discussed in light of the dual-process framework of moral judgments and the (dis)embodiment of bilingual processing, which may advance our understanding of the interplay between language and morality as well as between emotion and cognition.

Inducible gene expression systems are important for studying bacterial gene function, yet most exhibit leakage. In this study, we engineered a leakage-free hybrid system for precise gene expression controls in Fusobacterium nucleatum by integrating the xylose-inducible expression system with the theophylline-responsive riboswitch. This innovative method enables concurrent control of target gene expression at both transcription and translation initiation levels. Using luciferase and the indole-producing enzyme tryptophanase (TnaA) as reporters, we demonstrated that the hybrid system displays virtually no observable signal in the absence of inducers. We employed this system to express FtsX, a protein related to fusobacterial cytokinesis, in an ftsX mutant strain, unveiling a dose-dependent manner in FtsX production. Without inducers, cells form long filaments, while increasing FtsX levels by increasing inducer concentrations led to a gradual reduction in cell length until normal morphology was restored. Crucially, this system facilitated essential gene investigation, identifying the signal peptidase lepB gene as vital for F. nucleatum . LepB's essentiality stems from depletion, affecting outer membrane biogenesis and cell division. This novel hybrid system holds the potential for advancing research on essential genes and accurate gene regulation in F. nucleatum . IMPORTANCE Fusobacterium nucleatum , an anaerobic bacterium prevalent in the human oral cavity, is strongly linked to periodontitis and can colonize areas beyond the oral cavity, such as the placenta and gastrointestinal tract, causing adverse pregnancy outcomes and promoting colorectal cancer growth. Given F. nucleatum 's clinical significance, research is underway to develop targeted therapies to inhibit its growth or eradicate the bacterium specifically. Essential genes, crucial for bacterial survival, growth, and reproduction, are promising drug targets. A leak-free-inducible gene expression system is needed for studying these genes, enabling conditional gene knockouts and elucidating the importance of those essential genes. Our study identified lepB as the essential gene by first generating a conditional gene mutation in F. nucleatum . Combining a xylose-inducible system with a riboswitch facilitated the analysis of essential genes in F. nucleatum , paving the way for potential drug development targeting this bacterium for various clinical applications., Competing Interests: The authors declare no conflict of interest.

Although the age of acquisition (AoA) effect has been established in numerous studies, how emotion word processing is modulated by AoA, along with affective factors, such as valence and arousal, is not well understood. Hence, the influence of age of acquisition (AoA), valence, and arousal on Chinese emotion word recognition was investigated through two experiments. Experiment 1 ( N = 30) adopted a valence judgment task to explore the roles of valence and AoA in emotion word recognition, whereas Experiment 2 ( N = 30) used a lexical decision task to examine AoA and arousal effects. A mixed linear effects model was used to examine the fixed effects of AoA, arousal, and valence and random effects of participants and items. The findings provided confirmation of the effects of AoA, valence, and arousal. Notably, AoA and valence had independent influences on emotion word recognition, as evidenced by longer reaction times for later-acquired words and negative words compared to early-acquired words and positive words (all ps < 0.05). On the other hand, AoA and arousal demonstrated interdependent effects on emotion word recognition. Specifically, a larger AoA effect was observed for low-arousing words (all ps < 0.05), whereas the influence of AoA on high-arousing words was insignificant. These results underscored the significance of AoA in processing emotion words and highlighted the interplay between AoA and arousal. Additionally, it is plausible to suggest that the AoA effect was primarily perceptual rather than semantic in nature.

The study of fusobacterial virulence factors has dramatically benefited from the creation of various genetic tools for DNA manipulation, including galK- based counterselection for in-frame deletion mutagenesis in Fusobacterium nucleatum, which was recently developed. However, this method requires a host lacking the galK gene, which is an inherent limitation. To circumvent this limitation, we explored the possibility of using the hicA gene that encodes a toxin consisting of a HicAB toxin-antitoxin module in Fusobacterium periodonticum as a new counterselective marker. Interestingly, the full-length hicA gene is not toxic in F. nucleatum, but a truncated hicA gene version lacking the first six amino acids is functional as a toxin. The toxin expression is driven by an rpsJ promoter and is controlled at its translational level by using a theophylline-responsive riboswitch unit. As a proof of concept, we created markerless in-frame deletions in the fusobacterial adhesin radD gene within the F. nucleatum rad operon and the tnaA gene that encodes the tryptophanase for indole production. After vector integration, plasmid excision after counterselection appeared to have occurred in 100% of colonies grown on theophylline-added plates and resulted in in-frame deletions in 50% of the screened isolates. This hicA -based counterselection system provides a robust and reliable counterselection in wild-type background F. nucleatum and should also be adapted for use in other bacteria. IMPORTANCE Fusobacterium nucleatum is an indole-producing human oral anaerobe associated with periodontal diseases, preterm birth, and several cancers. Little is known about the mechanisms of fusobacterial pathogenesis and associated factors, mainly due to the lack of robust genetic tools for this organism. Here, we showed that a mutated hicA gene from Fusobacterium periodonticum expresses an active toxin and was used as a counterselection marker. This hicA -based in-frame deletion system efficiently creates in-frame deletion mutations in the wild-type background of F. nucleatum. This is the first report to use the hicA gene as a counterselection marker in a bacterial genetic study.

Extensive research has explored the perception of English lexical stress by Chinese EFL learners and tried to unveil the underlying mechanism of the prosodic transfer from a native tonal language to a non-native stress language. However, the role of the pitch as the shared cue by lexical stress and lexical tone during the transfer remains controversial when the segmental cue (i.e., reduced vowel) is absent. By employing event-related potential (ERP) measurements, the current study aimed to further investigate the role of the pitch during the prosodic transfer from L1 lexical tone to L2 lexical stress and the underlying neural responses. Two groups of adult Chinese EFL learners were compared, as both Mandarin and Cantonese are tonal languages with different levels of complexity. The results showed that Cantonese speakers relied more than Mandarin speakers on pitch cues, not only in their processing of English lexical stress but also in word recognition. Our findings are consistent with the arguments of Cue Weighting and attest to the influence of native tonal language experience on second language acquisition. The results may have implications on pedagogical methods that pitch could be an important clue in second language teaching.

Rehabilitation movement assessment often requires patients to wear expensive and inconvenient sensors or optical markers. To address this issue, we propose a non-contact and real-time approach using a lightweight pose detection algorithm-Sports Rehabilitation-Pose (SR-Pose), and a depth camera for accurate assessment of rehabilitation movement. Our approach utilizes an E-Shufflenet network to extract underlying features of the target, a RLE-Decoder module to directly regress the coordinate values of 16 key points, and a Weight Fusion Unit (WFU) module to output optimal human posture detection results. By combining the detected human pose information with depth information, we accurately calculate the angle between each joint in three-dimensional space. Furthermore, we apply the DTW algorithm to solve the distance measurement and matching problem of video sequences with different lengths in rehabilitation evaluation tasks. Experimental results show that our method can detect human joint nodes with an average detection speed of 14.32ms and an average detection accuracy for pose of 91.2%, demonstrating its computational efficiency and effectiveness for practical application. Our proposed approach provides a low-cost and user-friendly alternative to traditional sensor-based methods, making it a promising solution for rehabilitation movement assessment.

The Gram-negative anaerobe Fusobacterium nucleatum is a major producer of hydrogen sulfide (H 2 S), a volatile sulfur compound that causes halitosis. Here, we dissected the genetic determinants of H 2 S production and its role in bacterial fitness and virulence in this important member of the oral microbiome. F. nucleatum possesses four enzymes, CysK1, CysK2, Hly, and MegL, that presumably metabolize l-cysteine to H 2 S, and CysK1 was previously shown to account for most H 2 S production in vitro , based on correlations of enzymatic activities with gene expression at mid-log phase. Our molecular studies showed that cysK1 and megL were highly expressed at the late exponential growth phase, concomitant with high-level H 2 S production, while the expression levels of the other genes remained substantially lower during all growth phases. Although the genetic deletion of cysK1 without supplementation with a CysK1-catalyzed product, lanthionine, caused cell death, the conditional Δ cysK1 mutant and a mutant lacking hly were highly proficient in H 2 S production. In contrast, a mutant devoid of megL showed drastically reduced H 2 S production, and a cysK2 mutant showed only minor deficiencies. Intriguingly, the exposure of these mutants to various antibiotics revealed that only the megL mutant displayed altered susceptibility compared to the parental strain: partial sensitivity to nalidixic acid and resistance to kanamycin. Most significantly, the megL mutant was attenuated in virulence in a mouse model of preterm birth, with considerable defects in the spread to amniotic fluid and the colonization of the placenta and fetus. Evidently, the l-methionine γ-lyase MegL is a major H 2 S-producing enzyme in fusobacterial cells that significantly contributes to fusobacterial virulence and antibiotic susceptibility. IMPORTANCE Fusobacterium nucleatum is a key commensal anaerobe of the human oral cavity that plays a significant role in oral biofilm development and contributes to additional pathologies at extraoral sites, such as promoting preterm birth and colorectal cancer. Although F. nucleatum is known as a major producer of hydrogen sulfide (H 2 S), its genetic determinants and physiological functions are not well understood. By a combination of bacterial genetics, biochemical methods, and in vivo models of infection, here, we demonstrate that the l-methionine γ-lyase MegL not only is a major H 2 S-producing enzyme of F. nucleatum but also significantly contributes to the antibiotic susceptibility and virulence of this organism.

Actinomyces oris plays an important role in oral biofilm development. Like many gram-positive bacteria, A. oris produces a sizable number of surface proteins that are anchored to bacterial peptidoglycan by a conserved transpeptidase named the housekeeping sortase SrtA; however, the biological role of many A. oris surface proteins in biofilm formation is largely unknown. Here, we report that the glycoprotein GspA-a genetic suppressor of srtA deletion lethality-not only promotes biofilm formation but also maintains cell membrane integrity under cation stress. In comparison to wild-type cells, under elevated concentrations of mono- and divalent cations the formation of mono- and multi-species biofilms by mutant cells devoid of gspA was significantly diminished, although planktonic growth of both cell types in the presence of cations was indistinguishable. Because gspA overexpression is lethal to cells lacking gspA and srtA, we performed a genetic screen to identify GspA determinants involving cell viability. DNA sequencing and biochemical characterizations of viable clones revealed that mutations of two critical cysteine residues and a serine residue severely affected GspA glycosylation and biofilm formation. Furthermore, mutant cells lacking gspA were markedly sensitive to sodium dodecyl sulfate, a detergent that solubilizes the cytoplasmic membranes, suggesting the cell envelope of the gspA mutant was altered. Consistent with this observation, the gspA mutant exhibited increased membrane permeability, independent of GspA glycosylation, compared to the wild-type strain. Altogether, the results support the notion that the cell wall-anchored glycoprotein GspA provides a defense mechanism against cation stress in biofilm development promoted by A. oris., (© 2022 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Most Actinobacteria encode a small transmembrane protein, whose gene lies immediately downstream of the housekeeping sortase coding for a transpeptidase that anchors many extracellular proteins to the Gram-positive bacterial cell wall. Here, we uncover the hitherto unknown function of this class of conserved proteins, which we name SafA, as a topological modulator of sortase in the oral Actinobacterium Actinomyces oris . Genetic deletion of safA induces cleavage and excretion of the otherwise predominantly membrane-bound SrtA in wild-type cells. Strikingly, the safA mutant, although viable, exhibits severe abnormalities in cell morphology, pilus assembly, surface protein localization, and polymicrobial interactions-the phenotypes that are mirrored by srtA depletion. The pleiotropic defect of the safA mutant is rescued by ectopic expression of safA from not only A. oris , but also Corynebacterium diphtheriae or Corynebacterium matruchotii . Importantly, the SrtA N terminus harbors a tripartite-domain feature typical of a bacterial signal peptide, including a cleavage motif AXA, mutations in which prevent SrtA cleavage mediated by the signal peptidase LepB2. Bacterial two-hybrid analysis demonstrates that SafA and SrtA directly interact. This interaction involves a conserved motif FPW within the exoplasmic face of SafA, since mutations of this motif abrogate SafA-SrtA interaction and induce SrtA cleavage and excretion as observed in the safA mutant. Evidently, SafA is a membrane-imbedded antagonist of signal peptidase that safeguards and maintains membrane homeostasis of the housekeeping sortase SrtA, a central player of cell surface assembly.

Fusobacterium nucleatum, an anaerobic Gram-negative bacterium frequently found in the human oral cavity and some extra-oral sites, is implicated in several important diseases: periodontitis, adverse pregnancy outcomes, and colorectal cancer. To date, how this obligate anaerobe copes with oxidative stress and host immunity within multiple human tissues remains unknown. Here, we uncovered a critical role in this process of a multigene locus encoding a single, fused methionine sulfoxide reductase (MsrAB), a two-component signal transduction system (ModRS), and thioredoxin (Trx)- and cytochrome c (CcdA)-like proteins, which are induced when fusobacterial cells are exposed to hydrogen peroxide. Comparative transcriptome analysis revealed that the response regulator ModR regulates a large regulon that includes trx , ccdA , and many metabolic genes. Significantly, specific mutants of the msrAB locus, including msrAB , are sensitive to reactive oxygen species and defective in adherence/invasion of colorectal epithelial cells. Strikingly, the msrAB mutant is also defective in survival in macrophages, and it is severely attenuated in virulence in a mouse model of preterm birth, consistent with its failure to spread to the amniotic fluid and colonize the placenta. Clearly, the MsrAB system regulated by the two-component system ModRS represents a major oxidative stress defense pathway that protects fusobacteria against oxidative damage in immune cells and confers virulence by enabling attachment and invasion of multiple target tissues. IMPORTANCE F. nucleatum colonizes various human tissues, including oral cavity, placenta, and colon. How this obligate anaerobe withstands oxidative stress in host immune cells has not been described. We report here that F. nucleatum possesses a five-gene locus encoding a fused methionine sulfoxide reductase (MsrAB), a two-component signal transduction system (ModRS), and thioredoxin- and cytochrome c -like proteins. Regulated by ModRS, MsrAB is essential for resistance to reactive oxygen species, adherence/invasion of colorectal epithelial cells, and survival in macrophage. Unable to colonize placenta and spread to amniotic fluid, the msrAB mutant failed to induce preterm birth in a murine model.

Although the close positive relationship between well-being and exercise adherence has been confirmed by numerous studies, it is still unclear whether this relationship exists for children and adolescents, because previous research mainly focuses on adults. The present review systematically explored the relationship between well-being ranging from individual to social aspects and exercise adherence based on extant studies. Seven studies including both quantitative and qualitative studies were analyzed. The results showed that well-being was not related to exercise adherence as strongly as expected. In some cases, well-being was even negatively associated with exercise adherence. Limited sample size, insensitive measurement of exercise adherence, gender, and mental and physical condition of children and adolescents might partially influence the relationship between well-being and exercise studies. However, the studies at hand are still in their infancy. More studies on the relationship between well-being and exercise adherence are needed for children and adolescents, especially in non-western countries., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Chen and Wu.)

The Porphyromonas gingivalis type IX secretion system (T9SS) promotes periodontal disease by secreting gingipains and other virulence factors. By in situ cryoelectron tomography, we report that the P. gingivalis T9SS consists of 18 PorM dimers arranged as a large, caged ring in the periplasm. Near the outer membrane, PorM dimers interact with a PorKN ring complex of ∼52 nm in diameter. PorMKN translocation complexes of a given T9SS adopt distinct conformations energized by the proton motive force, suggestive of different activation states. At the inner membrane, PorM associates with a cytoplasmic complex that exhibits 12-fold symmetry and requires both PorM and PorL for assembly. Activated motors deliver substrates across the outer membrane via one of eight Sov translocons arranged in a ring. The T9SSs are unique among known secretion systems in bacteria and eukaryotes in their assembly as supramolecular machines composed of apparently independently functioning translocation motors and export pores.

In previous word reading studies, lexicality has been used as a variable to examine the impacts of word form and meaning information on the many stages of word recognition process. Yet the neural dynamics associated with lexicality effect of various information processing for Chinese visual word recognition has not been well elucidated. In this study, Chinese native speakers were instructed to read Chinese disyllabic compound words, morphological legal (pseudo-words) and illegal non-words with their brain potentials recorded. Event-related potentials (ERP) results showed that N200 was related to Chinese orthographic processing, where three lexical conditions elicited comparable patterns. A semantic discrimination was found for N400 between pseudo-words/non-words and real words, which is in favor of the lexical view of the N400 effect. Further, a later ERP component P600 exhibited the difference between the non-words and pseudo-words, reflecting a re-analysis of word meaning or grammatical operation on Chinese morphological legality. Therefore, we argue that Chinese morphological information might have an independent representation (the P600 effect) in mental lexicon., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest., (© The Author(s), under exclusive licence to Springer Nature B.V. 2021.)

The present event-related potential (ERP) study explored whether masked emotion-laden words could facilitate the processing of both emotion-label words and emotion-laden words in a valence judgment task. The results revealed that emotion-laden words as primes failed to influence target emotion-label word processing, whereas emotion-laden words facilitated target emotion-laden words in the congruent condition. Specifically, decreased late positivity complex (LPC) was elicited by emotion-laden words primed by emotion-laden words of the same valence than those primed by emotion-laden words of different valence. Nevertheless, no difference was observed for emotion-label words as targets. These findings supported the mediated account that claimed emotion-laden words engendered emotion via the mediation of emotion-label words and hypothesized that emotion-laden words could not prime emotion-label words in the masked priming paradigm. Moreover, this study provided additional evidence showing the distinction between emotion-laden words and emotion-label words., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Wu, Zhang and Yuan.)

In Caenorhabditis elegans, ROS generated in response to intestinal infection induces SKN-1, a protective transcription factor homologous to nuclear factor erythroid 2-related factor 1 or 2 (NRF1/2) in mammals. Many factors regulate SKN-1, including the p38 mitogen-activated protein kinase (MAPK) cascade that activates SKN-1 by phosphorylation. In this work, another positive regulator of SKN-1 is identified: NIPI-3, a Tribbles pseudokinase. NIPI-3 has been reported to protect against intestinal infection by negatively regulating the CCAT enhancer binding protein (C/EBP) bZIP transcription factor CEBP-1. Here we demonstrate that CEBP-1 positively regulates the vhp-1 transcript, which encodes a phosphatase that dephosphorylates the p38 MAPK called PMK-1. The increased levels of VHP-1 caused by CEBP-1 transcriptional enhancement result in less PMK-1 phosphorylation, affecting SKN-1 activity and intestinal resistance to the pathogen. The data support a model in which NIPI-3's negative regulation of CEBP-1 decreases VHP-1 phosphatase activity, allowing increased stimulation of SKN-1 activity by the p38 MAPK phosphorylation cascade in the intestine., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)

A gram-negative colonizer of the oral cavity, Fusobacterium nucleatum not only interacts with many pathogens in the oral microbiome but also has the ability to spread to extraoral sites including placenta and amniotic fluid, promoting preterm birth. To date, however, the molecular mechanism of interspecies interactions-termed coaggregation-by F. nucleatum and how coaggregation affects bacterial virulence remain poorly defined. Here, we employed genome-wide transposon mutagenesis to uncover fusobacterial coaggregation factors, revealing the intertwined function of a two-component signal transduction system (TCS), named CarRS, and a lysine metabolic pathway in regulating the critical coaggregation factor RadD. Transcriptome analysis shows that CarR modulates a large regulon including radD and lysine metabolic genes, such as kamA and kamD , the expression of which are highly up-regulated in the Δ carR mutant. Significantly, the native culture medium of Δ kamA or Δ kamD mutants builds up abundant amounts of free lysine, which blocks fusobacterial coaggregation with streptococci. Our demonstration that lysine-conjugated beads trap RadD from the membrane lysates suggests that lysine utilizes RadD as its receptor to act as a metabolic inhibitor of coaggregation. Lastly, using a mouse model of preterm birth, we show that fusobacterial virulence is significantly attenuated with the Δ kamA and Δ carR mutants, in contrast to the enhanced virulence phenotype observed upon diminishing RadD (Δ radD or Δ carS mutant). Evidently, F. nucleatum employs the TCS CarRS and environmental lysine to modulate RadD-mediated interspecies interaction, virulence, and nutrient acquisition to thrive in the adverse environment of oral biofilms and extraoral sites., Competing Interests: The authors declare no competing interest.

In order to explore the affective priming effect of emotion-label words and emotion-laden words, the current study used unmasked (Experiment 1) and masked (Experiment 2) priming paradigm by including emotion-label words (e.g., sadness, anger) and emotion-laden words (e.g., death, gift) as primes and examined how the two kinds of words acted upon the processing of the target words (all emotion-laden words). Participants were instructed to decide the valence of target words, and their electroencephalogram was recorded at the same time. The behavioral and event-related potential (ERP) results showed that positive words produced a priming effect whereas negative words inhibited target word processing (Experiment 1). In Experiment 2, the inhibition effect of negative emotion-label words on emotion word recognition was found in both behavioral and ERP results, suggesting that modulation of emotion word type on emotion word processing could be observed even in the masked priming paradigm. The two experiments further supported the necessity of defining emotion words under an emotion word type perspective. The implications of the findings are proffered. Specifically, a clear understanding of emotion-label words and emotion-laden words can improve the effectiveness of emotional communications in clinical settings. Theoretically, the emotion word type perspective awaits further explorations and is still at its infancy.

In the present study, we examined modulations of the second language (L2) positive emotion-label words, positive emotion-laden words, and neutral words on conflict processing in a flanker task. Twenty Chinese-English bilinguals were instructed to decide the color of the central words that were vertically surrounded by the same words with the same or different color. During the task, their cortical activation was recorded. The result showed that L2 positive emotion-laden words elicited different brain activations from emotion-label words and neutral words at both early and late stages. Differential modulations on conflict processing between positive emotion-label words and positive emotion-laden words in the L2 existed even after approach-motivation intensity was controlled. These results suggest emotion word type affects conflict processing, even in L2.

Assembly of pili on the gram-positive bacterial cell wall involves 2 conserved transpeptidase enzymes named sortases: One for polymerization of pilin subunits and another for anchoring pili to peptidoglycan. How this machine controls pilus length and whether pilus length is critical for cell-to-cell interactions remain unknown. We report here in Actinomyces oris , a key colonizer in the development of oral biofilms, that genetic disruption of its housekeeping sortase SrtA generates exceedingly long pili, catalyzed by its pilus-specific sortase SrtC2 that possesses both pilus polymerization and cell wall anchoring functions. Remarkably, the srtA- deficient mutant fails to mediate interspecies interactions, or coaggregation, even though the coaggregation factor CafA is present at the pilus tip. Increasing ectopic expression of srtA in the mutant progressively shortens pilus length and restores coaggregation accordingly, while elevated levels of shaft pilins and SrtC2 produce long pili and block coaggregation by SrtA + bacteria. With structural studies, we uncovered 2 key structural elements in SrtA that partake in recognition of pilin substrates and regulate pilus length by inducing the capture and transfer of pilus polymers to the cell wall. Evidently, coaggregation requires proper positioning of the tip adhesin CafA via modulation of pilus length by the housekeeping sortase SrtA., Competing Interests: The authors declare no conflict of interest.

Despite recent increased attention to emotion conflict, little is known about whether emotion-label words (e.g., sadness, happiness) and emotion-laden words (e.g., death, birthday) function similarly in emotion conflict (i.e., a conflict between the target and distractor in emotion involvement), because the majority of the previous studies implicitly mixed the two. The present study aimed to compare emotion-label words and emotion-laden words in emotion conflict using a flanker task. Specifically, participants (N = 21) were asked to judge the valence of the target words that were vertically surrounded by the words with same (congruent) or different (incongruent) valence as being negative or positive. The behavioral results suggested that negative emotion-laden words were processed faster and more accurately than negative emotion-label words. ERP data further showed that negative emotion-label words elicited larger N200 than negative emotion-laden words on the left hemisphere, while such a difference was found for positive words on the right hemisphere. Moreover, emotion-laden words elicited smaller N200 in the incongruent condition than in the congruent condition, whereas no such a distinction was observed for emotion-label words. The findings suggest different cognitive and neural correlates of emotion-label words and emotion-laden words in emotion conflict.

The widely conserved LytR-CpsA-Psr (LCP) family of enzymes in Gram-positive bacteria is known to attach glycopolymers, including wall teichoic acid, to the cell envelope. However, it is undetermined if these enzymes are capable of catalyzing glycan attachment to surface proteins. In the actinobacterium Actinomyces oris , an LCP homolog here named LcpA is genetically linked to GspA, a glycoprotein that is covalently attached to the bacterial peptidoglycan by the housekeeping sortase SrtA. Here we show by X-ray crystallography that LcpA adopts an α-β-α structural fold, akin to the conserved LCP domain, which harbors characteristic catalytic arginine residues. Consistently, alanine substitution for these residues, R149 and R266, abrogates GspA glycosylation, leading to accumulation of an intermediate form termed GspA LMM , which is also observed in the lcpA mutant. Unlike other LCP proteins characterized to date, LcpA contains a stabilizing disulfide bond, mutations of which severely affect LcpA stability. In line with the established role of disulfide bond formation in oxidative protein folding in A. oris , deletion of vkor , coding for the thiol-disulfide oxidoreductase VKOR, also significantly reduces LcpA stability. Biochemical studies demonstrated that the recombinant LcpA enzyme possesses pyrophosphatase activity, enabling hydrolysis of diphosphate bonds. Furthermore, this recombinant enzyme, which weakly interacts with GspA in solution, catalyzes phosphotransfer to GspA LMM Altogether, the findings support that A. oris LcpA is an archetypal LCP enzyme that glycosylates a cell wall-anchored protein, a process that may be conserved in Actinobacteria , given the conservation of LcpA and GspA in these high-GC-content organisms. IMPORTANCE In Gram-positive bacteria, the conserved LCP family enzymes studied to date are known to attach glycopolymers, including wall teichoic acid, to the cell envelope. It is unknown if these enzymes catalyze glycosylation of surface proteins. We show here in the actinobacterium Actinomyces oris by X-ray crystallography and biochemical analyses that A. oris LcpA is an LCP homolog, possessing pyrophosphatase and phosphotransferase activities known to belong to LCP enzymes that require conserved catalytic Arg residues, while harboring a unique disulfide bond critical for protein stability. Importantly, LcpA mediates glycosylation of the surface protein GspA via phosphotransferase activity. Our studies provide the first experimental evidence of an archetypal LCP enzyme that promotes glycosylation of a cell wall-anchored protein in Gram-positive bacteria., (Copyright © 2019 Siegel et al.)

Fusobacterium nucleatum is a key member of the human oral biofilm. It is also implicated in preterm birth and colorectal cancer. To facilitate basic studies of fusobacterial virulence, we describe here a versatile transposon mutagenesis procedure and a pilot screen for mutants defective in biofilm formation. Out of 10 independent biofilm-defective mutants isolated, the affected genes included the homologs of the Escherichia coli cell division proteins FtsX and EnvC, the electron transport protein RnfA, and four proteins with unknown functions. Next, a facile new gene deletion method demonstrated that nonpolar, in-frame deletion of ftsX or envC produces viable bacteria that are highly filamentous due to defective cell division. Transmission electron and cryo-electron microscopy revealed that the Δ ftsX and Δ envC mutant cells remain joined with apparent constriction, and scanning electron microscopy (EM) uncovered a smooth cell surface without the microfolds present in wild-type cells. FtsX and EnvC proteins interact with each other as well as a common set of interacting partners, many with unknown function. Last, biofilm development is altered when cell division is blocked by MinC overproduction; however, unlike the phenotypes of Δ ftsX and Δ envC mutants, a weakly adherent biofilm is formed, and the wild-type rugged cell surface is maintained. Therefore, FtsX and EnvC may perform novel functions in Fusobacterium cell biology. This is the first report of an unbiased approach to uncover genetic determinants of fusobacterial biofilm development. It points to an intriguing link among cytokinesis, cell surface dynamics, and biofilm formation, whose molecular underpinnings remain to be elucidated. IMPORTANCE Little is known about the virulence mechanisms and associated factors in F. nucleatum , due mainly to the lack of convenient genetic tools for this organism. We employed two efficient genetic strategies to identify F. nucleatum biofilm-defective mutants, revealing FtsX and EnvC among seven biofilm-associated factors. Electron microscopy established cell division defects of the Δ ftsX and Δ envC mutants, accompanied with a smooth cell surface, unlike the microfold, rugged appearance of wild-type bacteria. Proteomic studies demonstrated that FtsX and EnvC interact with each other as well as a set of common and unique interacting proteins, many with unknown functions. Importantly, blocking cell division by MinC overproduction led to formation of a weakly adherent biofilm, without alteration of the wild-type cell surface. Thus, this work links cell division and surface dynamics to biofilm development and lays a foundation for future genetic and biochemical investigations of basic cellular processes in this clinically significant pathogen., (Copyright © 2018 Wu et al.)

Sortase is a cysteine-transpeptidase that anchors LPXTG-containing proteins on the Gram-positive bacterial cell wall. Previously, sortase was considered to be an important factor for bacterial pathogenesis and fitness, but not cell growth. However, the Actinomyces oris sortase is essential for cell viability, due to its coupling to a glycosylation pathway. In this chapter, we describe the methods to generate conditional srtA deletion mutants and identify srtA suppressors by Tn5 transposon mutagenesis. We also provide procedures for analyzing cell morphology of this mutant by thin-section electron microscopy. These techniques can be applied for analyses of other essential genes in A. oris.