An Autoinhibited Dimeric Form of BAX Regulates the BAX Activation Pathway.

Pro-apoptotic BAX is a cell fate regulator playing an important role in cellular homeostasis and pathological cell death. BAX is predominantly localized in the cytosol, where it has a quiescent monomer conformation. Following a pro-apoptotic trigger, cytosolic BAX is activated and translocates to the mitochondria to initiate mitochondrial dysfunction and apoptosis. Here, cellular, biochemical, and structural data unexpectedly demonstrate that cytosolic BAX also has an inactive dimer conformation that regulates its activation. The full-length crystal structure of the inactive BAX dimer revealed an asymmetric interaction consistent with inhibition of the N-terminal conformational change of one protomer and the displacement of the C-terminal helix α9 of the second protomer. This autoinhibited BAX dimer dissociates to BAX monomers before BAX can be activated. Our data support a model whereby the degree of apoptosis induction is regulated by the conformation of cytosolic BAX and identify an unprecedented mechanism of cytosolic BAX inhibition.

The effectiveness of a psychoeducational intervention on health outcomes of patients undergoing haemodialysis: A randomized controlled trial.

AIM: The aim of this work is to examine the effectiveness of a psychoeducational intervention on self-efficacy (primary outcome), anxiety, depression, treatment adherence, and health-related quality of life (HRQoL) of patients undergoing haemodialysis. METHODS: A two-group randomized controlled trial of 124 patients (65 and 59 patients in the intervention and control groups, respectively) recruited from a tertiary hospital in Singapore was conducted. Data were collected from January 2015 to June 2016. Outcomes were measured at baseline and 1, 3, and 6 months after the intervention. General linear model was used to analyse data. RESULTS: Our findings showed significant group effect on HRQoL (effects of kidney disease on daily life; p = 0.041), time effect on all outcomes (p < 0.05; except for treatment adherence behaviours and HRQoL [burden of kidney disease]), and group * time interaction effect on anxiety (p = 0.040) and depression (p = 0.003), with the intervention group reporting better outcomes. CONCLUSIONS: The positive effects of our intervention on patients' self-efficacy, psychological well-being, treatment adherence attitudes, and HRQoL implied its potential use in dialysis/renal centres to improve patients' self-care and health outcomes.

Experiences and Needs of Families Caring for Children and Adolescents With Chronic Kidney Disease: A meta-Synthesis.

PROBLEM: Families struggle to care for children and adolescents with chronic kidney disease (CKD). They face extensive burden of care and altered family dynamics. ELIGIBILITY CRITERIA: A meta-synthesis review was conducted to explore the experiences and needs of families caring for children and adolescents with CKD using seven electronic databases (CINAHL, EMBASE, ProQuest Dissertations and Theses, PsycINFO, PubMed, Scopus, and Web of Science). The inclusion criteria are (1) qualitative English studies from January 2010 to December 2020 that (2) report personal experiences or needs of (3) family members caring for children and adolescents aged 19 years and below who have been diagnosed with CKD of any stage (4) across all settings. Quality appraisal was done using the Critical Appraisal Skill Program checklist. Data was synthesised using Sandelowski & Barroso's (2007) method. SAMPLE: 2,236 records were identified and 13 eligible studies were included. Family members involved mothers (n = 190), fathers (n = 83), siblings (n = 5), and grandparents (n = 2). RESULTS: Three themes emerged: (1) demands of caregiving, (2) support systems, and (3) defining and making sense of new reality. CONCLUSIONS: Family caregivers experience overwhelming demands of caregiving and unmet support needs to cope. Appropriate interventions are needed to alleviate their burden. IMPLICATIONS: Knowledge of learning and support needs of families caring for children with CKD may shape nursing education and practice to cultivate more effective communication for better psychosocial family support.

TRIM37 Promotes Pancreatic Cancer Progression through Modulation of Cell Growth, Migration, Invasion, and Tumor Immune Microenvironment.

TRIM37 dysregulation has been observed in several cancer types, implicating its possible role in tumorigenesis. However, the role of TRIM37 in pancreatic cancer progression remains unclear. In the present study, we observed that TRIM37 knockdown resulted in reduced proliferation, clonogenicity, migration, and invasion ability of pancreatic cancer cells. Furthermore, an in vivo study using an orthotopic syngeneic animal model further confirmed that reduced expression of TRIM37 in cancer cells suppressed tumor growth in vivo. Moreover, in mice bearing TRIM37 knockdown pancreatic cancer cells, the proportion of CD11b+F4/80+MHCIIlow immunosuppressive macrophages was significantly reduced in tumor milieu, which might be due to the regulatory role of TRIM37 in cytokine production by pancreatic cancer cells. Collectively, these findings suggest a key role of TRIM37 in promoting pancreatic cancer progression.

Advantages and challenges of fostering cognitive integration through virtual collaborative learning: a qualitative study.

BACKGROUND: The drastic shift from face-to-face classes to online learning due to the COVID-19 pandemic has enabled educators to ensure the continuity of learning for health professions students in higher education. Collaborative learning, a pedagogy used to facilitate knowledge integration by helping students translate theory from basic sciences to clinical application and practice, has thus been transformed from a face-to-face to a virtual strategy to achieve the learning objectives of a multi-disciplinary and integrated module. OBJECTIVES: This study aimed to describe and evaluate, through focus group discussions, a virtual collaborative learning activity implemented to assist first year undergraduate nursing students to develop cognitive integration in a module consisting of pathophysiology, pharmacology, and nursing practice. METHODS: Fourteen first year undergraduate students and four faculty involved in facilitating the virtual collaboration participated in the study. Focus group discussions were conducted to elicit the perceptions of students and staff on the virtual collaborative learning session conducted at the end of the semester. RESULTS: Three themes were generated from the thematic analysis of the students' focus group scripts. These were: (1) achieving engagement and interaction, (2) supporting the collaborative process, and (3) considering practical nuances. The three themes were further subdivided into subthemes to highlight noteworthy elements captured during focus group discussions. Three themes also emerged from the focus group discussion scripts of faculty participants: (1) learning to effectively manage, (2) facing engagement constraints, and (3) achieving integration. These themes were further sectioned into salient subthemes. CONCLUSION: The virtual collaborative learning pedagogy is valuable in fostering cognitive integration. However, meticulous planning considering various variables prior to implementation is needed. With better planning directed at addressing the learners' needs and the faculty's capabilities and readiness for online learning pedagogies, and with a strong institutional support to help mitigate the identified constraints of virtual collaborative learning, students and faculty will benefit.

miR-1975 serves as an indicator of clinical severity upon influenza infection.

Emerging evidence highlights the role of non-coding small RNAs in host-influenza interaction. We have identified a Y RNA-derived small RNA, miR-1975, which is upregulated upon influenza A virus infection in A549 cells. The aim of this study is to investigate whether miR-1975 serves as an indicator of clinical severity upon influenza infection. We investigate the abundance of miR-1975 in sera from clinical patients and its correlation with hypoxemia status. We quantified its amounts in sera from influenza virus-infected patients and healthy volunteers by means of stem-loop RT-PCR. Median values of miR-1975 were significantly higher in influenza virus-infected patients, especially in hypoxemic patients. miR-1975 levels at the acute stage of the disease were highly correlated with the fraction of inspired oxygen used by the patients and total ventilator days. Receiver operator characteristic curve analysis revealed that miR-1975 levels in combination with days of fever before presenting to hospital had significant predictive value for hypoxemia and respiratory failure for patients infected with influenza virus. Our results reveal that circulating miR-1975 has great potential to serve as a biomarker for predicting prognosis in patients infected with influenza virus.

Effects of psychosocial interventions on psychological outcomes among caregivers of advanced cancer patients: a systematic review and meta-analysis.

PURPOSE: This systematic review aimed to synthesize the effectiveness of psychosocial interventions on caregivers of advanced cancer patients, in comparison with usual care, on caregivers' quality of life (QoL), anxiety, and depression symptoms. METHODS: Comprehensive searches for published and unpublished studies were performed using nine electronic databases, two trial registers, and reference lists of included studies. Two reviewers independently screened, appraised, and extracted data. The Cochrane risk of bias tool was used to appraise the methodological quality of included studies, while the Cochrane data extraction tool was used to elicit relevant information. Meta-analysis, narrative analysis, and sensitivity analysis were conducted to synthesize data. Standardized mean differences (SMD) represented effects of psychosocial interventions. RESULTS: Fifteen randomized controlled trials were included in this review. At post-intervention, findings revealed a significant small pooled effect size (SMD = 0.45) on QoL and significant moderate effect on depression (SMD = - 0.65). However, a small non-significant pooled effect size was observed on anxiety (SMD = - 0.24). At follow-up assessments, effect sizes of all outcomes were small and non-significant. Overall quality of evidence was rated very low for all outcomes and most studies had unclear or high risk of bias. Thus, results should be interpreted with caution. CONCLUSION: Psychosocial interventions were effective in improving QoL and depression among caregivers of persons with advanced cancer. However, future randomized control trials with lower risk of bias, larger sample size, detailed participant characteristics, and informative interventions are desirable.

Critical involvement of atypical chemokine receptor CXCR7 in allergic airway inflammation.

Trafficking and recruitment of immune cells to the site of inflammation with spatial and temporal synchronization is crucial for the development of allergic airway inflammation. Particularly, chemokines are known to be key players in these processes. Previous studies revealed that the CXCL12/CXCR4 axis plays an important role in regulating allergic airway inflammation. However, the role of CXCR7, a recently discovered second receptor for CXCL12, in regulating airway inflammation has not been explored. Initially, CXCR7 was considered as a decoy receptor; however, numerous subsequent studies revealed that engagement of CXCR7 triggered its own signalling or modulated CXCR4-mediated signalling. In the present study, we detected the expression of CXCR7 in airway epithelial cells. Use of a lentiviral delivery system to knock down the expression of CXCR7 in the lung of sensitized mice abrogated the cardinal features of asthma, indicating that CXCR7 plays a role in regulating allergic airway inflammation. The activation of mitogen-activated protein kinase and Akt signalling in response to CXCL12 in the mouse epithelial cell line MLE-12 was reduced when CXCR7 expression was knocked down. However, either knockdown or overexpression of CXCR7 in MLE-12 did not affect CXCL12-mediated calcium influx, indicating that CXCR7 does not modulate CXCR4-mediated signalling, and that it functions as a signalling receptor rather than a decoy receptor. Finally, we found that the expression of chemokine CCL2 is regulated by CXCR7/CXCL12-mediated signalling through ß-arrestin in airway epithelial cells. Hence, regulating the expression of CCL2 in airway epithelial cells may be one mechanism by which CXCR7 participates in regulating allergic airway inflammation.

Dietary and fluid restriction perceptions of patients undergoing haemodialysis: an exploratory study.

AIMS AND OBJECTIVES: To explore the perspectives of patients undergoing haemodialysis in Singapore on an imposed dietary and fluid restriction regime. BACKGROUND: Adherence to prescribed dietary and fluid restriction constructs the fundamental basis of self-care with improved morbidity and mortality. However, most patients have struggled to adhere in this aspect. Existing studies have presented limited understanding on the facilitators and barriers of dietary and fluid adherence among haemodialysis patients. DESIGN: An exploratory qualitative study. METHODS: A purposive sample of 14 patients undergoing haemodialysis was recruited from a renal unit of a tertiary hospital in Singapore. Data were collected through face-to-face individual interviews and subsequently analysed by thematic analysis. RESULTS: Four themes emerged: (1) Pessimism, (2) Existing struggles, (3) Perceived quality of support, and (4) Immensity of self-discipline. CONCLUSIONS: The imposed dietary and fluid restriction is a constant struggle and a cause of suffering among haemodialysis patients in Singapore. Nonetheless, they are generally submissive to their fluid restrictions for the sake of survival or to meet the expectations of their loved ones. The imposed dietary restrictions are generally neglected. RELEVANCE TO CLINICAL PRACTICE: The findings from this study can provide useful information in reviewing existing educational strategies, policies and nursing care. This is especially important because most patients exhibit high reliance on healthcare professionals.

The inducible nitric-oxide synthase (iNOS)/Src axis mediates Toll-like receptor 3 tyrosine 759 phosphorylation and enhances its signal transduction, leading to interferon-ß synthesis in macrophages.

Double-stranded RNA (dsRNA) induces phosphorylation of Toll-like receptor 3 (TLR3) at tyrosine 759 and subsequently triggers signaling pathways to promote interferon-ß (IFN-ß) production. In this study, we found that dsRNA stimulation induces biphasic TLR3 Tyr-759 phosphorylation in macrophages. In addition to the immediate TLR3 Tyr-759 phosphorylation, we identified a second wave of Tyr-759 phosphorylation accompanied by an increase of both Src and ifn-ß transcription in the later phase of dsRNA stimulation. Interestingly, Src phosphorylated TLR3 Tyr-759 in vitro and in vivo. However, knockdown of Src abolished the late phase of TLR3 Tyr-759 phosphorylation and decreased the nuclear accumulation of interferon regulatory factors 3 and 7 (IRF3 and -7) and IFN-ß production. Reintroduction of Src restored all of these molecular changes. Notably, via down-regulation of Src, dsRNA-elicited TLR3 Tyr-759 phosphorylation, the nuclear accumulation of IRF3/IRF7, and IFN-ß generation were inhibited in inducible nitric-oxide synthase (iNOS)-null macrophages. TLR3 knockdown destabilized Src and reduced the nuclear level of IRF3/IRF7 and IFN-ß production in macrophages exposed to LPS (a TLR4 ligand known to induce Src and IFN-ß expression). Ectopic expression of wild type TLR3, but not its 759-phenylalanine mutant, restored Src activity and ifn-ß transcription. Taken together, these results suggested an essential role of the iNOS/Src/TLR3 axis in IFN-ß production in macrophages.

The iNOS/Src/FAK axis contributes to lithium chloride-mediated macrophage migration.

Lithium chloride (LiCl) has long been used as a mood stabilizer for bipolar mood depression patients. However, its biological effects on immune cells are unclear yet. In this study, we observed that upon LiCl stimulation, the motility and the content of total protein tyrosine phosphorylation in RAW264.7 macrophages and murine peritoneal macrophages (PEMs) were significantly increased. The inhibition of LiCl-induced macrophage migration by PP2 (an inhibitor for Src family kinases (SFKs)) suggested the involvement of SFKs in this process. Interestingly, LiCl induced NF-kB activation, and while Src was greatly induced, the expression of its myeloid relatives (i.e. Lyn, Fgr, Hck) was almost unaltered in RAW264.7 cells. Knockdown of Src suppressed LiCl-elicited movement and the level of FAK Pi-Tyr861, which could be reversed by siRNA-resistant Src. Consistent with Src and migration increment was iNOS-dependent in macrophages, markedly reduced Src expression, activity and cell migration were observed in iNOS-null PEMs treated with LiCl. Moreover, FAK knockdown suppressed LiCl-stimulated macrophage motility, suggesting the involvement of FAK in this process. Remarkably, similar increment of iNOS, Src, FAK Pi-Tyr861 and migration ability could also be detected in RAW264.7 treated with other GSK3ß inhibitors (i.e. SB216763 and Kenpaullone). These results corroborate that through inhibition of GSK3ß, the iNOS/Src/FAK axis occupies a critical role in macrophage locomotion in response to LiCl.

Postpartum social support of women in Singapore: A pilot study.

This pilot study aimed to identify the levels of support expected and received by postpartum women in Singapore. Another objective was to compare the different subdomains of social support. A descriptive design was adopted using a self-administered questionnaire comprising demographics and the Postpartum Support Questionnaire. The study took place at a subsidized obstetrics and gynaecology clinic at a restructured hospital in Singapore from the end of December 2009 to the end of February 2010. The 25 participants were 6 to 8 weeks postpartum and had delivered healthy term infants. They were recruited via convenience sampling. Data were analysed using descriptive statistics and inferential statistics. The overall support needs of the participants were met. Only their informational support needs were unmet. Assessing the needs of postpartum women and teaching them how to convey and manage expectations are recommended initiatives for health-care practitioners to consider.

The pathological effects of CCR2+ inflammatory monocytes are amplified by an IFNAR1-triggered chemokine feedback loop in highly pathogenic influenza infection.

BACKGROUND: Highly pathogenic influenza viruses cause high levels of morbidity, including excessive infiltration of leukocytes into the lungs, high viral loads and a cytokine storm. However, the details of how these pathological features unfold in severe influenza infections remain unclear. Accumulation of Gr1 + CD11b + myeloid cells has been observed in highly pathogenic influenza infections but it is not clear how and why they accumulate in the severely inflamed lung. In this study, we selected this cell population as a target to investigate the extreme inflammatory response during severe influenza infection. RESULTS: We established H1N1 IAV-infected mouse models using three viruses of varying pathogenicity and noted the accumulation of a defined Gr1 + CD11b + myeloid population correlating with the pathogenicity. Herein, we reported that CCR2+ inflammatory monocytes are the major cell compartments in this population. Of note, impaired clearance of the high pathogenicity virus prolonged IFN expression, leading to CCR2+ inflammatory monocytes amplifying their own recruitment via an interferon-α/ß receptor 1 (IFNAR1)-triggered chemokine loop. Blockage of IFNAR1-triggered signaling or inhibition of viral replication by Oseltamivir significantly suppresses the expression of CCR2 ligands and reduced the influx of CCR2+ inflammatory monocytes. Furthermore, trafficking of CCR2+ inflammatory monocytes from the bone marrow to the lung was evidenced by a CCR2-dependent chemotaxis. Importantly, leukocyte infiltration, cytokine storm and expression of iNOS were significantly reduced in CCR2-/- mice lacking infiltrating CCR2+ inflammatory monocytes, enhancing the survival of the infected mice. CONCLUSIONS: Our results indicated that uncontrolled viral replication leads to excessive production of inflammatory innate immune responses by accumulating CCR2+ inflammatory monocytes, which contribute to the fatal outcomes of high pathogenicity virus infections.

Efficacy of relaxation intervention on pain, self-efficacy, and stress-related variables in patients following total knee replacement surgery.

Following total knee replacement (TKR) surgery, patients frequently experience intense levels of pain, stress, and anxiety that may reduce their self-efficacy and thus affect their postoperative recovery. Relaxation intervention is beneficial to help patients manage physical pain and emotional tension. However, evidence for the efficacy of relaxation intervention on patients following TKR is still inconclusive. This study aimed to investigate whether a relaxation intervention helped to reduce pain, stress, and anxiety, and whether it helped to increase perceived relaxation and self-efficacy in patients following TKR. A single-group, pretest-posttest quasi-experimental study was carried out at a tertiary hospital in Singapore. A convenience sampling of 18 participants was recruited. Patients received three-session, individual-based relaxation interventions comprised of breathing exercises, muscle relaxation, and guided imagery. Data were collected by self-reported questionnaires and physiologic measures and were analyzed using descriptive statistics, paired t test, and repeated measure analysis of variance. Intent-to-treat analyses were used to deal with missing data. Following the intervention, participants reported significantly lower pain, stress, and anxiety and greater perceived relaxation and self-efficacy. Findings from this study contribute to both nursing science and clinical practice. The relaxation intervention can be offered as part of standard care for patients following TKR in hospitals.

Decreased PLUNC expression in nasal polyps is associated with multibacterial colonization in chronic rhinosinusitis patients.

PLUNC (palate, lung, and nasal epithelium clone) is an epithelium-secreted protein that plays a crucial role in the host's defense against bacterial infection. The function of PLUNC in the sinus remains poorly understood. To examine whether the expression levels of PLUNC could serve as a predictive outcome biomarker for patients with CRSwNP and bacterial colonization, we investigated the association of PLUNC expression levels with bacterial colonization in the sinuses. A total of 174 patients who underwent sinus surgery for chronic rhinosinusitis with nasal polyps (CRSwNP) were enrolled in this study. The tissue samples obtained from patients were examined using preoperative sinus computed tomography (CT) scans, postoperative bacterial cultures, and nasal polyp examinations. PLUNC mRNA and protein expression were quantified using RT-PCR and immunohistochemistry. We identified that decreased PLUNC expression is associated with multibacterial colonization (P = 0.0001), specifically those mediated by Staphyloccocus aureus (P = 0.037) and Pseudomonas aeruginosa (P = 0.002). The patients who required repeated sinus surgeries for recurrent or persistent sinusitis also presented much lower PLUNC expression than those who did not require repeated sinus surgery (P = 0.001). However, gender, age, and CT scores were not associated with PLUNC expression. These results suggest that reduced PLUNC expression is associated with bacterial colonization as well as treatment outcome in CRSwNP patients. Investigation of the association between PLUNC expressions and chronic rhinosinusitis may lead to the development of a novel biomarker for treatment outcome in CRSwNP patients.

Eps8 protein facilitates phagocytosis by increasing TLR4-MyD88 protein interaction in lipopolysaccharide-stimulated macrophages.

Toll-like receptors (TLRs) are crucial in macrophage phagocytosis, which is pivotal in host innate immune response. However, the detailed mechanism is not fully defined. Here, we demonstrated that the induction of Src and Eps8 in LPS-treated macrophages was TLR4- and MyD88-dependent, and their attenuation reduced LPS-promoted phagocytosis. Confocal microscopy indicated the colocalization of Eps8 and TLR4 in the cytosol and at the phagosome. Consistently, both Eps8 and TLR4 were present in the same immunocomplex regardless of LPS stimulation. Inhibition of this complex formation by eps8 siRNA or overexpression of pleckstrin homology domain-truncated Eps8 (i.e. 261-p97(Eps8)) decreased LPS-induced TLR4-MyD88 interaction and the following activation of Src, focal adhesion kinase, and p38 MAPK. Importantly, attenuation of Eps8 impaired the bacterium-killing ability of macrophages. Thus, Eps8 is a key regulator of the LPS-stimulated TLR4-MyD88 interaction and contributes to macrophage phagocytosis.

Novel c-di-GMP recognition modes of the mouse innate immune adaptor protein STING.

The mammalian ER protein STING (stimulator of interferon genes; also known as MITA, ERIS, MPYS or TMEM173) is an adaptor protein that links the detection of cytosolic dsDNA to the activation of TANK-binding kinase 1 (TBK1) and its downstream transcription factor interferon regulatory factor 3 (IFN3). Recently, STING itself has been found to be the direct receptor of bacterial c-di-GMP, and crystal structures of several human STING C-terminal domain (STING-CTD) dimers in the apo form or in complex with c-di-GMP have been published. Here, a novel set of structures of mouse STING-CTD (mSTING(137-344)) in apo and complex forms determined from crystals obtained under different crystallization conditions are reported. These novel closed-form structures exhibited considerable differences from previously reported open-form human STING-CTD structures. The novel mSTING structures feature extensive interactions between the two monomers, a unique asymmetric c-di-GMP molecule with one guanine base in an unusual syn conformation that is well accommodated in the dimeric interface with many direct specific interactions and two unexpected equivalent secondary peripheral c-di-GMP binding sites. Replacement of the amino acids crucial for specific c-di-GMP binding in mSTING significantly changes the ITC titration profiles and reduces the IFN-ß reporter luciferase activity. Taken together, these results reveal a more stable c-di-GMP binding mode of STING proteins that could serve as a template for rational drug design to stimulate interferon production by mammalian cells.

A bovine whey protein extract can induce the generation of regulatory T cells and shows potential to alleviate asthma symptoms in a murine asthma model.

The number of people with asthma has dramatically increased over the past few decades and the cost of care is more than $11·3 billion per year. The use of steroids is the major treatment to control asthma symptoms, but the side effects are often devastating. Seeking new drugs or new strategies to reduce the dose of steroid taken has always been an important task. A bovine whey protein extract (WPE), which is enriched in transforming growth factor-ß (TGF-ß), has been demonstrated to have the potential for reducing symptoms associated with mild-to-moderate T-helper cell type 1-mediated psoriasis in human subjects. However, whether WPE also has potential for inhibiting T-helper cell type 2 (Th2)-mediated disease remains unclear. In the present study, using a murine asthma model, we found that sensitised mice fed WPE daily, before they were challenged, resulted in reducing airway inflammation, serum ovalbumin-specific IgE, Th2-related cytokine production and airway hyperresponsiveness. Increase in the regulatory T cell (Treg) population in vitro and in vivo was observed when treated with WPE. According to the results from the TGF-ß-blocking antibody study, we suggest that TGF-ß is the main component that endows WPE with the potential to reduce the generation of Treg. Thus, the present data suggest that WPE has the potential to alleviate the symptoms of asthma by inducing the generation of Treg. Therefore, regular administration of WPE might be potentially beneficial for patients with asthma.

The iNOS/Src/FAK axis is critical in Toll-like receptor-mediated cell motility in macrophages.

The Toll-like receptors (TLRs) play a pivotal role in innate immunity for the detection of highly conserved, pathogen-expressed molecules. Previously, we demonstrated that lipopolysaccharide (LPS, TLR4 ligand)-increased macrophage motility required the participation of Src and FAK, which was inducible nitric oxide synthase (iNOS)-dependent. To investigate whether this iNOS/Src/FAK pathway is a general mechanism for macrophages to mobilize in response to engagement of TLRs other than TLR4, peptidoglycan (PGN, TLR2 ligand), polyinosinic-polycytidylic acid (polyI:C, TLR3 ligand) and CpG-oligodeoxynucleotides (CpG, TLR9 ligand) were used to treat macrophages in this study. Like LPS stimulation, simultaneous increase of cell motility and Src (but not Fgr, Hck, and Lyn) was detected in RAW264.7, peritoneal macrophages, and bone marrow-derived macrophages exposed to PGN, polyI:C and CpG. Attenuation of Src suppressed PGN-, polyI:C-, and CpG-elicited movement and the level of FAK Pi-Tyr861, which could be reversed by the reintroduction of siRNA-resistant Src. Besides, knockdown of FAK reduced the mobility of macrophages stimulated with anyone of these TLR ligands. Remarkably, PGN-, polyI:C-, and CpG-induced Src expression, FAK Pi-Tyr861, and cell mobility were inhibited in macrophages devoid of iNOS, indicating the importance of iNOS. These findings corroborate that iNOS/Src/FAK axis occupies a central role in macrophage locomotion in response to engagement of TLRs.