Plasminogen activator urokinase interacts with the fusion protein and antagonizes the growth of Peste des petits ruminants virus.

Peste des petits ruminants is an acute and highly contagious disease caused by the Peste des petits ruminants virus (PPRV). Host proteins play a crucial role in viral replication. However, the effect of fusion (F) protein-interacting partners on PPRV infection is poorly understood. In this study, we found that the expression of goat plasminogen activator urokinase (PLAU) gradually decreased in a time- and dose-dependent manner in PPRV-infected goat alveolar macrophages (GAMs). Goat PLAU was subsequently identified using co-immunoprecipitation and confocal microscopy as an F protein binding partner. The overexpression of goat PLAU inhibited PPRV growth and replication, whereas silencing goat PLAU promoted viral growth and replication. Additionally, we confirmed that goat PLAU interacted with a virus-induced signaling adapter (VISA) to antagonize F-mediated VISA degradation, increasing the production of type I interferon. We also found that goat PLAU reduced the inhibition of PPRV replication in VISA-knockdown GAMs. Our results show that the host protein PLAU inhibits the growth and replication of PPRV by VISA-triggering RIG-I-like receptors and provides insight into the host protein that antagonizes PPRV immunosuppression.IMPORTANCEThe role of host proteins that interact with Peste des petits ruminants virus (PPRV) fusion (F) protein in PPRV replication is poorly understood. This study confirmed that goat plasminogen activator urokinase (PLAU) interacts with the PPRV F protein. We further discovered that goat PLAU inhibited PPRV replication by enhancing virus-induced signaling adapter (VISA) expression and reducing the ability of the F protein to degrade VISA. These findings offer insights into host resistance to viral invasion and suggest new strategies and directions for developing PPR vaccines.

Tankyrases inhibit innate antiviral response by PARylating VISA/MAVS and priming it for RNF146-mediated ubiquitination and degradation.

During viral infection, sensing of viral RNA by retinoic acid-inducible gene-I-like receptors (RLRs) initiates an antiviral innate immune response, which is mediated by the mitochondrial adaptor protein VISA (virus-induced signal adaptor; also known as mitochondrial antiviral signaling protein [MAVS]). VISA is regulated by various posttranslational modifications (PTMs), such as polyubiquitination, phosphorylation, O-linked ß-d-N-acetylglucosaminylation (O-GlcNAcylation), and monomethylation. However, whether other forms of PTMs regulate VISA-mediated innate immune signaling remains elusive. Here, we report that Poly(ADP-ribosyl)ation (PARylation) is a PTM of VISA, which attenuates innate immune response to RNA viruses. Using a biochemical purification approach, we identified tankyrase 1 (TNKS1) as a VISA-associated protein. Viral infection led to the induction of TNKS1 and its homolog TNKS2, which translocated from cytosol to mitochondria and interacted with VISA. TNKS1 and TNKS2 catalyze the PARylation of VISA at Glu137 residue, thereby priming it for K48-linked polyubiquitination by the E3 ligase Ring figure protein 146 (RNF146) and subsequent degradation. Consistently, TNKS1, TNKS2, or RNF146 deficiency increased the RNA virus-triggered induction of downstream effector genes and impaired the replication of the virus. Moreover, TNKS1- or TNKS2-deficient mice produced higher levels of type I interferons (IFNs) and proinflammatory cytokines after virus infection and markedly reduced virus loads in the brains and lungs. Together, our findings uncover an essential role of PARylation of VISA in virus-triggered innate immune signaling, which represents a mechanism to avoid excessive harmful immune response.

MAVS signaling shapes microglia responses to neurotropic virus infection.

Viral encephalitis is characterized by a series of immunological reactions that can control virus infection in the brain, but dysregulated responses may cause excessive inflammation and brain damage. Microglia are brain-resident myeloid cells that are specialized in surveilling the local CNS environment and in case of viral brain infection they contribute to the control of the infection and to restriction of viral dissemination. Here, we report that after exposure to neurotropic vesicular stomatitis virus (VSV), murine in vitro microglia cultures showed rapid upregulation of a broad range of pro-inflammatory and antiviral genes, which were stably expressed over the entire 8 h infection period. Additionally, a set of immunomodulatory genes was upregulated between 6 and 8 h post infection. In microglia cultures, the induction of several immune response pathways including cytokine responses was dependent on mitochondrial antiviral-signaling protein (MAVS). Consequently, in Mavs-deficient microglia the control of virus propagation failed as indicated by augmented virus titers and the accumulation of viral transcripts. Thus, in the analyzed in vitro system, MAVS signaling is critically required to achieve full microglia activation and to mediate profound antiviral effects. In Mavs-deficient mice, intranasal VSV instillation caused higher disease severity than in WT mice and virus dissemination was noticed beyond the olfactory bulb. Virus spread to inner regions of the olfactory bulb, i.e., the granular cell layer, correlated with the recruitment of highly inflammatory non-microglia myeloid cells into the olfactory bulb in Mavs-/- mice. Furthermore, increased cytokine levels were detected in the nasal cavity, the olfactory bulb and in other brain regions. Thus, microglial MAVS signaling is critically needed for virus sensing, full microglia activation, and for orchestration of protective immunity in the virus-infected CNS.

Comparative proteomic analysis of vancomycin-sensitive and vancomycin-intermediate resistant Staphylococcus aureus.

PURPOSE: The extensive use of vancomycin has led to the development of Staphylococcus aureus strains with varying degrees of resistance to vancomycin. The present study aimed to explore the molecular causes of vancomycin resistance by conducting a proteomics analysis of subcellular fractions isolated from vancomycin-intermediate resistant S. aureus (VISA) and vancomycin-sensitive S. aureus (VSSA) strains. METHODS: We conducted proteomics analysis of subcellular fractions isolated from 2 isogenic S. aureus strains: strain 11 (VSSA) and strain 11Y (VISA). We used an integrated quantitative proteomics approach assisted by bioinformatics analysis, and comprehensively investigated the proteome profile. Intensive bioinformatics analysis, including protein annotation, functional classification, functional enrichment, and functional enrichment-based cluster analysis, was used to annotate quantifiable targets. RESULTS: We identified 128 upregulated proteins and 21 downregulated proteins in strain 11Y as compared to strain 11. The largest group of differentially expressed proteins was composed of enzymatic proteins associated with metabolic and catalytic activity, which accounted for 32.1% and 50% of the total proteins, respectively. Some proteins were indispensable parts of the regulatory networks of S. aureus that were altered with vancomycin treatment, and these proteins were related to cell wall metabolism, cell adhesion, proteolysis, and pressure response. CONCLUSION: Our proteomics study revealed regulatory proteins associated with vancomycin resistance in S. aureus. Some of these proteins were involved in the regulation of cell metabolism and function, which provides potential targets for the development of strategies to manage vancomycin resistance in S. aureus.

Visa Opportunities for International Medical Graduates Applying for U.S. Academic Radiology Department Faculty Positions: A National Survey.

BACKGROUND. International medical graduates (IMGs) are a source of physicians who could help alleviate radiologist workforce shortages in the United States. However, IMGs may face barriers in obtaining appropriate visas (e.g., H-1B or O-1 visas) to allow faculty employment. OBJECTIVE. The purpose of this study was to assess the policies and experiences of U.S. academic radiology departments in offering visas to IMGs applying for faculty positions. METHODS. A web-based survey on policies and experiences in offering visas to IMG faculty candidates was distributed to chairs of U.S. radiology departments with a diagnostic radiology training program recognized by the National Resident Matching Program. Individual survey questions were optional. The initial survey and subsequent reminders were sent from October 7, 2022, through November 7, 2022. RESULTS. The survey response rate was 81% (143/177). A total of 24% (28/115), 38% (44/115), 17% (20/115), and 20% (23/115) of departments offered H-1B visas to IMG faculty frequently, sometimes, rarely, and never, respectively; 3% (3/113), 27% (31/113), 22% (25/113), and 48% (54/113) of departments offered O-1 visas frequently, sometimes, rarely, and never, respectively. However, 41% (46/113) and 5% (6/113) of departments had default policies of offering H-1B and O-1 visas for IMG faculty candidates, respectively. The most common reasons given for why departments did not offer visas included, for both H-1B and O-1 visas, the time-consuming process, lack of reliability of candidates' starting time, and the expense of the visa application; for O-1 visas, the reasons given also included lack of expertise. A total of 15% (16/108) of departments set their own visa policies, 75% (81/108) followed institutional policies, and 10% (11/108) followed policies set by other entities (e.g., state government). CONCLUSION. Although to at least some extent most U.S. academic radiology departments offer H-1B and O-1 visas for IMGs seeking faculty positions, use of such visas typically is not the departments' default policy. A variety of barriers contributed to visas not being offered. The departments' visa policies were primarily determined at the institutional level. CLINICAL IMPACT. The identified barriers faced by U.S. academic radiology departments in offering visas to IMG faculty candidates impact the role of IMGs in helping to address radiologist workforce shortages.

Diagnostic value of BHI-V4 for heterogeneous and vancomycin-intermediate Staphylococcus aureus isolates: a systematic review and meta-analysis.

BACKGROUND: Brain-heart infusion agar supplemented with 4 µg/mL of vancomycin (BHI-V4) was commonly used for the detection of heterogeneous (hVISA) and vancomycin-intermediate Staphylococcus aureus (VISA). However, its diagnostic value remains unclear. This study aims to compare the diagnostic accuracy of BHI-V4 with population analysis profiling with area under the curve (PAP-AUC) in hVISA/VISA. METHODS: The protocol of this study was registered in INPLASY (INPLASY2023120069). The PubMed and Cochrane Library databases were searched from inception to October 2023. Review Manager 5.4 was used for data visualization in the quality assessment, and STATA17.0 (MP) was used for statistical analysis. RESULTS: In total, eight publications including 2153 strains were incorporated into the meta-analysis. Significant heterogeneity was evident although a threshold effect was not detected across the eight studies. The summary receiver operating characteristic (SROC) was 0.77 (95% confidence interval [CI], 0.74-0.81). The pooled sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, diagnostic score and diagnostic odds ratio were 0.59 (95% CI: 0.46-0.71), 0.96 (95%CI: 0.83-0.99), 14.0 (95% CI, 3.4-57.1), 0.43 (95%CI, 0.32-0.57), 3.48(95%CI, 2.12-4.85) and 32.62 (95%CI, 8.31-128.36), respectively. CONCLUSION: Our study showed that BHI-V4 had moderate diagnostic accuracy for diagnosing hVISA/VISA. However, more high-quality studies are needed to assess the clinical utility of BHI-V4.

Tran slation , cultural adap tation an d psycho metric anal ysis of U rd u ver sion of Vic torian; institu te of sp or ts ass es sme nt: Achill es quest i onnaire fo r athletes with achilles tendinopathy.

OBJECTIVE: To translate, culturally adapt and psychometrically analyse the Urdu version of the Victorian Institute of Spor t Assessment-Achilles questionnaire. METHODS: The cross-sec tional study was conducted at the Pak istan Sports Board, Lahore, Pakistan, from June 17, 2021, to February 15, 2022, and comprised patients with Achilles tendinopathy in group A and healthy controls in group B. Beaton's guidelines for cultural adaptation and validation for self-repor ted measures were followed to translate and validate the Victorian Institute of Sport Assessment-Achilles questionnaire in Urdu language. Data was analysed using SPSS 23. RESULTS: Of the 180 subjects with mean age 28.06±5.95 years, 125(69.6%) were males. There were 130(72.2%) patients in group A and 50(27.8%) controls in group B. The overall mean score of the Victorian Institute of Sport Assessment- Achille s questionnaire was 55.99±25.43; group A 41.14±9.54 and group B 94.60±4.22. The Urdu version exhibited excellent internal consistency with Cronbach's alpha values 0.95, and excellent test-retest reliability (p<0.001). Absolute reliability was expressed by standard error of measurement 5.317 and minimal detectable change (6.38). Convergent validity demonstrated strong correlation with the physical domain (r=0.81) of the Urdu version of the World Health Organisation Quality of Life Brief Version. CONCLUSIONS: The Victorian Institute of Sport Assessment-Achilles questionnaire could be utilised for assessing severity of Achilles tendinopathy among Urdu-speaking population for clinical as well as research purposes.

Validation and reliability of the Simple Achille Value (SAV) in Achilles tendon disorders.

BACKGROUND: Achilles tendon (AT) disorders significantly impact patient life, necessitating accurate assessment tools. Current methods are often complex and time-consuming. This study aims to validate the Simple Ankle Value (SAV) for AT disorders. METHODS: A multicenter study was conducted involving 101 participants, including a surgical, a conservative, and a control group. Participants completed the SAV, VISA-A, and EFAS scores. The study assessed correlations among scores, reliability, responsiveness to change, threshold and ceiling effect and discriminative ability of the SAV. RESULTS: There was a significant strong correlation with the EFAS and a significant moderate to strong correlation with the VISA-A. The score showed excellent reliability (ρ = 0.95, 95 %CI: [0.913; 0.976], p < 0.001). Responsiveness to change was significant between preoperative and postoperative SAV (37.99 ± 25.73 vs 70.86 ± 21.26). The SAV discriminated between patient and controls with no threshold or ceiling effect. CONCLUSION: The SAV provides a valid, reliable, and responsive method for assessing AT disorders. It offers a simplified and effective alternative to more complex scores. STUDY DESIGN: Cohort study (Diagnosis); Level of evidence, 2.

Breaching the family walls: Modelling the impact of prolonged visa insecurity on asylum-seeking children.

OBJECTIVE: Children in asylum-seeking families are increasingly subject to deterrent host nation policies that undermine security in the post-migration context, however, little is known on the mental health consequences of such policy. This study examined the impact of prolonged visa insecurity on child mental health, by comparing two cohorts of refugee children entering Australia between 2010 and 2013, distinguished by visa security. METHODS: The insecure visa sample comprised children from Tamil asylum-seeking families, while the secure visa sample was drawn from refugee families participating in the multi-ethnic 'Building a New Life in Australia' cohort study. Children in each sample were assessed for current mental health problems and trauma exposure. Mothers were assessed for trauma exposure, post-migration family stressors and post-traumatic stress disorder (PTSD). The effects of prolonged visa insecurity on child mental health via family-and child-level variables were modelled using multi-level path analysis. RESULTS: Data comprised 361 children, aged 10-18, and 242 mothers across three levels of visa insecurity: permanent protection (n = 293), temporary protection (n = 40) and bridging visa (n = 28). Modelling showed that (1) visa insecurity was associated with poorer child mental health, (2) the association was mediated sequentially by post-migration family stressors and maternal PTSD and (3) the association was moderated by maternal PTSD. CONCLUSION: Our findings suggest that when government policy persistently undermines post-migration security, the capacity of families to protect children from accrued stressors is lowered, leaving a significantly higher proportion of children developing along trajectories of risk rather than resilience.

A Genetically Engineered Bivalent Vaccine Coexpressing a Molecular Adjuvant against Classical Swine Fever and Porcine Epidemic Diarrhea.

Classical swine fever (CSF) and porcine epidemic diarrhea (PED) are highly contagious viral diseases that pose a significant threat to piglets and cause substantial economic losses in the global swine industry. Therefore, the development of a bivalent vaccine capable of targeting both CSF and PED simultaneously is crucial. In this study, we genetically engineered a recombinant classical swine fever virus (rCSFV) expressing the antigenic domains of the porcine epidemic diarrhea virus (PEDV) based on the modified infectious cDNA clone of the vaccine strain C-strain. The S1N and COE domains of PEDV were inserted into C-strain cDNA clone harboring the mutated 136th residue of Npro and substituted 3'UTR to generate the recombinant chimeric virus vC/SM3'UTRN-S1NCOE. To improve the efficacy of the vaccine, we introduced the tissue plasminogen activator signal (tPAs) and CARD domain of the signaling molecule VISA into vC/SM3'UTRN-S1NCOE to obtain vC/SM3'UTRN-tPAsS1NCOE and vC/SM3'UTRN-CARD/tPAsS1NCOE, respectively. We characterized three vaccine candidates in vitro and investigated their immune responses in rabbits and pigs. The NproD136N mutant exhibited normal autoprotease activity and mitigated the inhibition of IFN-ß induction. The introduction of tPAs and the CARD domain led to the secretory expression of the S1NCOE protein and upregulated IFN-ß induction in infected cells. Immunization with recombinant CSFVs expressing secretory S1NCOE resulted in a significantly increased in PEDV-specific antibody production, and coexpression of the CARD domain of VISA upregulated the PEDV-specific IFN-γ level in the serum of vaccinated animals. Notably, vaccination with vC/SM3'UTRN-CARD/tPAsS1NCOE conferred protection against virulent CSFV and PEDV challenge in pigs. Collectively, these findings demonstrate that the engineered vC/SM3'UTRN-CARD/tPAsS1NCOE is a promising bivalent vaccine candidate against both CSFV and PEDV infections.

Antibiotic Exposure Leads to Reduced Phage Susceptibility in Vancomycin Intermediate Staphylococcus aureus (VISA).

In the time of antimicrobial resistance, phage therapy is frequently suggested as a possible solution for such difficult-to-treat infections. Vancomycin-intermediate Staphylococcus aureus (VISA) remains a relatively rare yet increasing occurrence in the clinic for which phage therapy may be an option. However, the data presented herein suggest a potential cross-resistance mechanism to phage following vancomycin exposure in VISA strains. When comparing genetically similar strains differing in their susceptibility to vancomycin, those with intermediate levels of vancomycin resistance displayed decreased sensitivity to phage in solid and liquid assays. Serial passaging with vancomycin induced both reduced vancomycin susceptibility and phage sensitivity. As a consequence, the process of phage infection was shown to be interrupted after DNA ejection from adsorbed phage but prior to phage DNA replication, as demonstrated through adsorption assays, lysostaphin sensitivity assays, electron microscopy, and quantitative PCR (qPCR). At a time when phage products are being used for experimental treatments and tested in clinical trials, it is important to understand possible interference between mechanisms underlying antibiotic and phage resistance in order to design effective therapeutic regimens.

The Victorian Sports Assessment Institute-Achilles Tendinopathy Questionnaire (VISA-A): Chinese cross-cultural adaptation and psychometric validation.

PURPOSE: We aimed to create a standardized cross-cultural adaptation of the simplified Chinese version of VISA-A, test its reliability and validity and conduct exploratory factor analysis on the correlation between items. METHODS: According to international recommendations for the cross-cultural adaptation of questionnaires, after considering the opinions of patients, we translated and revised the English version to create a simplified Chinese version of the questionnaire. We recruited healthy subjects in the general specialty of one university (n = 90) and the physical education specialty of another university (n = 89), and we recruited patients with Achilles tendinopathy in a third group (n = 85). Reliability was evaluated by calculating test-retest reliability and internal consistency, validity was evaluated by exploring structural and criterion validity (correlation with the physical function and body pain items of the SF-36), and responsiveness was evaluated by calculating area under the receiver operating characteristic curve (AUC). RESULTS: The simplified Chinese version of the VISA-A had no ceiling or floor effects. Four common factors were extracted and explained by the exploratory factor analysis. The test-retest reliability (ICC = 0.97) and internal consistency (Cronbach's alpha = 0.84) were adequate. The questionnaire had moderate correlations with the physical function and body pain items of the SF-36. The AUC was 0.9407. CONCLUSION: The simplified Chinese version of the VISA-A had good reliability and validity and excellent responsiveness, but the factorial structure is not inconsistent with the dimensions of the original version. It can be used to assess and manage patients with Achilles tendinitis in the Chinese culture.

The impact of changing nonimmigrant visa policies on international students' psychological adjustment and well-being in the United States during the COVID-19 pandemic: a qualitative study.

BACKGROUND: Since the Immigration and Nationality Act of 1952, the number of international students in the United States had been gradually increasing. However, the total numbers have begun to decrease since 2019-2020 school year due to the Trump administration's policy and COVID-19. Still, little is known about how international students' psychological adjustment and well-being have been affected by changing nonimmigrant visa policy and the COVID-19 pandemic.  METHODS: We conducted a total of 34 online semi-structured in-depth interviews with international students from 18 countries of origin studying in the San Francisco Bay Area, California. More than 60% of the participants (21 out of 34) were aged 21 to 25. Among our 34 participants, gender and 18 were male and 16 were female, and 19 were undergraduate students and 15 were master's students. The majority of the participants were first-generation college students (22/34, 64.71%). Verbatim transcription was done for all interviews. NVivo was used for both deductive and inductive approaches to the qualitative analysis. RESULTS: Overall, the recent political climate negatively impacted participants' psychology of adjustment and well-being. July 6, 2020 Policy Directive for international students caused severe uncertainty about whether they can continue studying in the United States. There were many resources or services needed to overcome this period, such as extended mental and emotional support from the counseling services as well as financial and informational support from the international student office and university. Although international students had the benefit of the university's food assistance program, they were not eligible to receive any external support outside of the university and financial aid at the local and federal levels. Whether maintaining F-1 visa status was one of their major concerns. Due to COVID-19, job opportunities were limited, which made international students difficult to obtain Curricular Practical Training (CPT) and secure a job in the United States within the 90-day unemployment limit of Optical Practical Training (OPT). H-1B visa and permanent residency were other challenges to go through, but participants saw positive perspectives from the Biden administration. CONCLUSIONS: Uncertain policy changes due to COVID-19 and presidential transitions impacted international students' psychological well-being and adjustment. International students are important populations in the United States who have supported jobs that are high in demand and economically contributed to the United States. It is expected that future policies at various levels support international students' life and improve their health equity and mental health.

Consequences of Patellar Tendinopathy on Isokinetic Knee Strength and Jumps in Professional Volleyball Players.

Patellar tendinopathy (PT) in professional volleyball players can have an impact on their careers. We evaluated the impact of this pathology in this specific population in terms of isokinetic strength and jumping performances. Thirty-six professional male volleyball players (mean age: 24.8 ± 5.2) performed isokinetic knee assessments, single-leg countermovement jumps and one leg hop test. They filled out the Victorian Institute of Sport Assessment-Patella (VISA-P) score. Two groups were assessed: "PT group" (n = 15) and "control group" (n = 21). The VISA-P score was lower in the PT group (p < 0.0001). No difference was found between the isokinetic strength limb symmetry index and the jump performance limb symmetry index. The healthy legs of the control group were compared with the affected (PT+) and the unaffected legs (PT−) of the PT group. Compared with the healthy legs, both PT+ and PT− legs showed decreased values of quadriceps and hamstring strengths. Only PT+ legs scored lower than healthy legs in countermovement jumps and hop tests. No differences were found between PT+ and PT− legs for muscle strengths and jumps. A low correlation existed between quadriceps strength and jumping performances (r > 0.3; p < 0.001). Volleyball players with PT showed a decrease in the isokinetic knee strength. This strength deficit was found both on the symptomatic legs and the asymptomatic ones. Jumps were only significantly altered on the pathological legs. Highlighting that the unaffected limbs were also impaired in addition to the affected limbs may help provide a better adaptation of the rehabilitation management.

Hyperosmolar dextrose injection for Osgood-Schlatter disease: a double-blind, randomized controlled trial.

OBJECTIVE: Osgood-Schlatter disease (OSD) is one of the common causes of long-term knee pain, leading to functional limitations, long-term deformity of the tubercle interfering with kneeling, and impaired peer-important sport participation. Nonetheless, patient management continues to rely on the usual conservative methods. This study examined the use of hyperosmolar dextrose injection in patients with OSD. METHODS: We conducted a randomized, double-blind clinical trial involving 70 patients with OSD. One group received a hyperosmolar dextrose injection (12.5%), while the other received a saline injection. The injections were conducted under ultrasound guidance. The Victorian Institute of Sport Assessment (VISA) score was used to assess each patient's pain and sport level. RESULTS: The dextrose group outperformed the saline group in improvement in the VISA-Patella (VISA-P) score from baseline to 3 months (27.1 ± 6.5 vs. 1.4 ± 2.6; mean difference 25.4 (22.4 to 28.3); p < .0001), 6 months (31.7 ± 7.9 vs. 25.2 ± 7.8; mean difference 6.2 (3.2 to 9.4); p < .0001), and 12 months (34 ± 9.0 vs. 28.2 ± 7.5; mean difference 5.5 (1.9 to 9.1); p = .0026). The changes in both groups were clinically important, suggesting that both therapies were active treatments. The dextrose group improved too rapidly for spontaneous improvement to explain much of this change. CONCLUSION: After three injections, at the 6-month and 12-month follow-up visits, the VISA-P scores of the two groups were significantly improved; the dextrose group score was better than the saline group score, and there were significant differences between the two groups.

Health-related quality of life in patients with Achilles tendinopathy: Comparison to the general population of the United Kingdom.

BACKGROUND: There is little evidence available regarding the impact of Achilles Tendinopathy (AT) on health-related quality of life (HRQOL). The primary aim of this study was to quantify the clinical and health-related quality-of-life patient-reported outcome measures for a population presenting with either mid-substance or insertional Achilles tendinopathy. METHODS: A prospective comparative observational study of consecutive patients with AT presenting for extracorporeal shockwave therapy (ESWT) at a large teaching hospital. The primary outcome was assessment of a validated health-related quality of life PROMs (Euroqol EQ-5D-5L) and comparison to 2 general UK population datasets. The secondary outcomes were Visual Analogue Pain Scale (VAS-Pain) and two validated foot-specific patient reported outcome measures (Foot Function Index (FFI) and Victorian Institute of Sports Assessment-Achilles (VISA-A)). RESULTS: Between March 2014 and June 2021, 320 consecutive patients (125 male; 195 female) were diagnosed with AT and referred for a first course of ESWT. EQ-5D-5L PROMs were prospectively collected for 303 of these patients (94.7%). The mean age (± standard deviation(SD)) was 52.1 ± 11.4 years. The mean EQ-5D-5L Index score (mean±SD) for the AT cohort was 0.783 ± 0.131. Patients less than 55 years with AT had a statistically significantly worse quality of life compared with members of the same age group in the general population. The mean VAS-Pain, FFI, VISA-A clinical outcome scores were 6.0 ± 2.3, 49.5 ± 21.2 and 34.1 ± 14.4 respectively. There was a statistically significant moderate correlation between HRQOL and clinical PROMs (VAS-Pain and FFI vs EQ-5D) however there was no correlation with age. CONCLUSION: This study demonstrates that patients under the age of 55 with AT have a significantly reduced quality of life compared with the general population. LEVEL OF EVIDENCE: III.

Protein Biomarker Discovery for Methicillin-Sensitive, Heterogeneous Vancomycin-Intermediate and Vancomycin-Intermediate Staphylococcus aureus Strains Using Label-Free Data-Independent Acquisition Proteomics.

Rapid identification of methicillin-sensitive Staphylococcus aureus (MSSA), heterogeneous vancomycin-intermediate S. aureus (hVISA), and vancomycin-intermediate S. aureus (VISA) is important for accurate treatment, timely intervention, and prevention of outbreaks. Here, 90 S. aureus isolates were analyzed for protein biomarker discovery, including MSSA, vancomycin-susceptible S. aureus (VSSA), hVISA, and VISA strains. Label-free data-independent acquisition proteomics was used to identify protein biomarkers that allow for discrimination among MSSA, hVISA, and VISA strains. There were 8786 nonredundant peptides identified, corresponding to 418 different annotated nonredundant proteins. Two VISA protein biomarkers, two hVISA protein biomarkers, and one MSSA protein biomarker with high sensitivities and specificities were discovered and verified. Data are available via MassIVE with identifier MSV000085776.

Thioredoxin 2 Negatively Regulates Innate Immunity to RNA Viruses by Disrupting the Assembly of the Virus-Induced Signaling Adaptor Complex.

The virus-induced signaling adaptor (VISA) complex plays a critical role in the innate immune response to RNA viruses. However, the mechanism of VISA complex formation remains unclear. Here, we demonstrate that thioredoxin 2 (TRX2) interacts with VISA at mitochondria both in vivo and in vitro Knockdown and knockout of TRX2 enhanced the formation of the VISA-associated complex, as well as virus-triggered activation of interferon regulatory factor 3 (IRF3) and transcription of the interferon beta 1 (IFNB1) gene. TRX2 inhibits the formation of VISA aggregates by repressing reactive oxygen species (ROS) production, thereby disrupting the assembly of the VISA complex. Furthermore, our data suggest that the C93 residue of TRX2 is essential for inhibition of VISA aggregation, whereas the C283 residue of VISA is required for VISA aggregation. Collectively, these findings uncover a novel mechanism of TRX2 that negatively regulates VISA complex formation.IMPORTANCE The VISA-associated complex plays pivotal roles in inducing type I interferons (IFNs) and eliciting the innate antiviral response. Many host proteins are identified as VISA-associated-complex proteins, but how VISA complex formation is regulated by host proteins remains enigmatic. We identified the TRX2 protein as an important regulator of VISA complex formation. Knockout of TRX2 increases virus- or poly(I·C)-triggered induction of type I IFNs at the VISA level. Mechanistically, TRX2 inhibits the production of ROS at its C93 site, which impairs VISA aggregates at its C283 site, and subsequently impedes the assembly of the VISA complex. Our findings suggest that TRX2 plays an important role in the regulation of VISA complex assembly.

Retention of visa-trainee post-graduate residents in Canada: a retrospective cohort study.

BACKGROUND: Visa trainees (international medical graduates [IMG] who train in Canada under a student or employment visa) are expected to return home after completing their training. We examine the retention patterns of visa trainee residents funded by Canadian (regular ministry and other), foreign, or mixed sources. METHODS: We linked data from the Canadian Post-MD Medical Education Registry with Scott's Medical Database for a retrospective cohort study. Eligible trainees were IMG visa trainees as of their first year of training, started their residency program no earlier than 2000, and exited training between 2006 and 2016. We used Cox regression to compare the retention of visa trainees by funding source. RESULTS: Of 1,913 visa trainees, 431(22.5%), 1353 (70.7%) and 129 (6.8%) had Canadian, foreign, or mixed funding, respectively. The proportion of trainees remaining in Canada decreased over time, with 35.5% (679/1913); 17.7% (186/1052); 10.8% (11/102) in Canada one, five, and ten years, respectively after their exit from PGME training. Trainees who remained on visas (HR: 1.91; [95% CI 1.59, 2.30]), were funded exclusively by foreign sources (HR: 1.46; [95% CI 1.25, 1.69]), and who had graduated from 'Western' countries (HR: 1.39; [95% CI 1.06, 1.84]) were more likely to leave Canada compared to trainees who became citizens/permanent residents, were funded by Canadian sources, or were visa graduates of Canadian medical schools, respectively. CONCLUSIONS: Most visa trainees leave Canada following their training. Trainees with Canadian connections (funding and/or change in legal status) were more likely to remain in Canada.

Understanding the Mechanism of Action of NAI-112, a Lanthipeptide with Potent Antinociceptive Activity.

NAI-112, a glycosylated, labionine-containing lanthipeptide with weak antibacterial activity, has demonstrated analgesic activity in relevant mouse models of nociceptive and neuropathic pain. However, the mechanism(s) through which NAI-112 exerts its analgesic and antibacterial activities is not known. In this study, we analyzed changes in the spinal cord lipidome resulting from treatment with NAI-112 of naive and in-pain mice. Notably, NAI-112 led to an increase in phosphatidic acid levels in both no-pain and pain models and to a decrease in lysophosphatidic acid levels in the pain model only. We also showed that NAI-112 can form complexes with dipalmitoyl-phosphatidic acid and that Staphylococcus aureus can become resistant to NAI-112 through serial passages at sub-inhibitory concentrations of the compound. The resulting resistant mutants were phenotypically and genotypically related to vancomycin-insensitive S. aureus strains, suggesting that NAI-112 binds to the peptidoglycan intermediate lipid II. Altogether, our results suggest that NAI-112 binds to phosphate-containing lipids and blocks pain sensation by decreasing levels of lysophosphatidic acid in the TRPV1 pathway.