Test-retest reliability of wireless inertial-sensor derived measurements of knee joint kinematics.

Advances in sensor technology have provided an opportunity to measure gait characteristics using body-worn inertial measurement units (IMUs). Whilst research investigating the validity of IMUs in reporting gait characteristics is extensive, research investigating the reliability of IMUs is limited. This study aimed to investigate the inter-session reliability of wireless IMU derived measures of gait (i.e., knee angle, range of motion) taking multiple test administrators into account. Fifteen healthy volunteers (43 ± 15 years) completed two visits. Within each visit, participants were required to perform two sets of 6 gait trials (6-metre walk tests). IMUs were placed on the participant in 7 locations on the lower limbs and waist. A different test administrator (n = 3) applied the IMUs at each set. At visit 2, this procedure was repeated with the same test administrators as visit 1. Kinematic measures of maximum angle (Knee_Max), minimum angle (Knee_Min), and range of motion (RoM) are reported for the left and right knee. The intraclass correlation coefficients (ICC), standard error of measurement (SEM) and minimum detectable change (MDC) are reported to determine IMU reliability. The results confirmed moderate to good inter-session reliability across all features (0.73-0.87). SEM values ranged from 1.21-3.32° and MDC values ranged from 3.37 - 9.21°. Therefore, IMUs appear to be a reliable method to determine inter-session gait characteristics across multiple test administrators.

An Automatic Foot and Shank IMU Synchronization Algorithm: Proof-of-concept.

When using wearable sensors for measurement and analysis of human performance, it is often necessary to integrate and synchronise data from separate sensor systems. This paper describes a synchronization technique between IMUs attached to the shanks and insoles attached at the feet and aims to solve the need to compute the ankle joint angle, which relies on synchronized sensor data. This will additionally enable concurrent analysis using gait kinematic and kinetic features. A proof-of-concept of the algorithm, which relies on cross-correlation of gyroscope sensor data from the shank and foot, to align the sensor systems is demonstrated. The algorithm output is validated against those signals synchronized using manually annotated heel-strike and toe-off ground-truth signal landmarks, identified in both the shank and feet signals using previously published definitions. Results demonstrate that the developed algorithm is capable of synchronizing both sensor systems, based on IMU data from both healthy participants and participants suffering from knee osteoarthritis, with a mean lag time bias of 25.56ms when compared to the ground truth. A proof-of-concept of technique to synchronise IMUs attached to the shanks and insoles attached at the feet is demonstrated and offers an alternative approach to sensor system synchronisation.

BVD seroprevalence in the Irish cattle population as the national BVD programme progresses toward eradication.

BACKGROUND: Bovine Viral Diarrhoea Virus (BVDV) infection remains endemic in many countries worldwide. Ireland, in common with several other European counties, commenced an BVDV eradication programme in the last decade, Managing eradication programmes requires careful monitoring of diseases prevalence and understanding factors associated with disease exposure to ensure eradication programmes remain evidence based and tailored to the evolving epidemiological situation. METHODS: In this study, we explore the seroprevalence of BVDV exposure over a four-year period (2017 to 2020) in Ireland from a cohort of animals (n = 6,449) under 30 months of age sampled at slaughter, who were born subsequent to the commencement of a compulsory national eradication programme. Temporal trends and risk factor analysis were undertaken using multilevel logit regression models. RESULTS: There was a declining temporal trend in seroprevalence over the sample years of the study, and risk varied at both county- and herd-levels. The unadjusted marginal animal-level seroprevalence reduced from 9.1% in 2017 (95%; CI: 7.2-10.9) to 3.9% in 2020 (95%; CI: 3.2-4.6). The final model suggested that seropositivity in study cattle was strongly related with the presence of a PI animal in the herd during the year of the animal's birth, and to a lesser extent the status of the herd from which the animal was slaughtered. The risk of seroconversion increased significantly with increasing size of the herd of slaughter, in females relative to males, and in dairy relative to suckler herds. CONCLUSIONS: This study has shown that the BVDV serostatus of cattle at slaughter is correlated to the BVD infection history of the herd into which the animal was born and the herd from which it was slaughtered. Herd location, increased herd size and dairy production were associated with increased probability of serconversion. These findings will be used to inform the targeting of surveillance strategies once BVDV freedom has been achieved.

Self-Selected Motivational Music Enhances Physical Performance in Normoxia and Hypoxia in Young Healthy Males.

Humans exposed to hypoxia are susceptible to physiological and psychological impairment. Music has ergogenic effects through enhancing psychological factors such as mood, emotion, and cognition. This study aimed to investigate music as a tool for mitigating the performance decrements observed in hypoxia. Thirteen males (mean ± SD; 24 ± 4 years) completed one familiarization session and four experimental trials; (1) normoxia (sea level, 0.209 FiO2) and no music; (2) normoxia (0.209 FiO2) with music; (3) normobaric hypoxia (∼3800 m, 0.13 FiO2) and no music; and (4) normobaric hypoxia (0.13 FiO2) with music. Experimental trials were completed at 21°C with 50% relative humidity. Music was self-selected prior to the familiarization session. Each experimental trial included a 15-min time trial on an arm bike, followed by a 60-s isometric maximal voluntary contraction (MVC) of the biceps brachii. Supramaximal nerve stimulation quantified central and peripheral fatigue with voluntary activation (VA%) calculated using the doublet interpolation method. Average power output (W) was reduced with a main effect of hypoxia (p = 0.02) and significantly increased with a main effect of music (p = 0.001). When combined the interaction was additive (p = 0.87). Average MVC force (N) was reduced in hypoxia (p = 0.03) but VA% of the biceps brachii was increased with music (p = 0.02). Music reduced subjective scores of mental effort, breathing discomfort, and arm discomfort in hypoxia (p < 0.001). Music increased maximal physical exertion through enhancing neural drive and diminishing detrimental mental processes, enhancing performance in normoxia (6.3%) and hypoxia (6.4%).

Mental fatigue independent of boredom and sleepiness does not impact self-paced physical or cognitive performance in normoxia or hypoxia.

This study investigated the individual and combined effects of mental fatigue (MF) and hypoxia (HYP) on physical and cognitive performance. Fifteen males (24 ± 3 years) completed one familiarization session and six experimental trials, including: 1) normoxia (0.209 FiO2) and no MF; 2) normoxia (0.209 FiO2) with MF; 3) mild normobaric HYP (0.13 FiO2) and no MF; 4) mild normobaric HYP (0.13 FiO2) with MF; 5) severe normobaric HYP (0.10 FiO2) and no MF; 6) severe normobaric HYP (0.10 FiO2) with MF. Each condition included a 15-min self-paced time trial, followed by a 60-s isometric maximal voluntary contraction of the biceps brachii. MF was induced using a 16-min individualized cognitive test prior to exercise performance. Following each time trial, participants performed the Tower of Hanoi cognitive test. A main effect of HYP was observed on average power output, oxygen consumption and muscle oxygenation (P ≤ 0.004), with no effect of MF (P ≥ 0.599). Voluntary activation of the biceps brachii was also reduced in HYP (68.42 ± 5.64%, P = 0.039). No effect of MF or HYP was observed on cognitive performance (P ≥ 0.138). HYP impacted physical performance, whilst MF had no effect on self-paced physical or cognitive performance.

Manipulation of Autophagy and Apoptosis Facilitates Intracellular Survival of Staphylococcus aureus in Human Neutrophils.

Polymorphonuclear neutrophils (PMN) are critical for first line innate immune defence against Staphylococcus aureus. Mature circulating PMN maintain a short half-life ending in constitutive apoptotic cell death. This makes them unlikely candidates as a bacterial intracellular niche. However, there is significant evidence to suggest that S. aureus can survive intracellularly within PMN and this contributes to persistence and dissemination during infection. The precise mechanism by which S. aureus parasitizes these cells remains to be established. Herein we propose a novel mechanism by which S. aureus subverts both autophagy and apoptosis in PMN in order to maintain an intracellular survival niche during infection. Intracellular survival of S. aureus within primary human PMN was associated with an accumulation of the autophagic flux markers LC3-II and p62, while inhibition of the autophagy pathway led to a significant reduction in intracellular survival of bacteria. This intracellular survival of S. aureus was coupled with a delay in neutrophil apoptosis as well as increased expression of several anti-apoptotic factors. Importantly, blocking autophagy in infected PMN partially restored levels of apoptosis to that of uninfected PMN, suggesting a connection between the autophagic and apoptotic pathways during intracellular survival. These results provide a novel mechanism for S. aureus intracellular survival and suggest that S. aureus may be subverting crosstalk between the autophagic and apoptosis pathways in order to maintain an intracellular niche within human PMN.

A comparison of methods used for inducing mental fatigue in performance research: individualised, dual-task and short duration cognitive tests are most effective.

Despite research indicating the negative impact that mental fatigue has on physical and cognitive performance, whether this is a result of mental fatigue or a state of under-arousal remains unclear. The current research aimed to explore the effectiveness of the methods being used to induce mental fatigue. Twelve participants attended six sessions in which two cognitive tests, the AX-continuous performance test (AX-CPT) and the TloadDback test, were compared for their effectiveness in inducing mental fatigue. Both tests were set at a standard processing speed (1.2 ms) for two conditions, and a further condition involved the individualisation of the TloadDback test. Participants presented significantly higher physiological and psychological arousal (p < 0.05) in the individualised dual-task test compared to the AX-CPT. The individualised TloadDback test is a more effective method of inducing mental fatigue compared to the AX-CPT, as it sustains physiological arousal whilst inducing measurable reductions in mental resources. Practitioner summary: Mental fatigue negatively impacts physical and cognitive performance. It is unclear whether the current methods being used to induce mental fatigue are effective. This study compared different methods and confirmed that short, individualised and dual-task tests are most effective for inducing mental fatigue whilst maintaining arousal.

Memory Th1 Cells Are Protective in Invasive Staphylococcus aureus Infection.

Mechanisms of protective immunity to Staphylococcus aureus infection in humans remain elusive. While the importance of cellular immunity has been shown in mice, T cell responses in humans have not been characterised. Using a murine model of recurrent S. aureus peritonitis, we demonstrated that prior exposure to S. aureus enhanced IFNγ responses upon subsequent infection, while adoptive transfer of S. aureus antigen-specific Th1 cells was protective in naïve mice. Translating these findings, we found that S. aureus antigen-specific Th1 cells were also significantly expanded during human S. aureus bloodstream infection (BSI). These Th1 cells were CD45RO+, indicative of a memory phenotype. Thus, exposure to S. aureus induces memory Th1 cells in mice and humans, identifying Th1 cells as potential S. aureus vaccine targets. Consequently, we developed a model vaccine comprising staphylococcal clumping factor A, which we demonstrate to be an effective human T cell antigen, combined with the Th1-driving adjuvant CpG. This novel Th1-inducing vaccine conferred significant protection during S. aureus infection in mice. This study notably advances our understanding of S. aureus cellular immunity, and demonstrates for the first time that a correlate of S. aureus protective immunity identified in mice may be relevant in humans.

Manipulation of Autophagy in Phagocytes Facilitates Staphylococcus aureus Bloodstream Infection.

The capacity for intracellular survival within phagocytes is likely a critical factor facilitating the dissemination of Staphylococcus aureus in the host. To date, the majority of work on S. aureus-phagocyte interactions has focused on neutrophils and, to a lesser extent, macrophages, yet we understand little about the role played by dendritic cells (DCs) in the direct killing of this bacterium. Using bone marrow-derived DCs (BMDCs), we demonstrate for the first time that DCs can effectively kill S. aureus but that certain strains of S. aureus have the capacity to evade DC (and macrophage) killing by manipulation of autophagic pathways. Strains with high levels of Agr activity were capable of causing autophagosome accumulation, were not killed by BMDCs, and subsequently escaped from the phagocyte, exerting significant cytotoxic effects. Conversely, strains that exhibited low levels of Agr activity failed to accumulate autophagosomes and were killed by BMDCs. Inhibition of the autophagic pathway by treatment with 3-methyladenine restored the bactericidal effects of BMDCs. Using an in vivo model of systemic infection, we demonstrated that the ability of S. aureus strains to evade phagocytic cell killing and to survive temporarily within phagocytes correlated with persistence in the periphery and that this effect is critically Agr dependent. Taken together, our data suggest that strains of S. aureus exhibiting high levels of Agr activity are capable of blocking autophagic flux, leading to the accumulation of autophagosomes. Within these autophagosomes, the bacteria are protected from phagocytic killing, thus providing an intracellular survival niche within professional phagocytes, which ultimately facilitates dissemination.

TRAM is required for TLR2 endosomal signaling to type I IFN induction.

Detection of microbes by TLRs on the plasma membrane leads to the induction of proinflammatory cytokines such as TNF-α, via activation of NF-κB. Alternatively, activation of endosomal TLRs leads to the induction of type I IFNs via IFN regulatory factors (IRFs). TLR4 signaling from the plasma membrane to NF-κB via the Toll/IL-1R (TIR) adaptor protein MyD88 requires the TIR sorting adaptor Mal, whereas endosomal TLR4 signaling to IRF3 via the TIR domain-containing adaptor-inducing IFN-ß (TRIF) requires the TRIF-related adaptor molecule (TRAM). Similar to TLR4 homodimers, TLR2 heterodimers can also induce both proinflammatory cytokines and type I IFNs. TLR2 plasma membrane signaling to NF-κB is known to require MyD88 and Mal, whereas endosomal IRF activation by TLR2 requires MyD88. However, it was unclear whether TLR2 requires a sorting adaptor for endosomal signaling, like TLR4 does. In this study, we show that TLR2-dependent IRF7 activation at the endosome is both Mal- and TRAM-dependent, and that TRAM is required for the TLR2-dependent movement of MyD88 to endosomes following ligand engagement. TRAM interacted with both TLR2 and MyD88, suggesting that TRAM can act as a bridging adapter between these two molecules. Furthermore, infection of macrophages lacking TRAM with herpes viruses or the bacterium Staphylococcus aureus led to impaired induction of type I IFN, indicating a role for TRAM in TLR2-dependent responses to human pathogens. Our work reveals that TRAM acts as a sorting adaptor not only for TLR4, but also for TLR2, to facilitate signaling to IRF7 at the endosome, which explains how TLR2 is capable of causing type I IFN induction.

Staphylococcus aureus infection of mice expands a population of memory γδ T cells that are protective against subsequent infection.

The development of vaccines against Staphylococcus aureus has consistently failed in clinical trials, likely due to inefficient induction of cellular immunity. T cell-derived IL-17 is one of the few known correlates of antistaphylococcoal immunity, conferring protection against S. aureus infections through its ability to promote phagocytic cell effector functions. A comprehensive understanding of the discrete T cell subsets critical for site-specific IL-17-mediated bacterial clearance will therefore be necessary to inform the development of vaccines that efficiently target cellular immunity. In this study, we have identified a population of CD44+ CD27- memory γδ T cells, expanded upon infection of C57BL/6 mice with S. aureus, which produce high levels of IL-17 and mediate enhanced bacterial clearance upon reinfection with the bacterium. These cells are comprised largely of the Vγ4+ subset and accumulate at the site of infection subsequent to an initial Vγ1.1+ and Vγ2+ T cell response. Moreover, these Vγ4+ T cells are retained in the peritoneum and draining mediastinal lymph nodes for a prolonged period following bacterial clearance. In contrast to its critical requirement for γδ T cell activation during the primary infection, IL-1 signaling was dispensable for activation and expansion of memory γδ T cells upon re-exposure to S. aureus. Our findings demonstrate that a γδ T cell memory response can be induced upon exposure to S. aureus, in a fashion analogous to that associated with classical αß T cells, and suggest that induction of IL-17-expressing γδ T cells may be an important property of a protective vaccine against S. aureus.

Targeted nasal vaccination provides antibody-independent protection against Staphylococcus aureus.

Despite showing promise in preclinical models, anti-Staphylococcus aureus vaccines have failed in clinical trials. To date, approaches have focused on neutralizing/opsonizing antibodies; however, vaccines exclusively inducing cellular immunity have not been studied to formally test whether a cellular-only response can protect against infection. We demonstrate that nasal vaccination with targeted nanoparticles loaded with Staphylococcus aureus antigen protects against acute systemic S. aureus infection in the absence of any antigen-specific antibodies. These findings can help inform future developments in staphylococcal vaccine development and studies into the requirements for protective immunity against S. aureus.

Nlrp-3-driven interleukin 17 production by γδT cells controls infection outcomes during Staphylococcus aureus surgical site infection.

Recent work has identified T cells and the cytokines they produce as important correlates of immune protection during Staphylococcus aureus infections through the ability of these T cells to regulate local neutrophil responses. However, the specific T-cell subsets that are involved in coordinating protection at distinct sites of infection remains to be established. In this study, we identify for the first time an important role for γδT cells in controlling S. aureus surgical site infection (SSI). γδT cells are recruited to the wound site following S. aureus challenge, where they represent the primary source of interleukin 17 (IL-17), with a small contribution from other non-γδT cells. The IL-17 response is entirely dependent upon IL-1 receptor signaling. Using IL-17 receptor-deficient mice, we demonstrate that IL-17 is required to control bacterial clearance during S. aureus SSI. However, we demonstrate a strain-dependent requirement for γδT cells in this process due to the differential abilities of individual strains to activate IL-1ß production. IL-1ß processing relies upon activation of the Nlrp3 inflammasome complex, and we demonstrate that Nlrp3-deficient and IL-1 receptor-deficient mice have an impaired ability to control S. aureus SSI due to reduced production of IL-17 by γδT cells at the site of infection. Given that IL-17 has been identified as an important correlate of immune protection during S. aureus infection, it is vital that the unique cellular sources of this cytokine and mechanisms inducing its activation are identified at distinct sites of infection. Our study demonstrates that while IL-17 may be critically important for mediating immune protection during S. aureus SSI, the relative contribution of γδT cells to these protective effects may be strain dependent.

Nasal colonisation by Staphylococcus aureus depends upon clumping factor B binding to the squamous epithelial cell envelope protein loricrin.

Staphylococcus aureus asymptomatically colonises the anterior nares, but the host and bacterial factors that facilitate colonisation remain incompletely understood. The S. aureus surface protein ClfB has been shown to mediate adherence to squamous epithelial cells in vitro and to promote nasal colonisation in both mice and humans. Here, we demonstrate that the squamous epithelial cell envelope protein loricrin represents the major target ligand for ClfB during S. aureus nasal colonisation. In vitro adherence assays indicated that bacteria expressing ClfB bound loricrin most likely by the "dock, lock and latch" mechanism. Using surface plasmon resonance we showed that ClfB bound cytokeratin 10 (K10), a structural protein of squamous epithelial cells, and loricrin with similar affinities that were in the low µM range. Loricrin is composed of three separate regions comprising GS-rich omega loops. Each loop was expressed separately and found to bind ClfB, However region 2 bound with highest affinity. To investigate if the specific interaction between ClfB and loricrin was sufficient to facilitate S. aureus nasal colonisation, we compared the ability of ClfB⁺S. aureus to colonise the nares of wild-type and loricrin-deficient (Lor⁻/⁻) mice. In the absence of loricrin, S. aureus nasal colonisation was significantly impaired. Furthermore a ClfB⁻ mutant colonised wild-type mice less efficiently than the parental ClfB⁺ strain whereas a similar lower level of colonisation was observed with both the parental strain and the ClfB⁻ mutant in the Lor⁻/⁻ mice. The ability of ClfB to support nasal colonisation by binding loricrin in vivo was confirmed by the ability of Lactococcus lactis expressing ClfB to be retained in the nares of WT mice but not in the Lor⁻/⁻ mice. By combining in vitro biochemical analysis with animal model studies we have identified the squamous epithelial cell envelope protein loricrin as the target ligand for ClfB during nasal colonisation by S. aureus.