The characteristics of the implicit body model of the trunk.

Knowing where the body is in space requires reference to a stored model of the size and shape of body parts, termed the body model. This study sought to investigate the characteristics of the implicit body model of the trunk by assessing the position sense of midline and lateral body landmarks. Sixty-nine healthy participants localised midline and lateral body landmarks on their thorax, waist and hips, with perceived positions of these landmarks compared to actual positions. This study demonstrates evidence of a significant distortion of the implicit body model of the trunk, presenting as a squatter trunk, wider at the waist and hips. A significant difference was found between perceived and actual location in the horizontal (x) and vertical (y) directions for the majority of trunk landmarks. Evidence of a rightward bias was noted in the perception of six of the nine body landmarks in the horizontal (x) direction, including all midline levels. In the vertical (y) direction, a substantial inferior bias was evident at the thorax and waist. The implicit body model of the trunk is shown to be distorted, with the lumbar spine (waist-to-hip region) held to be shorter and wider than reality.

Analysing oxygen reduction electrocatalysis on transition metal doped niobium oxide(110).

A good oxygen reduction reaction (ORR) catalyst should be stable and active under electrochemical reaction conditions. Niobium pentaoxide (Nb2O5) is known to be stable under ORR conditions. However it has a large band gap, which makes conductivity a challenge during the reaction. In this work, we aim to understand if surface modification of the 110 facet of niobium pentaoxide with transition metal doping has any effect on its ORR activity and conductivity. While the problem of conductivity in the case of transition metal oxides (TMOs) can be partially rectified by transition metal doping, it has negligible influence on the ORR activity of the doped systems.

The commercial pig as a model of spontaneously-occurring osteoarthritis.

BACKGROUND: Preclinical osteoarthritis models where damage occurs spontaneously may better reflect the initiation and development of human osteoarthritis. The aim was to assess the commercial pig as a model of spontaneous osteoarthritis development by examining pain-associated behaviour, joint cartilage integrity, as well as the use of porcine cartilage explants and isolated chondrocytes and osteoblasts for ex vivo and in vitro studies. METHODS: Female pigs (Large white x Landrace x Duroc) were examined at different ages from 6 weeks to 3-4 years old. Lameness was assessed as a marker of pain-associated behaviour. Femorotibial joint cartilage integrity was determined by chondropathy scoring and histological staining of proteoglycan. IL-6 production and proteoglycan degradation was assessed in cartilage explants and primary porcine chondrocytes by ELISA and DMMB assay. Primary porcine osteoblasts from damaged and non-damaged joints, as determined by chondropathy scoring, were assessed for mineralisation, proliferative and mitochondrial function as a marker of metabolic capacity. RESULTS: Pigs aged 80 weeks and older exhibited lameness. Osteoarthritic lesions in femoral condyle and tibial plateau cartilage were apparent from 40 weeks and increased in severity with age up to 3-4 years old. Cartilage from damaged joints exhibited proteoglycan loss, which positively correlated with chondropathy score. Stimulation of porcine cartilage explants and primary chondrocytes with either IL-1ß or visfatin induced IL-6 production and proteoglycan degradation. Primary porcine osteoblasts from damaged joints exhibited reduced proliferative, mineralisation, and metabolic capacity. CONCLUSION: In conclusion, the commercial pig represents an alternative model of spontaneous osteoarthritis and an excellent source of tissue for in vitro and ex vivo studies.

Prevalence of frailty among community dwelling older adults in receipt of low level home support: a cross-sectional analysis of the North Dublin Cohort.

BACKGROUND: There is increasing demand for formal government funded home help services to support community-dwelling older people in Ireland, yet limited information exists on the health profiles of this group, especially regarding frailty. Our aim was to profile a large cohort of adults in receipt of low level home help and to determine the prevalence of frailty. METHODS: A total 1312 older adults, (≥ 65 years) in receipt of low level home help (< 5 h per week) were reviewed by community nurses and frailty was assessed using the Clinical Frailty Scale (CFS) in this cross-sectional study. Characteristics of the group were compared between males and females and prevalence of frailty was reported according to gender and principal care. Associations between frailty and a number of variables were explored using bivariate and regression analysis. RESULTS: The cohort of low level home-help users was a mean age of 82.1 (SD 7.3) years, predominantly female (70.6%) and over half (69.2%) lived alone. The prevalence of frailty in this population was 41.5%, with subjects primarily considered mildly (23.2%) or moderately frail (14.5%) by the CFS. A further 38.4% were classed as vulnerable. The degree of frailty did not differ significantly across the younger categories aged 65-84 years. However, in the oldest age groups, namely 90-94 and >95 years, moderate frailty was significantly higher relative to the younger groups (21% and 34%, p < 0.05, p < 0.01 respectively). Home help hours significantly correlated with frailty (rs = 0.371, p < 0.001) and functional dependency (rs = 0.609, p < 0.001), but only weakly with age (rs = 0.101, p = 0.034). Based on regression analysis, determinants of frailty included greater dependency (Barthel score), higher home help hours, non-self-caring and communication difficulty, all of which significantly contributed to the model, with a r squared value of 0.508. CONCLUSION: A high prevalence of frailty (41.5%) was documented in this population which associated with higher home help utilisation. Frailty was associated with greater functional dependency, but not strongly with chronological age, until after 90 years. These findings highlight opportunities for developing intervention strategies targeted at ageing in place among home help users.

Tissue-specific transcriptome sequencing analysis expands the non-human primate reference transcriptome resource (NHPRTR).

The non-human primate reference transcriptome resource (NHPRTR, available online at http://nhprtr.org/) aims to generate comprehensive RNA-seq data from a wide variety of non-human primates (NHPs), from lemurs to hominids. In the 2012 Phase I of the NHPRTR project, 19 billion fragments or 3.8 terabases of transcriptome sequences were collected from pools of ∼ 20 tissues in 15 species and subspecies. Here we describe a major expansion of NHPRTR by adding 10.1 billion fragments of tissue-specific RNA-seq data. For this effort, we selected 11 of the original 15 NHP species and subspecies and constructed total RNA libraries for the same ∼ 15 tissues in each. The sequence quality is such that 88% of the reads align to human reference sequences, allowing us to compute the full list of expression abundance across all tissues for each species, using the reads mapped to human genes. This update also includes improved transcript annotations derived from RNA-seq data for rhesus and cynomolgus macaques, two of the most commonly used NHP models and additional RNA-seq data compiled from related projects. Together, these comprehensive reference transcriptomes from multiple primates serve as a valuable community resource for genome annotation, gene dynamics and comparative functional analysis.

Calcitonin gene-related peptide in the joint: contributions to pain and inflammation.

Arthritis is the commonest cause of disabling chronic pain, and both osteoarthritis (OA) and rheumatoid arthritis (RA) remain major burdens on both individuals and society. Peripheral release of calcitonin gene-related peptide (CGRP) contributes to the vasodilation of acute neurogenic inflammation. Contributions of CGRP to the pain and inflammation of chronic arthritis, however, are only recently being elucidated. Animal models of arthritis are revealing the molecular and pathophysiological events that accompany and lead to progression of both arthritis and pain. Peripheral actions of CGRP in the joint might contribute to both inflammation and joint afferent sensitization. CGRP and its specific receptors are expressed in joint afferents and up-regulated following arthritis induction. Peripheral CGRP release results in activation of synovial vascular cells, through which acute vasodilatation is followed by endothelial cell proliferation and angiogenesis, key features of chronic inflammation. Local administration of CGRP to the knee also increases mechanosensitivity of joint afferents, mimicking peripheral sensitization seen in arthritic joints. Increased mechanosensitivity in OA knees and pain behaviour can be reduced by peripherally acting CGRP receptor antagonists. Effects of CGRP pathway blockade on arthritic joint afferents, but not in normal joints, suggest contributions to sensitization rather than normal joint nociception. CGRP therefore might make key contributions to the transition from normal to persistent synovitis, and the progression from nociception to sensitization. Targeting CGRP or its receptors within joint tissues to prevent these undesirable transitions during early arthritis, or suppress them in established disease, might prevent persistent inflammation and relieve arthritis pain.

The non-human primate reference transcriptome resource (NHPRTR) for comparative functional genomics.

RNA-based next-generation sequencing (RNA-Seq) provides a tremendous amount of new information regarding gene and transcript structure, expression and regulation. This is particularly true for non-coding RNAs where whole transcriptome analyses have revealed that the much of the genome is transcribed and that many non-coding transcripts have widespread functionality. However, uniform resources for raw, cleaned and processed RNA-Seq data are sparse for most organisms and this is especially true for non-human primates (NHPs). Here, we describe a large-scale RNA-Seq data and analysis infrastructure, the NHP reference transcriptome resource (http://nhprtr.org); it presently hosts data from12 species of primates, to be expanded to 15 species/subspecies spanning great apes, old world monkeys, new world monkeys and prosimians. Data are collected for each species using pools of RNA from comparable tissues. We provide data access in advance of its deposition at NCBI, as well as browsable tracks of alignments against the human genome using the UCSC genome browser. This resource will continue to host additional RNA-Seq data, alignments and assemblies as they are generated over the coming years and provide a key resource for the annotation of NHP genomes as well as informing primate studies on evolution, reproduction, infection, immunity and pharmacology.

Cytokine systems approach demonstrates differences in innate and pro-inflammatory host responses between genetically distinct MERS-CoV isolates.

BACKGROUND: The recent emergence of a novel coronavirus in the Middle East (designated MERS-CoV) is a reminder of the zoonotic and pathogenic potential of emerging coronaviruses in humans. Clinical features of Middle East respiratory syndrome (MERS) include atypical pneumonia and progressive respiratory failure that is highly reminiscent of severe acute respiratory syndrome (SARS) caused by SARS-CoV. The host response is a key component of highly pathogenic respiratory virus infection. Here, we computationally analyzed gene expression changes in a human airway epithelial cell line infected with two genetically distinct MERS-CoV strains obtained from human patients, MERS-CoV SA 1 and MERS-CoV Eng 1. RESULTS: Using topological techniques, including persistence homology and filtered clustering, we performed a comparative transcriptional analysis of human Calu-3 cell host responses to the different MERS-CoV strains, with MERS-CoV Eng 1 inducing early kinetic changes, between 3 and 12 hours post infection, compared to MERS-CoV SA 1. Robust transcriptional changes distinguished the two MERS-CoV strains predominantly at the late time points. Combining statistical analysis of infection and cytokine-stimulated Calu-3 transcriptomics, we identified differential innate responses, including up-regulation of extracellular remodeling genes following MERS-CoV Eng 1 infection and differential pro-inflammatory responses. CONCLUSIONS: Through our genomics-based approach, we found topological differences in the kinetics and magnitude of the host response to MERS-CoV SA 1 and MERS-CoV Eng 1, with differential expression of innate immune and pro-inflammatory responsive genes as a result of IFN, TNF and IL-1α signaling. Predicted activation for STAT3 mediating gene expression relevant for epithelial cell-to-cell adherens and junction signaling in MERS-CoV Eng 1 infection suggest that these transcriptional differences may be the result of amino acid differences in viral proteins known to modulate innate immunity during MERS-CoV infection.

Tactile localization accuracy at the low back.

Localizing tactile stimulation is an important capability for everyday function and may be impaired in people with persistent pain. This study sought to provide a detailed description of lumbar spine tactile localization accuracy in healthy individuals. Sixty-nine healthy participants estimated where they were touched at nine different points, labelled in a 3 × 3 grid over the lumbar spine. Mislocalization between the perceived and actual stimulus was calculated in horizontal (x) and vertical (y) directions, and a derived hypotenuse (c) mislocalization was calculated to represent the direct distance between perceived and actual points. In the horizontal direction, midline sites had the smallest mislocalization. Participants exhibited greater mislocalization for left- and right-sided sites, perceiving sites more laterally than they actually were. For all vertical values, stimulated sites were perceived lower than reality. A greater inaccuracy was observed in the vertical direction. This study measured tactile localization for the low back utilizing a novel testing method. The large inaccuracies point to a possible distortion in the underlying perceptual maps informing the superficial schema; however, further testing comparing this novel method with an established tactile localization task, such as the point-to-point method, is suggested to confirm these findings.

Increased viral loads and exacerbated innate host responses in aged macaques infected with the 2009 pandemic H1N1 influenza A virus.

In contrast to seasonal influenza virus infections, which typically cause significant morbidity and mortality in the elderly, the 2009 H1N1 virus caused severe infection in young adults. This phenomenon was attributed to the presence of cross-protective antibodies acquired by older individuals during previous exposures to H1N1 viruses. However, this hypothesis could not be empirically tested. To address this question, we compared viral replication and the development of the immune response in naïve young adult and aged female rhesus macaques infected with A/California/04/2009 H1N1 (CA04) virus. We show higher viral loads in the bronchoalveolar lavage (BAL) fluid and nasal and ocular swabs in aged animals, suggesting increased viral replication in both the lower and upper respiratory tracts. T cell proliferation was higher in the BAL fluid but delayed and reduced in peripheral blood in aged animals. This delay in proliferation correlated with a reduced frequency of effector CD4 T cells in old animals. Aged animals also mobilized inflammatory cytokines to higher levels in the BAL fluid. Finally, we compared changes in gene expression using microarray analysis of BAL fluid samples. Our analyses revealed that the largest difference in host response between aged and young adult animals was detected at day 4 postinfection, with a significantly higher induction of genes associated with inflammation and the innate immune response in aged animals. Overall, our data suggest that, in the absence of preexisting antibodies, CA04 infection in aged macaques is associated with changes in innate and adaptive immune responses that were shown to correlate with increased disease severity in other respiratory disease models.

Functional genomics reveals the induction of inflammatory response and metalloproteinase gene expression during lethal Ebola virus infection.

Ebola virus is the etiologic agent of a lethal hemorrhagic fever in humans and nonhuman primates with mortality rates of up to 90%. Previous studies with Zaire Ebola virus (ZEBOV), mouse-adapted virus (MA-ZEBOV), and mutant viruses (ZEBOV-NP(ma), ZEBOV-VP24(ma), and ZEBOV-NP/VP24(ma)) allowed us to identify the mutations in viral protein 24 (VP24) and nucleoprotein (NP) responsible for acquisition of high virulence in mice. To elucidate specific molecular signatures associated with lethality, we compared global gene expression profiles in spleen samples from mice infected with these viruses and performed an extensive functional analysis. Our analysis showed that the lethal viruses (MA-ZEBOV and ZEBOV-NP/VP24(ma)) elicited a strong expression of genes 72 h after infection. In addition, we found that although the host transcriptional response to ZEBOV-VP24(ma) was nearly the same as that to ZEBOV-NP/VP24(ma), the contribution of a mutation in the NP gene was required for a lethal phenotype. Further analysis indicated that one of the most relevant biological functions differentially regulated by the lethal viruses was the inflammatory response, as was the induction of specific metalloproteinases, which were present in our newly identify functional network that was associated with Ebola virus lethality. Our results suggest that this dysregulated proinflammatory response increased the severity of disease. Consequently, the newly discovered molecular signature could be used as the starting point for the development of new drugs and therapeutics. To our knowledge, this is the first study that clearly defines unique molecular signatures associated with Ebola virus lethality.

Exploring Workplace Testing with Real-Time Polymerase Chain Reaction SARS-CoV-2 Testing.

BACKGROUND: Molecular tests (ie, real-time polymerase chain reaction [RT-PCR]) and antigen tests are used to detect SARS-CoV-2. RT-PCR tests are generally considered to be the standard for clinical diagnosis of SARS-CoV-2 due to accuracy and reliability but can take longer to return results than antigen tests. Our aim was to examine if point-of-care (POC) testing for SARS-CoV-2 infection would provide a flexible resource to help achieve workplace safety. We compared test results and time-to-test results between a POC RT-PCR test and a send-out PCR test in a program implemented in summer 2020. RESULTS: POC testing shortened the time to results to 110 minutes in the POC setting from the 754 minutes for send-out tests. The specificity of POC RT-PCR single POC testing was 98.7% compared with send-out RT-PCR testing and was confirmed at 99.8% in a validation analysis. The sensitivity of the POC testing was 100% compared with send-out RT-PCR, although in a validation analysis, sensitivity appeared as 0% because only the 12 positive or indeterminate samples on the first analysis were retested and the majority were false-positives that were correctly ruled out. CONCLUSIONS: POC testing for SARS-CoV-2 with RT-PCR technology is possible at reduced time compared with send-out PCR testing.

Compliance with ECT NICE guidance by the John Connolly ECT clinic: January 2010 - July 2010.

OBJECTIVES: To review current practice at the John Connolly Wing ECT clinic and to explore compliance with NICE ECT guidance. Standards used included the ECT TA59 guidelines of 2003 with the updated depression guidance CG90 of 2009. To recommend a programme of action to the Trust which would ensure that clinical practice and service delivery within the Trust complies with NICE guidance. METHOD: A retrospective baseline Trust wide audit was conducted between the period of January 2010 to July 2010 inclusive. Cases were identified using ECT clinic record then computer Rio notes explored for evidence of compliance with NICE guidelines as set out in the audit standards. All data was extracted from the case notes on the Rio system. An audit tool was completed for each case. The data recorded on the audit tool was explored and entered onto an Excel spreadsheet for analysis. RESULTS: A total of 14 patients were identified. Of these, 6 were male and 8 were female. They comprised of 8 inpatients and 6 outpatients. The majority of patients had a diagnosis a severe depressive episode. 13 patients received bilateral ECT. In 1 case the first 3 sessions were unilateral and the rest were bilateral due to patient choice. 9 patients consented to ECT; 5 lacked capacity to consent and 1 of those was treated under Section 62 of the Mental Health Act. The number of treatments ranged from 0-15 with an average number of 7. This included 1 patient who did not receive ECT at all due to concerns raised by anaesthetist once at the ECT clinic. Reasons for stopping ECT included a response being achieved in 5 patients; anaesthetic risk in 3; withdrawal of consent in 2; T6 no longer valid in 1; no reason documented in 3 patients. Compliance with NICE guidelines was particularly good regarding the indications for ECT. An adequate trial of treatment was evidenced prior to consideration of ECT. Documentation of the exploration of the risk to benefit ratio both amongst the team and with the patient was poor. Assessment of the patient after each ECT and on-going cognitive assessment was poor. CONCLUSION: This audit highlights the need for sound documentation of our practice. It also stresses the need for further clarity regarding the roles and responsibilities of the RMO and their team and the ECT team. RECOMMENDATIONS: An ECT Care Pathway document has been produced to improve compliance with NICE guidance and improve documentation of practice. This document has been introduced for use in the Trust. We plan to re-audit for improvement in compliance.

Ethical issues in using ambient intelligence in health-care settings.

Ambient intelligence is increasingly finding applications in health-care settings, such as helping to ensure clinician and patient safety by monitoring staff compliance with clinical best practices or relieving staff of burdensome documentation tasks. Ambient intelligence involves using contactless sensors and contact-based wearable devices embedded in health-care settings to collect data (eg, imaging data of physical spaces, audio data, or body temperature), coupled with machine learning algorithms to efficiently and effectively interpret these data. Despite the promise of ambient intelligence to improve quality of care, the continuous collection of large amounts of sensor data in health-care settings presents ethical challenges, particularly in terms of privacy, data management, bias and fairness, and informed consent. Navigating these ethical issues is crucial not only for the success of individual uses, but for acceptance of the field as a whole.

Impact of COVID-19 on Primary Care Practice Sites and Their Vulnerable Patients.

Purpose: While COVID-19 has upended lives, it has also catalyzed innovation with potential to advance health delivery. Yet, we know little about how the delivery system, and primary care in particular, has responded and how this has impacted vulnerable patients. We aimed to understand the impact of COVID-19 on primary care practice sites and their vulnerable patients and to identify explanations for variation. Approach: We developed and administered a survey to practice managers and physician leaders from 173 primary care practice sites, October-November 2020. We report and graphically depict results from univariate analysis and examine potential explanations for variation in practices' process innovations in response to COVID-19 by assessing bivariate relationships between seven dependent variables and four independent variables. Findings: Among 96 (55.5%) respondents, primary care practice sites on average took more safety (8.5 of 12) than financial (2.5 of 17) precautions in response to COVID-19. Practice sites varied in their efforts to protect patients with vulnerabilities, providing care initially postponed, and experience with virtual visits. Financial risk, practice size, practitioner age, and emergency preparedness explained variation in primary care practices' process innovations. Many practice sites plan to sustain virtual visits, dependent mostly on patient and provider preference and continued reimbursement. Value: While findings indicate rapid and substantial innovation, conditions must enable primary care practice sites to build on and sustain innovations, to support care for vulnerable populations, including those with multiple chronic conditions and socio-economic barriers to health, and to prepare primary care for future emergencies.

The use of metal-catalyzed oxidation to suppress background staining caused by marker amplification reagents and the effects of this oxidation on the stability of antibody-antigen complexes in immunohistochemistry.

Marker amplification is a powerful technique for visualizing immunohistochemically deposited markers that otherwise would be invisible. Amplification usually is achieved with physical developers, which are solutions that contain a source of silver(I) plus a reducing agent. When the marker is present in extremely small quantities, prolonged incubation in the developer is required and unwanted background staining in the form of type III argyrophilia becomes problematic. Suppression of type III argyrophilia can be achieved by metal-catalyzed oxidation using the copper/H2O2 system, which normally is applied immediately prior to amplification. Because there is no reason, in principle, why metal-catalyzed oxidation should not be employed at earlier stages in the immunohistochemical staining procedure, we investigated whether earlier oxidation might confer any advantages over the traditional methodology. Immunocolloidal gold combined with two light insensitive physical developers was chosen as the model system, because visualization by light microscopy requires extended periods in the developers. Moreover, the system does not suffer from problems concerning endogenous enzyme- or non-enzyme-catalyzed marker deposition. Applying metal-catalyzed oxidation at each stage of the immunohistochemical procedure revealed that the technique could be employed successfully prior to staining, but not following the primary or secondary antibodies. In the latter cases, specific immunolocalization was lost entirely and only generalized nonspecific staining was seen. A limited investigation into the mechanism of metal-catalyzed oxidation of aldehyde fixed tissue sections suggested that it involved the formation of aldehyde groups. We suggest that the application of metal-catalyzed oxidation prior to immunohistochemical staining would have the advantages of both suppressing type III argyrophilia and inhibiting unwanted endogenous peroxidase activity. We also suggest that metal-catalyzed oxidation might reduce the affinity of tissue for other transition metals, such as copper, whose potential for improving marker amplification techniques has been demonstrated previously in dot-blot model systems.

Inflammation reduces mechanical thresholds in a population of transient receptor potential channel A1-expressing nociceptors in the rat.

Inflammatory hypersensitivity is characterized by behavioural reductions in withdrawal thresholds to noxious stimuli. Although cutaneous primary afferent neurones are known to have lowered thermal thresholds in inflammation, whether their mechanical thresholds are altered remains controversial. The transient receptor potential channel A1 (TRPA1) is a receptor localized to putative nociceptive neurones and is implicated in mechanical and thermal nociception. Herein, we examined changes in the properties of single primary afferents in normal and acutely inflamed rats and determined whether specific nociceptive properties, particularly mechanical thresholds, are altered in the subpopulation of afferents that responded to the TRPA1 agonist cinnamaldehyde (TRPA1-positive afferents). TRPA1-positive afferents in normal animals belonged to the mechanonociceptive populations, many of which also responded to heat or capsaicin but only a few of which responded to cold. In acute inflammation, a greater proportion of afferents responded to cinnamaldehyde and an increased proportion of dorsal root ganglion neurones expressed TRPA1 protein. Functionally, in inflammation, TRPA1-positive afferents showed significantly reduced mechanical thresholds and enhanced activity to agonist stimulation. Inflammation altered thermal thresholds in both TRPA1-positive and TRPA1-negative afferents. Our data show that a subset of afferents is sensitized to mechanical stimulation by inflammation and that these afferents are defined by expression of TRPA1.

Peripheral cannabinoid CB1 receptors inhibit evoked responses of nociceptive neurones in vivo.

We investigated the effect of peripheral administration of a selective cannabinoid CB1 receptor agonist arachidonyl-2-choroethylamide (ACEA), on evoked responses of primary afferents in vivo. Extracellular recordings were made from filaments of the saphenous nerve that responded to noxious mechanical stimulation of their receptive fields and effects of ACEA (30 and 50 microg/100 microl, i.a.) were studied. ACEA significantly inhibited evoked responses, effects that were blocked by co-administration of the cannabinoid CB1 receptor antagonist AM251 (30 microg/100 microl). These results demonstrate a cannabinoid CB1 receptor-mediated inhibition of primary afferent nociceptor excitability and provide further support for a peripheral site of action of cannabinoids.