Expression of semaphorin-3A in the joint and role in osteoarthritis.

Osteoarthritis (OA) is characterised by the deterioration of cartilage in the joints and pain. We hypothesise that semaphorin-3A (sema-3A), a chemorepellent for sensory nerves, plays a role in joint degradation and pain. We used the mechanical joint loading (MJL) model of OA to investigate sema-3A expression in the joint and examine its association with the development of OA and pain. We also analyse its effect on chondrocyte differentiation using the ATDC5 cell line. We demonstrate that sema-3A is present in most tissues in the healthy joint and its expression increases in highly innervated tissues, such as cruciate ligaments, synovial lining and subchondral bone, in loaded compared to nonloaded control joints. In contrast, sema-3A expression in cartilage was decreased in the severe OA induced by the application of high loads. There was a significant increase in circulating sema-3A, 6 weeks after MJL compared to the nonloaded mice. mRNA for sema-3A and its receptor Plexin A1 were upregulated in the dorsal root ganglia of mice submitted to MJL. These increases were supressed by zoledronate, an inhibitor of bone pain. Sema-3A was expressed at all stages of Chondrocyte maturation and, when added exogenously, stimulated expression of markers of chondrocyte differentiation. This indicates that sema-3A could affect joint tissues distinctively during the development of OA. In highly innervated joint tissues, sema-3A could control innervation and/or induce pain-associated neuronal changes. In cartilage, sema-3A could favour its degeneration by modifying chondrocyte differentiation.

An update of murine models and their methodologies in immune-mediated joint damage and pain research.

Murine models have played an indispensable role in the understanding of rheumatic and musculoskeletal disorders (RMD), elucidating the genetic, endocrine and biomechanical pathways involved in joint pathology and associated pain. To date, the available models in RMD can be classified as induced or spontaneous, both incorporating transgenic alternatives that improve specific insights. It is worth noting that the selection of the most appropriate model together with the evaluation of their specific characteristics and technical capabilities are crucial when designing the experiments. Furthermore, it is also imperative to consistently adhere to the ethical standards concerning animal experimentation. Recognizing the inherent limitation that any model can entirely encapsulates the complexity of the pathophysiology of these conditions, the aim of this review is to provide an updated overview on the methodology of current murine models in major arthropathies and their immune-mediated pathways, addressing to basic, translational and pharmacological research in joint damage and pain.

The Measure of an Outcome: Comparing Norming and Stacking to Benchmark the Effectiveness of Brain Injury Rehabilitation Services.

Practitioners have a clinical, ethical, academic, and economic responsibility to dispassionately consider how effective their services are. Approaches to measure how "good" or "bad" healthcare is include clinical audit, satisfaction surveys, and routine outcome measurement. However, the process of comparing the clinical outcomes of a specific service against the 'best' services in the same specialism, also known as benchmarking, remains challenging, and it is unclear how it affects quality improvement. This paper piloted and compared two different approaches to benchmarking to assess clinical outcomes in neurorehabilitation. Norming involved comparing routine measures of clinical outcome with external validators. Stacking involved pooling and comparing internal data across several years. The analyses of routine clinical outcome data from 167 patients revealed significant differences in the patient characteristics of those admitted to the same service provider over time, but no differences in outcomes achieved when comparing with historical data or with external reference data. These findings illustrate the potential advantages and limitations of using stacking and norming to benchmark clinical outcomes, and how the results from each approach might be used to evaluate service effectiveness and inform quality improvement within the field of brain injury rehabilitation.

What do we know about pseudodementia?

Depression and dementia can lead to generalised cognitive and memory dysfunction. Thus, differentiating these disorders is important and challenging. Pseudodementia is a term used clinically to describe symptoms that resemble dementia but are caused by other conditions (most frequently depression), rather than being recognised as an official diagnosis. Pseudodementia is characterised by a cognitive impairment that mimics dementia but which does not have its origin in neurological degeneration, deriving instead from functional psychiatric conditions. This condition is more commonly observed in older adults (particularly those over the age of 50 or 60 years), and its risk factors overlap with those for depression. Pseudodementia is essentially characterised by deficits in memory, executive function and speech and, therefore, can easily be confused with dementia, although there are aspects that allow its differentiation. Diagnosing pseudodementia can be difficult, especially as there is significant overlap between its symptoms and those of other conditions. However, it is important to recognise characteristic aspects of this disorder, as its correct identification is essential for proper treatment.

Enhanced meristem development, tolerance to oxidative stress and hyposensitivity to nitric oxide in the hypermorphic vq10-H mutant in AtVQ10 gene.

Nitric oxide (NO) and reactive oxygen are common factors in multiple plant responses to stress, and their involvement in hypoxia-triggered responses is key to ensure growth under adverse environmental conditions. Here, we analyse the regulatory functions exerted by hypoxia-, NO- and oxidative stress-inducible Arabidopsis gene coding for the VQ motif-containing protein 10 (VQ10). A hypermorphic vq10-H mutant allowed identifying VQ10-exerted regulation on root and shoot development as well as its role in regulating responses to NO and oxidative stress. Enhanced VQ10 expression in vq10-H plants led to enhanced elongation of the primary root, and increased root cell division and meristem size during early postgermination development. In shoots, VQ10 activation of cell division was counteracted by WRKY33-exerted repression, thus leading to a dwarf bushy phenotype in plants with enhanced VQ10 expression in a wrky33 knock-out background. Low number of differentially expressed genes were identified when vq10-H versus Col-0 plants were compared either under normoxia or hypoxia. vq10-H and VQ10ox plants displayed less tolerance to submergence but, in turn, were more tolerant to oxidative stress and less sensitive to NO than wild-type plants. VQ10 could be a node integrating redox-related regulation on development and stress responses.

Evaluation of residual antibacterial effects on canine skin surface and hair following treatment with five commercial mousse products against Staphylococcus pseudintermedius.

BACKGROUND: Antibacterial effect studies of commercial antiseptics typically have evaluated hair and not the skin. OBJECTIVES: To evaluate the antibacterial effects of mousse products on both canine skin and hair. ANIMALS: Fifteen short-haired and eight long-haired dogs without skin disease. MATERIALS AND METHODS: Five mousses were applied once: (1) 2% chlorhexidine and 2% miconazole; (2) 0.05% phytosphingosine; (3) 2% salicylic acid and 10% ethyl lactate; (4) 3% chlorhexidine and 0.5% climbazole; and (5) 2% chlorhexidine and 1% ketoconazole. Skin swabs and hair were collected from application sites before treatment, and at 1 h and at Day (D)2, D4, D8, D10 and D14 post-treatment. Skin swabs and hair were placed on Mueller-Hinton plates inoculated with Staphylococcus pseudintermedius inoculum suspension. Inhibition zones were measured after incubation. RESULTS: Inhibition was not noted with mousses 2 and 3. In mousse 5, inhibition zone sizes produced by swabs from long- and short-haired dogs were not significantly different (p = 0.105), and all swabs and hair produced inhibition until D14, regardless of hair length. By contrast, in mousse 1, inhibition zones produced by swabs from long-haired dogs were smaller than those from short-haired dogs (p < 0.001), and swabs from long-haired dogs produced a shorter duration of bacterial inhibition than hair. CONCLUSIONS AND CLINICAL RELEVANCE: The antibacterial effects of mousse 5 were not affected by hair length. Hair may be acceptable for evaluating effects on the skin in short-haired dogs. However, long hair may interfere with product distribution and duration of bacterial inhibition. Therefore, the evaluation of hair alone may overestimate clinically relevant antibacterial effects.

Protecting Police Officers Against Burnout: Overcoming a Fragmented Research Field.

This study aims to identify the determinants of burnout in police officers. We considered a wide range of psychosocial risk factors, individual variables that have been previously found to be associated with burnout in police officers (affective and cognitive empathy, self-care), and variables whose unique impact on burnout of police officers needs further clarification (organizational justice and organizational identification). The study was conducted in Portugal, and the sample was constituted by 573 members of the National Republican Guard (GNR-Guarda Nacional Republicana). The participants were invited to answer an online anonymous survey, which included previously validated measures of the following variables: burnout (exhaustion and disengagement), psychosocial risk factors, self-care, empathy (cognitive and affective), organizational justice, and organizational identification. Furthermore, we controlled for the potential impact of demographic variables (age, gender, years of professional experience, religiosity, political orientation, and income). Multiple regression analysis showed that when taken together, only a few of the variables associated with burnout had a unique impact on both exhaustion and disengagement: quantitative demands and affective empathy were burnout risk factors; meaningful work, organizational justice (distributive justice, procedural justice, and interactional justice), and organizational identification were burnout protective factors. Our results highlight the importance of developing theoretical models and planning interventions to prevent burnout in police officers, focusing mainly on the above-mentioned variables.

Subclinical Hypothyroidism is Not Associated with Femoral Osteoporosis in Individuals Aged 50 Years or Older.

BACKGROUND: Thyroid dysfunction and osteoporosis are conditions strongly associated with aging, and the prevalence of both conditions is expected to increase in the coming decades. Thyroid hormones regulate bone metabolism, and the role of subclinical hypothyroidism on bone mineral density (BMD) is still controversial. Hence, this study aims to assess the association of subclinical hypothyroidism with femoral osteopenia and osteoporosis in individuals aged 50 years or older. METHODOLOGY: This retrospective cohort study was carried out with 864 outpatients having at least one result for TSH levels before the first record of dual-energy X-ray absorptiometry (DXA). The primary endpoints were osteopenia (-2.5 standard deviation (SD)

Nutritional Assessment and Support in Children with Chronic Kidney Disease: The Benefits of Working with a Registered Dietitian.

BACKGROUND: An unbalanced dietary pattern, characterized by high animal protein content: may worsen metabolic control, accelerate renal deterioration and consequently aggravate the stage of the chronic kidney disease (CKD) in pediatric patients with this condition. AIM: to assess the effect of a registered dietitian (RD) intervention on the CKD children's eating habits. METHODS: Anthropometric and dietetic parameters, obtained at baseline and 12 months after implementing healthy eating and nutrition education sessions, were compared in 16 patients (50% girls) of 8.1 (1-15) years. On each occasion, anthropometry, 3-day food records and a food consumption frequency questionnaire were carried out. The corresponding relative intake of macro- and micronutrients was contrasted with the current advice by the European Food Safety Authority (EFSA) and with consumption data obtained using the Spanish dietary guidelines. Student's paired t-test, Wilcoxon test and Mc Nemar test were used. RESULTS: At Baseline 6% were overweight, 69% were of normal weight and 25% were underweight. Their diets were imbalanced in macronutrient composition. Following nutritional education and dietary intervention 63%, 75% and 56% met the Dietary Reference Values requirements for fats, carbohydrates and fiber, respectively, but not significantly. CKD children decreased protein intake (p < 0.001), increased dietary fiber intake at the expense of plant-based foods consumption (p < 0.001) and a corresponding reduction in meat, dairy and processed food intake was noticed. There were no changes in the medical treatment followed or in the progression of the stages. CONCLUSIONS: RD-led nutrition intervention focused on good dieting is a compelling helpful therapeutic tool to improve diet quality in pediatric CKD patients.

Fungal Endocarditis: A Rare Case of Multiple Arterial Embolization.

Fungal endocarditis is a rare and fatal condition, with a mortality of up to 75%, affecting immunocompromised hosts with a predisposing condition, namely, a history of previous cardiac or noncardiac surgery. Embolization is frequent, accounting for 44% of cases, and as the most common site is the brain, it can cause leptomeningitis, parenchymal granulomas, or abscesses. This case report describes a man with aortic valve replacement one year ago and a recent carotid endarterectomy who was admitted with fever and neurological deficits. The workup permitted a diagnosis of fungal endocarditis, and the patient underwent a combined and aggressive treatment approach with antifungal therapy and surgery, with a successful replacement of the aortic valve. During hospitalization, the patient's neurological status deteriorated, and a cerebral abscess was discovered on the left frontal lobe. Despite the poor prognosis, the patient recovered slowly and was discharged from the hospital three months later. The present case highlights the high index of suspicion needed for the diagnosis and the need for a multidisciplinary team to approach these patients to achieve a positive outcome.

HOXA5 is a key regulator of class 3 semaphorins expression in the synovium of rheumatoid arthritis patients.

OBJECTIVE: Class 3 semaphorins are reduced in the synovial tissue of RA patients and these proteins are involved in the pathogenesis of the disease. The aim of this study was to identify the transcription factors involved in the expression of class 3 semaphorins in the synovium of RA patients. METHODS: Protein and mRNA expression in synovial tissue from RA and individuals at risk (IAR) patients, human umbilical vein endothelial cells (HUVEC) and RA fibroblast-like synoviocytes (FLS) was determined by ELISA, immunoblotting and quantitative PCR. TCF-3, EBF-1 and HOXA5 expression was knocked down using siRNA. Cell viability, migration and invasion were determined using MTT, calcein, wound closure and invasion assays, respectively. RESULTS: mRNA expression of all class 3 semaphorins was significantly lower in the synovium of RA compared with IAR patients. In silico analysis suggested TCF-3, EBF-1 and HOXA5 as transcription factors involved in the expression of these semaphorins. TCF-3, EBF-1 and HOXA5 silencing significantly reduced the expression of several class 3 semaphorin members in FLS and HUVEC. Importantly, HOXA5 expression was significantly reduced in the synovium of RA compared with IAR patients and was negatively correlated with clinical disease parameters. Additionally, TNF-α down-regulated the HOXA5 expression in FLS and HUVEC. Finally, HOXA5 silencing enhanced the migratory and invasive capacities of FLS and the viability of HUVEC. CONCLUSION: HOXA5 expression is reduced during the progression of RA and could be a novel therapeutic strategy for modulating the hyperplasia of the synovium, through the regulation of class 3 semaphorins expression.

Type I Interferons induce endothelial destabilization in Systemic Lupus Erythematosus in a Tie2-dependent manner.

Endothelial cell (EC) dysfunction is a hallmark of Systemic Lupus Erythematosus (SLE) and Tie2 is a receptor essential for vascular stability. Inflammatory processes promote inhibition of Tie2 homeostatic activation, driving vascular dysfunction. In this work we determined whether type I Interferons (IFN) induce Tie2 signalling-mediated endothelial dysfunction in patients with SLE. Serum levels of Angiopoietin (Ang)-1, Ang-2 and soluble (s)Tie1 in patients with SLE and healthy controls were measured by ELISA. Monocytes from patients with SLE and Human Umbilical Vein EC (HUVEC) were stimulated with IFN-α, IFN-ß (1000 I.U.) or SLE serum (20%). mRNA and protein expression, phosphorylation and translocation were determined by quantitative PCR, ELISA, Western Blot, flow cytometry and confocal microscopy. Viability and angiogenic capacity were determined by calcein and tube formation assays. We found that sTie1 and Ang-2 serum levels were increased and Ang-1 decreased in patients with SLE and were associated with clinical characteristics. Type I IFN significantly decreased Ang-1 and increased Ang-2 in monocytes from patients with SLE. Type I IFN increased sTie1 and Ang-2 secretion and reduced Tie2 activation in HUVEC. Functionally, type I IFN significantly reduced EC viability and impaired angiogenesis in a Tie2 signalling-dependent manner. Finally, SLE serum increased Ang-2 and sTie1 secretion and significantly decreased tube formation. Importantly, Tie1 and IFNAR1 knockdown reversed these effects in tube formation. Overall, type I IFN play an important role in the stability of EC by inhibiting Tie2 signalling, suggesting that these processes may be implicated in the cardiovascular events observed in patients with SLE.

Semaphorin3B promotes an anti-inflammatory and pro-resolving phenotype in macrophages from rheumatoid arthritis patients in a MerTK-dependent manner.

Previous works from our group show that Semaphorin3B (Sema3B) is reduced in RA and plays a protective role in a mouse arthritis model. In turn, MerTK plays a protective function in murine arthritis models, is expressed by synovial tissue macrophages and is linked to remission in patients with RA. In this study, we examined the role of Sema3B in the phenotypic characteristics of RA macrophages and the implication of MerTK. Peripheral blood monocytes from RA patients were differentiated into IFN-γ (RA MØIFN-γ) or M-CSF (RA MØM-CSF) macrophages and stimulated with LPS, Sema3B or their combination. Alternatively, RA fibroblast like synoviocytes (FLS) were stimulated with RA MØIFN-γ and RA MØM-CSF supernatants. Gene expression was determined by qPCR and protein expression and activation by flow cytometry, ELISA and western blot. Sema3B down-regulated the expression of pro-inflammatory mediators, in both RA MØIFN-γ and RA MØM-CSF. We observed a similar reduction in RA FLS stimulated with the supernatant of Sema3B-treated RA MØIFN-γ and RA MØM-CSF. Sema3B also modulated cell surface markers in macrophages towards an anti-inflammatory phenotype. Besides, MerTK expression and activation was up-regulated by Sema3B, just as GAS6 expression, Resolvin D1 secretion and the phagocytic activity of macrophages. Importantly, the inhibition of MerTK and neuropilins 1 and 2 abrogated the anti-inflammatory effect of Sema3B. Our data demonstrate that Sema3B modulates the macrophage characteristics in RA, inducing a skewing towards an anti-inflammatory/pro-resolving phenotype in a MerTK-dependant manner. Therefore, here we identify a new mechanism supporting the protective role of Sema3B in RA pathogenesis.

Mechanical regulation of the early stages of angiogenesis.

Favouring or thwarting the development of a vascular network is essential in fields as diverse as oncology, cardiovascular disease or tissue engineering. As a result, understanding and controlling angiogenesis has become a major scientific challenge. Mechanical factors play a fundamental role in angiogenesis and can potentially be exploited for optimizing the architecture of the resulting vascular network. Largely focusing on in vitro systems but also supported by some in vivo evidence, the aim of this Highlight Review is dual. First, we describe the current knowledge with particular focus on the effects of fluid and solid mechanical stimuli on the early stages of the angiogenic process, most notably the destabilization of existing vessels and the initiation and elongation of new vessels. Second, we explore inherent difficulties in the field and propose future perspectives on the use of in vitro and physics-based modelling to overcome these difficulties.

A neurobehavioral therapy approach to the rehabilitation and support of persons with brain injury: Practice-based evidence from a UK charitable rehabilitation provider.

Background: The treatment and rehabilitation for people with acquired brain injury is continually evolving, with increasing recognition of the importance of approaches that adopt a multi-disciplinary biopsychosocial perspective focused on improving adjustment, social participation, and wellbeing. However, there is significant variability as to how such approaches are delivered, across the various stages of recovery, rehabilitation settings, and within different healthcare systems. Objective: This paper had three aims. The first was to describe the neurobehavioral therapy (NBT) approach to brain injury rehabilitation adopted in our charitable organization. The second aim was to report how the NBT approach evolved in response to changes in referral patterns, and patient needs within a broader, longer-term clinical pathway. The third aim was to assess the effectiveness of the NBT approach by analyzing outcome data. Methods: Retrospective analyses of standardized outcome data were completed to investigate the effectiveness of our approach. Case vignettes are provided to illustrate the key components of the approach. Results: Outcome data suggested that the approach is effective in delivering positive outcomes for patients. Furthermore, the data show differences in presentation between three clinical streams (restoration, compensation, and scaffolding) within the NBT approach. Conclusions: This paper describes the adaption of the 'traditional' neurobehavioral approach to brain injury rehabilitation into a model of delivery that can benefit a more diverse range of people living with the heterogenous and long-term consequences of brain injury.

A 3D Digital Analysis of the Hard Palate Wound Healing after Free Gingival Graft Harvest: A Pilot Study in the Short Term.

Purpose: Within this context, this pilot study aimed to evaluate the healing dynamics process of the hard palate after free gingival graft harvesting in the short term (3 months), utilizing digital imaging technology and tridimensional analysis software. Furthermore, assessing the results found to verify the existence of a relationship between gender or age with tissue loss. Materials and Methods: For connective-tissue harvesting, fifteen patients with gingival recessions type (RT) 1 and RT2 were selected. On the surgery day (before the procedure) and after three months, palatal impressions were taken in all patients, and cast models were done for posterior model scanning. The following variables were analyzed: mean thickness alterations (x¯ TA), maximum thickness loss (MTL), mean maximum thickness loss (x¯ MTL), and volume alterations (VA). A descriptive and bivariate analysis of the data was done. The data were submitted for statistical evaluation and were significant if p < 0.05. Results: Fifteen patients were analyzed, 11 females (73.3%) and four males (26.7%). The patients' average age was 28 ± 8.52 years (ranging between 16 and 48 years old). The palatal wound region's mean thickness and volume changes were −0.26 mm (±0.31) and 46.99 mm3 (±47.47 mm3) at three months. There was no statistically significant result correlating age/gender with any variable evaluated. Conclusions: Connective tissue graft harvesting promoted changes with a standard volume and thickness loss of palatal soft tissue. A 3D digital evaluation was a non-invasive method with a reproducible technique for measuring thickness or volume after connective tissue is collected. There was no relationship between age/gender and any variables analyzed.

Nature and Mindfulness to Cope with Work-Related Stress: A Narrative Review.

In recent years, work-related stress has grown exponentially and the negative impact that this condition has on people's health is considerable. The effects of work-related stress can be distinguished in those that affect workers (e.g., depression and anxiety) and those that affect the company (e.g., absenteeism and productivity). It is possible to distinguish two types of prevention interventions. Individual interventions aim at promoting coping and individual resilience strategies with the aim of modifying cognitive assessments of the potential stressor, thus reducing its negative impact on health. Mindfulness techniques have been found to be effective stress management tools that are also useful in dealing with stressful events in the workplace. Organizational interventions modify the risk factors connected to the context and content of the work. It was found that a restorative workplace (i.e., with natural elements) reduces stress and fatigue, improving work performance. Furthermore, practicing mindfulness in nature helps to improve the feeling of wellbeing and to relieve stress. In this paper, we review the role of mindfulness-based practices and of contact with nature in coping with stressful situations at work, and we propose a model of coping with work-related stress by using mindfulness in nature-based practices.

Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.