Salivary biglycan-neo-epitope-BGN262: A novel surrogate biomarker for equine osteoarthritic sub-chondral bone sclerosis and to monitor the effect of short-term training and surface arena.

Objective: We aimed to delineate a novel soluble Biglycan Neo-epitope-BGN262 in saliva from young reference and osteoarthritic horses in conjunction with the influence of short-term training exercise, riding surface hardness, circadian rhythm, and feeding on its soluble levels. Design: A custom-made inhibition ELISA was used for the quantification of BGN262 in saliva. Cohort 1: A cross-sectional study comprising reference (N â€‹= â€‹19) and OA horses (N â€‹= â€‹9) with radiographically classified subchondral bone sclerosis. Receiver operating characteristic curve analysis was performed to evaluate the robustness of BGN262. Cohorts 2 (N â€‹= â€‹5) & 3 (N â€‹= â€‹7): Longitudinal studies of sampling during a short-term training exercise (sand-fibre) and a cross-over design of short-training exercise on 2 different riding arenas (sand and sand-fibre), respectively. Capillary western immunoassay was used to determine the BGN262 molecular size in a selection of saliva samples collected from cohort 1. Results: Cohort 1: Salivary BGN262 levels were significantly higher in the OA group. The Receiver operating characteristic curve analysis showed an area under the curve of 0.8304 [0.6386 to 1.022], indicating a good separation from the reference group. Cohorts 2 & 3: Salivary BGN262 levels significantly changed during the exercise on sand and sand-fibre arena, with a trend towards higher levels for sand-fibre. The size of the BGN262 fragment determined by Capillary western assay was 18 â€‹kDa. Conclusions: The data presented show saliva BGN262 levels as a novel biomarker in evaluating the influence of exercise, and interaction with riding arenas alongside assessing osteoarthritis severity.

Learning in lockdown: Using the COVID-19 crisis to teach children about food and climate change.

Food systems are significant sources of global greenhouse gas emissions (GHGE). Since emission intensity varies greatly between different foods, changing food choices towards those with lower GHGE could make an important contribution to mitigating climate change. Public engagement events offer an opportunity to communicate these multifaceted issues and raise awareness about the climate change impact of food choices. An interdisciplinary team of researchers was preparing food and climate change educational activities for summer 2020. However, the COVID-19 pandemic and lockdown disrupted these plans. In this paper, we report on shifting these events online over the month of June 2020. We discuss what we did and the reception to our online programme. We then reflect on and highlight issues that arose. These relate to: (1) the power dynamics of children, diet and climate change; (2) mental health, diet and COVID-19; (3) engaging the wider science, agriculture and food communities; (4) the benefits of being unfunded and the homemade nature of this programme; (5) the food system, STEAM (science, technology, engineering, arts and mathematics) and diversity; and (6) how our work fits into our ongoing journey of food and climate change education.

Profiling of donor-specific immune effector signatures in response to rituximab in a human whole blood loop assay using blood from CLL patients.

Rituximab is widely used in the treatment of haematological malignancies, including chronic lymphocytic leukaemia (CLL), the most common leukaemia in adults. However, some patients, especially those with high tumour burden, develop cytokine release syndrome (CRS). It is likely that more patients will develop therapy-linked CRS in the future due to the implementation of other immunotherapies, such as CAR T-cell, for many malignancies. Current methods for CRS risk assessment are limited, hence there is a need to develop new methods. To better recapitulate an in vivo setting, we implemented a unique human whole blood "loop" system to study patient-specific immune responses to rituximab in blood derived from CLL patients. Upon rituximab infusion, both complement-dependent cytotoxicity (CDC) and antibody-dependent cellular cytotoxicity (ADCC) profiles were evident in CLL patient blood, coincident with CLL cell depletion. Whereas B cell depletion is induced in healthy persons in the blood loop, only patients display B cell depletion coupled with CRS. With the exception of one donor who lacked NK cells, all other five patients displayed variable B cell depletion along with CRS profile. Additionally, inhibition of CDC or ADCC via either inhibitors or antibody Fc modification resulted in skewing of the immune killing mechanism consistent with published literature. Herein we have shown that the human whole blood loop model can be applied using blood from a specific indication to build a disease-specific CRS and immune activation profiling ex vivo system. Other therapeutic antibodies used for other indications may benefit from antibody characterization in a similar setting.

Development and performance of a biomimetic artificial perilymph for in vitro testing of medical devices.

OBJECTIVE: Cochlear implants interface with the fluid in the cochlea called perilymph. The volume of this fluid present in human and animal model cochlea is prohibitively low for isolation for in vitro studies. Thus, there is a need for an artificial perilymph that reflects the complexity of this fluid in terms of competitive protein adsorption. APPROACH: This study established a biomimetic artificial perilymph (BAP) comprising serum albumin, immunoglobulin G, transferrin, inter-alpha-trypsin inhibitor, apolipoprotein A1 and complement C3 to represent the major components of human perilymph. Adsorption of the BAP components to platinum was analysed. MAIN RESULTS: It was established that this six component BAP provided competitive and complex adsorption behaviours consistent with biologically derived complex fluids. Additionally, adsorption of the BAP components to platinum cochlear electrodes resulted in a change in polarisation impedance consistent with that observed for the cochlear device in vivo. SIGNIFICANCE: This study established a BAP fluid suitable for furthering the understanding of the implant environment for electroactive devices that interface with the biological environment.

Harnessing chondroitin sulphate in composite scaffolds to direct progenitor and stem cell function for tissue repair.

The development of bioscaffolds that incorporate chondroitin sulphate (CS) and their applications with progenitor and stem cells in cartilage, bone, cornea, skin, and neural repair are reviewed. CS is a heterogeneous structure due to the organisation of multiple CS disaccharide sulphation motifs, giving rise to a vast range of CS chain structures, and hence the wide range of biological activity. The incorporation of this biological molecule represents a significant advance in bioscaffold design and performance in tissue repair strategies. The intrinsic stem-cell directive properties of CS are covered in the context of tissue development, and the differing CS disaccharide motifs, referred to as the 'glyco-code'. These structural motifs contribute to stem cell proliferation and differentiation in the scaffold environment and improve outcomes in terms of tissue repair or regeneration worthy of future research.

Coherent normalization for in vivo measurements of gadolinium in bone.

OBJECTIVE: Recent evidence of gadolinium (Gd) deposition in bones of healthy individuals who have previously received Gd-based contrast agents (GBCAs) for MRI has led to a demand for in vivo measurement techniques. The technique of x-ray fluorescence provides a low risk and painless method to assess Gd deposition in bone, and has the potential to be a useful clinical tool. However, interpatient variability creates a challenge while performing in vivo measurements. APPROACH: We explored the use of coherent normalization, which involves normalizing the Gd K x-rays to the coherent scattered γ-ray from the excitation source, for bone Gd measurements through a series of phantom-based experiments and Monte Carlo simulations. MAIN RESULTS: We found coherent normalization is able to correct for variation in overlying tissue thickness over a wide range (0-12.2 mm). The Gd signal to coherent signal ratio is independent of tissue thickness for both experiments and Monte Carlo simulations. SIGNIFICANCE: Coherent normalization has been demonstrated to be used in practice with normal healthy adults to improve in vivo bone Gd measurements.

Risk factors for human-directed canine aggression in a referral level clinical population.

Risk factors for human-directed aggression were investigated using retrospective analysis of data from a referral-level clinical behaviour population in the UK. A sample of 200 cases involving human-directed canine aggression and 200 control cases involving no instance of human-directed aggression were selected at random from a population of 746 cases. The final model suggested that clinical cases with human-directed aggression were significantly younger than those presenting with other undesired behaviours (P=0.008) and that male dogs were 1.4 times more likely to be aggressive towards human beings than female dogs (P=0.019). Dogs were 1.7 times more likely to be aggressive towards people if they had attended more than five puppy classes than if they had never attended puppy class (P=0.015) and that dogs were 2.8 times more likely to be aggressive towards human beings if there was another dog between 0 months and 24 months of age in the home (P=0.004). These factors only account for 7 per cent to 10 per cent of the variance between the human-directed aggression population and the control population, but factors such as attendance at puppy classes and numbers of dogs in the household suggest the need for longitudinal studies to investigate temporal relationships.

Confirming improved detection of gadolinium in bone using in vivo XRF.

The safety of using Gd in MRI contrast agents has recently been questioned, due to recent evidence of the retention of Gd in individuals with healthy renal function. Bone has proven to be a storage site for Gd, as unusually high concentrations have been measured in femoral heads of patients undergoing hip replacement surgery, as well as in autopsy samples. All previous measurements of Gd in bone have been invasive and required the bone to be removed from the body. X-ray fluorescence (XRF) offers a non-invasive and non-destructive method for carrying out in vivo measurements of Gd in humans. An updated XRF system provides improved detection limits in a short measurement time of 30-min. A new four-detector system and higher activity Cd-109 excitation source of 5GBq results in minimum detection limits (MDLs) of 1.64-1.72µgGd/g plaster for an average overlaying tissue thickness of the tibia. These levels are well within the range of previous in vitro Gd measurements. Additional validation through comparison with ICP-MS measurements has confirmed the ability of the XRF system for detecting Gd further, proving it is a feasible system to carry out human measurements.

Pericellular colocalisation and interactive properties of type VI collagen and perlecan in the intervertebral disc.

The aim of this study was to immunolocalise type VI collagen and perlecan and determine their interactive properties in the intervertebral disc (IVD). Confocal laser scanning microscopy co-localised perlecan with type VI collagen as pericellular components of IVD cells and translamellar cross-bridges in ovine and murine IVDs. These cross-bridges were significantly less abundant in the heparin sulphate deficient Hspg2 exon 3 null mouse IVD than in wild type. This association of type VI collagen with elastic components provides clues as to its roles in conveying elastic recoil properties to annular tissues. Perlecan and type VI collagen were highly interactive in plasmon resonance studies. Pericellular colocalisation of perlecan and type VI collagen provides matrix stabilisation and cell-matrix communication which allows IVD cells to perceive and respond to perturbations in their biomechanical microenvironment. Perlecan, at the cell surface, provides an adhesive interface between the cell and its surrounding extracellular matrix. Elastic microfibrillar structures regulate tensional connective tissue development and function. The 2010 Global Burden of Disease study examined 291 disorders and identified disc degeneration and associated low back pain as the leading global musculoskeletal disorder emphasising its massive socioeconomic impact and the need for more effective treatment strategies. A greater understanding of how the IVD achieves its unique biomechanical functional properties is of great importance in the development of such therapeutic measures.

Inspection of thick welded joints using laser-ultrasonic SAFT.

The detection of defects in thick butt joints in the early phase of multi-pass arc welding would be very valuable to reduce cost and time in the necessity of reworking. As a non-contact method, the laser-ultrasonic technique (LUT) has the potential for the automated inspection of welds, ultimately online during manufacturing. In this study, testing has been carried out using LUT combined with the synthetic aperture focusing technique (SAFT) on 25 and 50mm thick butt welded joints of steel both completed and partially welded. EDM slits of 2 or 3mm height were inserted at different depths in the multi-pass welding process to simulate a lack of fusion. Line scans transverse to the weld are performed with the generation and detection laser spots superimposed directly on the surface of the weld bead. A CCD line camera is used to simultaneously acquire the surface profile for correction in the SAFT processing. All artificial defects but also real defects are visualized in the investigated thick butt weld specimens, either completed or partially welded after a given number of passes. The results obtained clearly show the potential of using the LUT with SAFT for the automated inspection of arc welds or hybrid laser-arc welds during manufacturing.

A phantom-based feasibility study for detection of gadolinium in bone in-vivo using X-ray fluorescence.

Gadolinium (Gd) based contrast agents have been commonly used over the past three decades to improve contrast in magnetic resonance imaging. These complexes, originally thought to be stable and clear from the body shortly after administration, have been shown to dissociate to a small extent and deposit in organs such as bone. A safe and non-invasive method for measuring Gd in bone is necessary for further exploring Gd retention in the body following the administration of a contrast agent. A feasibility study using a K x-ray fluorescence (K-XRF) system to measure Gd in human tibias was investigated. Bone phantoms mimicking human tibia were created with Gd concentrations ranging from 0 to 120ppm. The minimum detection limit (MDL) was calculated from 20-hour and 7-hour phantom measurements with a source activity of 0.11GBq. All MDL values were scaled to a more realistic measurement time of 30-minutes with a stronger source. Scaling arguments were based on activity ratio, measurement time, and system dead time. The MDL for a 1GBq source was estimated to be 3.60-3.64ppm, for an average range of tissue thicknesses overlaying a human tibia. For a stronger source of 5GBq and a four detector cloverleaf system, the MDL was estimated to be 1.49-1.52ppm. Determined and predicted MDLs are within the range of previous in-vitro Gd measurement data. The K-XRF system shows promising results for detecting Gd in bone and should be seriously considered for in-vivo measurements.

Emergence of tan spot disease caused by toxigenic Pyrenophora tritici-repentis in Australia is not associated with increased deployment of toxin-sensitive cultivars.

The wheat disease tan (or yellow leaf) spot, caused by Pyrenophora tritici-repentis, was first described in the period 1934 to 1941 in Canada, India, and the United States. It was first noted in Australia in 1953 and only became a serious disease in the 1970s. The emergence of this disease has recently been linked to the acquisition by P. tritici-repentis of the ToxA gene from the wheat leaf and glume blotch pathogen, Stagonospora nodorum. ToxA encodes a host-specific toxin that interacts with the product of the wheat gene Tsn1. Interaction of ToxA with the dominant allele of Tsn1 causes host necrosis. P. tritici-repentis races lacking ToxA give minor indistinct lesions on wheat lines, whereas wheat lines expressing the recessive tsn1 are significantly less susceptible to the disease. Although the emergence and spread of tan spot had been attributed to the adoption of minimum tillage practices, we wished to test the alternative idea that the planting of Tsn1 wheat lines may have contributed to the establishment of the pathogen in Australia. To do this, wheat cultivars released in Australia from 1911 to 1986 were tested for their sensitivity to ToxA. Prior to 1941, 16% of wheat cultivars were ToxA-insensitive and hence, all other factors being equal, would be more resistant to the disease. Surprisingly, only one of the cultivars released since 1940 was ToxA insensitive, and the area planted to ToxA-insensitive cultivars varied from 0 to a maximum of only 14% in New South Wales. Thus, the majority of the cultivars were ToxA-sensitive both before and during the period of emergence and spread of the disease. We therefore conclude that the spread of P. tritici-repentis in Australia cannot be causally linked to the deployment of ToxA-sensitive cultivars.

The effect of silica nanoparticulate coatings on serum protein adsorption and cellular response.

Serum protein adsorption on colloidal silica surfaces was investigated using a quartz crystal microbalance with dissipation (QCM-D) monitoring. The amount of serum proteins adsorbed on colloidal silica-coated surfaces was not significantly different from the control silica surfaces, with the exception of 21nm colloidal silica which experienced significantly less (P<0.05) fibrinogen adsorption compared with control silica. The adhesion and proliferation of human endothelial cells (C11STH) on nano-scale colloidal silica surfaces were significantly reduced compared with control silica surfaces, suggesting that the conformation of adsorbed proteins on the colloidal silica surfaces plays a role in modulating the amount of cell binding. Fibronectin is one of the main extracellular matrix proteins involved in endothelial cell attachment to biomaterial surfaces. There was reduced binding of a monoclonal anti-fibronectin antibody, that reacted specifically with the cell-binding fragment, to fibronectin-coated colloidal silica surfaces compared with control silica surfaces. This suggests that the fibronectin adsorbed on the colloidal silica-coated surfaces was conformationally changed compared with control silica reducing the availability of the cell-binding domain of fibronectin.

Treatment of invasive squamous cell carcinoma with 5-percent imiquimod cream.

Skin cancer is a major problem in the elderly. Squamous cell carcinoma (SCC), the second most common skin cancer, typically occurs in this age group. Despite a number of modalities readily available for treatment (depending on the tumor site, and depth of invasion) there remains the problem of individuals with multiple lesions who may be unsuitable for existing treatments for SCC, particularly surgery. Consequently, the search for novel treatments continues. To our knowledge, there are only 6 published reports of invasive SCC treated with 5-percent imiquimod cream.

Papulosquamous dermatitis. Syphilis: a forgotten disease?

We report a case of secondary syphilis in a 54-year-old, bisexual, HIV positive man. Although syphilis is a well-known sexually-transmitted infection, it is not widely seen nowadays. However, a possible diagnosis of syphilis should not be overlooked in any individual, irrespective of HIV status.

Pelvic floor muscle functioning in women with vulvar vestibulitis syndrome.

Vaginal sEMG biofeedback and pelvic floor physical therapists' manual techniques are being increasingly included in the treatment of vulvar vestibulitis syndrome (VVS). Successful treatment outcomes have generated hypotheses concerning the role of pelvic floor pathology in the etiology of VVS. However, no data on pelvic floor functioning in women with VVS compared to controls are available. Twenty-nine women with VVS were matched to 29 women with no pain with intercourse. Two independent, structured pelvic floor examinations were carried out by physical therapists blind to the diagnostic status of the participants. Results indicated that therapists reached almost perfect agreement in their diagnosis of pelvic floor pathology. A series of significant correlations demonstrated the reliability of assessment results across muscle palpation sites. Women with VVS demonstrated significantly more vaginal hypertonicity, lack of vaginal muscle strength, and restriction of the vaginal opening, compared to women with no pain with intercourse. Anal palpation could not confirm generalized hypertonicity of the pelvic floor. We suggest that pelvic floor pathology in women with VVS is reactive in nature and elicited with palpations that result in VVS-type pain. Treatment interventions need to recognize the critical importance of addressing the conditioned, protective muscle guarding response in women with VVS.

Effects of acupuncture and care interventions on the outcome of inflammatory symptoms of the breast in lactating women.

OBJECTIVES: To compare modes of care and treatment for lactating women with inflammatory symptoms of the breast, with special focus on the use of acupuncture. METHOD: Eighty-eight mothers were randomized into three treatment groups. All three groups were given advice regarding emptying of the breasts and care in the form of comfort interventions. Acupuncture was included in the treatment regime for two of the groups. A severity index was created by adding together scores for signs and symptoms: breast tension, erythema and pain. FINDINGS: Mothers in all groups expressed relative satisfaction with the breastfeeding situation despite considerable discomfort. There was no significant difference between the groups for the number of mothers requiring more than three contact days for recovery nor for their severity index scores on day 3. These findings must be interpreted with care but may suggest that care interventions play as great a part in the recovery of these women as acupuncture treatment or the use of oxytocin spray. Antibiotic therapy was used in 9% of the study population, which is in contrast to other studies. PROPOSED FUTURE ACTION: Based on these results, a new study has been designed to test the hypothesis that acupuncture hastens recovery from inflammatory processes in the lactating breast and approximately 200 mothers will be randomized in a new expanded study.

On-line measurement of texture, thickness and plastic strain ratio using laser-ultrasound resonance spectroscopy.

Laser-ultrasound resonance spectroscopy, a non-contact ultrasonic technique, was used to determine reliably and rapidly the crystallographic texture, the average plastic strain ratio, and the thickness of sheet metal on the production line. As with laser-ultrasonics, a short laser pulse is used to generate a wide-band pulse of ultrasound and a laser interferometer is used for its detection. In this paper, a large number of echoes are collected and analyzed together using Fourier techniques to measure the natural resonance frequencies in the thickness of the sheet. One longitudinal and two shear resonance frequencies were measured together with their harmonics. From these frequencies, two crystallographic orientation distribution coefficients, W(400) and W(420), are obtained, as well as a highly accurate measurement of the sheet thickness that is corrected for changes in ultrasonic velocity caused by texture variations. Using these coefficients, the average and in-plane twofold and fourfold variations of the plastic strain ratio, respectively r delta(2)r, and delta(4)r, can be evaluated. These parameters are indications of the formability of metals sheets, which is of industrial interest. Measurements on 1 mm thick, low carbon steel sheets have shown the following measurement accuracies: r to within +/-0.08, delta(2)r, and delta(4)r to within +/-0.1, and thickness to better than +/-1 microm. On-line tests at LTV Steel Company showed that the sensitivity of the apparatus is sufficient to detect systematic variations in texture along the length of similar production coils and that the on-line repeatability for r was of order +/-0.02.