Galectin-3 as a novel biomarker in cats with hypertrophic cardiomyopathy.

INTRODUCTION/OBJECTIVES: Galectin-3 (Gal-3) is a circulating biomarker of fibrosis. In humans, increased Gal-3 is predictive of myocardial fibrosis and adverse cardiac events. The aim of this study was to evaluate the potential for Gal-3 as a cardiac biomarker in cats with hypertrophic cardiomyopathy (HCM). MATERIALS AND METHODS: Eighty cats were enrolled (25 healthy cats with normal hearts, 35 with HCM American College of Veterinary Internal Medicine (ACVIM) stage B, and 21 with HCM ACVIM stage C). Each cat received a full echocardiogram, health panel, and total thyroxin level. Galectin-3 levels were measured for each enrolled patient. Troponin I and N-terminal pro-brain natriuretic peptide (NT-proBNP) were obtained for the majority of cats. Additionally, 17 ACVIM stage B cats underwent cardiac-gated magnetic resonance (CMR) imaging to assess myocardial extracellular volume (ECV), a noninvasive measure of myocardial fibrosis. RESULTS: Galectin-3 levels are increased in cats with HCM ACVIM stage B and C compared to healthy cats; however, no significant differences were detected between ACVIM stage B and ACVIM stage C cats. In HCM-affected cats, Galectin-3 showed statistically significant correlations with left atrial dimensions, left atrial:aorta ratio, and CMR-derived ECV. Quantitative NT-proBNP showed excellent discrimination between all groups and troponin I was able to discriminate between ACVIM stage C and normal cats, but not between other groups. CONCLUSIONS: Circulating Gal-3 levels are increased in cats with HCM and is positively correlated with left atrial dimensions and ECV in affected cats. Further studies evaluating the relationship between Gal-3, myocardial fibrosis, and clinical outcomes are warranted.

Rehabilitation post-COVID-19: cross-sectional observations using the Stanford Hall remote assessment tool.

INTRODUCTION: The multisystem COVID-19 can cause prolonged symptoms requiring rehabilitation. This study describes the creation of a remote COVID-19 rehabilitation assessment tool to allow timely triage, assessment and management. It hypotheses those with post-COVID-19 syndrome, potentially without laboratory confirmation and irrespective of initial disease severity, will have significant rehabilitation needs. METHODS: Cross-sectional study of consecutive patients referred by general practitioners (April-November 2020). Primary outcomes were presence/absence of anticipated sequelae. Binary logistic regression was used to test association between acute presentation and post-COVID-19 symptomatology. RESULTS: 155 patients (n=127 men, n=28 women, median age 39 years, median 13 weeks post-illness) were assessed using the tool. Acute symptoms were most commonly shortness of breath (SOB) (74.2%), fever (73.5%), fatigue (70.3%) and cough (64.5%); and post-acutely, SOB (76.7%), fatigue (70.3%), cough (57.4%) and anxiety/mood disturbance (39.4%). Individuals with a confirmed diagnosis of COVID-19 were 69% and 63% less likely to have anxiety/mood disturbance and pain, respectively, at 3 months. CONCLUSIONS: Rehabilitation assessment should be offered to all patients suffering post-COVID-19 symptoms, not only those with laboratory confirmation and considered independently from acute illness severity. This tool offers a structure for a remote assessment. Post-COVID-19 programmes should include SOB, fatigue and mood disturbance management.

Enabling uptake and sustainability of supervision roles by women GPs in Australia: a narrative analysis of interviews.

BACKGROUND: Worldwide, the proportion of women entering careers in medicine is increasing. To ensure diversity and capacity in the general practice ("GP") supervision workforce, a greater understanding from the perspective of women GPs engaged in or considering the clinical supervision of trainee doctors is important. This narrative inquiry aims to explore the uptake and sustainability of supervision roles for women GPs in the Australian context. METHODS: Qualitative interviews with Australian women GPs were conducted between July and September 2021. Women GPs were selected to represent a range of demographics, practice contexts, and supervision experience to promote broad perspectives. Narrative analysis drew on participant perspectives, allowing emergent stories to be explored using story arcs based on the characters, settings, problems, actions, and resolutions. These stories were evaluated by a broad research team and a high level of agreement of the final narratives and counter-narratives was achieved. RESULTS: Of the 25 women who enrolled, 17 completed interviews. Six narratives emerged, including: power and control, pay, time, other life commitments, quality of supervision, and supervisor identity. These represented significant intersecting issues with the potential to impact the uptake and sustainability of supervision by women GPs. Some women GPs reported a lack of agency to make decisions about their role in supervision and were not remunerated for teaching. Uptake and sustainability of supervision was constrained by other life commitments, which could be buffered by team-sharing arrangements and a supportive practice. Although adding a burden of time atop their complex and sensitive consultations, women GPs were committed to being available to registrars and supervising at a high standard. To foster high quality supervision, women GPs were interested in up-skilling resources, building experience and harnessing support networks. Women sensed imposter syndrome when negotiating a supervisor identity, which could be managed by explicitly valuing their contribution. CONCLUSION: The findings can inform the development of more specific resources, supports and structures to enable women GPs in Australia to uptake and sustain the supervision of trainee doctors at a level they find both acceptable and rewarding.

Cannabidiol and refractory epilepsy: parental and caregiver perspectives of participation in a compassionate access scheme.

BACKGROUND: The Compassionate Access Scheme (CAS) being delivered through the Queensland Children's Hospital is designed to allow access to an investigational purified Cannabidiol oral solution to paediatric patients with severe refractory epilepsy. The objectives of this study were to conduct semi-structured interviews to: 1. Understand families' expectations and attitudes about the use of an investigational cannabinoid product for their child's seizures; 2. Understand families' perceptions of Cannabidiol's efficacy for their child's seizures; and other aspects of their child's behaviour, quality of life and/or cognition. METHODS: Children aged 2-18 years had been enrolled in, or were enrolled in a compassionate access scheme for Cannabidiol at the time of the study. Semi-structured interviews (n = 19) with parents or caregivers (n = 23) of children diagnosed with refractory epilepsy were voice-recorded, transcribed and analysed to generate common themes. RESULTS: Key themes emerged relating to seizure activity, family and school engagement, drug safety and legal access, efficacy, clinical support, social acceptance of the medication and program delivery. The use of Cannabidiol was perceived to have benefits in relation to reducing the severity and frequency of seizure activity for almost a third of patients experiencing refractory epilepsy. Participants described other benefits including improved social engagement, wakefulness and a reduction of side effects related to a reduction of conventional medication dosage. CONCLUSION: This study provided unique perspectives of families' experiences managing untreatable epilepsy, their experiences with conventional and experimental pharmacological treatments and health services. Whilst families' perceptions showed the use of Cannabidiol did not provide a therapeutic reduction in the seizure activity for all patients diagnosed with refractory epilepsy, it's use as an additional pharmacological agent was perceived to provide other benefits by some patient families.

Enhanced Antifibrinolytic Efficacy of a Plasmin-Specific Kunitz-Inhibitor (60-Residue Y11T/L17R with C-Terminal IEK) of Human Tissue Factor Pathway Inhibitor Type-2 Domain1.

Current antifibrinolytic agents reduce blood loss by inhibiting plasmin active sites (e.g., aprotinin) or by preventing plasminogen/tissue plasminogen activator (tPA) binding to fibrin clots (e.g., ε-aminocaproic acid and tranexamic acid); however, they have adverse side effects. Here, we expressed 60-residue (NH2NAE…IEKCOOH) Kunitz domain1 (KD1) mutants of human tissue factor pathway inhibitor type-2 that inhibit plasmin as well as plasminogen activation. A single (KD1-L17R-KCOOH) and a double mutant (KD1-Y11T/L17R- KCOOH) were expressed in Escherichia coli as His-tagged constructs, each with enterokinase cleavage sites. KD1-Y11T/L17R-KCOOH was also expressed in Pichia pastoris. KD1-Y11T/L17R-KCOOH inhibited plasmin comparably to aprotinin and bound to the kringle domains of plasminogen/plasmin and tPA with Kd of ~50 nM and ~35 nM, respectively. Importantly, compared to aprotinin, KD1-L17R-KCOOH and KD1-Y11T/L17R-KCOOH did not inhibit kallikrein. Moreover, the antifibrinolytic potential of KD1-Y11T/L17R-KCOOH was better than that of KD1-L17R-KCOOH and similar to that of aprotinin in plasma clot-lysis assays. In thromboelastography experiments, KD1-Y11T/L17R-KCOOH was shown to inhibit fibrinolysis in a dose dependent manner and was comparable to aprotinin at a higher concentration. Further, KD1-Y11T/L17R-KCOOH did not induce cytotoxicity in primary human endothelial cells or fibroblasts. We conclude that KD1-Y11T/L17R-KCOOH is comparable to aprotinin, the most potent known inhibitor of plasmin and can be produced in large amounts using Pichia.

Physical activity and the prevention, reduction, and treatment of alcohol and other drug use across the lifespan (The PHASE review): A systematic review.

The aim of this review is to systematically describe and quantify the effects of PA interventions on alcohol and other drug use outcomes, and to identify any apparent effect of PA dose and type, possible mechanisms of effect, and any other aspect of intervention delivery (e.g. key behaviour change processes), within a framework to inform the design and evaluation of future interventions. Systematic searches were designed to identify published and grey literature on the role of PA for reducing the risk of progression to alcohol and other drug use (PREVENTION), supporting individuals to reduce alcohol and other drug use for harm reduction (REDUCTION), and promote abstinence and relapse prevention during and after treatment of alcohol and other drug use (TREATMENT). Searches identified 49,518 records, with 49,342 excluded on title and abstract. We screened 176 full text articles from which we included 32 studies in 32 papers with quantitative results of relevance to this review. Meta-analysis of two studies showed a significant effect of PA on prevention of alcohol initiation (risk ratio [RR]: 0.72, 95%CI: 0.61 to 0.85). Meta-analysis of four studies showed no clear evidence for an effect of PA on alcohol consumption (Standardised Mean Difference [SMD]: 0.19, 95%, Confidence Interval -0.57 to 0.18). We were unable to quantitatively examine the effects of PA interventions on other drug use alone, or in combination with alcohol use, for prevention, reduction or treatment. Among the 19 treatment studies with an alcohol and other drug use outcome, there was a trend for promising short-term effect but with limited information about intervention fidelity and exercise dose, there was a moderate to high risk of bias. We identified no studies reporting the cost-effectiveness of interventions. More rigorous and well-designed research is needed. Our novel approach to the review provides a clearer guide to achieve this in future research questions addressed to inform policy and practice for different populations and settings.

Diagnostic development for parallel wave-number measurement of lower hybrid waves in EAST.

An eight-channel magnetic probe diagnostic system has been designed and installed adjacent to the 4.6 GHz lower hybrid (LH) grill antenna in the low-field side of the Experimental Advanced Superconducting Tokamak (EAST) in order to study the n∥ evolution of LH waves in the first pass from the launcher to the core plasma. The magnetic probes are separated by 6.6 mm, which allows measurement of the dominant parallel refractive index n∥ up to n∥ = 5 for 4.6 GHz LH waves. The magnetic probes are designed to be sensitive to the magnetic field component perpendicular to the background magnetic field with a slit on the casing that encloses the probe. The intermediate frequency stage, which consists of two mixing stages, down-coverts the frequency of the measured wave signals at 4.6 GHz to 20 MHz. A bench test demonstrates the phase stability of the magnetic probe diagnostic system. By evaluating the phase variation of the measured signals along the background magnetic field, the dominant n∥ of the LH wave in the scrape-off layer has been deduced during the 2019 experimental campaign. In the low density plasma, the measured dominant n∥ of the LH waves is about 2.1, corresponding to the main peak 2.04 of the launched n∥ spectrum. n∥ deduced by the least-squares linear fit method remains near this value in the low density plasma with a high spatial correlation magnitude of 0.9. With an eight-channel probe system, a wave-number spectrum has also been deduced, which has a peak near to the measured dominant n∥.

Observation of Efficient Lower Hybrid Current Drive at High Density in Diverted Plasmas on the Alcator C-Mod Tokamak.

Efficient lower hybrid current drive (LHCD) is demonstrated at densities up to n[over ¯]_{e}≈1.5×10^{20} m^{-3} in diverted plasmas on the Alcator C-Mod tokamak by operating at increased plasma current-and therefore reduced Greenwald density fraction. This density exceeds the nominal "LH density limit" at n[over ¯]_{e}≈1.0×10^{20} m^{-3} reported previously, above which an anomalous loss of current drive efficiency was observed. The recovery of current drive efficiency to a level consistent with engineering scalings is correlated with a reduction in density shoulders and turbulence levels in the far scrape-off layer. Concurrently, rf wave interaction with the edge and/or scrape-off-layer plasma is reduced, as indicated by a minimal broadening of the wave frequency spectrum measured at the plasma edge. These results have important implications for sustaining steady-state tokamak operation and indicate a pathway forward for implementing efficient LHCD in a reactor.

Cyberbiosecurity: An Emerging New Discipline to Help Safeguard the Bioeconomy.

Cyberbiosecurity is being proposed as a formal new enterprise which encompasses cybersecurity, cyber-physical security and biosecurity as applied to biological and biomedical-based systems. In recent years, an array of important meetings and public discussions, commentaries and publications have occurred that highlight numerous vulnerabilities. While necessary first steps, they do not provide a systematized structure for effectively promoting communication, education and training, elucidation and prioritization for analysis, research, development, test and evaluation and implementation of scientific, technological, standards of practice, policy, or even regulatory or legal considerations for protecting the bioeconomy. Further, experts in biosecurity and cybersecurity are generally not aware of each other's domains, expertise, perspectives, priorities, or where mutually supported opportunities exist for which positive outcomes could result. Creating, promoting and advancing a new discipline can assist with formal, beneficial and continuing engagements. Recent key activities and publications that inform the creation of Cyberbiosecurity are briefly reviewed, as is the expansion of Cyberbiosecurity to include biomanufacturing which is supported by a rigorous analysis of a biomanufacturing facility. Recommendations are provided to initialize Cyberbiosecurity and place it on a trajectory to establish a structured and sustainable discipline, forum and enterprise.

PEDOT doped with algal, mammalian and synthetic dopants: polymer properties, protein and cell interactions, and influence of electrical stimulation on neuronal cell differentiation.

Poly(3,4-ethylenedioxythiophene) (PEDOT) films were electrochemically polymerised with several synthetic (dodecylbenzosulfonic acid (DBSA)) and biological (dextran sulphate (DS), chondroitin sulphate (CS), alginic acid (ALG) and ulvan (ULV)) dopant anions, and their physical, mechanical and electrochemical properties characterised. PEDOT films incorporating the biological dopants ALG and ULV produced films of the greatest surface roughness (46 ± 5.1 and 31 ± 1.9 nm, respectively), and demonstrated significantly lower shear modulus values relative to all other PEDOT films (2.1 ± 0.1 and 1.2 ± 0.2 MPa, respectively). Quartz crystal microgravimetry was used to study the adsorption of the important extracellular matrix protein fibronectin, revealing protein adsorption to be greatest on PEDOT doped with DS, followed by DBSA, ULV, CS and ALG. Electrical stimulation experiments applying a pulsed current using a biphasic waveform (250 Hz) were undertaken using PEDOT doped with either DBSA or ULV. Electrical stimulation had a significant influence on cell morphology and cell differentiation for PEDOT films with either dopant incorporated, with the degree of branching per cell increased by 10.5× on PEDOT-DBSA and 6.5× on PEDOT-ULV relative to unstimulated cells, and mean neurite length per cell increasing 2.6× and 2.2× on stimulated vs. unstimulated PEDOT-DBSA and PEDOT-ULV, respectively. We demonstrate the cytocompatibility of synthetic and biologically doped PEDOT biomaterials, including the new algal derived polysaccharide dopant ulvan, which, along with DBSA doped PEDOT, is shown to significantly enhance the differentiation of PC12 neuronal cells under electrical stimulation.

Cyberbiosecurity: From Naive Trust to Risk Awareness.

The cyber-physical nature of biotechnology raises unprecedented security concerns. Computers can be compromised by encoding malware in DNA sequences, and biological threats can be synthesized using publicly available data. Trust within the biotechnology community creates vulnerabilities at the interface between cyberspace and biology. Awareness is a prerequisite to managing these risks.

Tissue engineering with gellan gum.

Engineering complex tissues for research and clinical applications relies on high-performance biomaterials that are amenable to biofabrication, maintain mechanical integrity, support specific cell behaviours, and, ultimately, biodegrade. In most cases, complex tissues will need to be fabricated from not one, but many biomaterials, which collectively fulfill these demanding requirements. Gellan gum is an anionic polysaccharide with potential to fill several key roles in engineered tissues, particularly after modification and blending. This review focuses on the present state of research into gellan gum, from its origins, purification and modification, through processing and biofabrication options, to its performance as a cell scaffold for both soft tissue and load bearing applications. Overall, we find gellan gum to be a highly versatile backbone material for tissue engineering research, upon which a broad array of form and functionality can be built.

Poor growth, thyroid dysfunction and vitamin D deficiency remain prevalent despite reduced intensity chemotherapy for hematopoietic stem cell transplantation in children and young adults.

Myeloablative conditioning regimens for hematopoietic stem cell transplant (HSCT) are known to affect endocrine function, but little is known regarding reduced intensity conditioning (RIC) regimens. We retrospectively reviewed 114 children and young adults after single RIC HSCT. The analysis was grouped by age (<2 and ⩾2 years) and diagnosis (hemophagocytic lymphohistiocystosis/X-linked lymphoproliferative syndrome (HLH/XLP), other immune disorders, metabolic/genetic disorders). All groups displayed short stature by mean height-adjusted Z-score (HAZ) before (-1.29) and after HSCT (HAZ -1.38, P=0.47). After HSCT, younger children with HLH/XLP grew better (HAZ -3.41 vs -1.65, P=0.006), whereas older subjects had decline in growth (HAZ -0.8 vs -1.01, P=0.06). Those with steroid therapy beyond standard GVHD prophylaxis were shorter than those without (P 0.04). After HSCT, older subjects with HLH/XLP became thinner with a mean body mass index (BMI) Z-score of 1.20 vs 0.64, P=0.02, and similar to metabolic/genetic disorders (BMI-Z= 0.59 vs -0.99, P<0.001). BMI increased among younger children in these same groups. Thyroid function was abnormal in 24% (18/76). 25-OH vitamin D levels were insufficient in 73% (49/65), with low bone mineral density in 8 of 19 evaluable subjects. Despite RIC, children and young adults still have significant late endocrine effects. Further research is required to compare post-transplant endocrine effects after RIC to those after standard chemotherapy protocols.

Production of Recombinant Chemokines and Validation of Refolding.

The diverse roles of chemokines in normal immune function and many human diseases have motivated numerous investigations into the structure and function of this family of proteins. Recombinant chemokines are often used to study how chemokines coordinate the trafficking of immune cells in various biological contexts. A reliable source of biologically active protein is vital for any in vitro or in vivo functional analysis. In this chapter, we describe a general method for the production of recombinant chemokines and robust techniques for efficient refolding that ensure consistently high biological activity. Considerations for initiating development of protocols consistent with Current Good Manufacturing Practices (cGMPs) to produce biologically active chemokines suitable for use in clinical trials are also discussed.

Probing the PEDOT:PSS/cell interface with conductive colloidal probe AFM-SECM.

Conductive colloidal probe Atomic Force-Scanning Electrochemical Microscopy (AFM-SECM) is a new approach, which employs electrically insulated AFM probes except for a gold-coated colloid located at the end of the cantilever. Hence, force measurements can be performed while biasing the conductive colloid under physiological conditions. Moreover, such colloids can be modified by electrochemical polymerization resulting, e.g. in conductive polymer-coated spheres, which in addition may be loaded with specific dopants. In contrast to other AFM-based single cell force spectroscopy measurements, these probes allow adhesion measurements at the cell-biomaterial interface on multiple cells in a rapid manner while the properties of the polymer can be changed by applying a bias. In addition, spatially resolved electrochemical information e.g., oxygen reduction can be obtained simultaneously. Conductive colloid AFM-SECM probes modified with poly(3,4-ethylenedioxythiophene) doped with polystyrene sulfonate ( PEDOT: PSS) are used for single cell force measurements in mouse fibroblasts and single cell interactions are investigated as a function of the applied potential.

Conductive composite fibres from reduced graphene oxide and polypyrrole nanoparticles.

Continuous composite fibres composed of polypyrrole (PPy) nanoparticles and reduced graphene oxide (rGO) at different mass ratios were fabricated using a single step wet-spinning approach. The electrical conductivity of the composite fibres increased significantly with the addition of rGO. The mechanical properties of the composite fibres also improved by the addition of rGO sheets compared to fibres containing only PPy. The ultimate tensile strength of the fibres increased with the proportion of rGO mass present. The elongation at break was greatest for the composite fibre containing equal mass ratios of PPy nanoparticles and rGO sheets. L929 fibroblasts seeded onto fibres showed no reduction in cell viability. To further assess toxicity, cells were exposed to media that had been used to extract any aqueous-soluble leachates from developed fibre. Overall, these composite fibres show promising mechanical and electrical properties while not significantly impeding cell growth, opening up a wide range of potential applications including nerve and muscle regeneration studies.