Stonefish toxin defines an ancient branch of the perforin-like superfamily

The lethal factor in stonefish venom is stonustoxin (SNTX), a heterodimeric cytolytic protein that induces cardiovascular collapse in humans and native predators. Here, using X-ray crystallography, we make the unexpected finding that SNTX is a pore-forming member of an ancient branch of the Membrane Attack Complex-Perforin/Cholesterol-Dependent Cytolysin (MACPF/CDC) superfamily. SNTX comprises two homologous subunits (α and β), each of which comprises an N-terminal pore-forming MACPF/CDC domain, a central focal adhesion-targeting domain, a thioredoxin domain, and a C-terminal tripartite motif family-like PRY SPla and the RYanodine Receptor immune recognition domain. Crucially, the structure reveals that the two MACPF domains are in complex with one another and arranged into a stable early prepore-like assembly. These data provide long sought after near-atomic resolution insights into how MACPF/CDC proteins assemble into prepores on the surface of membranes. Furthermore, our analyses reveal that SNTX-like MACPF/CDCs are distributed throughout eukaryotic life and play a broader, possibly immune-related function outside venom.Human envenoming by the tropical stonefish (Synanceia horrida and related species) results in extreme pain, edema, hypotension, respiratory distress, and on rare occasions, death (1). The lethal factor in stonefish venom is an ∼150-kDa protein termed stonustoxin (SNTX), an unusual example of a vertebrate cytolytic protein complex (2). SNTX is a soluble heterodimeric assembly of two closely related proteins termed SNTX-α and -β that share sequence identity of ∼50% (3). With the exception of a C-terminal PRY SPla and the RYanodine Receptor (PRYSPRY) domain in each protein (4), SNTX shares no obvious sequence similarity to any structurally or functionally characterized molecule. SNTX induces species-specific hemolytic activity (2) by an apparent pore-forming mechanism (5). It induces platelet aggregation (6), and like the closely related Trachynilysin (from Synanceia trachynis), SNTX exhibits activity suggesting that it may function as a neurotoxin (7, 8).Because eukaryote pore-forming toxins are relatively rare, we reasoned that SNTX might represent a new exemplar of a vertebrate pore-forming protein. Previous studies had shown that it was possible to purify and crystallize SNTX (9); however, no structure has been reported to date. Accordingly, to address the structural basis for SNTX activity, we determined its X-ray crystal structure.     

Relationship between obesity and pulse pressure in children: results of the National Health and Nutrition Survey (NHANES) 1988–1994

BackgroundObesity is a known cardiometabolic risk factor in children. In adults, pulse pressure (PP) is a known predictor and a risk factor of cardiovascular (CV) diseases. In this study, we examined the association between measures of obesity and PP in children.MethodsA retrospective analysis of 4667 children ages 6–17 years from the National Health and Nutrition Survey (1988–1994) was performed. We defined wide PP as 4th quartile PP and high waist circumference (WC) as >75th percentile based on age and gender.ResultsThere were 51% boys, 74% whites, 16% blacks, 10% Hispanics, 12% obese, 26% with high WC, 26% with wide PP, and 9% with high blood pressure (BP). Prevalence of wide PP was high among obese children. A significantly higher mean PP was observed in boys, Blacks, obese, those with high WC and high BP. The adjusted odds ratio (OR) for wide PP was higher in boys, Blacks, and those with high WC.ConclusionThere was a statistically significant independent association observed between wide PP and high WC, but not with obesity based on BMI. Further exploration of wide PP as a CV risk factor in childhood and its relationship to CV outcomes appears warranted.     

Simulation: Moving from Technology Challenge to Human Factors Success

Recognition of the many limitations of traditional apprenticeship training is driving new approaches to learning medical procedural skills. Among simulation technologies and methods available today, computer-based systems are topical and bring the benefits of automated, repeatable, and reliable performance assessments. Human factors research is central to simulator model development that is relevant to real-world imaging-guided interventional tasks and to the credentialing programs in which it would be used.     

Coexistence of ANCA-associated glomerulonephritis and anti-phospholipase A(2) receptor antibody-positive membranous nephropathy

Antibodies to myeloperoxidase (MPO) and proteinase 3 (PR3) have been demonstrated to mediate anti-neutrophil cytoplasmic antibody (ANCA)-associated disease. For membranous nephropathy, antibodies to the podocyte-expressed phospholipase A(2) receptor (anti-PLA(2)R) are highly associated with disease activity and have been reported in at least 70% of patients with idiopathic membranous nephropathy (IMN). We present a case of a 56-year-old male with a 1 year history of hypertension, leg edema, and proteinuria, who presented with advanced renal failure and was found to have both ANCA-associated glomerulonephritis (GN) and IMN on kidney biopsy. Consistent with the idea that this is due to the chance occurrence of two independent diseases, we found both anti-MPO and anti-PLA(2)R antibodies in the patient's sera. Treatment with methylprednisolone, plasmapheresis, and cyclophosphamide resulted in improvement in kidney function and proteinuria, together with the simultaneous decrease in both autoantibodies. This is the first demonstration of two pathogenic antibodies giving rise to ANCA-associated GN and IMN in the same patient. It confirms the importance of classifying disease based upon the underlying mechanism, in addition to renal histopathology, to both optimize therapy and predict prognosis.     

Antibody titres elicited by the 2018 seasonal inactivated influenza vaccine decline by 3 months post‐vaccination but persist for at least 6 months

BackgroundIn Australia, seasonal inactivated influenza vaccine is typically offered in April. However, the onset, peak and end of a typical influenza season vary, and optimal timing for vaccination remains unclear. Here, we investigated vaccine‐induced antibody response kinetics over 6 months in different age groups.MethodsWe conducted a prospective serosurvey among 71 adults aged 18–50 years, 15 community‐dwelling (‘healthy’) and 16 aged‐care facility resident (‘frail’) older adults aged ≥65 years who received the 2018 southern hemisphere vaccines. Sera were collected at baseline, and 1, 2, 4, and 6 months post‐vaccination. Antibody titres were measured by haemagglutination inhibition or microneutralisation assays. Geometric mean titres were estimated using random effects regression modelling and superimposed on 2014–2018 influenza season epidemic curves.ResultsAntibody titres peaked 1.2–1.3 months post‐vaccination for all viruses, declined by 3 months post‐vaccination but, notably, persisted above baseline after 6 months in all age groups by 1.3‐ to 1.5‐fold against A(H1N1)pdm09, 1.7‐ to 2‐fold against A(H3N2), 1.7‐ to 2.1‐fold against B/Yamagata and 1.8‐fold against B/Victoria. Antibody kinetics were similar among different age groups. Antibody responses were poor against cell‐culture grown compared to egg‐grown viruses.ConclusionsThese results suggest subtype‐specific antibody‐mediated protection persists for at least 6 months, which corresponds to the duration of a typical influenza season.