A Coxsackievirus B1-mediated nonlytic Extracellular Vesicle-to-cell mechanism of virus transmission and its possible control through modulation of EV release

Like most non-enveloped viruses, CVB1 mainly uses cell lysis to spread. Details of a nonlytic virus transmission remain unclear. Extracellular Vesicles (EVs) transfer biomolecules between cells. We show that CVB1 entry into HeLa cells results in apoptosis and release of CVB1-induced ‘medium-sized’ EVs (CVB1i-mEVs). These mEVs (100–300 nm) harbour CVB1 as shown by immunoblotting with anti-CVB1-antibody; viral capsids were detected by transmission electron microscopy and RT-PCR revealed CVB1 RNA. The percentage of mEVs released from CVB1-infected HeLa cells harbouring virus was estimated from TEM at 34 %. Inhibition of CVB1i-mEV production, with calpeptin or siRNA knockdown of CAPNS1 in HeLa cells limited spread of CVB1 suggesting these vesicles disseminate CVB1 virions to new host cells by a nonlytic EV-to-cell mechanism. This was confirmed by detecting CVB1 virions inside HeLa cells after co-culture with CVB1i-mEVs; EV release may also prevent apoptosis of infected cells whilst spreading apoptosis to secondary sites of infection

Validation of an Immunoassay for anti-thymidine phosphorylase antibodies in patients with MNGIE treated with enzyme replacement therapy

Erythrocyte encapsulated thymidine phosphorylase is recombinant Escherichia coli thymidine phosphorylase encapsulated within human autologous erythrocytes and is under development as an enzyme replacement therapy for the ultra-rare inherited metabolic disorder mitochondrial neurogastrointestinal encephalomyopathy. This study describes the method validation of a two-step bridging electrochemiluminescence immunoassay for the detection of anti-thymidine phosphorylase antibodies in human serum according to current industry practice and regulatory guidelines. The analytical method was assessed for screening cut point, specificity, selectivity, precision, prozone effect, drug tolerance, and stability. Key findings were a correction factor of 129 relative light units for the cut-point determination; a specificity cut point of 93% inhibition; confirmed intra-assay and inter-assay precision; assay sensitivity of 356 ng/mL; no matrix or prozone effects up to 25,900 ng/mL; a drug tolerance of 156 ng/mL; and stability at room temperature for 24 hr and up to five freeze-thaws. Immunogenicity evaluations of serum from three patients who received erythrocyte encapsulated thymidine phosphorylase under a compassionate treatment program showed specific anti-thymidine phosphorylase antibodies in one patient. To conclude, a sensitive, specific, and selective immunoassay has been validated for the measurement of anti-thymidine phosphorylase antibodies; this will be utilized in a phase II pivotal clinical trial of erythrocyte encapsulated thymidine phosphorylase

Autoantibodies against type I IFNs in patients with life-threatening COVID-19

Interindividual clinical variability in the course of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is vast. We report that at least 101 of 987 patients with life-threatening coronavirus disease 2019 (COVID-19) pneumonia had neutralizing immunoglobulin G (IgG) autoantibodies (auto-Abs) against interferon-w (IFN-w) (13 patients), against the 13 types of IFN-a (36), or against both (52) at the onset of critical disease; a few also had auto-Abs against the other three type I IFNs. The auto-Abs neutralize the ability of the corresponding type I IFNs to block SARS-CoV-2 infection in vitro. These auto-Abs were not found in 663 individuals with asymptomatic or mild SARS-CoV-2 infection and were present in only 4 of 1227 healthy individuals. Patients with auto-Abs were aged 25 to 87 years and 95 of the 101 were men. A B cell autoimmune phenocopy of inborn errors of type I IFN immunity accounts for life-threatening COVID-19 pneumonia in at least 2.6% of women and 12.5% of men

The development and assessment of a model for the study of placental transport involving the culture of human near-term placental trophoblast on amniotic membrane

SIGLEAvailable from British Library Document Supply Centre-DSC:DX204261 / BLDSC - British Library Document Supply CentreGBUnited Kingdo

Development and evaluation of a PCR-based test for detection of Asterias (Echinodermata : Asteroidea) larvae in Australian plankton samples from ballast water

The northern Pacific seastar, Asterias amurensis, spread to Tasmania in the 1980s from its native range in the North Pacific. The seastar has subsequently established in Port Phillip Bay on mainland Australia. Transportation of larvae in ballast water is one vector for these introductions and is likely to contribute to additional range expansion of this species. Larval identification methods are critical to assess risks of further transport to uninvaded ecosystems; however, morphological identification of larval asteroids is impractical and unreliable. Therefore, we have developed a sensitive PCR-based method that specifically detects Asterias DNA. The method works on isolated seastar larva, mixed plankton and ballast water samples. Trials using uninfected ballast water samples spiked with known numbers of A. amurensis larvae indicate that the technique can detect single larva in 200 mg of plankton. The test also detects other Asterias species; therefore, discrimination between seastars within the genus Asterias was accomplished using denaturing gradient gel electrophoresis (DGGE). Currently, this method is being used to facilitate research on ballast water transport, distribution and the ecology of A. amurensis larvae. The general approach can be expanded to provide a rapid and cost-effective approach for detecting a suite of marine species in ballast water and environmental samples

Grafting tomato (Solanum lycopersicum) onto the rootstock of a high-altitude accession of Solanum habrochaites improves suboptimal-temperature tolerance

Grafting is regarded as a promising tool to broaden the temperature optimum of elite tomato cultivars. However, suitable low-temperature tolerant tomato rootstocks are not yet available and its breeding is hampered by a lack of variation in low-temperature tolerance within the cultivated tomato. In this study, therefore, the impact of grafting tomato (Solanum lycopersicum Mill. cv. Moneymaker, Sl) onto the rootstock of a cold-tolerant high-altitude accession of a related wild species (Solanum habrochaites LA 1777 Humb. & Bonpl., Sh) was examined at different combinations of optimal (25 degrees C) and/or suboptimal (15 degrees C) air/root-zone temperatures (RZT), i.e. 25/25, 25/15, 15/25 and 15/15 degrees C. Self-grafted tomato plants were used as controls. Both scion/rootstock combinations, Sl/Sl and Sl/Sh, were grown hydroponically and compared for biomass production and partitioning, plant morphology, carbohydrate partitioning and leaf C and N status. Grafting tomato onto Sh increased the relative growth rate of shoots with 26 and 11% at 25/15 and 15/15 degrees C, respectively. This increase could be attributed to stimulation of the leaf expansion rate. Graft combinations with Sh rootstocks were characterized by higher root mass ratios, particularly at 15 degrees C RZT. Suboptimal RZT strongly reduced the relative growth rate of Sl roots but not of Sh. This was correlated to differences in inhibition of root elongation. In contrast to tomato grafted onto Sh, leaf total C and total N concentrations increased in self-grafted tomato plants in response to 15 degrees C RZT. The increase in leaf total C concentration of Sl/Sl graft combinations at 15 degrees C RZT could be ascribed largely to starch accumulation. This study illustrates that growth of vegetative tomato plants at suboptimal temperature is for a significant part inhibited by its poor root development. Grafting tomato onto a low-temperature rootstock provides an alternative tool to reduce, in part, the grow-limiting effects of suboptimal RZ temperature on the shoot. To improve the low-temperature tolerance of existing commercial tomato rootstocks, S. habrochaites LA 1777 appeared to be a valuable germplasm pool. (C) 2008 Elsevier B.V. All rights reserved

Port of Hastings National Demonstration Project - Verification of the Type II error rate of the Ballast Water Decision Support System (DSS).

In July 2001, Australia introduced a risk-based Decision Support System (DSS) to manage ballast water on international shipping. Up until now, however, the accuracy of the risk assessments made by the DSS have never been evaluated or tested. It was not until the DSS was considered as a mechanism to assist the management of domestically sourced ballast water, coupled with advances in genetic technologies, that the opportunity arose to evaluate the accuracy of the predictions of the DSS. The results of that evaluation are reported here..

Percutaneous Intervention for Concurrent Chronic Total Occlusions in Patients With STEMI: The EXPLORE Trial

BACKGROUND: In 10% to 15% of patients with ST-segment elevation myocardial infarction (STEMI), concurrent coronary chronic total occlusion (CTO) in a non-infarct-related artery is present and is associated with increased morbidity and mortality. OBJECTIVES: The EXPLORE (Evaluating Xience and Left Ventricular Function in Percutaneous Coronary Intervention on Occlusions After ST-Elevation Myocardial Infarction) trial evaluated whether patients with STEMI and concurrent CTO in a non-infarct-related artery benefit from additional percutaneous coronary intervention (PCI) of CTO shortly after primary PCI. METHODS: From November 2007 through April 2015, we enrolled 304 patients with acute STEMI who underwent primary PCI and had concurrent CTO in 14 centers in Europe and Canada. A total of 150 patients were randomly assigned to early PCI of the CTO (CTO PCI), and 154 patients were assigned to conservative treatment without PCI of the CTO (no CTO PCI). Primary outcomes were left ventricular ejection fraction (LVEF) and left ventricular end diastolic volume (LVEDV) on cardiac magnetic resonance imaging after 4 months. RESULTS: The investigator-reported procedural success rate in the CTO PCI arm of the trial was 77%, and the adjudicated success rate was 73%. At 4 months, mean LVEF did not differ between the 2 groups (44.1 +/- 12.2% vs. 44.8 +/- 11.9%, respectively; p = 0.60). Mean LVEDV at 4 months was 215.6 +/- 62.5 ml in the CTO PCI arm versus 212.8 +/- 60.3 ml in the no-CTO PCI arm (p = 0.70). Subgroup analysis revealed that patients with CTO located in the left anterior descending coronary artery who were randomized to the CTO PCI strategy had significantly higher LVEF compared with patients randomized to the no-CTO PCI strategy (47.2 +/- 12.3% vs. 40.4 +/- 11.9%; p = 0.02). There were no differences in terms of 4-month major adverse coronary events (5.4% vs. 2.6%; p = 0.25). CONCLUSIONS: Additional CTO PCI within 1 week after primary PCI for STEMI was feasible and safe. In patients with STEMI and concurrent CTO, we did not find an overall benefit for CTO PCI in terms of LVEF or LVEDV. The finding that early CTO PCI in the left anterior descending coronary artery subgroup was beneficial warrants further investigation. (Evaluating Xience and Left Ventricular Function in Percutaneous Coronary Intervention on Occlusions After ST-Segment Elevation Myocardial Infarction; NTR1108)