Purification and quantification of recombinant Epstein-Barr viral glycoproteins gp350/220 from Chinese hamster ovary cells

Truncated Epstein-Barr virus (EBV) membrane antigen gp350/220 (EBV-MA) lacking the membrane anchor was expressed and secreted into the medium of recombinant Chinese hamster ovary cells that had been cultured in Plasmapur hollow-fibre modules using defined serum-free medium. The EBV-MA in the medium was concentrated by 70% (w/v) ammonium sulphate precipitation and subsequently purified by immunoaffinity chromatography using an anti-EBV-MA (EBV.0T6) monoclonal antibody (mAb) column. Adsorbed antigen was eluted with 3 M MgCl2 in phosphate-buffered saline, concentrated by Mono Q anion-exchange chromatography and analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, silver staining and Western blotting using EBV-positive serum and anti-EBV-MA specific mAbs. Monospecific polyclonal rabbit antibodies against the purified EBV-MA were raised and purified by protein G affinity chromatography. For the measurement of EBV-MA antigen levels a sandwich enzyme-linked immunosorbent assay using rabbit polyclonal antibodies and a horseradish peroxidase-conjugated anti-MA mAb was developed having a detection level of 10 ng/ml

Imaging angiogenesis in patients with head and neck squamous cell carcinomas by [68Ga]Ga-DOTA-E-[c(RGDfK)]2 PET/CT

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The 1989 and 2015 outbursts of V404 Cygni: a global study of wind-related optical features

The black hole transient V404 Cygni exhibited a bright outburst in June 2015 that was intensively followed over a wide range of wavelengths. Our team obtained high time resolution optical spectroscopy (~90 s), which included a detailed coverage of the most active phase of the event. We present a database consisting of 651 optical spectra obtained during this event, that we combine with 58 spectra gathered during the fainter December 2015 sequel outburst, as well as with 57 spectra from the 1989 event. We previously reported the discovery of wind-related features (P-Cygni and broad-wing line profiles) during both 2015 outbursts. Here, we build diagnostic diagrams that enable us to study the evolution of typical emission line parameters, such as line fluxes and equivalent widths, and develop a technique to systematically detect outflow signatures. We find that these are present throughout the outburst, even at very low optical fluxes, and that both types of outflow features are observed simultaneously in some spectra, confirming the idea of a common origin. We also show that the nebular phases depict loop patterns in many diagnostic diagrams, while P-Cygni profiles are highly variable on time-scales of minutes. The comparison between the three outbursts reveals that the spectra obtained during June and December 2015 share many similarities, while those from 1989 exhibit narrower emission lines and lower wind terminal velocities. The diagnostic diagrams presented in this work have been produced using standard measurement techniques and thus may be applied to other active low-mass X-ray binaries.Comment: Accepted for publication in MNRAS. 23 pages paper, plus a 9 pages appendix with extra tables and figures. 18 figures are included in the paper and 8 in the appendi

Virtual reality cognitive-behavioural therapy versus cognitive-behavioural therapy for paranoid delusions:a study protocol for a single-blind multi-Centre randomised controlled superiority trial

Abstract Background Seventy per cent of patients with psychotic disorders has paranoid delusions. Paranoid delusions are associated with significant distress, hospital admission and social isolation. Cognitive-behavioural therapy for psychosis (CBTp) is the primary psychological treatment, but the median effect size is only small to medium. Virtual reality (VR) has a great potential to improve the effectiveness of CBTp. In a previous study, we found that VR based CBT (VRcbt) for paranoid delusions is superior to waiting list. As a next step, a direct comparison with CBTp is needed. The present study aims to investigate whether VRcbt is more effective and cost-effective than regular CBTp in treating paranoid delusions and improving daily life social functioning of patients with psychotic disorders. Methods A total of 106 patients with DSM-5 diagnosis of psychotic disorder and at least moderate level of paranoid ideations will be recruited for this multicentre randomized controlled trial (RCT). Patients will be randomized to either VRcbt or standard CBTp for paranoid delusions. VRcbt consists of maximum 16 sessions in virtual social situations that trigger paranoid ideations and distress, delivered in an 8–12 week time frame. Standard CBTp also consists of maximum 16 sessions including exposure and behavioural experiments, delivered in an 8–12 week time frame. The two groups will be compared at baseline, post-treatment and six months follow-up. Primary outcome is the level of paranoid ideations in daily life social situations, measured with ecological momentary assessments (EMA) at semi-random moments ten times a day during seven days, before and after treatment. Every session, participants and therapists will rate the level of paranoid ideation and global clinical impression. Discussion Comparison of VRcbt and CBTp will provide information about the relative (cost-) effectiveness of VRcbt for this population. VRcbt may become a preferred psychological treatment for paranoid delusions and social anxiety in patients with psychotic disorder. Trial registration Netherlands Trial Register, NL7758. Registered on 23 May 2019

Plant–soil feedback of native and range-expanding plant species is insensitive to temperature

Temperature change affects many aboveground and belowground ecosystem processes. Here we investigate the effect of a 5°C temperature increase on plant–soil feedback. We compare plant species from a temperate climate region with immigrant plants that originate from warmer regions and have recently shifted their range polewards. We tested whether the magnitude of plant–soil feedback is affected by ambient temperature and whether the effect of temperature differs between these groups of plant species. Six European/Eurasian plant species that recently colonized the Netherlands (non-natives), and six related species (natives) from the Netherlands were selected. Plant–soil feedback of these species was determined by comparing performance in conspecific and heterospecific soils. In order to test the effect of temperature on these plant–soil feedback interactions, the experiments were performed at two greenhouse temperatures of 20/15°C and 25/20°C, respectively. Inoculation with unconditioned soil had the same effect on natives and non-natives. However, the effect of conspecific conditioned soil was negative compared to heterospecific soil for natives, but was positive for non-natives. In both cases, plant–soil interactions were not affected by temperature. Therefore, we conclude that the temperature component of climate change does not affect the direction, or strength of plant–soil feedback, neither for native nor for non-native plant species. However, as the non-natives have a more positive soil feedback than natives, climate warming may introduce new plant species in temperate regions that have less soil-borne control of abundance

[F-18]FDG-PET/CT to prevent futile surgery in indeterminate thyroid nodules:a blinded, randomised controlled multicentre trial

Purpose To assess the impact of an [F-18]FDG-PET/CT-driven diagnostic workup to rule out malignancy, avoid futile diagnostic surgeries, and improve patient outcomes in thyroid nodules with indeterminate cytology. Methods In this double-blinded, randomised controlled multicentre trial, 132 adult euthyroid patients with scheduled diagnostic surgery for a Bethesda III or IV thyroid nodule underwent [F-18]FDG-PET/CT and were randomised to an [F-18] FDG-PET/CT-driven or diagnostic surgery group. In the [F-18]FDG-PET/CT-driven group, management was based on the [F-18]FDG-PET/CT result: when the index nodule was visually [F-18]FDG-positive, diagnostic surgery was advised; when [F-18]FDG-negative, active surveillance was recommended. The nodule was presumed benign when it remained unchanged on ultrasound surveillance. In the diagnostic surgery group, all patients were advised to proceed to the scheduled surgery, according to current guidelines. The primary outcome was the fraction of unbeneficial patient management in one year, i.e., diagnostic surgery for benign nodules and active surveillance for malignant/borderline nodules. Intention-to-treat analysis was performed. Subgroup analyses were performed for non-Hurthle cell and Hurthle cell nodules. Results Patient management was unbeneficial in 42% (38/91 [95% confidence interval [CI], 32-53%]) of patients in the [F-18] FDG-PET/CT-driven group, as compared to 83% (34/41 [95% CI, 68-93%]) in the diagnostic surgery group (p < 0.001). [F-18]FDG-PET/CT-driven management avoided 40% (25/63 [95% CI, 28-53%]) diagnostic surgeries for benign nodules: 48% (23/48 [95% CI, 33-63%]) in non-Hurthle cell and 13% (2/15 [95% CI, 2-40%]) in I-Liable cell nodules (p = 0.02). No malignant or borderline tumours were observed in patients under surveillance. Sensitivity, specificity, negative and positive predictive value, and benign call rate (95% CI) of [F-18]FDG-PET/CT were 94.1% (80.3-99.3%), 39.8% (30.0-50.2%), 95.1% (83.5-99.4%), 35.2% (25.4-45.9%), and 31.1% (23.3-39.7%), respectively. Conclusion An [F-18]FDG-PET/CT-driven diagnostic workup of indeterminate thyroid nodules leads to practice changing management, accurately and oncologically safely reducing futile surgeries by 40%. For optimal therapeutic yield, application should be limited to non-Hurthle cell nodules

Transcriptional repressor ZEB2 promotes terminal differentiation of CD8⁺ effector and memory T cell populations during infection

ZEB2 is a multi-zinc-finger transcription factor known to play a significant role in early neurogenesis and in epithelial-mesenchymal transition-dependent tumor metastasis. Although the function of ZEB2 in T lymphocytes is unknown, activity of the closely related family member ZEB1 has been implicated in lymphocyte development. Here, we find that ZEB2 expression is up-regulated by activated T cells, specifically in the KLRG1(hi) effector CD8(+) T cell subset. Loss of ZEB2 expression results in a significant loss of antigen-specific CD8(+) T cells after primary and secondary infection with a severe impairment in the generation of the KLRG1(hi) effector memory cell population. We show that ZEB2, which can bind DNA at tandem, consensus E-box sites, regulates gene expression of several E-protein targets and may directly repress Il7r and Il2 in CD8(+) T cells responding to infection. Furthermore, we find that T-bet binds to highly conserved T-box sites in the Zeb2 gene and that T-bet and ZEB2 regulate similar gene expression programs in effector T cells, suggesting that T-bet acts upstream and through regulation of ZEB2. Collectively, we place ZEB2 in a larger transcriptional network that is responsible for the balance between terminal differentiation and formation of memory CD8(+) T cells

D2.3 EuropaBON Proposal for an EU Biodiversity Observation Coordination Centre (EBOCC)

Observations are key to understanding the state of nature, the drivers of biodiversity loss and the impacts on ecosystem services and ultimately on people. Many EU policies and initiatives call for unbiased, integrated and regularly updated data on biodiversity and ecosystem services. However, biodiversity monitoring efforts are spatially and temporally fragmented, taxonomically biased and not integrated across Europe. EuropaBON has addressed this gap by developing an EU-wide framework for biodiversity monitoring. With this deliverable, EuropaBON proposes the terms of reference for an EU Biodiversity Observation Coordination Centre (EBOCC), a permanent infrastructure that could coordinate and foster the generation and use of high quality data to underpin the biodiversity knowledge-base used across EU policies, providing guidance and trainings when necessary. Such a centre represents one of the key solutions to overcome the critical challenges of biodiversity monitoring in Europe. Having this integrated and continuous monitoring capacity would allow more timely and efficient interventions that would optimise our capacity to revert biodiversity loss and prevent environmental degradation. It would also increase the value-added to the data flows, reaching high-value outputs with some existing low-value inputs. This deliverable offers a critical analysis of the existing monitoring landscape in Europe, extracting key messages about the main challenges, lessons learned and possible solutions. Based on a comprehensive analysis of needs and, most importantly, on an inclusive consultation process, the deliverable designs an EBOCC that tackles the key biodiversity monitoring challenges. The proposal specifies the mission, the tasks, the most urgent topics, the main policies and the key stakeholders that the EBOCC should serve and focus on during the first stage of its implementation. It also includes detailed analyses about governance models and potential costs. With this proposal, EuropaBON fosters the setting up and testing an operational EBOCC that could address the urgent need for coordination, integration, harmonisation and strengthening of biodiversity data collection and analysis, in order to inform policy-making at local, national, European and international level