Retargeting the \u3ci\u3eClostridium botulinum\u3c/i\u3e C2 toxin to the neuronal cytosol

Many biological toxins are known to attack specific cell types, delivering their enzymatic payloads to the cytosol. This process can be manipulated by molecular engineering of chimeric toxins. Using toxins with naturally unlinked components as a starting point is advantageous because it allows for the development of payloads separately from the binding/translocation components. Here the Clostridium botulinum C2 binding/translocation domain was retargeted to neural cell populations by deleting its non-specific binding domain and replacing it with a C. botulinum neurotoxin binding domain. This fusion protein was used to deliver fluorescently labeled payloads to Neuro-2a cells. Intracellular delivery was quantified by flow cytometry and found to be dependent on artificial enrichment of cells with the polysialoganglioside receptor GT1b. Visualization by confocal microscopy showed a dissociation of payloads from the early endosome indicating translocation of the chimeric toxin. The natural Clostridium botulinum C2 toxin was then delivered to human glioblastoma A172 and synchronized HeLa cells. In the presence of the fusion protein, native cytosolic enzymatic activity of the enzyme was observed and found to be GT1b-dependent. This retargeted toxin may enable delivery of therapeutics to peripheral neurons and be of use in addressing experimental questions about neural physiology

An evaluation of the relative efficacy of an open airway, an oxygen reservoir and continuous positive airway pressure 5 cmH2O on the non-ventilated lung

Publisher's copy made available with the permission of the publisher © Australian Society of AnaesthetistsThe aim of this study, during one-lung ventilation, was to evaluate if oxygenation could be improved by use of a simple oxygen reservoir or application of 5 cmH2O continuous positive airway pressure (CPAP) to the non-ventilated lung compared with an open airway. Twenty-three patients with lung malignancy, undergoing thoracotomy requiring at least 60 minutes of one-lung ventilation before lung lobe excision, were studied. After routine induction and establishment of one-lung ventilation, the three treatments were applied in turn to the same patient in a sequence selected randomly. The first treatment was repeated as a fourth treatment and these results of the repeated treatment averaged to minimize the effect of slow changes. Arterial oxygenation was measured by an arterial blood gas 15 minutes after the application of each treatment. Twenty patients completed the study. Mean PaO2 (in mmHg) was 210.3 (SD 105.5) in the 'OPEN' treatment, 186.0 (SD 109.2) in the 'RESERVOIR' treatment, and 240.5 (SD 116.0) in the 'CPAP' treatment. This overall difference was not quite significant (P=0.058, paired ANOVA), but comparison of the pairs showed that there was a significant better oxygenation only with the CPAP compared to the reservoir treatments (t=2.52, P=0.021). While the effect on the surgical field was not apparent in most patients, in one patient surgery was impeded during CPAP. Our results show that the use of a reservoir does not give oxygenation better than an open tube, and is less effective than the use of CPAP 5 cmH2O on the non-ventilated lung during one-lung ventilation.J. Slimani, W. J. Russell, C. Jurisevichttp://www.aaic.net.au/Article.asp?D=200404

Halothane potentiates the alcohol-adduct induced TNF-alpha release in heart endothelial cells

BACKGROUND: The possibility exists for major complications to occur when individuals are intoxicated with alcohol prior to anesthetization. Halothane is an anesthetic that can be metabolized by the liver into a highly reactive product, trifluoroacetyl chloride, which reacts with endogenous proteins to form a trifluoroacetyl-adduct (TFA-adduct). The MAA-adduct which is formed by acetaldehyde (AA) and malondialdehyde reacting with endogenous proteins, has been found in both patients and animals chronically consuming alcohol. These TFA and MAA-adducts have been shown to cause the release of inflammatory products by various cell types. If both adducts share a similar mechanism of cell activation, receiving halothane anesthesia while intoxicated with alcohol could exacerbate the inflammatory response and lead to cardiovascular injury. METHODS: We have recently demonstrated that the MAA-adduct induces tumor necrosis factor-α (TNF-α) release by heart endothelial cells (HECs). In this study, pair and alcohol-fed rats were randomized to receive halothane pretreatments intra peritoneal. Following the pretreatments, the intact heart was removed, HECs were isolated and stimulated with unmodified bovine serum albumin (Alb), MAA-modified Alb (MAA-Alb), Hexyl-MAA, or lipopolysaccharide (LPS), and supernatant concentrations of TNF-α were measured by ELISA. RESULTS: Halothane pre-treated rat HECs released significantly greater TNF-α concentration following MAA-adduct and LPS stimulation than the non-halothane pre-treated in both pair and alcohol-fed rats, but was significantly greater in the alcohol-fed rats. CONCLUSION: These results demonstrate that halothane and MAA-adduct pre-treatment increases the inflammatory response (TNF-α release). Also, these results suggest that halothane exposure may increase the risk of alcohol-induced heart injury, since halothane pre-treatment potentiates the HEC TNF-α release measured following both MAA-Alb and LPS stimulation

Defining the gap between research and practice in public relations programme evaluation - towards a new research agenda

The current situation in public relations programme evaluation is neatly summarized by McCoy who commented that 'probably the most common buzzwords in public relations in the last ten years have been evaluation and accountability' (McCoy 2005, 3). This paper examines the academic and practitioner-based literature and research on programme evaluation and it detects different priorities and approaches that may partly explain why the debate on acceptable and agreed evaluation methods continues. It analyses those differences and proposes a research agenda to bridge the gap and move the debate forward

Media justice: Madeleine McCann, intermediatization and "trial by media" in the British press

Three-year-old Madeleine McCann disappeared on 3 May 2007 from a holiday apartment in Portugal. Over five years and multiple investigations that failed to solve this abducted child case, Madeleine and her parents were subject to a process of relentless ‘intermediatization’. Across 24–7 news coverage, websites, documentaries, films, YouTube videos, books, magazines, music and artworks, Madeleine was a mediagenic image of innocence and a lucrative story. In contrast to Madeleine’s media sacralization, the representation of her parents, Kate and Gerry McCann, fluctuated between periods of vociferous support and prolonged and libellous ‘trial by media’. This article analyses how the global intermediatization of the ‘Maddie Mystery’ fed into and fuelled the ‘trial by media’ of Kate and Gerry McCann in the UK press. Our theorization of ‘trial by media’ is developed and refined through considering its legal limitations in an era of ‘attack journalism’ and unprecedented official UK inquiries into press misconduct and criminality

Molten core debris-sodium interactions: M-Series experiments. [LMFBR]

Five new kilogram-scale experiments have been carried out. Four of the experiments simulated the situation where molten core debris flows from a breached reactor vessel into a dry reactor cavity and is followed by a flow of sodium (Ex-vessel case) and one experiment simulated the flow of core debris into an existing pool of sodium (In-vessel case). The core debris was closely simulated by a thermite reaction which produced a molten mixture of UO/sub 2/, ZrO/sub 2/, and stainless steel. There was efficient fragmentation of the debris in all experiments with no explosive interactions observed

Complications from Surgeries Related to Ovarian Cancer Screening

The aim of this study was to evaluate complications of surgical intervention for participants in the Kentucky Ovarian Cancer Screening Program and compare results to those of the Prostate, Lung, Colorectal and Ovarian Cancer Screening trial. A retrospective database review included 657 patients who underwent surgery for a positive screen in the Kentucky Ovarian Cancer Screening Program from 1988–2014. Data were abstracted from operative reports, discharge summaries, and office notes for 406 patients. Another 142 patients with incomplete records were interviewed by phone. Complete information was available for 548 patients. Complications were graded using the Clavien–Dindo (C–D) Classification of Surgical Complications and considered minor if assigned Grade I (any deviation from normal course, minor medications) or Grade II (other pharmacological treatment, blood transfusion). C–D Grade III complications (those requiring surgical, endoscopic, or radiologic intervention) and C–D Grade IV complications (those which are life threatening) were considered “major”. Statistical analysis was performed using SAS 9.4 software. Complications were documented in 54/548 (10%) subjects. For women with malignancy, 17/90 (19%) had complications compared to 37/458 (8%) with benign pathology (p \u3c 0.003). For non-cancer surgery, obesity was associated with increased complications (p = 0.0028). Fifty patients had minor complications classified as C–D Grade II or less. Three of 4 patients with Grade IV complications had malignancy (p \u3c 0.0004). In the Prostate, Lung, Colorectal and Ovarian Cancer Screening trial, 212 women had surgery for ovarian malignancy, and 95 had at least one complication (45%). Of the 1080 women with non-cancer surgery, 163 had at least one complication (15%). Compared to the Prostate, Lung, Colorectal and Ovarian Cancer Screening trial, the Kentucky Ovarian Cancer Screening Program had significantly fewer complications from both cancer and non-cancer surgery (p \u3c 0.0001 and p = 0.002, respectively). Complications resulting from surgery performed as a result of the Kentucky Ovarian Cancer Screening Program were infrequent and significantly fewer than reported in the Prostate, Lung, Colorectal and Ovarian Cancer Screening trial. Complications were mostly minor (93%) and were more common in cancer versus non-cancer surgery

Present Status and Future Programs of the n_TOF Experiment

This is an Open Access article distributed under the terms of the Creative Commons Attribution-Noncommercial License 3.0, which permits unrestricted use, distribution, and reproduction in any noncommercial medium, provided the original work is properly citedThe neutron time-of-flight facility n_TOF at CERN, Switzerland, operational since 2001, delivers neutrons using the Proton Synchrotron (PS) 20 GeV/c proton beam impinging on a lead spallation target. The facility combines a very high instantaneous neutron flux, an excellent time of flight resolution due to the distance between the experimental area and the production target (185 meters), a low intrinsic background and a wide range of neutron energies, from thermal to GeV neutrons. These characteristics provide a unique possibility to perform neutron-induced capture and fission cross-section measurements for applications in nuclear astrophysics and in nuclear reactor technology.The most relevant measurements performed up to now and foreseen for the future will be presented in this contribution. The overall efficiency of the experimental program and the range of possible measurements achievable with the construction of a second experimental area (EAR-2), vertically located 20 m on top of the n_TOF spallation target, might offer a substantial improvement in measurement sensitivities. A feasibility study of the possible realisation of the installation extension will be also presented

Cross section measurements of 155,157Gd(n, γ) induced by thermal and epithermal neutrons

© SIF, Springer-Verlag GmbH Germany, part of Springer Nature 2019Neutron capture cross section measurements on 155Gd and 157Gd were performed using the time-of-flight technique at the n_TOF facility at CERN on isotopically enriched samples. The measurements were carried out in the n_TOF experimental area EAR1, at 185 m from the neutron source, with an array of 4 C6D6 liquid scintillation detectors. At a neutron kinetic energy of 0.0253 eV, capture cross sections of 62.2(2.2) and 239.8(8.4) kilobarn have been derived for 155Gd and 157Gd, respectively, with up to 6% deviation relative to values presently reported in nuclear data libraries, but consistent with those values within 1.6 standard deviations. A resonance shape analysis has been performed in the resolved resonance region up to 181 eV and 307 eV, respectively for 155Gd and 157Gd, where on average, resonance parameters have been found in good agreement with evaluations. Above these energies and up to 1 keV, the observed resonance-like structure of the cross section has been analysed and characterised. From a statistical analysis of the observed neutron resonances we deduced: neutron strength function of 2. 01 (28) × 10 - 4 and 2. 17 (41) × 10 - 4; average total radiative width of 106.8(14) meV and 101.1(20) meV and s-wave resonance spacing 1.6(2) eV and 4.8(5) eV for n + 155Gd and n + 157Gd systems, respectively.Peer reviewedFinal Accepted Versio