A study of the practical challenges associated with developing a new test for bovine tuberculosis

Bovine tuberculosis, a chronic disease caused by Mycobacterium bovis, is a major economic problem affecting farmers both in the UK and worldwide. In order to prevent and control bovine tuberculosis it is very important to be able to promptly and definitively diagnose infected animals. However, the current diagnostic tools show some limitations which can contribute to cause economic losses and to the transmission of the disease to other animals in a herd, to wildlife and potentially to humans. Therefore, there is an urgent need to have available new diagnostic methods that allow the definitive detection of bovine tuberculosis. In this work, the practical issues associated with the cost-effective production of two diagnostic kits that employ mycobacteriophage (ActiphageTM and ActiphageTM Rapid) were investigated. This technology exploits the ability of bacteriophage to infect live mycobacterial species in a strain specific manner. This technique was originally developed as a tool for the diagnosis of human tuberculosis (FastPlaqueTBTM, BIOTEC) and has now been refined to offer transformational tools with significant applications in veterinary, agriculture and - potentially – the human sector. These kits are manufactured and distributed by PBD Biotech Ltd. (Thurston, Suffolk, UK) and allow the detection of the presence of M. bovis and M. avium subsp. paratuberculosis in diverse sample types, such as blood and milk. The assays can therefore be used for infection control and as a part of a quality assurance programme to confirm the absence of these organisms in non-pasteurized dairy products. Furthermore, the methods can be adapted to detect other types of mycobacteria and applied to detect infection not only in cattle but also in other animal hosts including sheep, goats, deer, horses, alpacas, llamas as well as companion animals and exotic species. In order to develop the simplest method that gives a reproducible titre of at least 1010 pfu ml-1, three different ways of manufacturing the freeze-dried bacteriophage supplied with the kits were explored. With all of the three methods, there was no difference in the titre of the phage culture before and after freeze drying. This suggests that the process does not have a negative impact on the viability of the phage. With the first two methods employed, the titre obtained was normally 109 pfu ml-1. Only one time, using method 2, the titre obtained was 1010 pfu ml-1. This titre seems to have been routinely achieved modifying method 2 in order to ensure an optimal elution of the bacteriophage from the agar to the medium. Shelf life studies on the freeze dried phage are being performed and, so far, they revealed an average stability of about 3 months if stored at +5 / +8°C. The practical issues related to manufacturing of the virucide tablets supplied with the ActiphageTM Core kit were also explored and a softer formula led to jamming of the tablet press and broken tablets but also produced a tablet that was easier to dissolve. Once the production issues were solved by adjusting the machine settings, a procedure for checking the weights of the tablets was developed. Finally, work was carried out to identify the best source of the other kits components and to design and test packaging materials, and finally an evaluation of the cost of competitor tests was performed to better understand the market price that could be charged for ActiphageTM tests

Laboratory Ageing Thermal Cycles of MV Cable Joints: results of different diagnostic techniques

The article reports the results of a multi-year experimentation in progress at the RSE laboratories and aimed at an in-depth diagnostic analysis of the causes of failure responsible for the out-of-services which affect MV cable joints of most of the urban network in the summer periods. To this aim an experimental set-up consisting of twelve cables equipped with different types of joints were connected together to form a ring in short-circuit. The ring was energized at rated voltage and current cycles were applied by means of external coils in order to reproduce the daily load variation typical of a MV cable network. Felts were wrapped around the cables to simulate the reduced thermal conductivity of the soil during summer periods. Continuous monitoring of the degradation of the joints was carried out by Partial Discharge (PD, on-line and off-line) measurements, together with Frequency Dielectric Spectroscopy (FDS) and Dissipation Factor (DF) - measurements. Results of the experimentation are presented and discussed

A New Inference System for the Robust Identification of Defects Supporting PD in MV Cables

A method for the identification of a large number of defects supporting Partial Discharges (PD) in MV cables is presented in the paper. A specific set of parameters that summarize features related to the shape of the Phase-Resolved PD (PRPD) patterns, is organized in a tree structure on the assumption of a correlation between the PRPD shapes and the defect typologies. Tree inspection resorts to the Fuzzy-Logic to take into account the inherent uncertainty of the identification results. Predicates that address PRPD to the defect typologies and the relevant membership values are provided as the output taking into account even more than one option. Both training and working examples obtained testing artificial defects and real cables, are presented and discussed

CD40 activation of BCP-ALL cells generates IL-10-producing, IL-12-defective APCs that induce allogeneic T-cell anergy

The use of leukemia cells as antigenpresenting cells (APCs) in immunotherapy is critically dependent on their capacity to initiate and sustain an antitumor-specific immune response. Previous studies suggested that pediatric B-cell precursor acute lymphoblastic leukemia (BCP-ALL) cells could be manipulated in vitro through the CD40-CD40L pathway to increase their immunostimulatory capacity. We extended the APC characterization of CD40L-activated BCP-ALL for their potential use in immunotherapy in a series of 19 patients. Engaging CD40 induced the up-regulation of CCR7 in 7 of 11 patients and then the migration to CCL19 in 2 of 5 patients. As accessory cells, CD40Lactivated BCP-ALL induced a strong proliferation response of naive T lymphocytes. Leukemia cells, however, were unable to sustain proliferation over time, and T cells eventually became anergic. After CD40-activation, BCP-ALL cells released substantial amounts of interleukin-10 (IL-10) but were unable to produce bioactive IL-12 or to polarize TH1 effectors. Interestingly, adding exogenous IL-12 induced the generation of interferon- (IFN- )–secreting TH1 effectors and reverted the anergic profile in a secondary response. Therefore, engaging CD40 on BCP-ALL cells is insufficient for the acquisition of full functional properties of immunostimulatory APCs. These results suggest caution against the potential use of CD40L-activated BCP-ALL cells as agents for immunotherapy unless additional stimuli, such as IL-12, are provided.Fil: D'Amico, Giovanna. Università Milano Bicocca; ItaliaFil: Vulcano, Marisa. Universidad de Buenos Aires. Facultad de Medicina; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; ArgentinaFil: Bugarin, Cristina. Università Milano Bicocca; ItaliaFil: Bianchi, Giancarlo. Università Milano Bicocca; ItaliaFil: Pirovano, Gisella. Università Milano Bicocca; ItaliaFil: Bonamino, Martin. Università Milano Bicocca; ItaliaFil: Marin, Virna. Università Milano Bicocca; ItaliaFil: Allavena, Paola. Università Milano Bicocca; ItaliaFil: Biagi, Ettore. Università Milano Bicocca; ItaliaFil: Biondi, Andrea. Università Milano Bicocca; Itali

First Results of the 140^{140}Ce(n,γ)141^{141}Ce Cross-Section Measurement at n_TOF

An accurate measurement of the $^{140}$Ce(n,γ) energy-dependent cross-section was performed at the n_TOF facility at CERN. This cross-section is of great importance because it represents a bottleneck for the s-process nucleosynthesis and determines to a large extent the cerium abundance in stars. The measurement was motivated by the significant difference between the cerium abundance measured in globular clusters and the value predicted by theoretical stellar models. This discrepancy can be ascribed to an overestimation of the $^{140}$Ce capture cross-section due to a lack of accurate nuclear data. For this measurement, we used a sample of cerium oxide enriched in $^{140}$Ce to 99.4%. The experimental apparatus consisted of four deuterated benzene liquid scintillator detectors, which allowed us to overcome the difficulties present in the previous measurements, thanks to their very low neutron sensitivity. The accurate analysis of the p-wave resonances and the calculation of their average parameters are fundamental to improve the evaluation of the $^{140}$Ce Maxwellian-averaged cross-section

First Results of the 140^{140}Ce(n,γ)141^{141}Ce Cross-Section Measurement at n_TOF

An accurate measurement of the $^{140}$Ce(n,γ) energy-dependent cross-section was performed at the n_TOF facility at CERN. This cross-section is of great importance because it represents a bottleneck for the s-process nucleosynthesis and determines to a large extent the cerium abundance in stars. The measurement was motivated by the significant difference between the cerium abundance measured in globular clusters and the value predicted by theoretical stellar models. This discrepancy can be ascribed to an overestimation of the $^{140}$Ce capture cross-section due to a lack of accurate nuclear data. For this measurement, we used a sample of cerium oxide enriched in $^{140}$Ce to 99.4%. The experimental apparatus consisted of four deuterated benzene liquid scintillator detectors, which allowed us to overcome the difficulties present in the previous measurements, thanks to their very low neutron sensitivity. The accurate analysis of the p-wave resonances and the calculation of their average parameters are fundamental to improve the evaluation of the $^{140}$Ce Maxwellian-averaged cross-section

Lithium and GSK3-β promoter gene variants influence white matter microstructure in bipolar disorder

Lithium is the mainstay for the treatment of bipolar disorder (BD) and inhibits glycogen synthase kinase 3-β (GSK3-β). The less active GSK3-β promoter gene variants have been associated with less detrimental clinical features of BD. GSK3-β gene variants and lithium can influence brain gray matter structure in psychiatric conditions. Diffusion tensor imaging (DTI) measures of white matter (WM) integrity showed widespred disruption of WM structure in BD. In a sample of 70 patients affected by a major depressive episode in course of BD, we investigated the effect of ongoing long-term lithium treatment and GSK3-β promoter rs334558 polymorphism on WM microstructure, using DTI and tract-based spatial statistics with threshold-free cluster enhancement. We report that the less active GSK3-β rs334558*C gene-promoter variants, and the long-term administration of the GSK3-β inhibitor lithium, were associated with increases of DTI measures of axial diffusivity (AD) in several WM fiber tracts, including corpus callosum, forceps major, anterior and posterior cingulum bundle (bilaterally including its hippocampal part), left superior and inferior longitudinal fasciculus, left inferior fronto-occipital fasciculus, left posterior thalamic radiation, bilateral superior and posterior corona radiata, and bilateral corticospinal tract. AD reflects the integrity of axons and myelin sheaths. We suggest that GSK3-β inhibition and lithium could counteract the detrimental influences of BD on WM structure, with specific benefits resulting from effects on specific WM tracts contributing to the functional integrity of the brain and involving interhemispheric, limbic, and large frontal, parietal, and fronto-occipital connections

Burnout among surgeons before and during the SARS-CoV-2 pandemic: an international survey

Background: SARS-CoV-2 pandemic has had many significant impacts within the surgical realm, and surgeons have been obligated to reconsider almost every aspect of daily clinical practice. Methods: This is a cross-sectional study reported in compliance with the CHERRIES guidelines and conducted through an online platform from June 14th to July 15th, 2020. The primary outcome was the burden of burnout during the pandemic indicated by the validated Shirom-Melamed Burnout Measure. Results: Nine hundred fifty-four surgeons completed the survey. The median length of practice was 10 years; 78.2% included were male with a median age of 37 years old, 39.5% were consultants, 68.9% were general surgeons, and 55.7% were affiliated with an academic institution. Overall, there was a significant increase in the mean burnout score during the pandemic; longer years of practice and older age were significantly associated with less burnout. There were significant reductions in the median number of outpatient visits, operated cases, on-call hours, emergency visits, and research work, so, 48.2% of respondents felt that the training resources were insufficient. The majority (81.3%) of respondents reported that their hospitals were included in the management of COVID-19, 66.5% felt their roles had been minimized; 41% were asked to assist in non-surgical medical practices, and 37.6% of respondents were included in COVID-19 management. Conclusions: There was a significant burnout among trainees. Almost all aspects of clinical and research activities were affected with a significant reduction in the volume of research, outpatient clinic visits, surgical procedures, on-call hours, and emergency cases hindering the training. Trial registration: The study was registered on clicaltrials.gov "NCT04433286" on 16/06/2020