Identification of host cell factors required for intoxication through use of modified cholera toxin

We describe a novel labeling strategy to site-specifically attach fluorophores, biotin, and proteins to the C terminus of the A1 subunit (CTA1) of cholera toxin (CTx) in an otherwise correctly assembled and active CTx complex. Using a biotinylated N-linked glycosylation reporter peptide attached to CTA1, we provide direct evidence that ∼12% of the internalized CTA1 pool reaches the ER. We also explored the sortase labeling method to attach the catalytic subunit of diphtheria toxin as a toxic warhead to CTA1, thus converting CTx into a cytolethal toxin. This new toxin conjugate enabled us to conduct a genetic screen in human cells, which identified ST3GAL5, SLC35A2, B3GALT4, UGCG, and ELF4 as genes essential for CTx intoxication. The first four encode proteins involved in the synthesis of gangliosides, which are known receptors for CTx. Identification and isolation of the ST3GAL5 and SLC35A2 mutant clonal cells uncover a previously unappreciated differential contribution of gangliosides to intoxication by CTx.Fundação para a Ciência e a Tecnologia (Fellowship

E-Debitum: managing software energy debt

35th IEEE/ACM International Conference on Automated Software Engineering Workshops (ASEW ’20) - International Workshop on Sustainable Software Engineering (SUSTAIN-SE)This paper extends previous work on the concept of a new software energy metric: Energy Debt. This metric is a reflection on the implied cost, in terms of energy consumption over time, of choosing an energy flawed software implementation over a more robust and efficient, yet time consuming, approach. This paper presents the implementation a SonarQube tool called E-Debitum which calculates the energy debt of Android applications throughout their versions. This plugin uses a robust, well defined, and extendable smell catalogue based on current green software literature, with each smell defining the potential energy savings. To conclude, an experimental validation of E-Debitum was executed on 3 popular Android applications with various releases, showing how their energy debt fluctuated throughout releases.This work is financed by National Funds through the Portuguese funding agency, FCT -Fundação para a Ciência e a Tecnologia within project UIDB/50014/2020

Identification of genetic variants associated with Huntington's disease progression: a genome-wide association study

Background Huntington's disease is caused by a CAG repeat expansion in the huntingtin gene, HTT. Age at onset has been used as a quantitative phenotype in genetic analysis looking for Huntington's disease modifiers, but is hard to define and not always available. Therefore, we aimed to generate a novel measure of disease progression and to identify genetic markers associated with this progression measure. Methods We generated a progression score on the basis of principal component analysis of prospectively acquired longitudinal changes in motor, cognitive, and imaging measures in the 218 indivduals in the TRACK-HD cohort of Huntington's disease gene mutation carriers (data collected 2008–11). We generated a parallel progression score using data from 1773 previously genotyped participants from the European Huntington's Disease Network REGISTRY study of Huntington's disease mutation carriers (data collected 2003–13). We did a genome-wide association analyses in terms of progression for 216 TRACK-HD participants and 1773 REGISTRY participants, then a meta-analysis of these results was undertaken. Findings Longitudinal motor, cognitive, and imaging scores were correlated with each other in TRACK-HD participants, justifying use of a single, cross-domain measure of disease progression in both studies. The TRACK-HD and REGISTRY progression measures were correlated with each other (r=0·674), and with age at onset (TRACK-HD, r=0·315; REGISTRY, r=0·234). The meta-analysis of progression in TRACK-HD and REGISTRY gave a genome-wide significant signal (p=1·12 × 10−10) on chromosome 5 spanning three genes: MSH3, DHFR, and MTRNR2L2. The genes in this locus were associated with progression in TRACK-HD (MSH3 p=2·94 × 10−8 DHFR p=8·37 × 10−7 MTRNR2L2 p=2·15 × 10−9) and to a lesser extent in REGISTRY (MSH3 p=9·36 × 10−4 DHFR p=8·45 × 10−4 MTRNR2L2 p=1·20 × 10−3). The lead single nucleotide polymorphism (SNP) in TRACK-HD (rs557874766) was genome-wide significant in the meta-analysis (p=1·58 × 10−8), and encodes an aminoacid change (Pro67Ala) in MSH3. In TRACK-HD, each copy of the minor allele at this SNP was associated with a 0·4 units per year (95% CI 0·16–0·66) reduction in the rate of change of the Unified Huntington's Disease Rating Scale (UHDRS) Total Motor Score, and a reduction of 0·12 units per year (95% CI 0·06–0·18) in the rate of change of UHDRS Total Functional Capacity score. These associations remained significant after adjusting for age of onset. Interpretation The multidomain progression measure in TRACK-HD was associated with a functional variant that was genome-wide significant in our meta-analysis. The association in only 216 participants implies that the progression measure is a sensitive reflection of disease burden, that the effect size at this locus is large, or both. Knockout of Msh3 reduces somatic expansion in Huntington's disease mouse models, suggesting this mechanism as an area for future therapeutic investigation

Concomitant changes in clinical and posturographic data in elderly fallers during the course of an in-home anti-falling multimodal program – A preliminary investigation

International audienceSummaryAim of the study The objective of this preliminary study was to evaluate the effectiveness, in terms of fall reduction, of an in-home strategy that we have developed for elderly fallers. We also aimed at finding links between the expected changes in the data obtained in static posturography and in clinical balance tests through our program. Patients and methods Twelve elderly patients living at home who were diagnosed as fallers (5 males and 7 females; 77.9 ± 4.1 years) participated in the study. Our multimodal intervention lasted 6 months. Before this period, and one year later, an evaluation was conducted using cognitive (MMSE), clinical balance tests (i.e. Berg Balance Scale, Balance One leg, Timed Up and Go, and Functional Reach tests) and static posturography (where the area of body sway, velocity and medio–lateral and antero–posterior amplitudes were recorded twice, first with eyes open and then with eyes closed). Results Among the 12 patients who were diagnosed as fallers, eight became non-fallers. When comparing data obtained after the intervention with those obtained beforehand, we found significant changes in all of the clinical balance tests and in the posturographic-derived variables indicating improvements in the balance control in our group of subjects. We also found significant correlations between the changes in the Berg Balance Scale scores and the changes in the area of body sway data, in antero–posterior amplitude both with eyes open and with eyes closed, and also in the medio–lateral amplitude in the eyes closed condition. Conclusions We prospectively demonstrated the relevance of our anti-falling intervention at home and of the use of posturography for clinical follow-up

Monitoring the risk for a “valley of inadequate anesthesia” using bispectral index and the noxious stimulation response index during different inflow speeds of sevoflurane.

Title: Monitoring the risk for a “valley of inadequate anesthesia” using bispectral index and the noxious stimulation response index during different inflow speeds of sevoflurane. Authors: Mira Van Thielen MD1 2, Rik Carette MD1 , Jan F.A. Hendrickx MD, PhD1 2 , Andre M. De Wolf MD, PhD3 , Hugo E.M. Vereecke MD, PhD4 5 Affilliations: 1 OLV Hospital, Aalst, Belgium; 2 Ghent University, Ghent, Belgium; 3 Northwestern University, Chicago, USA; 4 AZ Sint-Jan Brugge-Oostende AV, Brugge, Belgium; 5 University Medical Center Groningen and University of Groningen, Groningen, The Netherlands Abstract Introduction: The “valley of inadequate anesthesia” refers to an increased risk for awareness, movement or hemodynamic instability evoked by an insufficient inflow of volatile agents to compensate for the gradual elimination of intravenously administered boluses of induction agents. The combined drug effects of volatile and intravenous drugs should ensure a sufficient hypnotic drug effect and immobility (as respectively quantified by bispectral index (BIS, Medtronic) and the noxious stimulation response index (NSRI, Dräger)), while maintaining stability in arterial blood pressure (ABP) and heart rate (HR). NSRI is a derivative of the probability of tolerance of laryngoscopy, as estimated by Hannivoort et al. for most combinations of propofol, opioids and volatile agents.1 NSRI of 20 equals an estimated probability of immobility in response to laryngoscopy of 90%. We compare the time course of BIS, NSRI, ABP and HR after a bolus of propofol and sufentanil, followed by three different inflow speeds of sevoflurane. We hypothesize that BIS and NSRI should stay below 60 and 20, respectively, at all times between the start of sevoflurane and reaching a target of 1.0 MAC. We also compare ABP and HR between groups. Methods: 33 ASA score I-III patients (age range: 27-86 years; BMI range 22-34 kg/m2 ) presenting for abdominal surgery received sufentanil (0.2 µg/kg) and propofol (1 or 2 mg/kg, depending on age) followed by intubation of the trachea, 2.5 min after loss of responsiveness and rocuronium (0.6 mg/kg). Sevoflurane was administered (after randomization) in a ‘slow’(n = 9), ‘medium’(n = 8), and ‘fast’(n = 9) group; defined by a time constant of respectively 10.9, 5.7, and 2.6 min wash-in time towards 1.0 MAC. These inflow speeds are similar to those used in a commercialized automatic controller of the inflow of volatile agents (Flow-I, Maquet). BIS and NSRI were blinded to the anesthesiologist. For all measures, the 95% confidence interval (CI 95%) of the difference of means was calculated at one minute intervals between minutes 0 to 25 after propofol (p < 0.05 if zero is outside 95% CI). An escape bolus of sufentanil (0.1 µg/kg) was allowed per protocol (and included in the NSRI calculations) to counter movement, tachycardia or hypertension indicating insufficient anesthesia at any time. Results and discussion: Extra sufentanil was needed in all groups, for movement (1 in slow, 1 in medium, 1 in fast), for tachycardia (1 in slow, 1 in medium and 1 in fast), and for hypertension (3 in medium, 2 in fast). Figure 1 shows the time course of BIS, NSRI, HR and MAP. All groups included cases with BIS > 60 and/or NSRI > 20 (n in slow > medium > fast). High BIS and NSRI were not always consistent. Figure 2 shows similar BIS and NSRI during the first 8 minutes in all groups, confirming equipotency for these measures till the start of sevoflurane. Mean NSRI differs between slow and medium, slow and fast and medium and fast (respectively from minute 10 to 18, 10 to 20 and 10 to 14) evoked by the deliberate differences in inflow speed of sevoflurane. Mean BIS differs between slow and medium and slow and fast (respectively from minute 15 to 16 and 11 to 24). Heart rate and blood pressure did not differ between groups. A Brice questionnaire found no cases of explicit recall. Conclusion: After a bolus dose of propofol, sufentanil, and rocuronium, both BIS>60 and NSRI>20 warn the anesthesiologist for an increased risk of a “valley of inadequate anesthesia”. ABP and HR don’t identify differences in drug potency between groups. A higher initial dose of sufentanil (e.g. 0,3 µg/kg) might reduce the need for escape treatment. Reference 1. Hannivoort LN, et al., Br J Anaesth. 2016 May;116(5):624-31