The effect of quarantine due to Covid-19 pandemic on seizure frequency in 102 adult people with epilepsy from Apulia and Basilicata regions, Southern Italy

Objective: following the COVID-19 pandemic, a quarantine was imposed to all of regions Italy by 9th March until 3rd May 2020. We investigated the effect of COVID-19 infection and quarantine on seizure frequency in adult people with epilepsy (PwE) of Apulia and Basilicata regions, Southern Italy. Methods: This is an observational, retrospective study based on prospective data collection of 102 successive PWE. The frequency of seizures was evaluated during pre-quarantine (January- February), quarantine (March-April), and post-quarantine period (May-June), while PwE were divided into A) cases responding to treatment with ≤ 1 seizure per year; B) cases responding to treatment with 2-5 seizure per year; C) cases with drug-resistant epilepsy with ≤ 4 seizures per month; D) cases with drug-resistant epilepsy with 5-10 seizures per month. PwE underwent several self-report questionnaires regarding therapeutic compliance, mood, stress and sleep during quarantine period. Results: Approximately 50 % of PwE showed seizure frequency changes (22.55 % an increase and 27.45 % a reduction) during quarantine. Seizure frequency significantly (p < 0.05) increased in PwE responding to treatment with ≤ 1 seizure per year, while significantly (p < 0.05) reduced in PwE with drug-resistant epilepsy with 5-10 seizures per month. The data was not influenced by therapeutic adherence, sleep and depression. The analysis of anxiety showed a moderate level of anxiety in PwE responding to treatment with < 1 seizure per year, while moderate stress was perceived by all PwE. Seizure frequency changes were related to quarantine, but not to COVID-19 infection. In fact, unlike other regions of Italy, particularly Northern Italy, Apulia and Basilicata regions were less affected by COVID-19 infection, and almost all PwE recognized the quarantine as a stressful event. Emotional distress and anxiety due to social isolation, but also the relative reduction of triggers for epileptic seizures were the most important factors for changes in seizure frequency. Conclusions: Our study adds to the growing concern that the indirect effects of COVID-19 pandemic will far outstrip the direct consequences of the infection

Tegument Glycoproteins and Cathepsins of Newly Excysted Juvenile Fasciola hepatica Carry Mannosidic and Paucimannosidic N-glycans

Recently, the prevalence of Fasciola hepatica in some areas has increased considerably and the availability of a vaccine to protect livestock from infection would represent a major advance in tools available for controlling this disease. To date, most vaccine-target discovery research on this parasite has concentrated on proteomic and transcriptomic approaches whereas little work has been carried out on glycosylation. As the F. hepatica tegument (Teg) may contain glycans potentially relevant to vaccine development and the Newly Excysted Juvenile (NEJ) is the first lifecycle stage in contact with the definitive host, our work has focused on assessing the glycosylation of the NEJTeg and identifying the NEJTeg glycoprotein repertoire. After in vitro excystation, NEJ were fixed and NEJTeg was extracted. Matrix-assisted laser desorption ionisation-time of flight-mass spectrometry (MALDI-TOF-MS) analysis of released N-glycans revealed that oligomannose and core-fucosylated truncated N-glycans were the most dominant glycan types. By lectin binding studies these glycans were identified mainly on the NEJ surface, together with the oral and ventral suckers. NEJTeg glycoproteins were affinity purified after targeted biotinylation of the glycans and identified using liquid chromatography and tandem mass spectrometry (LC-MS/MS). From the total set of proteins previously identified in NEJTeg, eighteen were also detected in the glycosylated fraction, including the F. hepatica Cathepsin B3 (FhCB3) and two of the Cathepsin L3 (FhCL3) proteins, among others. To confirm glycosylation of cathepsins, analysis at the glycopeptide level by LC-ESI-ion-trap-MS/MS with collision-induced dissociation (CID) and electron-transfer dissociation (ETD) was carried out. We established that cathepsin B1 (FhCB1) on position N80, and FhCL3 (BN1106_s10139B000014, scaffold10139) on position N153, carry unusual paucimannosidic Man2GlcNAc2 glycans. To our knowledge, this is the first description of F. hepatica NEJ glycosylation and the first report of N-glycosylation of F. hepatica cathepsins. The significance of these findings for immunological studies and vaccine development is discussed

Performance of the Crowbar of the LHC High Power RF System

The counter-rotating proton beams in the Large Hadron Collider (LHC) are captured and accelerated to their final energies by two identical 400 MHz Radio Frequency (RF) systems. The RF power source required for each beam comprises eight 300 kW klystrons. The output power of each klystron is fed via a circulator and a waveguide line to the input coupler of a single-cell superconducting (SC) cavity. Each unit of four klystrons is powered by a -100kV/40A AC/DC power converter. A fast protection system (crowbar) protects the four klystrons in each of these units. Although the LHC RF system has shown has very good performance, operational experience has shown that the five-gap double-ended thyratrons used in the crowbar system suffer, from time to time, from auto-firing, which result in beam dumps. This paper presents the recent results obtained with an alternative solution based on solid state thyristors. Comparative measurements with the thyratron are shown

The periodontal risk score (PRS): initiation and model validation for 6762 teeth

Tooth-level prognostic systems are valuable tools for treatment planning and risk assessment of periodontally involved teeth. Recently the Miller-McEntire prognosis index was found to outperform comparable systems. However, it had some limitations. The present study aimed to develop and evaluate the prognostic performance of a modified version that address most limitations of the previous model, called the periodontal Risk Score (PRS)

Characterisation of <i>F</i>. <i>hepatica</i> A-BB-NEJTeg glycoprotein composition detected following glycoprotein biotinylation, affinity chromatography isolation and LC-MS/MS analysis.

<p>Characterisation of <i>F</i>. <i>hepatica</i> A-BB-NEJTeg glycoprotein composition detected following glycoprotein biotinylation, affinity chromatography isolation and LC-MS/MS analysis.</p

List of the fragment ions after CID-MS/MS of parent ions of tryptic peptide of FhCL3 (BN1106_s10139B000014).

<p>List of the fragment ions after CID-MS/MS of parent ions of tryptic peptide of FhCL3 (BN1106_s10139B000014).</p

Glycosylation of <i>F</i>. <i>hepatica</i> NEJSom.

<p>The total protein profile of <i>F</i>. <i>hepatica</i> NEJSom (Som) was revealed after SDS-PAGE fractionation and visualised in-gel by silver staining <b>(A).</b> NEJSom preparations were SDS-PAGE fractioned, transferred to nitrocellulose membranes and incubated with the biotinylated-labelled lectins GSL-I, SBA, PNA and Jacalin (<b>B</b>). Negative controls consisted of nitrocellulose membranes without lectin incubation (1) and nitrocellulose membranes incubated with biotinylated-conjugate lectins that were previously incubated with their specific inhibitors (2). The list of lectins used and their corresponding inhibitors are detailed in <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004688#pntd.0004688.t001" target="_blank">Table 1</a> (<b>C</b>). An additional incubation with IRDye-labelled streptavidin was used to detect positive lectin binding at different molecular weights (red markers). Glycoproteins were revealed by infrared imaging.</p

Glycoprotein detection and localisation on <i>F</i>. <i>hepatica</i> NEJTeg by lectin blot and lectin fluorescence staining.

<p>The total protein profile of <i>F</i>. <i>hepatica</i> NEJTeg (Teg) was revealed after SDS-PAGE fractionation and visualised in-gel by silver staining (<b>A</b>). Fixed NEJ and nitrocellulose membranes were incubated with FITC-labelled or biotinylated-conjugate lectins respectively. Specific lectin binding to glycans with terminal Fuc- (AAL, LCA, PSA), Man- (ConA, GNL), Galβ1-3GalNAc- (Jacalin, PNA), Gal/GalNAc- (RCA-120, GSL-I, SBA, DBA), Lac- (ECL), oligosaccharides- (PHA-E, PHA-L) or GlcNAc- (WGA, sWGA, GSL-II) motifs is shown. Oral sucker (or) and ventral sucker (vs) are identified with arrows (<b>B</b>). Negative controls consisted of nitrocellulose membranes without lectin incubation (1), nitrocellulose membranes incubated with biotinylated-conjugate lectins that were previously incubated with their specific inhibitors (2) and fixed NEJ incubated with FITC-labelled lectins that were previously incubated with their specific inhibitors (3). The list of lectins used and their corresponding inhibitors are detailed in <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004688#pntd.0004688.t001" target="_blank">Table 1</a> (<b>C</b>). Nitrocellulose membranes were exposed to IRDye-labelled streptavidin to reveal positive lectin binding (green) at various molecular weights (markers in red). Glycoprotein profiles were visualised by IR system. Epifluorescence microscope was employed to detect glycan localisation (green) and DNA was counterstained with DAPI (blue). Merged image of both micrographs is included for each lectin. Scale bar = 100 μm</p

Identification of short <i>N-</i>glycans occupying an <i>N-</i>glycosylation site of FhCB1.

<p>Nano-RP-LC-ESI-ion trap-MS/MS with collision-induced dissociation (<b>A</b> and <b>B</b>) and with electron transfer dissociation (<b>C</b>) of the tryptic glycopeptide Y₇₉NVSENDLPESFDAR₉₃ from FhCB1 (BN1106_s6570B000050). The [M+3H]³⁺ parent ions (blue diamonds) at <i>m/z</i> 829.50 (<b>A</b>) and 878.70 (<b>B</b>) of the glycopeptide carrying a glycan of composition H2N2 and F1H2N2 respectively were selected. Fragment ions are indicated. The residual signals at the <i>m/z</i> corresponding to [H+3H]³⁺ (<b>Δ</b>), to the doubly charged ions that result from capture of 1 electron without dissociation [H+3H]^2+· (<b>X</b>) and to the singly charged ions that result from capture of 2 electrons without dissociation [H+3H]^+·· (*) are indicated (<b>C</b>). Monoisotopic masses are given. Man (green circle), GlcNAc (blue square) and Fuc (red triangle), pep (peptide moiety).</p

Identification of <i>F</i>. <i>hepatica</i> cathepsins in NEJTeg.

<p>NEJTeg was loaded in SDS-PAGE and stained with Coomassie Blue in order to stain protein bands (<b>A</b>). Bands 1 (B1), 2 (B2), 3 (B3) and 4 (B4), were excised, tryptic digested and identified with LC-MS/MS analysis. Potential <i>N-</i>glycosylation sites (Pot <i>N-</i>gly) and monoisotopic mass of the tryptic peptides containing glycosylation were predicted (<b>B</b>). UP (unique peptide).</p