1H, 13C, 15N backbone resonance assignment of apo and ADP-ribose bound forms of the macro domain of Hepatitis E virus through solution NMR spectroscopy

International audienceAbstract The genome of Hepatitis E virus (HEV) is 7.2 kilobases long and has three open reading frames. The largest one is ORF1, encoding a non-structural protein involved in the replication process, and whose processing is ill-defined. The ORF1 protein is a multi-modular protein which includes a macro domain (MD). MDs are evolutionarily conserved structures throughout all kingdoms of life. MDs participate in the recognition and removal of ADP-ribosylation, and specifically viral MDs have been identified as erasers of ADP-ribose moieties interpreting them as important players at escaping the early stages of host-immune response. A detailed structural analysis of the apo and bound to ADP-ribose state of the native HEV MD would provide the structural information to understand how HEV MD is implicated in virus-host interplay and how it interacts with its intracellular partner during viral replication. In the present study we present the high yield expression of the native macro domain of HEV and its analysis by solution NMR spectroscopy. The HEV MD is folded in solution and we present a nearly complete backbone and sidechains assignment for apo and bound states. In addition, a secondary structure prediction by TALOS + analysis was performed. The results indicated that HEV MD has a α/β/α topology very similar to that of most viral macro domains

Shock waves in strongly coupled plasmas

Shock waves are supersonic disturbances propagating in a fluid and giving rise to dissipation and drag. Weak shocks, i.e., those of small amplitude, can be well described within the hydrodynamic approximation. On the other hand, strong shocks are discontinuous within hydrodynamics and therefore probe the microscopics of the theory. In this paper we consider the case of the strongly coupled N=4 plasma whose microscopic description, applicable for scales smaller than the inverse temperature, is given in terms of gravity in an asymptotically $AdS_5$ space. In the gravity approximation, weak and strong shocks should be described by smooth metrics with no discontinuities. For weak shocks we find the dual metric in a derivative expansion and for strong shocks we use linearized gravity to find the exponential tail that determines the width of the shock. In particular we find that, when the velocity of the fluid relative to the shock approaches the speed of light $v\to 1$ the penetration depth $\ell$ scales as $\ell\sim (1-v^2)^{1/4}$. We compare the results with second order hydrodynamics and the Israel-Stewart approximation. Although they all agree in the hydrodynamic regime of weak shocks, we show that there is not even qualitative agreement for strong shocks. For the gravity side, the existence of shock waves implies that there are disturbances of constant shape propagating on the horizon of the dual black holes.Comment: 47 pages, 8 figures; v2:typos corrected, references adde

Prevalence and determinants of sex-specific dietary supplement use in a greek cohort

We describe the profile of dietary supplement use and its correlates in the Epirus Health Study cohort, which consists of 1237 adults (60.5% women) residing in urban north-west Greece. The association between dietary supplement use and demographic characteristics, lifestyle behaviors, personal medical history and clinical measurements was assessed using logistic regression models, separately for women and men. The overall prevalence of dietary supplement use was 31.4%, and it was higher in women (37.3%) compared to men (22.4%; p-value = 4.2−08). Based on multivariable logistic regression models, dietary supplement use in women was associated with age (positively until middle-age and slightly negatively afterwards), the presence of a chronic health condition (OR = 1.71; 95% CI, 1.18–2.46), lost/removed teeth (OR = 0.52; 95% CI, 0.35–0.78) and diastolic blood pressure (OR per 5 mmHg increase =0.84; 95% CI, 0.73–0.96); body mass index and worse general health status were borderline inversely associated. In men, dietary supplement use was positively associated with being employed (OR = 2.53; 95% CI, 1.21–5.29). A considerable proportion of our sample used dietary supplements, and the associated factors differed between women and men

Health status outcomes after spontaneous coronary artery dissection and comparison with other acute myocardial infarction: The VIRGO experience.

Data on health status outcomes after spontaneous coronary artery dissection (SCAD) are limited. Using the Variation in Recovery: Role of Gender on Outcomes of Young AMI Patients (VIRGO) study we compared patients with SCAD and other acute myocardial infarction (AMI) at presentation (baseline), 1-month, and-12 months using standardized health status instruments. Among 3572 AMI patients ≤ 55 years, 67 had SCAD. SCAD patients were younger (median age (IQR) 45 (40.5-51) years vs. 48 (44-52) in other AMI, p = 0.003), more often female (92.5% vs. 66.6%), have college education (73.1% vs. 51.7%) and household income >$100,000 (43.3% vs. 17.7% (All p<0.001). SCAD patients at baseline had higher mean ± SD Short Form-12 [SF-12] physical component scores [PCS] (48.7±10.2 vs. 43.8±12.1, p<0.001) and mental component scores [MCS] (49.6±12.4 vs. 45.4±12.5, p = 0.008), and at 12-months [PCS (50.1±9.0 vs. 44.3±12.3, p<0.001) and MCS (53±10.1 vs 50.2±11.0, p = 0.045)]. The Euro-Quality of Life Scale [EQ-5D] VAS and EQ-5D index scores were similar at baseline, but higher at 12-months for SCAD (EQ-5D VAS: 82.2±10.2 vs. 72.3±21.0, p<0.001; EQ-5D index scores; 90.2±15.3 vs. 83.7±19.8, p = 0.012). SCAD patients had better baseline Seattle Angina Questionnaire [SAQ] physical limitation (88.8±20.1 vs. 81.2±25.4, p = 0.017). At 12-months SCAD patients had better physical limitation (98.0±8.5 vs. 91.4±18.8, p = 0.007), angina frequency (96.4±8.8 vs. 91.3±16.8, p = 0.018) and quality of life scores (80.7±14.7 vs 72.2±23.2, p = 0.005). Magnitude of change in health status from baseline to 12-months was not statistically different between the groups. After adjustment for time and comorbidities there remained no difference in most health status outcomes. SCAD patients fare marginally better than other AMI patients on most health status instruments and have similar 12-month health status recovery. Better pre-event health status suggests a need to modify exercise prescriptions and cardiac rehabilitation protocols to better assist this physically active population to recover.The VIRGO study was supported by a 4-year National Heart, Lung, and Blood Institute grant [number 5R01HL081153). IMJOVEN was supported in Spain by grant PI 081614 from the Fondo de Investigaciones Sanitarias del Instituto Carlos III, Ministry of Science and Technology, and additional funds from the Centro Nacional de Investigaciones Cardiovasculares (CNIC).S

A case-control validation of Type D personality in Greek patients with stable coronary heart disease

BACKGROUND: Type D personality has been associated with a variety of emotional and social difficulties as well as with poor prognosis in patients with established coronary heart disease (CHD). We examined the psychometric properties and validity of the Type D Scale-14 (DS14) and the prevalence of Type D personality among Greek patients with CHD while taking into account demographic; clinical, such as diabetes mellitus, hypertension, and hypercholesterolemia; as well as psychological variables such as depression, anxiety, and psychological stress. METHODS: Ninety-six patients with stable coronary heart disease and 80 healthy participants from the general population completed the Greek version of the DS14 and the Hospital Anxiety and Depression Scale (HADS). RESULTS: Cronbach's α coefficient for the negative affectivity (NA) and social inhibition (SI) subscales was 0.83 and 0.72 for the CHD and 0.88 and 0.76 for the control group, respectively. Internal-structural validity was assessed by a factor analysis (two-factor solution), and the factor structure of the original DS14 was replicated. Using the standardized cutoff point of NA ≥10 and SI ≥10, instead of the median scores, in order to have compatible results with the majority of studies, the prevalence of Type D personality was 51% for the CHD patients and 13% for the control group. Higher NA and SI were connected with higher anxiety, depression, and total psychological stress. Finally, more patients with CHD and Type D personality than those without were diagnosed with type 2 diabetes; however, no differences were observed in hypertension or hypercholesterolemia. CONCLUSIONS: These results indicate that the Type D construct is reliable and valid in a Greek population. The prevalence of Type D personality was higher in patients with stable coronary heart disease than in people from the general population. The DS14 subscales were positively correlated with higher anxiety, depression, and total psychological stress. Regarding other CHD risk factors, only diabetes mellitus was found more frequently in CHD patients with Type D personality

Bridging the gaps:Bole and Terra Sigillata as artefacts, as simples and as antibacterial clays

Medicinal earths are an important and yet, so far, little scientifically explored archaeological resource. They are almost always identified by their source locality. Our work over the last few years has focused on their chemical and mineralogical characterization and their testing as anti-bacterials. This paper presents the results of the mineralogical analysis and antibacterial testing of six medicinal earths, bole or Terra Sigillata (stamped earth) of unknown date and provenance in the Pharmacy Museum of the University of Basel. Only one of them, a red (Armenian?) ‘bole’, was found to be antibacterial against both Gram-positive and Gram-negative bacteria. A yellow powder of Terra Tripolitania was mildly antibacterial and against one pathogen only. We argue that medicinal earths are in a pivotal place to bridge the gap between currently dispersed pieces of information. This information relates to: (a) their nature, attributes, and applications as described in the texts of different periods, (b) the source of their clays and how best to locate them in the field today, and (c) the methods employed for their beneficiation, if known. We propose that work should be focused primarily onto those medicinal earths whose clay sources can be re-discovered, sampled and assessed. From then on, a parallel investigation should be initiated involving both earths and their natural clays (mineralogy at bulk and nano-sized levels, bio-geochemistry, microbiological testing). We argue that the combined study can shed light into the parameters driving antibacterial action in clays and assist in the elucidation of the mechanisms involved

The impact of accute homocysteinemia to inflammatory and thrombotic factors among healthy individuals and hypersensive patients

Plasma homocysteine (Hcy) is associated with increased cardiovascular risk and endothelial dysfunction, however the underlying mechanisms are still obscure. Aim of the study was to examine the impact of acute homocysteinemia to vascular function and evaluate the role ET-1, ADMA and oxidative stress in hypertensive patients and healthy individuals. Methods : In this double-blind placebo controlled study, 39 hypertensive subjects and 49 healthy volunteers underwent methionine-loading (100mg met/Kg BW) after they received vitamins C (2g) plus E (800IU) (16 hypertensives (HTN+vit) and 27 healthy (C+vit)) or placebo (23 hypertensives (HTN+placebo) and 22 healthy (C+placebo)). Endothelial function was evaluated by gauge-strain plethysmography (to determine endothelium dependent dilation (EDD)), at baseline and 4 hours post loading (4hPML). ET-1, ADMA, IL-6, vWf were measured by ELISA, while total lipid hydroperoxides (per-ox) levels were measured by a commercially available photometric technique. Endothelial function in forearm resistance vessels and concentrations of total cholesterol, LDL, HDL, VLDL, ApoA1, ApoB, triglycerides, CRP, IL-6, vWF, fibrinogen, PER-OX, ADMA, tHcy, ET-1, IL-6, vWf, ADMA were determined at baseline and 4 hours post methionine loading. Results : Homocysteine was similarly increased in both hypertensives (p<0.0001) and healthy controls (p<0.0001), and was not affected by pre-treatment with antioxidants. EDD was significantly decreased in HTN+placebo (p<0.05) and C+placebo (p<0.0001), while antioxidant treatment did not prevent this effect in either HTN+vit (p<0.01) or C+vit (p<0.001). Importantly, ET-1 was increased 4hPML only in hypertensive individuals (P<0.05 in HTN+placebo) an effect not prevented by antioxidants (p<0.01 in HTN+vit). PEROX was significantly decreased in the HTN+vit (p<0.05) but not in the HTN-placebo (p=NS). PEROX remained unchanged in C+Vit (p=NS) and were increased in C+placebo (p<0.05). No effect of methionine-loading was observed on ET-1 levels in healthy individuals (p=NS for all groups). Also, in controls, plasma ADMA was also increased in both vitamin- and placebo groups (p<0.01 for both) at 4h-PML. Importantly, hypertensives had higher ADMA at baseline (p<0.01 vs healthy) and remained unchanged 4hMPL (p=NS for both placebo and vitamins treated subjects). Finally, healthy subjects with high baseline fasting Hcy (fHcy) had higher vWf and IL-6 than did subjects with low fHcy ( p<0.05) and they were both correlated with fHcy (vWf : r=0.506, p=0.004, IL-6: r=0.380, p=0.048). Conclusions : Experimental homocysteinemia rapidly blunts endothelial function in both hypertensive subjects and healthy individuals. The rapid elevation of ET-1 levels observed only in hypertensives, suggests that ET-1 may be the key mediator of homocysteine-induced endothelial dysfunction, independently of oxidative stress. Also, that plasma ADMA levels are elevated in hypertensives, but remain unchanged after methionine, suggesting that ADMA plays an important role in endothelial dysfunction in hypertensives, but it is not responsible for methionine-induced endothelial dysfunction in this group of patients. Finally, in healthy subjects fHcy concentrations are correlated with proinflammatory cytokines (IL-6) and endothelial damage index (vWf).Η ομοκυστεϊναιμία αποτελεί ένα νεότερο παράγοντα καρδιαγγειακού κινδύνου και ειδικότερα αντικείμενο μελέτης στη συμμετοχή της πορείας της αθηρογένεσης. Σκοπός της παρούσας μελέτης είναι η διερεύνηση της παθοφυσιολογίας των αγγειακών βλαβών που ενδεχομένως να προκαλούνται κατά τις οξείες μεταβολές των επιπέδων ομοκυστεΐνης που παρατηρούνται κατά τη φόρτιση με μεθειονίνη, σε υγιή άτομα και σε άτομα με ιδιοπαθή υπέρταση καθώς και οι επιπτώσεις της οξείας πειραματικής ομοκυστεϊναιμίας στην ενδοθηλιακή λειτουργία, θρόμβωση και φλεγμονή στα άτομα αυτά. Επίσης, διερευνάται ο ρόλος του οξειδωτικού στρες και των αντιοξειδωτικών βιταμινών Ε και C στην παθοφυσιολογία των αγγειακών βλαβών της οξείας ομοκυστεϊναιμίας. Επιπλέον, αναζητείται η πιθανότητα μέρος της ενδοθηλιακής δυσλειτουργίας που παρατηρείται στην οξεία ομοκυστεϊναιμία να οφείλεται και στην εκτροπή της μεθειονίνης προς παραγωγή της ασυμμετρικής διμέθυλο-αργινίνης (ADMA), και ο βαθμός στον οποίο η παραγωγή ADMA συμμετέχει στην ελάττωση της βιοδιαθεσιμότητας του ΝΟ κατά την ομοκυστεϊναιμία. Τέλος, μελετάται ο ρόλος της ενδοθηλίνης-1 στην ανάπτυξη ενδοθηλιακής δυσλειτουργίας και ο μηχανισμός συμμετοχής της στην παθογένεση της αρτηριακής υπέρτασης. Μέθοδοι : Πραγματοποιήθηκε μια διπλή τυφλή μελέτη ελεγχόμενη με εικονικό φάρμακο, στην οποία ο πληθυσμός της περιελάμβανε 49 υγιή άτομα χωρίς κλασσικούς επιβαρυντικούς παράγοντες αθηροσκλήρυνσης και 39 ασθενείς νεοδιαγνωσθέντες για αρτηριακή υπέρταση (ΑΥ). Οι συμμετέχοντες τυχαιοποιήθηκαν σε 4 ομάδες και έλαβαν είτε μεθειονίνη (σε δόση φόρτισης 100mg/kg βάρος σώματος) και βιταμίνη Ε (800 IU) και βιταμίνη C (2gr) διαλυμένα σε σταθερής σύνθεσης χυμό πορτοκαλιού (16 υπερτασικοί και 27 υγιείς) ή μεθειονίνη (σε δόση φόρτισης 100mg/kg βάρος σώματος) διαλυμένη σε 150ml χυμό πορτοκαλιού χωρίς αντιοξειδωτικές βιταμίνες ( 23 υπερτασικοί και 22 υγιείς (ΥΓΙΕΙΣ+PLACEBO). Πραγματοποιήθηκε έλεγχος της λειτουργικότητας του ενδοθηλίου σε όλους τους συμμετέχοντες με πληθυσμογραφία ροής της βραχιονίου αρτηρίας. Συγκεκριμένα μελετήθηκε η ενδοθηλιο-εξαρτώμενη αγγειοδιαστολή (EDD), μετά από 5λεπτη ισχαιμία του άνω άκρου την οποία ακολούθησε η αντιδραστική υπεραιμία, και η ενδοθηλιο-ανεξάρτητη αγγειοδιαστολή (EID), μετά από χορήγηση 0,4mg νιτρογλυκερίνης υπογλωσσίως. Επίσης, πραγματοποιήθηκε αιμοληψία για την εκτίμηση των εξής παραγόντων : χοληστερόλης, LDL, HDL, VLDL, ApoA1, ApoB, τριγλυκεριδίων, CRP, IL-6, vWF, ινωδογόνο, PEROX, ADMA, ολική ομοκυστεΐνη, ενδοθηλίνη-1. Χρησιμοποιήθηκε ενζυμική ανοσοπροσροφητική ανάλυση (ELISA) για τον καθορισμό των επιπέδων ορού της ενδοθηλίνης-1 (ET-1), της ιντερλευκίνης-6 (IL-6), του παράγοντα von Willebrand (vWf) και της ADMA. Πραγματοποιήθηκε επανάληψη της πληθυσμογραφίας και της αιμοληψίας 4 ώρες μετά τη φόρτιση με μεθειονίνη.