45 research outputs found

    Towards a metabolomic approach to investigate iron-sulfur cluster biogenesis

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    Iron–sulfur clusters are prosthetic groups that are assembled on their acceptor proteins through a complex machine centered on a desulfurase enzyme and a transient scaffold protein. Studies to establish the mechanism of cluster formation have so far used either in vitro or in vivo methods, which have often resulted in contrasting or non‐comparable results. We suggest, here, an alternative approach to study the enzymatic reaction, that is based on the combination of genetically engineered bacterial strains depleted of specific components, and the detection of the enzymatic kinetics in cellular extracts through metabolomics. Our data prove that this ex vivo approach closely reproduces the in vitro results while retaining the full complexity of the system. We demonstrate that co‐presence of bacterial frataxin and iron is necessary to observe an inhibitory effect of the enzymatic activity of bacterial frataxin. Our approach provides a new powerful tool for the study of iron–sulfur cluster biogenesis

    Evaluation of an Active Humidification System for Inspired Gas

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    ObjectivesThe effectiveness of the active humidification systems (AHS) in patients already weaned from mechanical ventilation and with an artificial airway has not been very well described. The objective of this study was to evaluate the performance of an AHS in chronically tracheostomized and spontaneously breathing patients.MethodsMeasurements were quantified at three levels of temperature (T°) of the AHS: level I, low; level II, middle; and level III, high and at different flow levels (20 to 60 L/minute). Statistical analysis of repeated measurements was performed using analysis of variance and significance was set at a P<0.05.ResultsWhile the lowest temperature setting (level I) did not condition gas to the minimum recommended values for any of the flows that were used, the medium temperature setting (level II) only conditioned gas with flows of 20 and 30 L/minute. Finally, at the highest temperature setting (level III), every flow reached the minimum absolute humidity (AH) recommended of 30 mg/L.ConclusionAccording to our results, to obtain appropiate relative humidity, AH and T° of gas one should have a device that maintains water T° at least at 53℃ for flows between 20 and 30 L/m, or at T° of 61℃ at any flow rate

    Non-invasive diagnostic tests for Helicobacter pylori infection

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    BACKGROUND: Helicobacter pylori (H pylori) infection has been implicated in a number of malignancies and non-malignant conditions including peptic ulcers, non-ulcer dyspepsia, recurrent peptic ulcer bleeding, unexplained iron deficiency anaemia, idiopathic thrombocytopaenia purpura, and colorectal adenomas. The confirmatory diagnosis of H pylori is by endoscopic biopsy, followed by histopathological examination using haemotoxylin and eosin (H & E) stain or special stains such as Giemsa stain and Warthin-Starry stain. Special stains are more accurate than H & E stain. There is significant uncertainty about the diagnostic accuracy of non-invasive tests for diagnosis of H pylori. OBJECTIVES: To compare the diagnostic accuracy of urea breath test, serology, and stool antigen test, used alone or in combination, for diagnosis of H pylori infection in symptomatic and asymptomatic people, so that eradication therapy for H pylori can be started. SEARCH METHODS: We searched MEDLINE, Embase, the Science Citation Index and the National Institute for Health Research Health Technology Assessment Database on 4 March 2016. We screened references in the included studies to identify additional studies. We also conducted citation searches of relevant studies, most recently on 4 December 2016. We did not restrict studies by language or publication status, or whether data were collected prospectively or retrospectively. SELECTION CRITERIA: We included diagnostic accuracy studies that evaluated at least one of the index tests (urea breath test using isotopes such as13C or14C, serology and stool antigen test) against the reference standard (histopathological examination using H & E stain, special stains or immunohistochemical stain) in people suspected of having H pylori infection. DATA COLLECTION AND ANALYSIS: Two review authors independently screened the references to identify relevant studies and independently extracted data. We assessed the methodological quality of studies using the QUADAS-2 tool. We performed meta-analysis by using the hierarchical summary receiver operating characteristic (HSROC) model to estimate and compare SROC curves. Where appropriate, we used bivariate or univariate logistic regression models to estimate summary sensitivities and specificities. MAIN RESULTS: We included 101 studies involving 11,003 participants, of which 5839 participants (53.1%) had H pylori infection. The prevalence of H pylori infection in the studies ranged from 15.2% to 94.7%, with a median prevalence of 53.7% (interquartile range 42.0% to 66.5%). Most of the studies (57%) included participants with dyspepsia and 53 studies excluded participants who recently had proton pump inhibitors or antibiotics.There was at least an unclear risk of bias or unclear applicability concern for each study.Of the 101 studies, 15 compared the accuracy of two index tests and two studies compared the accuracy of three index tests. Thirty-four studies (4242 participants) evaluated serology; 29 studies (2988 participants) evaluated stool antigen test; 34 studies (3139 participants) evaluated urea breath test-13C; 21 studies (1810 participants) evaluated urea breath test-14C; and two studies (127 participants) evaluated urea breath test but did not report the isotope used. The thresholds used to define test positivity and the staining techniques used for histopathological examination (reference standard) varied between studies. Due to sparse data for each threshold reported, it was not possible to identify the best threshold for each test.Using data from 99 studies in an indirect test comparison, there was statistical evidence of a difference in diagnostic accuracy between urea breath test-13C, urea breath test-14C, serology and stool antigen test (P = 0.024). The diagnostic odds ratios for urea breath test-13C, urea breath test-14C, serology, and stool antigen test were 153 (95% confidence interval (CI) 73.7 to 316), 105 (95% CI 74.0 to 150), 47.4 (95% CI 25.5 to 88.1) and 45.1 (95% CI 24.2 to 84.1). The sensitivity (95% CI) estimated at a fixed specificity of 0.90 (median from studies across the four tests), was 0.94 (95% CI 0.89 to 0.97) for urea breath test-13C, 0.92 (95% CI 0.89 to 0.94) for urea breath test-14C, 0.84 (95% CI 0.74 to 0.91) for serology, and 0.83 (95% CI 0.73 to 0.90) for stool antigen test. This implies that on average, given a specificity of 0.90 and prevalence of 53.7% (median specificity and prevalence in the studies), out of 1000 people tested for H pylori infection, there will be 46 false positives (people without H pylori infection who will be diagnosed as having H pylori infection). In this hypothetical cohort, urea breath test-13C, urea breath test-14C, serology, and stool antigen test will give 30 (95% CI 15 to 58), 42 (95% CI 30 to 58), 86 (95% CI 50 to 140), and 89 (95% CI 52 to 146) false negatives respectively (people with H pylori infection for whom the diagnosis of H pylori will be missed).Direct comparisons were based on few head-to-head studies. The ratios of diagnostic odds ratios (DORs) were 0.68 (95% CI 0.12 to 3.70; P = 0.56) for urea breath test-13C versus serology (seven studies), and 0.88 (95% CI 0.14 to 5.56; P = 0.84) for urea breath test-13C versus stool antigen test (seven studies). The 95% CIs of these estimates overlap with those of the ratios of DORs from the indirect comparison. Data were limited or unavailable for meta-analysis of other direct comparisons. AUTHORS' CONCLUSIONS: In people without a history of gastrectomy and those who have not recently had antibiotics or proton ,pump inhibitors, urea breath tests had high diagnostic accuracy while serology and stool antigen tests were less accurate for diagnosis of Helicobacter pylori infection.This is based on an indirect test comparison (with potential for bias due to confounding), as evidence from direct comparisons was limited or unavailable. The thresholds used for these tests were highly variable and we were unable to identify specific thresholds that might be useful in clinical practice.We need further comparative studies of high methodological quality to obtain more reliable evidence of relative accuracy between the tests. Such studies should be conducted prospectively in a representative spectrum of participants and clearly reported to ensure low risk of bias. Most importantly, studies should prespecify and clearly report thresholds used, and should avoid inappropriate exclusions

    A gap analysis for threatened bat populations on Sardinia

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    Sardinia is home to three bat species of chief conservation importance, the endemic Plecotus sardus as well as Myotis punicus and Rhinolophus mehelyi, for which Sardinia is the Italian stronghold. We carried out two gap analyses overlapping the network of protected areas present on the island respectively with 1) the occurrence records of all species and 2) the binarized maps obtained from maximum entropy models. Unlike the other two species, P. sardus known records are confined to the central sector of the island and its suitable habitat partly overlaps with that of M. punicus but not with R. mehelyi. Due to its uniqueness and restricted range P. sardus requires a very specific management strategy for its protection to be successful. Both analyses led to the conclusion that all species considered require more extensive protection than is currently granted so that urgent measures should be taken to improve the current situation

    Shedding light on the effects of climate and anthropogenic pressures on the disappearance of Fagus sylvatica in the Italian lowlands: evidence from archaeo-anthracology and spatial analyses

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    Fagus sylvatica is one of the most representative trees of the European deciduous broadleaved forests, yet the impact of changing climatic conditions and anthropogenic pressures (anthromes) on its presence and distribution in the coastal and lowland areas of the Mediterranean Basin has long been overlooked. Here, we first analysed the local forest composition in two different time intervals (350-300 Before Current Era, BCE and 150-100 BCE) using charred wood remains from the Etruscan site of Cetamura (Tuscany, central Italy). Addition-ally, we reviewed all the relevant publications and the wood/charcoal data obtained from anthracological analysis in F. sylvatica, focusing on samples that date back to 4000 years before present, to better understand the drivers of beech presence and distribution during the Late Holocene (LH) in the Italian Peninsula. Then, we combined charcoal and spa-tial analyses to test the distribution of beech woodland at low elevation during LH in Italy and to evaluate the effect of climate change and/or anthrome on the disappearance of F. sylvatica from the lowlands. We collected 1383 charcoal fragments in Cetamura belonging to 21 woody taxa, with F. sylvatica being the most abun-dant species (28 %), followed by other broadleaved trees. We identified 25 sites in the Italian Peninsula with beech charcoals in the last 4000 years. Our spatial analyses showed a marked decrease in habitat suitability of F. sylvatica from LH to the present (ca. 48 %), particularly in the lowlands (0-300 m above sea level, a.s.l.) and at higher altitudes (&gt;900 m a.s.l). In the lowland areas, where F. sylvatica has disappeared, climate had a more uniform effect on beech distribution patterns across the entire elevation range analysed, whereas climate + anthrome and anthrome alone in-fluenced 69 % and 84 % of the lowland areas, respectively. Our results highlight the advantage of combining different approaches, such as charcoal analysis and spatial analyses, to explore biogeographic questions about the past and current distribution of F. sylvatica, with important implications for today's forest management and conservation policies and practices, as well as for future research projects

    Severe outbreak of Fusarium solani on Quercus ilex vectored by Xylosandrus compactus

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    The first European record of an outbreak of Fusarium solani on Quercus ilex following a massive attack by Xylosandrus compactus from southern Italy is reported
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