Mass spectrometry based proteomics for the molecular fingerprinting of Fiano, Greco and Falanghina cultivars

Abstract The official methodologies used for the identification and comparison of vine cultivars are ampelography and ampelometry. These methodologies are essentially based on qualitative assessments or biometric dependent morphological features of the plant. The heterogeneity of cultivars and consequently the increasing demand for a more detailed product typization, led to the introduction of new methodologies for the varietal characterization. In this scenario, proteomics has already proved to be a very useful discipline for the typization of many kinds of edible products. In this paper, we present a proteomic study carried out on three cultivars of Vitis vinifera peculiar of south Italy (Campania) used for white wine production (Fiano, Greco and Falanghina) by advanced biomolecular mass spectrometry approach. Our data highlight variations in the proteomic profiles during ripening for each cultivar and between analyzed cultivars, thus suggesting a new way to outline the biomolecular signature of vines

NEUROCOGNITIVE MANAGEMENT OF THE PRIMARY NEGATIVE SYMPTOMS OF SCHIZOPHRENIA: A ROLE OF ATYPICAL ANTIPSYCHOTICS

Patients with schizophrenia have profound and disabling cognitive deficits while negative symptoms represent a separate symptom domain, with respect to depression, neurocognition, and social cognition. Particularly, primary negative symptoms of schizophrenia represent a diagnostic and therapeutic challenge, In this study we try to evaluate the cognitive symptoms in 51 primary negative schizophrenic inpatients by the administration of simple, fast and understandable scales (MMSE, DSST, EpiTrack, PANSS cognitive factor). We also evaluate the correlation with some SGAs (aripiprazole, quetiapine, olanzapine, paliperidone). Our results support the evidence of the use of simple, rapid and acceptable scales for cognitive evaluation in clinical practice. Overall data indicate no statistically significant variations of the negative symptomatology in all the examined sample, although a reduction of the statistical averages in each group is observed (paliperidone and olanzapine, particularly)

Quantification of Polyphenols and Metals in Chinese Tea Infusions by Mass Spectrometry

Chemical compounds within tea (Camellia sinensis) are characterized by an extensive heterogeneity; some of them are crucial for their protective and defensive role in plants, and are closely connected to the benefits that the consumption of tea can provide. This paper is mainly focused on the characterization of polyphenols (secondary metabolites generally involved in defense against ultraviolet radiation and aggression by pathogens) and metals, extracted from nine Chinese tea samples, by integrating different mass spectrometry methodologies, LC-MS/MS in multiple reaction monitoring (MRM) and inductively coupled plasma mass spectrometry (ICP-MS). Our approach allowed to identify and compare forty polyphenols differently distributed in tea infusions at various fermentation levels. The exploration of polyphenols with nutraceutical potential in tea infusions can widely benefit especially tea-oriented populations. The worldwide consumption of tea requires at the same time a careful monitoring of metals released during the infusion of tea leaves. Metal analysis can provide the identification of many healthy minerals such as potassium, sodium, calcium, magnesium, differently affected by the fermentation of leaves. Our results allowed us: (i) to draw up a polyphenols profile of tea leaves subjected to different fermentation processes; (ii) to identify and quantify metals released from tea leaves during infusion. In this way, we obtained a molecular fingerprint useful for both nutraceutical applications and food control/typization, as well as for frauds detection and counterfeiting

NEUROCOGNITIVE MANAGEMENT OF THE PRIMARY NEGATIVE SYMPTOMS OF SCHIZOPHRENIA: A ROLE OF ATYPICAL ANTIPSYCHOTICS

Patients with schizophrenia have profound and disabling cognitive deficits while negative symptoms represent a separate symptom domain, with respect to depression, neurocognition, and social cognition. Particularly, primary negative symptoms of schizophrenia represent a diagnostic and therapeutic challenge, In this study we try to evaluate the cognitive symptoms in 51 primary negative schizophrenic inpatients by the administration of simple, fast and understandable scales (MMSE, DSST, EpiTrack, PANSS cognitive factor). We also evaluate the correlation with some SGAs (aripiprazole, quetiapine, olanzapine, paliperidone). Our results support the evidence of the use of simple, rapid and acceptable scales for cognitive evaluation in clinical practice. Overall data indicate no statistically significant variations of the negative symptomatology in all the examined sample, although a reduction of the statistical averages in each group is observed (paliperidone and olanzapine, particularly)

A Hyperthermoactive-Cas9 Editing Tool Reveals the Role of a Unique Arsenite Methyltransferase in the Arsenic Resistance System of Thermus thermophilus HB27.

Arsenic detoxification systems can be found in a wide range of organisms, from bacteria to humans. In a previous study, we discovered an arsenic-responsive transcriptional regulator in the thermophilic bacterium Thermus thermophilus HB27 (TtSmtB). Here, we characterize the arsenic resistance system of T. thermophilus in more detail. We employed TtSmtB-based pulldown assays with protein extracts from cultures treated with arsenate and arsenite to obtain an S-adenosyl-l-methionine (SAM)-dependent arsenite methyltransferase (TtArsM). In vivo and in vitro analyses were performed to shed light on this new component of the arsenic resistance network and its peculiar catalytic mechanism. Heterologous expression of TtarsM in Escherichia coli resulted in arsenite detoxification at mesophilic temperatures. Although TtArsM does not contain a canonical arsenite binding site, the purified protein does catalyze SAM-dependent arsenite methylation with formation of monomethylarsenites (MMAs) and dimethylarsenites (DMAs). In addition, in vitro analyses confirmed the unique interaction between TtArsM and TtSmtB. Next, a highly efficient ThermoCas9-based genome-editing tool was developed to delete the TtArsM-encoding gene on the T. thermophilus genome and to confirm its involvement in the arsenite detoxification system. Finally, the TtarsX efflux pump gene in the T. thermophilus ΔTtarsM genome was substituted by a gene encoding a stabilized yellow fluorescent protein (sYFP) to create a sensitive genome-based bioreporter system for the detection of arsenic ions. IMPORTANCE We here describe the discovery of an unknown protein by using a proteomics approach with a transcriptional regulator as bait. Remarkably, we successfully obtained a novel type of enzyme through the interaction with a transcriptional regulator controlling the expression of this enzyme. Employing this strategy, we isolated TtArsM, the first thermophilic prokaryotic arsenite methyltransferase, as a new enzyme of the arsenic resistance mechanism in T. thermophilus HB27. The atypical arsenite binding site of TtArsM categorizes the enzyme as the first member of a new arsenite methyltransferase type, exclusively present in the Thermus genus. The enzyme methylates arsenite-producing MMAs and DMAs. Furthermore, we developed an hyperthermophilic Cas9-based genome-editing tool, active up to 65°C. The tool allowed us to perform highly efficient, marker-free modifications (either gene deletion or insertion) in the T. thermophilus genome. With these modifications, we confirmed the critical role of TtArsM in the arsenite detoxification system and developed a sensitive whole-cell bioreporter for arsenic ions. We anticipate that the developed tool can be easily adapted for editing the genomes of other thermophilic bacteria, significantly boosting fundamental and metabolic engineering in hyperthermophilic microorganisms

Archaeal glycosylation: analysis of the glycome and identification of the enzymes involved

Glycosylation is one of the most prevalent post-translational modifications of proteins that expand the diversity of the proteome by the addition of different glycan moieties. Long believed a prerogative of eukaryotes, it is now clear that both N-glycosylation and O-glycosylation also transpire in Bacteria and Archaea. In recent years, substantial progresses are being made in understanding bacterial glycosylation while much less is known of this post-translational modification in Archaea, despite the fact that the first prokaryotic N-glycosylated protein was discovered over three decades ago in the haloarchaeon Halobacterium salinarum1 and that N-glycosylated proteins are more prevalent in Archaea than in Bacteria. Archaeal glycosylation displays bacterial- and eukaryal-like traits, such as monomeric oligosyltransferases and dolichol phosphate carrier respectively, in addition to unique features2. Insights into the biosynthesis and the nature of N-linked glycans decorating archaeal glycoproteins arise from euryarchaeal model species3. In Crenarchaea, a phylum evolutionarily distant from Euryarchaea, the study of the steps and the components of the machinery involved in glycosylation and of the nature of the glycosylated proteins is still in its infancy. However, preliminary results indicate that glycosylation in these organisms is indispensable for cell survival4 and even more widespread than in Euryarchaea5. Here, we report our recent studies on the enzymes from the hyperthermophilic crenarchaeon Sulfolobus solfataricus involved in the synthesis and in the maturation of the glycan component of the glycoproteins6. Moreover, sugar composition and structure of the sulfolobales glycoproteins are explored by glycoproteomic analysis. A better understanding of crenarchaeal glycosylation will provide new insights into this post-translational modification across evolution as well as protein processing under extreme conditions

Analysis of the glycome and identification of the enzymes involved in the glycosylation of Sulfolobus solfataricus

Glycosylation is one of the most prevalent post-translational modifications in proteins. Long believed a prerogative of eukaryotes, it is now clear that also Bacteria and Archaea show both N- and O-glycosylation. In recent years, substantial progresses are being made in understanding bacterial glycosylation while much less is known of this in Archaea despite the fact that the first prokaryotic N-glycosylated protein was discovered over three decades ago in the haloarchaeon Halobacterium salinarum1. N-glycosylated proteins in Archaea are more prevalent than in Bacteria, and, in the formers, glycosylation mechanism displays unique features and a sort of mixture of bacterial- and eukaryal-like traits2, such as monomeric oligosyltransferases and dolichol phosphate carrier respectively. Most of the studies on the biosynthesis and the nature of N-glycosylation has been performed mostly in Euryarchaea3. In Crenarchaea the study of the steps and the components of the machinery involved in glycosylation and of the nature of the glycosylated proteins is still in its infancy. However, preliminary results indicate that glycosylation in these organisms is indispensable for cell survival4 and even more widespread than in Euryarchaea5. Here, we report our recent studies on the enzymes from the hyperthermophilic crenarchaeon Sulfolobus solfataricus involved in the synthesis, maturation and degradation of the glycan components of the glycoproteins6. Moreover, sugar composition and structure of the glycoproteins from S. solfataricus are analysed by glycoproteomic analysis

Association of maternal hypertension and chorioamnionitis with preterm outcomes

OBJECTIVES: We compared the relative effect of hypertensive disorders of pregnancy and chorioamnionitis on adverse neonatal outcomes in very preterm neonates, and studied whether gestational age (GA) modulates these effects. METHODS: A cohort of neonates 23 to 30 weeks' GA, born in 2008 to 2011 in 82 hospitals adhering to the Italian Neonatal Network, was analyzed. Infants born from mothers who had hypertensive disorders (N = 2096) were compared with those born after chorioamnionitis (N = 1510). Statistical analysis employed logistic models, adjusting for GA, hospital, and potential confounders. RESULTS: Overall mortality was higher after hypertension than after chorioamnionitis (odds ratio [OR], 1.39; 95% confidence interval [CI], 1.08-1.80), but this relationship changed across GA weeks; the OR for hypertension was highest at low GA, whereas from 28 weeks' GA onward, mortality was higher for chorioamnionitis. For other outcomes, the relative risks were constant across GA; infants born after hypertension had an increased risk for bronchopulmonary dysplasia (OR, 2.20; 95% CI, 1.68-2.88) and severe retinopathy of prematurity (OR, 1.48; 95% CI, 1.02-2.15), whereas there was a lower risk for early-onset sepsis (OR, 0.25; 95% CI, 0.19-0.34), severe intraventricular hemorrhage (OR, 0.65; 95% CI, 0.48-0.88), periventricular leukomalacia (OR, 0.70; 95% CI, 0.48-1.01), and surgical necrotizing enterocolitis or gastrointestinal perforation (OR, 0.47; 95% CI, 0.31-0.72). CONCLUSIONS: Mortality and other adverse outcomes in very preterm infants depend on antecedents of preterm birth. Hypertension and chorioamnionitis are associated with different patterns of outcomes; for mortality, the effect changes across GA weeks. Copyright \uc2\ua9 2014 by the American Academy of Pediatrics

Survey of neonatal respiratory care and surfactant administration in very preterm infants in the Italian neonatal network

Introduction: Variation of respiratory care is described between centers around the world.The Italian Neonatal Network (INN), as a national group of the Vermont-Oxford Network (VON) allows to perform a wide analysis of respiratory care in very low birth weight infants. Methods:We analyzed the dataset of infants enrolled in the INN in 2009 and 2010 and, for surfactant administration only, from 2006 to 2010 from 83 participating centers. All definitions are those of the (VON). A questionnaire analysis was also performed with a questionnaire on centers practices. Results: We report data for 8297 infants. Data on ventilator practices and outcomes are outlined. Variation for both practices and outcome is found. Trend in surfactant administration is also analyzed. Conclusions. The great variation across hospitals in all the surveyed techniques points to the possibility of implementing potentially better practices with the aim of reducing unwanted variation. These data also show the power of large neonatal networks in identifying areas for potential improvement. © Mattioli 1885

Survey of neonatal respiratory care and surfactant administration in very preterm infants in the Italian neonatal network

Introduction: Variation of respiratory care is described between centers around the world.The Italian Neonatal Network (INN), as a national group of the Vermont-Oxford Network (VON) allows to perform a wide analysis of respiratory care in very low birth weight infants. Methods:We analyzed the dataset of infants enrolled in the INN in 2009 and 2010 and, for surfactant administration only, from 2006 to 2010 from 83 participating centers. All definitions are those of the (VON). A questionnaire analysis was also performed with a questionnaire on centers practices. Results: We report data for 8297 infants. Data on ventilator practices and outcomes are outlined. Variation for both practices and outcome is found. Trend in surfactant administration is also analyzed. Conclusions. The great variation across hospitals in all the surveyed techniques points to the possibility of implementing potentially better practices with the aim of reducing unwanted variation. These data also show the power of large neonatal networks in identifying areas for potential improvement. \ua9 Mattioli 1885