Reinvestigation of the tautomerism of some substituted 2-hydroxypyridines

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Could SARS-CoV-2 Have Bacteriophage Behavior or Induce the Activity of Other Bacteriophages?

SARS-CoV-2 has become one of the most studied viruses of the last century. It was assumed that the only possible host for these types of viruses was mammalian eukaryotic cells. Our recent studies show that microorganisms in the human gastrointestinal tract affect the severity of COVID-19 and for the first time provide indications that the virus might replicate in gut bacteria. In order to further support these findings, in the present work, cultures of bacteria from the human microbiome and SARS-CoV-2 were analyzed by electron and fluorescence microscopy. The images presented in this article, in association with the nitrogen (15N) isotope-labeled culture medium experiment, suggest that SARS-CoV-2 could also infect bacteria in the gut microbiota, indicating that SARS-CoV-2 could act as a bacteriophage. Our results add new knowledge to the understanding of the mechanisms of SARS-CoV-2 infection and fill gaps in the study of the interactions between SARS-CoV-2 and non-mammalian cells. These findings could be useful in suggesting specific new pharmacological solutions to support the vaccination campaign

Development of new methodologies for the mass spectrometry study of bioorganic macromolecules

In recent years, mass spectrometry has been increasingly used for the analysis of various macromolecules of biological, biomedical, and biochemical interest. This increase has been made possible by two key developments: the advent of electrospray ionization (ESI) and matrix-assisted laser desorption ionization (MALDI) sources. The two new techniques produce a significant increase in mass range and in sensitivity that led to the development of new applications and of new analyzer designs, software, and robotics. This review, apart from the description of the status of mass spectrometry in the analysis of bioorganic macromolecules, is mainly devoted to the illustration of the more recent promising techniques and on their possible future evolution

Bioinformatics in mass spectrometry data analysis for proteomics studies

Mass spectrometry is a technique widely employed for the identification and characterization of proteins. The role of bioinformatics is fundamental for the elaboration of mass spectrometry data due to the amount of data that this technique can produce. To process data efficiently, new software packages and algorithms are continuously being developed to improve protein identification and characterization in terms of high-throughput and statistical accuracy. However, many limitations exist concerning bioinformatics spectral data elaboration. This review aims to critically cover the recent and future developments of new bioinformatics approaches in mass spectrometry data analysis for proteomics studies

Effects of triclosan in the freshwater mussel Dreissena polymorpha: a proteomic investigation

Triclosan (TCS, 5-chloro-2-(2,4-dichlorophenoxy)phenol) is commonly used in several personal care products, textiles, and children's toys. Because the removal of TCS by wastewater treatment plants is incomplete, its environmental fate is to be discharged into freshwater ecosystems, where its ecotoxicological impact is still largely unexplored. Previously, we began a structured multi-tiered approach in order to evaluate TCS toxicity in the freshwater mussel Dreissena polymorpha. The results of our previous studies, based on in vitro and in vivo experiments, highlighted a pronounced cytogenotoxic effect exerted by TCS, and showed that an increase in oxidative stress was likely to be one of its main toxic mechanisms. In this work, in order to investigate TCS toxicity mechanisms in aquatic non-target species in greater depth, we decided to use a proteomic approach, analysing changes in protein expression profiles in gills of D. polymorpha exposed for seven days to TCS. Moreover, thiobarbituric acid reactive substances (TBARS) were measured to investigate further the role played by TCS in inducing oxidative stress. Finally, TCS bioaccumulation in mussel tissues was also assessed, to ensure an effective accumulation of the toxicant. Our results not only confirmed the role played by TCS in inducing oxidative stress, but furthered knowledge about the mechanism exerted by TCS in inducing toxicity in an aquatic non-target organisms. TCS induced significant alterations in protein expression profiles in gills of D. polymorpha. The wide range of proteins affected suggested that this chemical has marked effects on various biological processes, especially those involved in calcium binding or stress response. We also confirmed that the proteomic analysis, using 2-DE and de novo sequencing, is a reliable and powerful approach to investigate cellular responses to pollutants in a non-model organism with few genomic sequences available in databases

Cambiamenti del proteoma della bacca di vite (Vitis vinifera) durante la maturazione

La maturazione dell\u2019uva coinvolge molteplici eventi biochimici e fisiologici che modificano profondamente le caratteristiche della bacca. Durante questo processo, che inizia all\u2019invaiatura, si osservano, fra l\u2019altro, cambiamenti strutturali della parete, l\u2019aumento delle dimensioni del frutto, ascrivibile ad una crescita per distensione cellulare, ed il rammollimento del pericarpo. A questi eventi si accompagnano modificazioni nella composizione dei metaboliti, quali l\u2019accumulo di zuccheri, amminoacidi e potassio, la riduzione degli acidi organici, principalmente dell\u2019acido malico, e la comparsa di metaboliti secondari, quali tannini ed antociani.(Robinson SP and Davies C \u201cMolecular biology of grape berry ripening\u201d Australian journal of grape and wine research 6: 175-188; 2000). A dispetto del grande interesse per questa specie, poco conosciuti rimangono i processi molecolari e biochimici attivi durante la maturazione di questo frutto non climaterico. La presenza di elevate concentrazioni di alcuni metaboliti, quali ad esempio i composti fenolici, rende infatti particolarmente difficoltosa l\u2019individuazione di metodiche adeguate per questo materiale sperimentale. Mentre alcune informazioni, seppure incomplete, sull\u2019attivit\ue0 trascrizionale sono tuttavia ora disponibili, il profilo d\u2019espressione proteica nelle diverse fasi di maturazione rimane pressoch\ue9 sconosciuto. L\u2019incremento delle conoscenze sul genoma di vite ottenute in questi ultimi anni ha comunque creato i presupposti necessari per avviare studi di proteomica. In questo contesto va evidenziato come la fase d\u2019estrazione richieda una particolare attenzione. Precedenti ricerche, condotte su frutti di altre specie vegetali, hanno infatti riscontrato che molte proteine possono legarsi alla componente glucosidica della parete con conseguente creazione di artefatti; in questo contesto appare quindi prioritario studiare il profilo d\u2019espressione proteica nelle diverse frazioni cellulari (Rose JKC et al. \u201cTackling the plant proteome: practical approaches, hurdles and experimental tools\u201d The Plant Journal, 39, 715-733, 2004). Primo obbiettivo di questo lavoro \ue8 stato quello di individuare il protocollo pi\uf9 idoneo per l\u2019estrazione delle proteine del protoplasto e della parete dell\u2019esocarpo della bacca di vite, tessuto in cui viene sintetizzata la maggiore parte dei metaboliti secondari. I risultati ottenuti dall\u2019analisi elettroforetica bidimensionale (2D-PAGE) hanno evidenziato che i campioni estratti in tampone e purificati mediante ripartizione con fenolo, precipitazione con ammonio acetato in metanolo e successivi lavaggi in acetone presentano, per entrambe le frazioni, il maggiore numero di spot rispetto ad altri protocolli testati. Circa il 15% dei peptidi sono risultati presenti in entrambe le frazioni, confermando come la procedura di estrazione incida sulla composizione dei diversi proteomi. La ricerca \ue8 quindi proseguita studiando i profili proteici nelle diverse fasi di maturazione. L\u2019analisi 2D-PAGE ha rivelato evidenti cambiamenti d\u2019espressione di alcuni peptidi che sono stati quindi analizzati mediante spettrometria di massa (LC-ESI-MS/MS, liquid chromatography \u2013 Electrospray - tandem mass spectrometry). I risultati ottenuti, seppure ancora preliminari, evidenziano le potenzialit\ue0 di questo approccio sperimentale per la comprensione dei meccanismi molecolari alla base dei i profondi cambiamenti biochimici e fisiologici che caratterizzano la maturazione della bacca di vite