An FTIR Spectroscopy Investigation on Different Methods of Lipid Extraction from HepG2 Cells

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Environmental bisphenol A exposure triggers trained immunity-related pathways in monocytes

IntroductionTrained Immunity represents a novel revolutionary concept of the immunological response involving innate immune cells. Bisphenol A is a well-known endocrine disrupter, widely disseminated worldwide and accumulated in the human body. Due to the increased interest regarding the effects of plastic-derived compounds on the immune system, our purpose was to explore whether BPA was able to induce trained immunity in human primary monocytes in vitro using low environmental concentrations.Materials and methodsWe extracted BPA from the serum of 10 healthy individuals through a liquid-liquid extraction followed by a solid phase extraction and measured the concentration using an HPLC system coupled to a triple quadrupole mass spectrometer. In parallel, monocytes were isolated from whole blood and acutely stimulated or trained with BPA at three different concentrations (1 nM, 10 nM, 20 nM). Pro- and anti-inflammatory cytokines (IL-1β, TNF-α, IL-6, and IL-10) production were assessed after 24 hours of acute stimulation and after Lipopolysaccharide (LPS) rechallenge. A comprehensive overview of the metabolic changes after BPA acute stimulation and trained immunity induction was assessed through extracellular lactate measurements, Seahorse XFb metabolic flux analysis and ROS production.ResultsMonocytes primed with BPA showed increased pro- and anti-inflammatory cytokine responses upon restimulation, sustained by the modulation of the immunometabolic circuits. Moreover, we proved the non-toxic effect of BPA at each experimental concentration by performing an MTT assay. Additionally, correlation analysis were performed between pro- and anti-inflammatory cytokines production after LPS acute stimulation or BPA-mediated trained immunity and BPA serum concentrations showing a significant association between TNF-α and BPA circulating levels.DiscussionOverall, this study pointed out for the first time the immunological effects of an environmental chemical and plastic-derived compound in the induction of trained immunity in a healthy cohort

Clinical and molecular features of an infant patient affected by Leigh Disease associated to m.14459G > A mitochondrial DNA mutation: a case report

<p>Abstract</p> <p>Background</p> <p>Leigh Syndrome (LS) is a severe neurodegenerative disorder characterized by bilateral symmetrical necrotic lesions in the basal ganglia and brainstem. Onset is in early infancy and prognosis is poor. Causative mutations have been disclosed in mitochondrial DNA and nuclear genes affecting respiratory chain subunits and assembly factors.</p> <p>Case presentation</p> <p>Here we report the clinical and molecular features of a 15-month-old female LS patient. Direct sequencing of her muscle-derived mtDNA revealed the presence of two apparently homoplasmic variants: the novel m.14792C > G and the already known m.14459G > A resulting in p.His16Asp change in cytochrome b (MT-CYB) and p.Ala72Val substitution in ND6 subunit, respectively. The m.14459G > A was heteroplasmic in the mother's blood-derived DNA.</p> <p>Conclusions</p> <p>The m.14459G > A might lead to LS, complicated LS or Leber Optic Hereditary Neuropathy. A comprehensive re-evaluation of previously described 14459G > A-mutated patients does not explain this large clinical heterogeneity.</p

EVALITA Evaluation of NLP and Speech Tools for Italian - December 17th, 2020

Welcome to EVALITA 2020! EVALITA is the evaluation campaign of Natural Language Processing and Speech Tools for Italian. EVALITA is an initiative of the Italian Association for Computational Linguistics (AILC, http://www.ai-lc.it) and it is endorsed by the Italian Association for Artificial Intelligence (AIxIA, http://www.aixia.it) and the Italian Association for Speech Sciences (AISV, http://www.aisv.it)

FTIR Spectroscopy for Evaluation and Monitoring of Lipid Extraction Efficiency for Murine Liver Tissues Analysis

Over the past several decades, growing research on lipids and lipidomic technologies have shown the important role played by lipids in many different situations. A powerful technique used for lipids detection and characterization in biological tissues is Fourier Transform Infrared (FTIR) spectroscopy. The main goal of the present work is to exploit FTIR spectroscopy as a tool for monitoring lipid extraction efficiency by evaluating three different lipid extraction methods in murine liver tissues. In particular, infrared spectra have been obtained in the 4000&ndash;600 cm&minus;1 wavenumber region and the contributions of different functional groups have been evidenced. The ratio values estimated using the absorbance of selected bands related to different liver constituents have been used for a quantitative comparison of the efficiency of the different extraction methods

Valutazione della vulnerabilità sismica e interventi di riqualificazione di una scuola primaria a Piazza al Serchio

L’elevata pericolosità sismica del territorio italiano comporta la necessità di conoscere la vulnerabilità del patrimonio edilizio esistente ed in particolar modo di quello pubblico. Lo studio di tale problematica è importante per la determinazione del livello di sicurezza di queste strutture a seguito di un evento sismico sia per poter effettuare degli studi di scenario, individuando gli edifici più a rischio sul territorio e pianificare gli interventi utili al ripristino della sicurezza, sia per indirizzare i primi soccorsi post evento sismico verso le aree più vulnerabili. La gran parte delle costruzioni esistenti in Italia è vulnerabile perchè ricade per la quasi totalità in un periodo temporale in cui la conoscenza delle strutture, dei materiali e delle azioni era molto limitata ed al contempo sorretta da basi normative non sufficientemente adeguate sia dal punto di vista tecnico che istituzionale. A causa del crescente deterioramento delle infrastrutture, frequentemente combinato con il bisogno, per il miglioramento strutturale, di soddisfare requisiti di progetto più rigidi (specialmente a riguardo dei carichi sismici), l’adeguamento strutturale è diventato sempre più importante e riscuote oggi rilievo considerevole in tutto il mondo. L’ingegneria sismica è un campo giovane della scienza applicata multidisciplinare, che si è sviluppata negli ultimi cinquanta anni con notevole progresso. Risultati significativi nella sismologia, nella geotecnica e nell’ingegneria strutturale sono stati conseguiti nell’accumulo dell’ampia conoscenza di base per produrre la sicurezza delle strutture contro i rischi sismici. Gli edifici esistenti sono forse, presumibilmente, più numerosi dei nuovi fabbricati. Pertanto, il recupero sismico è emerso come il principale tema nell’ingegneria sismica ed è diventato un campo di ricerca di spicco nelle azioni sottoposte a considerevole rischio sismico. L’obiettivo base del recupero sismico è l’identificazione di solidità e fragilità degli edifici esistenti e l’innalzamento a livelli prestazionali compatibili con gli obiettivi suddetti

A Sphingolipidomic Profiling Approach for Comparing X-ray-Exposed and Unexposed HepG2 Cells

: An analytical method based on tandem mass spectrometry-shotgun is presently proposed to obtain sphingolipidomic profiles useful for the characterization of lipid extract from X-ray-exposed and unexposed hepatocellular carcinoma cells (HepG2). To obtain a targeted lipidic profile from a specific biological system, the best extraction method must be identified before instrumental analysis. Accordingly, four different classic lipid extraction protocols were compared in terms of efficiency, specificity, and reproducibility. The performance of each procedure was evaluated using the Fourier-transform infrared spectroscopic technique; subsequently, the quality of extracts was estimated using electrospray ionization tandem mass spectrometry. The selected procedure based on chloroform/methanol/water was successfully used in mass spectrometry-based shotgun sphingolipidomics, allowing for evaluation of the response of cells to X-ray irradiation, the most common anticancer therapy. Using a relative quantitative approach, the changes in the sphingolipid profiles of irradiated cell extracts were demonstrated, confirming that lipidomic technologies are also useful tools for studying the key sphingolipid role in regulating cancer growth during radiotherapy

An FTIR Spectroscopy Investigation on Different Methods of Lipid Extraction from HepG2 Cells

Fourier transform infrared (FTIR) spectroscopy is a non-invasive technique that is largely used for studying lipidomics. Lipids are a primary class of biological molecules that play numerous vital roles in various processes. In the present work, we adopted FTIR spectroscopy for monitoring the lipid extraction efficiency of different methods used for extracting lipids from hepatocarcinoma cells. The spectra acquired from samples obtained with the selected methods showed the contributions of different functional groups. A qualitative comparison indicated that all the spectra exhibited similar lipid species profiles. The peak intensity attributed to the CH2 asymmetric stretching mode has been used for a quantitative comparison of the efficiency of the different extraction methods