Publications by authors named "Angela Amoresano"

129 Publications

Therapeutic targeting of Lyn kinase to treat chorea-acanthocytosis.

Acta Neuropathol Commun 2021 May 3;9(1):81. Epub 2021 May 3.

Department of Neuroscience, Biomedicine and Movement Sciences, University of Verona, Verona, Italy.

Chorea-Acanthocytosis (ChAc) is a devastating, little understood, and currently untreatable neurodegenerative disease caused by VPS13A mutations. Based on our recent demonstration that accumulation of activated Lyn tyrosine kinase is a key pathophysiological event in human ChAc cells, we took advantage of Vps13a mice, which phenocopied human ChAc. Using proteomic approach, we found accumulation of active Lyn, γ-synuclein and phospho-tau proteins in Vps13a basal ganglia secondary to impaired autophagy leading to neuroinflammation. Mice double knockout Vps13a Lyn showed normalization of red cell morphology and improvement of autophagy in basal ganglia. We then in vivo tested pharmacologic inhibitors of Lyn: dasatinib and nilotinib. Dasatinib failed to cross the mouse brain blood barrier (BBB), but the more specific Lyn kinase inhibitor nilotinib, crosses the BBB. Nilotinib ameliorates both Vps13a hematological and neurological phenotypes, improving autophagy and preventing neuroinflammation. Our data support the proposal to repurpose nilotinib as new therapeutic option for ChAc patients.
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http://dx.doi.org/10.1186/s40478-021-01181-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8091687PMC
May 2021

Nutritional Controlled Preparation and Administration of Different Tomato Purées Indicate Increase of β-Carotene and Lycopene Isoforms, and of Antioxidant Potential in Human Blood Bioavailability: A Pilot Study.

Nutrients 2021 Apr 17;13(4). Epub 2021 Apr 17.

CEINGE-Biotecnologie Avanzate, Via G. Salvatore, 486, 80145 Naples, Italy.

The isoforms of lycopene, carotenoids, and their derivatives including precursors of vitamin A are compounds relevant for preventing chronic degenerative diseases such as cardiovascular diseases and cancer. Tomatoes are a major source of these compounds. However, cooking and successive metabolic processes determine the bioavailability of tomatoes in human nutrition. To evaluate the effect of acute/chronic cooking procedures on the bioavailability of lycopene and carotene isoforms in human plasma, we measured the blood levels of these compounds and of the serum antioxidant potential in volunteers after a meal containing two different types of tomato sauce ( or ). Using a randomized cross-over administration design, healthy volunteers were studied, and the above indicated compounds were determined by HPLC. The results indicate an increased bioavailability of the estimated compounds and of the serum antioxidant potential with both types of tomato purée and the subsequently derived sauces (the increase was greater with purée). This study sheds light on the content of nutrient precursors of vitamin A and other antioxidant compounds derived from tomatoes cooked with different strategies. Lastly, our study indicates that strained purée should be preferred over rustic purée.
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http://dx.doi.org/10.3390/nu13041336DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8073136PMC
April 2021

Effects of a Lifestyle Change Intervention on Semen Quality in Healthy Young Men Living in Highly Polluted Areas in Italy: The FASt Randomized Controlled Trial.

Eur Urol Focus 2021 Feb 10. Epub 2021 Feb 10.

Department of Food Safety, Nutrition and Veterinary Public Health, Italian National Institute of Health (ISS), Rome, Italy.

Background: Human semen quality is affected by lifestyle and environmental factors.

Objective: To evaluate the short-term effects of a diet and physical activity intervention on semen quality of healthy young men living in highly polluted areas of Italy.

Design, Setting, And Participants: A randomized controlled trial was conducted. Healthy young men were assigned to an intervention or a control group.

Intervention: A 4-mo Mediterranean diet and moderate physical activity program.

Outcome Measurements And Statistical Analysis: The primary outcomes were sperm concentration, motility and morphology, concentration of round cells, and semen total antioxidant capacity. Secondary outcomes were adherence to Mediterranean diet and physical activity. All outcomes were measured twice, at the enrollment (t0) and at the end of the intervention (t4).

Results And Limitations: A total of 263 individuals attended all visits, and underwent examinations and laboratory analyses: 137 in the intervention group and 126 in the control group. The adherence to Mediterranean diet and physical activity level increased more in the intervention group than in the control group from t0 to t4. Sperm concentration, total and progressive motility, and proportion of normal morphology cells increased in the intervention group but decreased in the control group, with statistically significant differences between the two groups at t4. The total antioxidant capacity increased in the intervention group but decreased in the control group, from t0 to t4.

Conclusions: Study results showed that an intervention based on Mediterranean diet and regular physical activity can determine an improvement of semen quality in healthy young men.

Patient Summary: Our study aimed to evaluate the effect of a lifestyle intervention on semen quality of healthy young men. We assigned the 263 enrolled individuals to an intervention or a control group. The intervention group followed a 4-mo Mediterranean diet and moderate physical activity program, at the end of which the participants showed an improvement of semen quality parameters.
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http://dx.doi.org/10.1016/j.euf.2021.01.017DOI Listing
February 2021

Follicular microenvironment: Oxidative stress and adiponectin correlated with steroids hormones in women undergoing in vitro fertilization.

Mol Reprod Dev 2021 02 18;88(2):175-184. Epub 2020 Dec 18.

Department of Biomedical, Experimental, and Clinical Sciences "Mario Serio", University of Florence, Florence, Italy.

Research has been focused on determining the follicular microenviroment produced by the theca and granulosa cells since the molecular characterisation of this body fluid could lead to the understanding of several fertility problems. Oxidative stress may be one of the factors involved in female infertility since it plays a key role in the modulation of oocyte maturation and finally pregnancy. An increase in oxidative stress is correlated with inflammation and intense research was developed to understand the interaction between inflammation and adiponectin, based on the fact that many adipokines are inflammation related proteins linked to reactive oxygen species production. The aim of this study is to investigate the correlation between total adiponectin levels and oxidative stress amount in the serum and follicular fluid (FF) of women who undergone in vitro fertilization. Moreover we verified the expression of adiponectin in granulosa and cumulus cells. To clarify the predictive value of steroid hormones in human assisted reproduction, twelve steroid hormones in FF and serum, were quantified in a single run liquid chromatography/mass spectrometry, by using a multiple reaction monitoring mode and we related the serum and follicular fluids adiponectin levels with the concentration of the investigated steroid hormones.
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http://dx.doi.org/10.1002/mrd.23447DOI Listing
February 2021

Oxidation of diclofenac in water by sodium hypochlorite: Identification of new degradation by-products and their ecotoxicological evaluation.

J Pharm Biomed Anal 2021 Feb 12;194:113762. Epub 2020 Nov 12.

Department of Chemical Sciences, University of Naples Federico II, Naples, Italy. Electronic address:

Diclofenac (DCF) is the most widely prescribed non-steroidal anti-inflammatory drug in the world and it has been detected in drinking and surface waters. In this paper, the effect of chlorination process on DCF in aqueous solutions was investigated and the structures of 14 isolated degradation by-products (DPs), of which nine are new, have been determined from combining mass spectrometry and nuclear magnetic resonance data and justified by a proposed mechanism of formation beginning from the parent drug. Some degradation by-products show only one phenyl, others are dimers or trimers of the parental compound, which has undergone oxidative decarboxylation of the side chain and/or chlorination of this or one or both aromatic rings. Ecotoxicological bioassays evidenced the following sensitivities D. magna < R. subcapitata < A. fischeri. The isolated DPs (DP1-8, except for DP9) exhibited effects ≥ 50 % in the exposed microalgae and crustaceans showing toxicities mainly ranked from slight to acute.
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http://dx.doi.org/10.1016/j.jpba.2020.113762DOI Listing
February 2021

From untargeted metabolomics to the multiple reaction monitoring-based quantification of polyphenols in chocolates from different geographical areas.

J Mass Spectrom 2021 Jan 7;56(1):e4651. Epub 2020 Sep 7.

Dipartimento di Scienze Chimiche, Università di Napoli 'Federico II', Via Cinthia, 26, Naples, 80126, Italy.

Plants, including cocoa bean, are the main source of metabolites with multiple biological functions. Polyphenol extracts are widely used as a nutraceutical supplement for their well-known health-promoting role. In this paper, a preliminary untargeted metabolic screening was carried out by matrix-assisted laser desorption/ionization (MALDI)-time of flight (TOF)/TOF on a pool of chocolate samples made by cocoa beans of different geographical areas. Then, a targeted approach was developed for polyphenol quantification by an optimized Liquid chromatography (LC)-tandem mass spectrometry (MS/MS) method multiple reaction monitoring (MRM) ion mode. Detection limit of polyphenol standard ranged between 1 and 25 pg/μl with variation coefficient lower than 15%. External calibration curves were used for quantification of polyphenols in 18 samples. Fifty polyphenols were detected in a single LC-MRM/MS run and quantified by monitoring almost 90 transitions in a 5-minute run. The polyphenols content of different cocoa beans from several countries was finally compared by principal component analysis (PCA) statistical analysis suggesting that the chocolate made by Ecuador cocoa beans showed the highest level of polyphenols.
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http://dx.doi.org/10.1002/jms.4651DOI Listing
January 2021

Relationship between the metabolic and lipid profile in follicular fluid of women undergoing in vitro fertilization.

Mol Reprod Dev 2020 09 3;87(9):986-997. Epub 2020 Sep 3.

Department of Experimental and Clinical Biomedical Sciences "Mario Serio", University of Florence, Florence, Italy.

Among the follicular fluid (FF) components promoting the development of the oocyte are included glycoproteins, several fatty acids, and steroid hormones synthesized by the dominant follicle. For this, the analysis of the metabolites present in FF can determine the quality of the oocyte. FF composition is in part determined by local follicular metabolic processes and in part a plasma transudate. Since the causes of impaired fertility may be due to a metabolic imbalance, metabolomics is useful to identify low molecular weight metabolites. Oxidative stress is involved in human infertility and the use of metabolomics can be crucial to identify which other metabolites besides reactive oxygen species are involved in oxidative stress correlated to infertility. To obtain new information on the study of signaling molecules in FF, the knowledge of the lipid content will be important to improve information on the understanding of follicular development. The objective of this study is to identify (a) a metabolic profile and a lipid profile of FF in women undergoing in vitro fertilization and (b) to correlate the previous information obtained regarding adiponectin and oxidative stress with the metabolic and lipid profile obtained in the present study. As result, we found an increase in oxidative stress due to both an increase of androgens and an accumulation of lipids in the follicular environment and we suggest that this might be one of the causes of reduced fertility.
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http://dx.doi.org/10.1002/mrd.23415DOI Listing
September 2020

Metabolomic profiling of food matrices: Preliminary identification of potential markers of microbial contamination.

J Food Sci 2020 Oct 4;85(10):3467-3477. Epub 2020 Sep 4.

Department of Biology, Laboratory of Hygiene, University of Naples "Federico II", Via Cintia 26, Naples, 80126, Italy.

The research aimed to generate an early warning system highlighting in real-time bacterial contamination of meat matrices and providing information which could support companies in accepting or rejecting batches. Current microorganisms' detection methods rely on techniques (plate counting), which provide retrospective values for microbial contamination. The purpose of this research was to evaluate the ability of the headspace solid-phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC/MS) methodologies to detect volatile organic carbons (VOCs), which may be associated to a peculiar microbiological contamination of food. The disposal of fast headspace gas chromatography-mass spectrometry (HS-SPME-GC/MS) able to accurately and rapidly (30 min per sample) detect pathogens in raw meat could replace the traditional and time-consuming (3 to 4 days) standardized microbiological analysis required by regulations. Experiments focused on qualitative and quantitative evaluations of VOCs produced by Salmonella Typhimurium, Campylobacter jejuni, and Staphylococcus aureus in different types of raw meat (beef, pork, chicken). HS-SPME-GC/MS allowed to use smaller sample volumes compared to traditional methods with no sample processing and the potentiality for its application on various food matrices for the detection of a wide variety of pathogens. Data analysis showed the identification of unique VOCs' profiles being possible markers of meat contamination due to their association to specific pathogens. The identification of VOCs markers in association to selected bacterial pathogens and their metabolites could support the rapid determination of specific meat samples contamination. Further research is required to outline-specific metabolic profiles for each microorganism responsible of meat contamination and prevent false positives.
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http://dx.doi.org/10.1111/1750-3841.15418DOI Listing
October 2020

Protein Glycosylation Investigated by Mass Spectrometry: An Overview.

Cells 2020 08 28;9(9). Epub 2020 Aug 28.

Department of Chemical Sciences, University of Naples Federico II, Via Cinthia 26, 80126 Naples, Italy.

The protein glycosylation is a post-translational modification of crucial importance for its involvement in molecular recognition, protein trafficking, regulation, and inflammation. Indeed, abnormalities in protein glycosylation are correlated with several disease states such as cancer, inflammatory diseases, and congenial disorders. The understanding of cellular mechanisms through the elucidation of glycan composition encourages researchers to find analytical solutions for their detection. Actually, the multiplicity and diversity of glycan structures bond to the proteins, the variations in polarity of the individual saccharide residues, and the poor ionization efficiencies make their detection much trickier than other kinds of biopolymers. An overview of the most prominent techniques based on mass spectrometry (MS) for protein glycosylation (glycoproteomics) studies is here presented. The tricks and pre-treatments of samples are discussed as a crucial step prodromal to the MS analysis to improve the glycan ionization efficiency. Therefore, the different instrumental MS mode is also explored for the qualitative and quantitative analysis of glycopeptides and the glycans structural composition, thus contributing to the elucidation of biological mechanisms.
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http://dx.doi.org/10.3390/cells9091986DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7564411PMC
August 2020

Loss of Function of the Gene Encoding the Histone Methyltransferase KMT2D Leads to Deregulation of Mitochondrial Respiration.

Cells 2020 07 13;9(7). Epub 2020 Jul 13.

Division of Medical Genetics, Fondazione IRCCS Casa Sollievo della Sofferenza, 71013 San Giovanni Rotondo (FG), Italy.

encodes a methyltransferase responsible for histone 3 lysine 4 (H3K4) mono-/di-methylation, an epigenetic mark correlated with active transcription. Here, we tested the hypothesis that pathogenic loss-of-function variants, which causes the Kabuki syndrome type 1, could affect the mitochondrial metabolic profile. By using Seahorse technology, we showed a significant reduction of the mitochondrial oxygen consumption rate as well as a reduction of the glycolytic flux in both knockout MEFs and skin fibroblasts of Kabuki patients harboring heterozygous pathogenic variants. Mass-spectrometry analysis of intermediate metabolites confirmed alterations in the glycolytic and TCA cycle pathways. The observed metabolic phenotype was accompanied by a significant increase in the production of reactive oxygen species. Measurements of the specific activities of the mitochondrial respiratory chain complexes revealed significant inhibition of CI (NADH dehydrogenase) and CIV (cytochrome c oxidase); this result was further supported by a decrease in the protein content of both complexes. Finally, we unveiled an impaired oxidation of glucose and larger reliance on long-chain fatty acids oxidation. Altogether, our findings clearly indicate a rewiring of the mitochondrial metabolic phenotype in the KMT2D-null or loss-of-function context that might contribute to the development of Kabuki disease, and represents metabolic reprogramming as a potential new therapeutic approach.
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http://dx.doi.org/10.3390/cells9071685DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7407568PMC
July 2020

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

Foods 2020 Jun 25;9(6). Epub 2020 Jun 25.

Department Chemical Sciences, University of Naples Federico II, Monte S. Angelo-Cinthia, 80126 Naples, Italy.

Chemical compounds within tea 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.
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http://dx.doi.org/10.3390/foods9060835DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7353651PMC
June 2020

Green synthesis of conductive polyaniline by laccase using a DNA template.

Eng Life Sci 2019 Sep 25;19(9):631-642. Epub 2019 Jul 25.

Dipartimento di Scienze chimiche Università di Napoli "Federico II" Napoli Italy.

The green synthesis of highly conductive polyaniline by using two biological macromolecules, i.e laccase as biocatalyst, and DNA as template/dopant, was achieved in this work. laccase B (TvB) was found effective in oxidizing both aniline and its less toxic/mutagenic dimer N-phenyl-p-phenylenediamine (DANI) to conductive polyaniline. Reaction conditions for synthesis of conductive polyanilines were set-up, and structural and electrochemical properties of the two polymers were extensively investigated. When the less toxic aniline dimer was used as substrate, the polymerization reaction was faster and gave less-branched polymer. DNA was proven to work as hard template for both enzymatically synthesized polymers, conferring them a semi-ordered morphology. Moreover, DNA also acts as dopant leading to polymers with extraordinary conductive properties (∼6 S/cm). It can be envisaged that polymer properties are magnified by the concomitant action of DNA as template and dopant. Herein, the developed combination of laccase and DNA represents a breakthrough in the green synthesis of conductive materials.
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http://dx.doi.org/10.1002/elsc.201900078DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6999394PMC
September 2019

Variation in prostaglandin metabolism during growth of the diatom Thalassiosira rotula.

Sci Rep 2020 03 25;10(1):5374. Epub 2020 Mar 25.

Stazione Zoologica Anton Dohrn, Villa Comunale, 80121, Napoli, Italy.

Prostaglandins (PGs) are hormone-like mediators in many physiological and pathological processes that are present in all vertebrates, in some terrestrial and aquatic invertebrates, and have also been identified in some macroalgae. They have recently been reported also in marine microalgae but their role as chemical mediators is largely unknown. Here we studied the expression pattern of the PG biosynthetic pathway during different growth phases of the centric diatom Thalassiosira rotula and assessed the release of PGs in the surrounding environment for the first time. We show that enzymes responsible for PGs formation such as cyclooxygenase, prostaglandin E synthase 2-like and prostaglandin F synthase are mainly expressed at the end of the exponential phase and that PGs are released especially during the stationary and senescent phases, suggesting a possible signaling function for these compounds. Phylogenetic analysis of the limiting enzyme, COX, indicate the presence in diatoms of more than one enzyme related to the oxidative metabolism of fatty acids belonging to the peroxidase-cyclooxygenase superfamily. These findings suggest a more complex evolution and diversity of metabolic pathways leading to the synthesis of lipid mediators in diatoms.
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http://dx.doi.org/10.1038/s41598-020-61967-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7096440PMC
March 2020

Synthetic Lethality Screening Identifies FDA-Approved Drugs that Overcome ATP7B-Mediated Tolerance of Tumor Cells to Cisplatin.

Cancers (Basel) 2020 Mar 6;12(3). Epub 2020 Mar 6.

Telethon Institute of Genetics and Medicine, Pozzuoli, 80078 Naples, Italy.

Tumor resistance to chemotherapy represents an important challenge in modern oncology. Although platinum (Pt)-based drugs have demonstrated excellent therapeutic potential, their effectiveness in a wide range of tumors is limited by the development of resistance mechanisms. One of these mechanisms includes increased cisplatin sequestration/efflux by the copper-transporting ATPase, ATP7B. However, targeting ATP7B to reduce Pt tolerance in tumors could represent a serious risk because suppression of ATP7B might compromise copper homeostasis, as happens in Wilson disease. To circumvent ATP7B-mediated Pt tolerance we employed a high-throughput screen (HTS) of an FDA/EMA-approved drug library to detect safe therapeutic molecules that promote cisplatin toxicity in the IGROV-CP20 ovarian carcinoma cells, whose resistance significantly relies on ATP7B. Using a synthetic lethality approach, we identified and validated three hits (Tranilast, Telmisartan, and Amphotericin B) that reduced cisplatin resistance. All three drugs induced Pt-mediated DNA damage and inhibited either expression or trafficking of ATP7B in a tumor-specific manner. Global transcriptome analyses showed that Tranilast and Amphotericin B affect expression of genes operating in several pathways that confer tolerance to cisplatin. In the case of Tranilast, these comprised key Pt-transporting proteins, including ATOX1, whose suppression affected ability of ATP7B to traffic in response to cisplatin. In summary, our findings reveal Tranilast, Telmisartan, and Amphotericin B as effective drugs that selectively promote cisplatin toxicity in Pt-resistant ovarian cancer cells and underscore the efficiency of HTS strategy for identification of biosafe compounds, which might be rapidly repurposed to overcome resistance of tumors to Pt-based chemotherapy.
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http://dx.doi.org/10.3390/cancers12030608DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7139527PMC
March 2020

Pt(II) versus Pt(IV) in Carbene Glycoconjugate Antitumor Agents: Minimal Structural Variations and Great Performance Changes.

Inorg Chem 2020 Mar 4;59(6):4002-4014. Epub 2020 Mar 4.

Dipartimento di Scienze Chimiche, Università di Napoli Federico II, Complesso Universitario di Monte S. Angelo, via Cintia 21, 80126 Napoli, Italy.

Octahedral Pt(IV) complexes () containing a glycoconjugate carbene ligand were prepared and fully characterized. These complexes are structural analogues to the trigonal bipyramidal Pt(II) species () recently described. Thus, an unprecedented direct comparison between the biological properties of Pt compounds with different oxidation states and almost indistinguishable structural features was performed. The stability profile of the novel Pt(IV) compounds in reference solvents was determined and compared to that of the analogous Pt(II) complexes. The uptake and antiproliferative activities of and were evaluated on the same panel of cell lines. DNA and protein binding properties were assessed using human serum albumin, the model protein hen egg white lysozyme, and double stranded DNA model systems by a variety of experimental techniques, including UV-vis absorption spectroscopy, fluorescence, circular dichroism, and electrospray ionization mass spectrometry. Although the compounds present similar structures, their in-solution stability, cellular uptake, and DNA binding properties are diverse. These differences may represent the basis of their different cytotoxicity and biological activity.
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http://dx.doi.org/10.1021/acs.inorgchem.9b03683DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7997382PMC
March 2020

A thermophilic C-phycocyanin with unprecedented biophysical and biochemical properties.

Int J Biol Macromol 2020 May 7;150:38-51. Epub 2020 Feb 7.

Department of Chemical Sciences, University of Naples Federico II, Naples, Italy. Electronic address:

C-phycoyanins are abundant light-harvesting pigments which have an important role in the energy transfer cascade of photosystems in prokaryotic cyanobacteria and eukaryotic red algae. These proteins have important biotechnological applications, since they can be used in food, cosmetics, nutraceutical, pharmaceutical industries and in biomedical research. Here, C-phycocyanin from the extremophilic red alga Galdieria phlegrea (GpPC) has been purified and characterized from a biophysical point of view by SDS-PAGE, mass spectrometry, UV-Vis absorption spectroscopy, circular dichroism and intrinsic fluorescence. Stability against pH variations, addition of the oxidizing agent hydrogen peroxide and the effects of temperature have been also investigated, together with its in cell antioxidant potential and antitumor activity. GpPC is stable under different pHs and unfolds at a temperature higher than 80 °C within the pH range 5.0-7.0. Its fluorescence spectra present a maximum at 650 nm, when excited at 589 nm. The protein exerts interesting in cell antioxidant properties even after high temperature treatments, like the pasteurization process, and is cytotoxic for A431 and SVT2 cancer cells, whereas it is not toxic for non-malignant cells. Our results assist in the development of C-phycocyanin as a multitasking protein, to be used in the food industry, as antioxidant and anticancer agent.
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http://dx.doi.org/10.1016/j.ijbiomac.2020.02.045DOI Listing
May 2020

Label-free quantitative proteomics of the MCF-7 cellular response to a ferritin-metallodrug complex.

Mol Omics 2020 04 4;16(2):165-173. Epub 2020 Feb 4.

Department of Chemical Sciences, University of Naples Federico II, via Cinthia, Napoli, 80126, Italy.

Auoxo3 is a gold(iii) compound endowed with cytotoxic activity towards a variety of malignant cells. Encapsulation of Auoxo3 within horse spleen ferritin (Ft) improves the selectivity of the gold compound towards cancer cells over normal cells. In the current work, the changes in protein expression are presented in response to MCF-7 stimulation with Auoxo3-encapsulated Ft versus the free Au(iii) compound by a label-free proteomics approach. A 159-protein dataset showed significant changes between the stimulations with Auoxo3 and Auoxo3-encapsulated Ft, suggesting that this cellular perturbation caused the alteration of different cellular processes. In detail, roughly 30% of proteins were downregulated mainly in the spliceosome complex (U2AF1, SF3B2, PRPF4, SNSRP200, EFTUD2, PRPF6, and PRPF8) in agreement with the cytostatic effect observed during cellular growth. Another 30% of proteins were upregulated primarily in glutathione biosynthesis, suggesting an alteration of the redox potential, as validated by Western blot analyses. To the best of our knowledge, this work represents the first large scale proteomics study on the effects of a gold-based drug encapsulated within the Ft nanocage on cancer cells.
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http://dx.doi.org/10.1039/c9mo00158aDOI Listing
April 2020

A cascade extraction of active phycocyanin and fatty acids from Galdieria phlegrea.

Appl Microbiol Biotechnol 2019 Dec 7;103(23-24):9455-9464. Epub 2019 Nov 7.

Department of Chemical Sciences, University of Naples Federico II, via Cinthia 4, 80126, Naples, Italy.

The setup of an economic and sustainable method to increase the production and commercialization of products from microalgae, beyond niche markets, is a challenge. Here, a cascade approach has been designed to optimize the recovery of high valuable bioproducts starting from the wet biomass of Galdieria phlegrea. This unicellular thermo-acidophilic red alga can accumulate high-value compounds and can live under conditions considered hostile to most other species. Extractions were performed in two sequential steps: a conventional high-pressure procedure to recover phycocyanins and a solvent extraction to obtain fatty acids. Phycocyanins were purified to the highest purification grade reported so far and were active as antioxidants on a cell-based model. Fatty acids isolated from the residual biomass contained high amount of PUFAs, more than those recovered from the raw biomass. Thus, a simple, economic, and high effective procedure was set up to isolate phycocyanin at high purity levels and PUFAs.
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http://dx.doi.org/10.1007/s00253-019-10154-0DOI Listing
December 2019

Biosensor surface functionalization by a simple photochemical immobilization of antibodies: experimental characterization by mass spectrometry and surface enhanced Raman spectroscopy.

Analyst 2019 Nov;144(23):6871-6880

Dipartimento di Fisica, Politecnico di Milano, Piazza Leonardo da Vinci 32, 20133 - Milano, Italy.

Surface functionalization is a key step in biosensing since it is the basis of an effective analyte recognition. Among all the bioreceptors, antibodies (Abs) play a key role thanks to their superior specificity, although the available immobilization strategies suffer from several drawbacks. When gold is the interacting surface, the recently introduced Photochemical Immobilization Technique (PIT) has been shown to be a quick, easy-to-use and very effective method to tether Abs oriented upright by means of thiols produced via tryptophan mediated disulphide bridge reduction. Although the molecular mechanism of this process is quite well identified, the detailed morphology of the immobilized antibodies is still elusive due to inherent difficulties related to the microscopy imaging of Abs. The combination of Mass Spectrometry, Surface-Enhanced Raman Spectroscopy and Ellman's assay demonstrates that Abs irradiated under the conditions in which PIT is realized show only two effective disulphide bridges available for binding. They are located in the constant region of the immunoglobulin light chain so that the most likely position Ab assumes is side-on, i.e. with one Fab (i.e. the antigen binding portion of the antibody) exposed to the solution. This is not a limitation of the recognition efficiency in view of the intrinsic flexibility of the Ab structure, which makes the free Fab able to sway in the solution, a feature of great importance in many biosensing applications.
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http://dx.doi.org/10.1039/c9an00443bDOI Listing
November 2019

A multi-scale time-resolved study of photoactivated dynamics in 5-benzyl uracil, a model for DNA/protein interactions.

Phys Chem Chem Phys 2019 Dec;21(48):26301-26310

Department of Physics "Ettore Pancini", University of Naples "Federico II", Naples, 80126, Italy.

We combine fluorescence up-conversion and time correlated single photon counting experiments to investigate the 5-benzyl uracil excited state dynamics in methanol from 100 fs up to several ns. This molecule has been proposed as a model for DNA/protein interactions. Our results show emission bands at about 310 and 350 nm that exhibit bi-exponential sub-ps decays. Calculations, including solvent effects by a mixed discrete-continuum model, indicate that the Franck Condon region is characterized by significant coupling between the excited states of the benzyl and the uracil moieties, mirrored by the short-lived emission at 310 nm. Two main ground state recovery pathways are identified, both contributing to the 350 nm emission. The first 'photophysical' decay path involves a ππ* excited state localized on the uracil and is connected to the ground electronic state by an easily accessible crossing with S0, accounting for the short lifetime component. Simulations indicate that a possible second pathway is characterized by exciplex formation, with partial benzene → uracil charge transfer character, that may lead instead to photocyclization. The relevance of our results is discussed in view of the photoactivated dynamics of DNA/protein complexes, with implications on their interaction mechanisms.
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http://dx.doi.org/10.1039/c9cp03839fDOI Listing
December 2019

Fiano, Greco and Falanghina grape cultivars differentiation by volatiles fingerprinting, a case study.

Heliyon 2019 Aug 22;5(8):e02287. Epub 2019 Aug 22.

Department of Chemical Sciences, University of Naples Federico II, Italy.

The biomolecular characterization of edible products is gaining an increasing importance in food chemistry. The characteristic aroma or bouquet of a wine is the result of complex interactions of volatile molecules and odor receptors. Its characterization is the subject of many different studies, aimed at the development of new methods to be used for the discovery of frauds and for the typization of Protected Designation of Origin (P.D.O.) or Protected Geographic Indication (P.G.I.) wines. We previously outlined the proteomic profile of three cultivars of from South Italy (Campania) used for white wine production (Fiano, Greco and Falanghina) during the ripening. In this work, we present a mass spectrometry based study aimed at obtaining the profile of volatiles on the same samples using solid phase micro extraction coupled to gas chromatography. We demonstrated that some of the main constituents of aroma (namely terpenes, alcohols, aldehydes, etc.) were characteristic of certain grapes and absent in others.
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http://dx.doi.org/10.1016/j.heliyon.2019.e02287DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6716974PMC
August 2019

A physicochemical investigation on the metal binding properties of TtSmtB, a thermophilic member of the ArsR/SmtB transcription factor family.

Int J Biol Macromol 2019 Oct 26;138:1056-1063. Epub 2019 Jul 26.

Department of Biology, University of Naples Federico II, Napoli, Italy. Electronic address:

The transcription factors of the ArsR/SmtB family are widespread within the bacterial and archaeal kingdoms. They are transcriptional repressors able to sense a variety of metals and undergo allosteric conformational changes upon metal binding, resulting in derepression of genes involved in detoxification. So far, the molecular determinants of specificity, selectivity, and metal binding mechanism have been scarcely investigated in thermophilic microorganisms. TtSmtB, the only ArsR/SmtB member present in the genome of Thermus thermophilus HB27, was chosen as a model to shed light into such molecular mechanisms at high temperature. In the present study, using a multidisciplinary approach, a structural and functional characterization of the protein was performed focusing on its metal interaction and chemical-physical stability. Our data demonstrate that TtSmtB has two distinct metal binding sites per monomer and interacts with di-tri-penta-valent ions with different affinity. Detailed knowledge at molecular level of protein-metal interaction is remarkable to design metal binding domains as scaffolds in metal-based therapies as well as in metal biorecovery or biosensing in the environment.
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http://dx.doi.org/10.1016/j.ijbiomac.2019.07.174DOI Listing
October 2019

Corrigendum to "Mathematical optimization of the green extraction of polyphenols from grape peels through a cyclic pressurization process" [Heliyon 5 (4) (April 2019) e01526].

Heliyon 2019 May 14;5(5):e01593. Epub 2019 May 14.

Department of Chemical Sciences, University of Naples Federico II, via Cintia, 80126, Naples, Italy.

[This corrects the article DOI: 10.1016/j.heliyon.2019.e01526.].
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http://dx.doi.org/10.1016/j.heliyon.2019.e01593DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6522654PMC
May 2019

Lanthionine and Other Relevant Sulfur Amino Acid Metabolites: Detection of Prospective Uremic Toxins in Serum by Multiple Reaction Monitoring Tandem Mass Spectrometry.

Methods Mol Biol 2019 ;2007:9-17

Department of Chemical Sciences, University of Naples "Federico II", Naples, Italy.

In the context of the vascular effects of hydrogen sulfide (HS), it is known that this gaseous endogenous biological modulator of inflammation, oxidative stress, etc. is a potent vasodilator. Chronic renal failure, a common disease affecting the aging population, is characterized by low levels of HS in plasma and tissues, which could mediate their typical hypertensive pattern, along with other abnormalities. Lanthionine and homolanthionine, natural non-proteinogenic amino acids, are formed as side products of HS production. Also in consideration of the intrinsic difficulties in HS measuring, these compounds have been proposed as reliable and stable markers of HS synthesis. However, in the setting of chronic renal failure patients on hemodialysis, they represent typical retention products (without ruling out the possibility of an increased intestinal synthesis) and prospective novel uremic toxins. Here, a method utilizing liquid chromatography-electrospray tandem mass spectrometry (LC-MS/MS) in multiple reaction monitoring ion mode has been developed and evaluated for the determination of these key HS metabolites in plasma, by using a triple quadrupole mass spectrometer.
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http://dx.doi.org/10.1007/978-1-4939-9528-8_2DOI Listing
March 2020

Mathematical optimization of the green extraction of polyphenols from grape peels through a cyclic pressurization process.

Heliyon 2019 Apr 17;5(4):e01526. Epub 2019 Apr 17.

Department of Chemical Sciences, University of Naples Federico II, via Cintia, 80126, Naples, Italy.

In the current era of high consumption and increasing waste, many products that are believed to be unusable can find a new purpose in the market. For example, the grape peel waste resulting from the production of wine contains numerous bioactive compounds. In reality, grape peels are by-products of winemaking that can be conveniently reused in many different ways, including agronomic use and cosmetic industry applications. Moreover, the by-products can also be used in the energy field as biomass for the production of biogas or in food plants for the production of energy. In this article, to extract polyphenols, grape peels were processed via a cyclically pressurized extraction method known as rapid solid-liquid dynamic extraction (RSLDE), which does not require the use of any organic solvent or include heating or cooling processes that can cause the loss of substances of interest. To better understand the cyclically pressurized extraction process, a numerical simulation was performed to evaluate the exchange between the grape piece solid matrix and water during the extraction process. Furthermore, a finite element model was used to numerically determine the time-dependent concentration distribution at specific times.
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http://dx.doi.org/10.1016/j.heliyon.2019.e01526DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6476133PMC
April 2019

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

Food Res Int 2019 06 10;120:26-32. Epub 2019 Feb 10.

Department of Chemical Sciences, University of Naples Federico II, Italy.

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.
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http://dx.doi.org/10.1016/j.foodres.2019.02.020DOI Listing
June 2019

A comparative assessment of metals and phthalates in commercial tea infusions: A starting point to evaluate their tolerance limits.

Food Chem 2019 Aug 2;288:193-200. Epub 2019 Mar 2.

Department of Biology, University of Naples Federico II, Botanical Garden, Via Foria 223, 80139 Naples, Italy; DNATech srl - Spin-off company of the University of Naples Federico II, Botanical Garden, Via Foria 223, 80139 Naples, Italy. Electronic address:

Tea is one of the most consumed beverages in the word. Here we report the concentrations of metals and phthalates in 32 commercial tea packages. The data were used to estimate the average daily intake of metals and phthalates, and associated Hazard Quotients (HQ) were calculated in order to determine risk of non-cancerous health effects for adults consuming tea on a daily basis. Tea samples were chosen based on the sales network, the price, the marketing quality and the presence of filters in the packages. Relatively high median concentrations of Al (5240 µg/L), Ni (44 µg/L), and Mn (2919 µg/L) were detected. No metals or phthalates quantified in the tea infusions and soluble tea showed an HQ greater than 1, indicating no risk of non-cancerous health effects. The data presented herein may serve as a starting point to evaluate tolerance limits of metals and phthalate in the tea infusion.
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http://dx.doi.org/10.1016/j.foodchem.2019.02.115DOI Listing
August 2019

Evolution of an insect immune barrier through horizontal gene transfer mediated by a parasitic wasp.

PLoS Genet 2019 03 5;15(3):e1007998. Epub 2019 Mar 5.

Department of Agricultural Sciences, University of Napoli Federico II, Portici (NA), Italy.

Genome sequencing data have recently demonstrated that eukaryote evolution has been remarkably influenced by the acquisition of a large number of genes by horizontal gene transfer (HGT) across different kingdoms. However, in depth-studies on the physiological traits conferred by these accidental DNA acquisitions are largely lacking. Here we elucidate the functional role of Sl gasmin, a gene of a symbiotic virus of a parasitic wasp that has been transferred to an ancestor of the moth species Spodoptera littoralis and domesticated. This gene is highly expressed in circulating immune cells (haemocytes) of larval stages, where its transcription is rapidly boosted by injection of microorganisms into the body cavity. RNAi silencing of Sl gasmin generates a phenotype characterized by a precocious suppression of phagocytic activity by haemocytes, which is rescued when these immune cells are incubated in plasma samples of control larvae, containing high levels of the encoded protein. Proteomic analysis demonstrates that the protein Sl gasmin is released by haemocytes into the haemolymph, where it opsonizes the invading bacteria to promote their phagocytosis, both in vitro and in vivo. Our results show that important physiological traits do not necessarily originate from evolution of pre-existing genes, but can be acquired by HGT events, through unique pathways of symbiotic evolution. These findings indicate that insects can paradoxically acquire selective advantages with the help of their natural enemies.
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http://dx.doi.org/10.1371/journal.pgen.1007998DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6420030PMC
March 2019

Encapsulation of the Dinuclear Trithiolato-Bridged Arene Ruthenium Complex Diruthenium-1 in an Apoferritin Nanocage: Structure and Cytotoxicity.

ChemMedChem 2019 03 11;14(5):594-602. Epub 2019 Feb 11.

Department of Chemical Sciences, University of Naples Federico II, Complesso Universitario di Monte Sant'Angelo, via Cinthia 21, 80126, Naples, Italy.

The effects of encapsulating the cytotoxic dinuclear trithiolato-bridged arene ruthenium complex [(η -p-MeC H iPr) Ru (μ -S-p-C H tBu) ]Cl (DiRu-1) within the apoferritin (AFt) nanocage were investigated. The DiRu-1-AFt nanocarrier was characterized by UV/Vis spectroscopy, ICP-MS, CD and X-ray crystallography. In contrast to previously reported Au- and Pt-based drug-loaded AFt carriers, we found no evidence of direct interactions between DiRu-1 and AFt. DiRu-1-AFt is cytotoxic toward immortalized murine BALB/c-3T3 fibroblasts transformed with SV40 virus (SVT2) and human epidermoid carcinoma A431 malignant cells, and exhibits moderate selectivity for these cancer cells over normal BALB/c-3T3 cells. DiRu-1-AFt triggers the production of reactive oxygen species, depolarization of mitochondrial membrane potential, and induces cell death via p53-mediated apoptosis. Comparison between our data and previous results suggests that the presence of specific interactions between a metal-based drug and AFt within the protein cage is not essential for drug encapsulation.
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http://dx.doi.org/10.1002/cmdc.201800805DOI Listing
March 2019