Leila Birolo

University of Naples Federico II | UNINA · Department of Chemical Sciences

This paper regards the preliminary characterization of wall paintings sampled in a Roman domus located in the city of Santa Maria Capua Vetere. Despite its history and archaeological remains, there is a lack of scientific studies on Roman materials and techniques adopted in this site. Samples were taken from different walls of the villa. Pigments,...

Exposing artworks and artifacts in museums is fundamental to disseminate culture and knowledge but is also risky since the exposure to environmental conditions can deteriorate exhibited objects.Light is crucial, being able to trigger photochemical effects, and jointly responsible (with tem-perature and humidity) for biological risk generation. In t...

The study applies both a minimal and an extended approach for a comprehensive picture of chemical components in wall paintings, including evidence of degradation. Pigments and ligands were characterized via a multi-methodological investigation, including optical microscopy, scanning electron microscopy, Raman micro-spectroscopy, GC-MS, and LC-MS/MS...

The use of multichannel LEDs is crucial in museum applications to design ad-hoc spectra able to combine fruition and protection needs, i.e., maximizing Colour Rendering Index and controlling energy content in risk wavelengths responsible for the photochemical degradation of the exposed objects. A similar approach could be applied also to prevent th...

Animal glues are widely used in restoration as adhesives, binders, and consolidants for organic and inorganic materials. Their variable performances are intrinsically linked to the adhesive properties of collagen, which determine the chemical, physical, and mechanical properties of the glue. We have molecularly characterized the protein components...

An extensive proteomic analysis was performed on a set of 12 bones of human victims of the eruption that in AD 79 rapidly buried Pompeii and Herculaneum, allowing the detection of molecular signatures imprinted in the surviving protein components. Bone collagen survived the heat of the eruption, bearing a piece of individual biological history enco...

Brewer’s spent grain (BSG), the solid residue of beer production, is attracting significant attention as raw material for the production of added value substances, since until recently it was mainly used as animal feed or deposited in landfills, causing serious environmental problems. Therefore, this work aimed at developing a bioprocess using BSG...

Inulin, a polydisperse fructan found as a common storage polysaccharide in the roots of several plants, represents a renewable non-food biomass resource for the synthesis of bio-based products. Exploitation of inulin-containing feedstocks requires the integration of different processes, including inulinase production, saccharification of inulin, an...

Identification and characterization of ancient proteins still require technical developments towards non-invasiveness, sensitivity, versatility and ease of use of the analyses. We report that the enzyme functionalized films, described in Cicatiello et al. (2018), can be used efficiently on the surface of different objects ranging from fixative-coat...

ICOSL/ICOS are costimulatory molecules pertaining to immune checkpoints; their binding transduces signals having anti-tumor activity. Osteopontin (OPN) is here identified as a ligand for ICOSL. OPN binds a different domain from that used by ICOS, and the binding induces a conformational change in OPN, exposing domains that are relevant for its func...

Two fungal strains, Aspergillus terreus MUT 271 and Trichoderma harzianum MUT 290, isolated from a Mediterranean marine site chronically pervaded by oil spills, can use crude oil as sole carbon source. Herein, these strains were investigated as producers of biosurfactants, apt to solubilize organic molecules as a preliminary step to metabolize them...

Marine microorganisms represent a reservoir of new promising secondary metabolites. Surface-active proteins with good emulsification activity can be isolated from fungal species that inhabit the marine environment and can be promising candidates for different biotechnological applications. In this study a novel surface-active protein, named Sap-Pc,...

In AD 79 the town of Herculaneum was suddenly hit and overwhelmed by volcanic ash-avalanches that killed all its remaining residents, as also occurred in Pompeii and other settlements as far as 20 kilometers from Vesuvius. New investigations on the victims' skeletons unearthed from the ash deposit filling 12 waterfront chambers have now revealed wi...

The Roman town of Herculaneum. A. Map showing approximate location of the site; B. View of the archaeological excavations; C. The seafront chambers crowded with different numbers of victims’ skeletons; D. A group of victims unearthed in chamber 12; E. Planimetry of chamber 10 (The authors created the image themselves. Photos [B, D] and images [C, E...

Thermal microscopic and ultra-structural modifications in human victims' bones from Herculaneum. Adult bone analyzed with a light microscope (scale bar 100 μm) and a scanning electron microscope (scale bar 10 μm, 1700×): Radius showing both linear and polygonal cracking (A) and incipient recrystallization (B). At 500–800°C the basic bone structure...

Type of samples analyzed and their source. (DOCX)

Planimetric sections of the town (A) and a boat-chamber (B). A. Section of the inferior IV cardo, the suburban area and the chambers facing the sea. Note the turbulent surge cloud passing through the town and emplacing on the beach and within the chambers; B. Section of the seafront area with the victims buried within the ash surge deposit (images...

Iron amounts resulting from inductively coupled plasma mass spectrometry performed on 103 specimens. (DOCX)

Representative Raman spectra. A. Samples 13, 21, 22, 25 and 45. Ap (apatite), C (amorphous carbon), Ca (Carbonate), He (hematite), Gl (SiO2 glasses), Io (mixture of multiple iron oxides), Ih (iron oxyhydroxides), Ma (Magnetite), Mg (maghemite), Pl (Plagioclase), Py (Pyroxene); B. samples 46, 49, 51, 75, 92. Ap (apatite), C (amorphous carbon), Ca (C...

Sex and average age at death of the 80 victims' skeletons found in chambers 5, 10, 11, and 12 in 1997–1999 archaeological investigations in the suburban area of Herculaneum. Individual A was one of the very few victims found in the town. (DOCX)

Details of the identification of human collagen proteins alpha-1(I) and alpha-2(I) in sample C5. (DOCX)

Identification of proteins in paintings and polychrome objects is a challenge, which requires the development of tailored analytical approaches. In the present study, a targeted proteomics approach was developed for discriminating among the three most common proteinaceous materials used as paint binders, i.e. milk, egg, and animal glue. In this stu...

An organic binder was identified in the painted fragments from the Canaanite palace of Tel Kabri, Israel. Recently dated to the late 18th C. B.C.E. by 14C, Tel Kabri is the most ancient of the Eastern Mediterranean sites in which Aegean style paintings have been found. The application of pigments was suspected to be using an organic binding medium,...

An organic binder was identified in the painted fragments from the Canaanite palace of Tel Kabri, Israel. Recently dated to the late 18th C. B.C.E. by 14C, Tel Kabri is the most ancient of the Eastern Mediterranean sites in which Aegean style paintings have been found. The application of pigments was suspected to be using an organic binding medium,...

A novel method for the analysis of proteinaceous materials present on painted surfaces was developed by taking advantage of the adhesive ability of some fungal proteins which can form a stable and homogeneous layer on flexible transparency sheets able to capture trypsin in a fully active form. We demonstrated that the bioactive sheets were able to...

The paper concerns the analyses of the fat that was found in the majority of the pottery excavated by John Allegro in the 1960s. It was also important to learn whether the Christmas Cave pottery is similar or not as that of Qumran in order to differentiate between the two groups of people who occupied the cave and the Qumran site.

We have studied two misfolded oligomeric forms of the protein HypF-N, which show similar morphologies but very different toxicities. We measured over 80 intermolecular distance-dependent parameters for each oligomer type using FRET, in conjunction with solution- and solid-state NMR and other biophysical techniques. The results indicate that the for...

The analysis of Pleurotus ostreatus genome reveals the presence of automatically annotated 53 lipase and 34 carboxylesterase putative coding-genes. Since no biochemical or physiological data are available so far, a functional approach was applied to identify lipases from P. ostreatus. In the tested growth conditions, four lipases were found express...

SDS-PAGE of recombinant lipases. Lane 1: Protein ladder; Lane 2: PleoLip369 crude extract; Lane 3: PleoLip241 crude extract; Lane 4: purified PleoLip369; Lane 5: purified PleoLip241. (DOCX)

Growth media conditions for extracellular lipase induction. The conditions that induce the production of extracellular lipase after five growth days are reported. All experiments have been conducted in triplicate. (DOCX)

Multiple alignment among between lipase coding sequences from P. ostreatus and sequences of lipases from Hypsizygus marmoreus and from Pleurotus eryngii. In the grey box the conserved pentapeptide is displayed. The red arrows indicate the three aminoacids of catalytic triade. (DOCX)

Determination of the chemical composition of different parts of the wall paintings (pigments, mortars and binders) allows to extract information about technology of preparation of the artefact. Herein we present a multimethodological characterization of wall paintings from a Roman Archeological site in Cuma, focusing on differences between a indoor...

A chemical investigation of the organic paint binders of the Giant Buddhas of Bāmiyān was performed using an analytical approach based on mass spectrometry, combining traditional gas chromatography/mass spectrometry protocols with advanced proteomics methodologies. The research was carried out on a selection of rescued fragments. The data revealed...

LC-MS/MS analyses. (DOCX)

GC/MS Saccharide fraction. (DOCX)

Fragments analysed from the Eastern Buddha. (DOCX)

Fragments analysed from the Western Buddha. (DOCX)

GC/MS Amino acidic fraction. (DOCX)

Twenty-four Actinobacteria strains, isolated from Arundo donax, Eucalyptus camaldulensis and Populus nigra biomass during natural biodegradation and with potential enzymatic activities specific for the degradation of lignocellulosic materials, were identified by a polyphasic approach. All strains belonged to the genus Streptomyces (S.) and in parti...

α-l-Rhamnosidases (α-RHAs) are a group of glycosyl hydrolases of biotechnological potential in industrial processes, which catalyze the hydrolysis of α-l-rhamnose terminal residues from several natural compounds. A novel α − RHA activity was identified in the crude extract of Novosphingobium sp. PP1Y, a marine bacterium able to grow on a wide range...

The recent scandal of poly implant prostheses (PIP), which were found in some cases to be made of non-medical grade silicone (as reported by the European Scientific Committee on Emerging and Newly Identified Health Risks), had a great social impact. Thousands of patients asked for implant removal with significant costs for public health care system...

The human flavoenzyme D-amino acid oxidase (hDAAO) degrades the NMDA-receptor modulator D-serine in the brain. Whereas hDAAO has been extensively characterized, little is known about its main modulator pLG72, a small protein encoded by the primate-specific gene G72 that has been associated with schizophrenia susceptibility. pLG72 interacts with neo...

A 24-amino acid leader peptide of a new human recombinant manganese superoxide dismutase can enter cells and carry molecules. Here, we demonstrated that six of the 24 amino acids penetrate cells through a particular gate represented by a specific amino acid sequence of the oestrogen receptor (ER). We analysed the internalization of the synthetic he...

In recent years, proteomics procedures have become increasingly popular for the characterization of proteinaceous materials in ancient samples of several cultural heritage objects. The knowledge of the materials used in a work of art is crucial, not only to give an insight in the historical context of objects and artists, but also to analyze degrad...

Rat seminal vesicle protein 4 (RSV4) is a member of the seminal vesicle protein family present in rats, 90 residues long. When secreted, it is involved in many functions related to the reproduction ranging from semen coagulation to sperm capacitation, but its fragments have also shown in vitro pharmacological properties such as anti-inflammatory an...

A deglycosylation step using Peptide -N-Glycosidase F (PNGaseF) has been introduced in a standard proteomic protocol to more confidently identify egg based binders. The ingenuity of introducing a PNGaseF digestion was aimed at removing the molecular hindrance, made up by the heavily glycosylated egg proteins, before the protease(s) hydrolysis. This...

A hallmark to decipher bioprocesses is to characterize protein-protein interactions in living cells. To do this, the development of innovative methodologies, which do not alter proteins and their natural environment, is particularly needed. Here, we report a method (LUCK, Laser UV Cross-linKing) to in vivo cross-link proteins by UV-laser irradiatio...

Gas chromatography/mass spectrometry, proteomic and scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM/EDS) analyses of residue on a stone flake from a 49,000 year-old layer of Sibudu (South Africa) indicate a mixture of ochre and casein from milk, likely obtained by killing a lactating wild bovid. Ochre powder production a...

Background Cellulases and xylanases are the key enzymes involved in lignocelluloses conversion into fermentable sugars. Western Ghat region (India) has been so far recognized as an active hot spot for new microorganisms isolation. The aim of this work was to isolate new microorganisms producing cellulases and xylanases to be applied in Brewer's Spe...

This manuscript describes the analysis of xylanase production by Aspergillus niger NRRL 328 in solid state fermentation (SSF) of soybean husks. A maximum value of extracellular xylanase activity of approximately 950 U g-1 was achieved after 96 h. Proteomic analyses performed on the enzymatic mixture responsible for the maximum value of xylanase act...

Pectinolytic enzymes play an important role in the processing of lignocellulosic materials because of their ability to improve the access of cellulases to their substrate by removing pectins. The strain Paenibacillus xylanolyticus 2-6L3 was isolated from mature compost obtained from agro-industrial wastes, and the enzyme pectate lyase from P. xylan...

In the Comment by Dr. I. Rabin from Bundesanstalt für Materialforschung und -prüfung in Berlin are stated many conjectures and apparent guesses contradicting several of our interpretations. Most of Rabin's ‘guesses’ and interpretations are unwarranted. Below we rebut some of the misinterpretations under separate headings.

The aim of this study was to demonstrate, and to characterize by high-resolution mass spectrometry that it is possible to preferentially induce covalent cross-links in peptides by using high-energy femtosecond ultraviolet (UV) laser pulses. The cross-link is readily formed only when aromatic amino acids are present in the peptide sequence. Three pe...

A protocol for a simple and reliable dot-blot immunoassay was developed and optimized to test work of art samples for the presence of specific proteinaceus material (i.e. ovalbumin-based). The analytical protocol has been extensively set up with respect, among the other, to protein extraction conditions, to densitometric analysis and to the colorim...

Myoglobin is an alpha-helical globular protein containing two highly conserved tryptophanyl residues at positions 7 and 14 in the N-terminal region. The simultaneous substitution of the two residues impairs the productive folding of the protein making the polypeptide chain highly prone to aggregate forming amyloid fibrils at physiological pH and ro...

Background The use of lignocellulosic materials for second generation ethanol production would give several advantages such as minimizing the conflict between land use for food and fuel production, providing less expensive raw materials than conventional agricultural feedstock, allowing lower greenhouse gas emissions than those of first generation...

90 bacteria isolated from raw composting materials were screened for their cellulolytic activity on solid medium containing carboxymethylcellulose. The bacteria producing the highest cellulolytic activity levels were identified by 16S rRNA sequencing as Bacillus licheniformis strain 1, Bacillus subtilis subsp. subtilis strain B7B, Bacillus subtilis...

Myoglobin is an alpha-helical globular protein containing two highly conserved tryptophanyl residues at positions 7 and 14 in the N-terminal region. The simultaneous substitution of the two residues increases the susceptibility of the polypeptide chain to misfold, causing amyloid aggregation under physiological condition, i.e., neutral pH and room...

Laccases are oxidative enzymes linked to biological degradation of lignin. The aim of this work was to evaluate the effect of inducers and different concentrations of nitrogen on production level of total laccase activity and pattern of laccase isoforms, produced in solid state fermentation of sugarcane bagasse by a selected strain of Pleurotus ost...

White-rot basidiomycetes, the most common wood-rotting organisms, are characterized by their ability to produce extracellular oxidative enzymes, among which laccases are regarded as promising catalysts for many biotechnological applications. A significant obstacle to the exploitation of laccase-based bioprocesses is the large amounts of enzyme requ...

An α-l-arabinofuranosidase produced by Pleurotus ostreatus (PoAbf) during solid state fermentation on tomato pomace was identified and the corresponding gene and cDNA were cloned and sequenced. Molecular analysis showed that the poabf gene carries 26 exons interrupted by 25 introns and has an open reading frame encoding a protein of 646 amino acid...

This study was aimed at developing a process of solid state fermentation (SSF) with the fungi Pleurotus ostreatus and Trametes versicolor on apple processing residues for wastewater decolorization. Both fungi were able to colonize apple residues without any addition of nutrients, material support or water. P. ostreatus produced the highest levels o...

Proteomic strategies are herein proved to be a complementary approach to the well established amino acid composition analysis for the characterization of the aging and deterioration phenomena occurring to proteinaceous materials in works-of-art. Amino acid analyses on several samples demonstrated that proteins in the frescoes from the Camposanto Mo...

Bacterial multicomponent monooxygenases (BMMs) are members of a wide family of diiron enzymes that use molecular oxygen to hydroxylate a variety of aromatic compounds. The presence of genes encoding for accessory proteins not involved in catalysis and whose role is still elusive, is a common feature of the gene clusters of several BMMs, including p...

IF(1), the natural inhibitor protein of F(O)F(1)ATP synthase able to regulate the ATP hydrolytic activity of both mitochondrial and cell surface enzyme, exists in two oligomeric states depending on pH: an inactive, highly helical, tetrameric form above pH 6.7 and an active, inhibitory, dimeric form below pH 6.7 [ Cabezon , E. , Butler , P. J. , Run...

Laccases (benzenediol:oxygen oxidoreductases, EC 1.10.3.2) are blue multicopper oxidases, catalyzing the oxidation of an array of aromatic substrates concomitantly with the reduction of molecular oxygen to water. Most of the known laccases have fungal or plant origins, although few laccases have been also identified in bacteria and insects. Most of...

It has been recently hypothesized that BAG3 protein, a co-chaperone of Hsp70/Hsc70, is involved in the regulation of several cell processes, such as apoptosis, autophagy and cell motility. Following the identification of Hsc70/Hsp70, further BAG3 molecular partners such as PLC-gamma and HspB8 were likewise identified, thus contributing to the chara...