Within the flavonoid class of natural products the prenylated sub-class is quite rich in structural variety and pharmacological activity. In the last twenty years a huge number of new structures has been reported, mostly from Leguminosae and Moraceae, with few coming from other genera. The presence, in different forms, of the isoprenoid chain can lead to impressive changes in biological activity, mostly attributed to an increased affinity for biological membranes and to an improved interaction with proteins. Molecules, such as xanthohumol and sophoraflavanone G, while being very structurally simple, show numerous pharmacological applications and are ideal candidates for SAR aimed to the discovery of new drugs. Only recently the biogenesis of these compounds has been more extensively studied and much attention has been focused on the enzymes involved in the modification and transfer of the prenyl unit.

Saliva contains a complex mixture of proteins and peptides as well as fragments derived from these molecules. By RP 1 -HPLC-ESI-MS analysis of the acidic soluble fraction of human whole saliva we have identified in the chromatographic pattern more than 120 different proteins and naturally occurring peptides (1-6). Their characterization was performed by a variety of mass spectrometric techniques coupled with different enzymatic treatments and amino acid sequencing. The proteins and naturally occurring peptides belong to families of well characterized salivary proteins including Histatins, Statherin, acidic and basic proline-rich proteins (aPRP and bPRP), Cystatins, and Defensins (1-6). Two-dimensional gel electrophoresis has also been used by other researchers for analysis of salivary proteins and peptides, but this technique is not well suited for identification of small peptides as illustrated by the difficulty in identifying Histatins and the majority of bPRPs and bPRP fragments (7-9). However, knowledge of salivary proteins and peptides as well as their naturally occurFrom the ‡Dipartimento di Scienze Applicate ai Biosistemi, Università di Cagliari,

Saliva is a body fluid of a unique composition devoted to protect the mouth cavity and the digestive tract. Our high performance liquid chromatography (HPLC)-electrospray ionization-MS analysis of the acidic soluble fraction of saliva from preterm human newborn surprisingly revealed more than 40 protein masses often undetected in adult saliva. We were able to identify the following proteins: stefin A and stefin B, S100A7 (two isoforms), S100A8, S100A9 (four isoforms), S100A11, S100A12, small proline-rich protein 3 (two isoforms), lysozyme C, thymosins ␤ 4 and ␤ 10 , antileukoproteinase, histone H1c, and ␣ and ␥ globins. The average mass value reported in international data banks was often incongruent with our experimental results mostly because of post-translational modifications of the proteins, e.g. acetylation of the N-terminal residue. A quantitative label-free MS analysis showed protein levels altered in relation to the postconceptional age and suggested coordinate and hierarchical functions for these proteins during development. In summary, this study shows for the first time that analysis of these proteins in saliva of preterm newborns might represent a noninvasive way to obtain precious information of the molecular mechanisms of development of human fetal oral structures. Molecular & Cellular Proteomics 10: 10.1074/mcp.M110.003467, 1-14, 2011.Saliva is a body fluid of a very complex and specific composition devoted to the protection and well-being of the oral cavity and, because it is swallowed, of the digestive tract (1). Protection is ensured by organic and inorganic solutes and specific peptides and proteins, such as acidic and basic proline-rich proteins, ␣-amylases, salivary cystatins, histatins, and statherin (2-5). In a previous study (6), we have established that some salivary proteins and peptides reach the levels typically observed in the adult around 18 years of age. Encouraged by the noninvasive specimen collection, we explored the salivary protein composition of at-term and preterm newborns, in order to establish the starting point of the secretion of the proteins and peptides specific of saliva. Our first study (7) showed that acidic proline-rich proteins secretion started, although at very low levels, at 7 months of postconceptional age. At this age the level of phosphorylation of these proteins was low and it increased reaching a value comparable with that of adults at about one year of age, in concomitance with the beginning of deciduous dentition. Other deep differences between human and preterm saliva were however evident. Highly abundant protein masses detected in preterm saliva were undetectable (at the sensitivity level of our MS apparatus) or at very low level in adult saliva. In a previous study (8) we identified, by different MS approaches, thymosin ␤ 4 (T␤ 4 ) and thymosin ␤ 10 (T␤ 10 ) in preterm newborn saliva and established by immunohistochemistry their presence in fetal salivary glands. This finding let us to suppose that in preterm newborns these peptides derived from glan...

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Proteomic platforms can be classified in bottom-up strategies, which analyze the sample after proteolytic digestion, and top-down strategies, which analyze the intact naturally occurring proteome. Bottom-up platforms are high-throughput because they can investigate a large number of proteins, regardless of their dimension. Nonetheless, information on post-translational modifications (PTMs) can be lost, especially those regarding naturally occurring cleavages and alternative splicing. Top-down platforms cannot cover vast proteomes, however, they can disclose subtle structural variations occurring during protein maturation and allow label-free relative quantifications in an unlimited number of samples. A repertoire of 256 masses belonging to naturally occurring proteins and peptides consistently detected by RP-HPLC-ESI-MS analysis of the acidic soluble fraction of human whole saliva is presented in this study. Of them, 233 have been identified, while 23 are still pending for the definitive characterization. The present review reports average and mono-isotopic masses of the peptides and proteins detected, RP-HPLC elution times, PTMs, origin and quali-quantitative variations observed in several physiological and pathological conditions. The information reported can be a reference for users of top-down RP-HPLC-ESI-MS proteomic platforms applied to the study of the human salivary proteome as well as of other human bodily fluids.

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Cystic fibrosis (CF) is a genetic disease affecting today nearly 70,000 patients worldwide and characterized by a hypersecretion of thick mucus difficult to clear arising from the defective CFTR protein. The overproduction of the mucus secreted in the lungs, along with its altered composition and consistency, results in airway obstruction that makes the lungs susceptible to recurrent and persistent bacterial infections and endobronchial chronic inflammation, which are considered the primary cause of bronchiectasis, respiratory failure, and consequent death of patients. Despite the difficulty of treating the continuous infections caused by pathogens in CF patients, various strategies focused on the symptomatic therapy have been developed during the last few decades, showing significant positive impact on prognosis. Moreover, nowadays, the discovery of CFTR modulators as well as the development of gene therapy have provided new opportunity to treat CF. However, the lack of effective methods for delivery and especially targeted delivery of therapeutics specifically to lung tissues and cells limits the efficiency of the treatments. Nanomedicine represents an extraordinary opportunity for the improvement of current therapies and for the development of innovative treatment options for CF previously considered hard or impossible to treat. Due to the peculiar environment in which the therapies have to operate characterized by several biological barriers (pulmonary tract, mucus, epithelia, bacterial biofilm) the use of nanotechnologies to improve and enhance drug delivery or gene therapies is an extremely promising way to be pursued. The aim of this review is to revise the currently used treatments and to outline the most recent progresses about the use of nanotechnology for the management of CF.