Phage display and experimental brain therapeutics

Phage display, a powerful polypeptide display technology, affords the rapid identification of peptides and proteins that interact with a target of interest The aims of the project were the phage display identification of peptides that interact with a druggable target in a brain disorder (glioblastoma multiforme) and the identification of peptides that serve as targeting vectors for brain delivery. Validation studies were undertaken to qualify the use of a cyclic 7-mer peptide phage library against targets including streptavidin and paracetamol chosen as examples of a large complex and small simple molecule, respectively. With the aim of identifying peptide phages that bind to the luminal surface of brain micro vasculature, a primary in-vitro porcine model of the blood-brain barrier (BBB) comprising primary brain capillary endothelial cells was established and characterised. An in-vivo phage display was undertaken in the rat with the aim of identifying peptide sequences that mediated translocation across the BBB into brain grey matter. A 7-mer cyclic peptide was identified with sequence AC-SYTSSTM-CGGGS that enhanced the uptake of phages into brain grey matter by 4-fold compared to control wild-type phages. This peptide may serve as a novel targeting vector for the delivery of a therapeutic cargo to the brain. Caveolin-1 was identified as a potential new therapeutic target in in-vitro models of grade IV astrocytomas (glioblastoma multiforme), with siRNA knockdown of caveolin-1 associated with reduced glioma cell proliferation and invasiveness. With the caveolin-1 scaffolding domain (aa 81-101 in the caveolin-1 protein) as a target, an in-vitro peptide phage selection was undertaken and identified a series of peptides that bind the scaffolding domain with high affinity. These peptides will serve as a template for the development of low molecular weight peptidomimetics that inhibit caveolin-1 function. In conclusion, the studies in this thesis have demonstrated the utility of phage display in experimental therapeutics of brain disorders.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Speciation Analysis of Mercury in Seawater from the Lagoon of Venice by on-line Pre-concentration HPLC-ICP-MS

A method based on the coupling of HPLC with ICP-MS with an on-line pre-concentration micro-column has been developed for the analysis of inorganic and methyl mercury in the dissolved phase of natural waters. This method allows the rapid pre-concentration and matrix removal of interferences in complex matrices such as seawater with minimal sampling handling. Detection limits of 0.07 ng L−1 for inorganic mercury and 0.02 ng L−1 for methyl mercury have been achieved allowing the determination of inorganic mercury and methyl mercury in filtered seawater fromtheVenice lagoon. Good accuracy and reproducibility was demonstrated by the repeat analysis of the certified reference material BCR-579 coastal seawater. The developed HPLC separation was shown to be also suitable for the determination of methyl mercury in extracts of the particulate phase

An evaluation of the PM2.5 trace elemental composition in the VeniceLagoon area and an analysis of the possible sources

Global emissions reported by many authors have shown as natural and anthropic sources can contribute to the principal aerosol classes, but values change according the local scenario. The Venice Lagoon is exposed to different anthropic source emissions like vehicular traffic, industrial thermoelectric power plant, petrochemical plant, incinerator plant, domestic heating, ship traffic, glass factories and airport. Samplings of PM2.5 were daily performed between March and November 2007 in Sacca San Biagio island (Venice), and values of PM2.5 concentration and element concentration were obtained. Monthly average concentrations (mgm3) during this period show higher values during the spring and the autumn. A good relationship between data obtained and concentration values from environmental local agencies is evidenced, both for PM2.5 from urban area (Venezia Mestre), and for PM10 sampled in the same area, as well as the influence of some meteorological parameters on PM2.5 concentration sampled. Trace elements samples were measured by an Inductively Coupled Plasma-Quadrupole Mass Spectrometry (ICP-QMS), and values (ng m3 and mg g1) for elements regulated by European directives (As, Cd, Ni, Pb), as well as, other elements (Na, Al, K, Ti, V, Mn, Fe, Co, Zn, Se, Ag) are also reported. Data analysis by mean of Principal Component Analysis (PCA) and Positive Matrix Factorization (PMF) pointed out four principal groups of elements like Mn–Fe–K, As–Se–Cd, V–Co, and Pb that could be assigned to specific sources of the Venetian wetland basin

Mary Hays's Female Biography : innovative, inspiring, radical, subversive

This thesis is a study of how Mary Hays's six volume Female Biography; or, Memoirs of Illustrious and Celebrated Women, of All Ages and Countries. Alphabetically Arranged (1803) extends Hays's earlier work. The volumes have often been dismissed as being apolitical and a retreat from the radical stance she had previously exhibited in, for instance, her Appeal to the Men of Great Britain in Behalf of Women (1798). Chapters investigate the increasing popularity of life writing in the eighteenth century; of how there was a strong interest in all types of lives; and of how Female Biography fits, and departs, from the genre's expectations. A discussion of a selected sample of conduct literature aimed at women is included, and Hays's rejection and polemic refute of the monolithic identity of the 'ideal' woman promulgated by such conduct literature is argued for as, in Female Biography, Hays is persistent in her insistence that women have distinct and individual characters; this is a theme that pervades all her work. One chapter is devoted to a discussion of the lengthy accounts that Hays apologises for in her Preface; these accounts enable her to thoroughly, effectively, and persistently pursue issues and themes she explored in her novels and polemical non-fiction: women's education; women's subjugation; and the abuse of power by those in authority. I suggest that Hays uses many of her accounts to expose entrenched ideas about sexual difference as prejudice and, as the genre of Biography was recommended to women as a source of moral exempla, something that Hays would have been acutely aware of, she is able insidiously to disseminate the radical and feminist politics exhibited in her earlier work to the readers of Female Biography.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Isotopic lead ratio analyzed by ICP-MS in some metallic slags coming from locri epizephiri (Magna Graecia, South Italy)

The lead isotopic composition of samples of iron slags recovered from two archaeological sites in Lokri Epizephyrii (Italy - Calabria) were analysed for the first time by ICP-MS. The 204Pb, 206Pb, 207Pb and 208Pb content of traces of lead present in the slags are reported. The analysis allows some preliminary conclusions on the different origins of iron slags and on the metallurgical processes that they had undergone to be made. The two archaeological sites are different by excavated extension, chronology and position in the ancient town: the first one, in loc. Dromo (in the middle of the ancient city), was a reduction area of iron minerals active in the 6th cent. B.C. and still little know; the slags from the second site (loc. Maras\ue0) were produced by a foundry discovered in 2002 in close proximity to one of the main sanctuaries of Locri Epizephiri, probably dedicated to Aphrodite, where the local Superintendency and the University of Udine are carrying out archaeological excavations since 1998. The analysis indicate that, in the 5th-4th cent. B.C., the metalworking used ingot or bloom, probably obtained from ores retrieved from the Stilaro mines located near the city (perhaps in the country of ancient Kaulonia, another Greek colony contemporary to Lokri). Some historical facts suggest that these mines came under Lokri jurisdiction in the mid of VI cent. B. C. and were finally conquered by Lokri in the beginning of IV cent. B.C., when the Maras\ue0 foundry was in use. The parts of this essay about history and archeology were written by Marina Rubinich, scientific director of the excavation at Maras\ue0 in behalf of the University of Udine. La composizione isotopica dei campioni di scorie di materiale ferroso, rinvenute in due scavi archeologici effettuati a Locri Epizephiri (Calabria), \ue8 stata analizzata per la prima volta con ICP-MS: in particolare si riporta il contenuto in 204Pb, 206Pb, 207Pb e 208Pb delle tracce di piombo presenti nelle scorie. Le analisi hanno permesso di fare alcune considerazioni preliminari sulla diversa origine delle scorie in ferro e sui processi metallurgici che hanno portato alla loro produzione. I due siti archeologici scelti si differenziano per l\u2019estensione dello scavo, per la cronologia e per la loro posizione all\u2019interno dell\u2019antica citt\ue0: il primo, situato al centro della citt\ue0 antica (localit\ue0 Dromo) e noto soltanto in parte, \ue8 un\u2019area di riduzione del minerale attiva nel VI secolo a.C.; le scorie provenienti dal secondo sito (localit\ue0 Maras\ue0) sono state prodotte da un\u2019officina metallurgica scoperta nel 2002 nelle strette vicinanze di uno dei principali santuari di Locri Epizephiri, probabilmente dedicato ad Afrodite, dove la locale Soprintendenza e l\u2019Universit\ue0 di Udine conducono scavi archeologici dal 1998. Le analisi indicano che nel V-IV secolo a.C. gli artigiani utilizzavano lingotti o blume di ferro, probabilmente ottenuti da minerali estratti dalle miniere dello Stilaro situate nei pressi della citt\ue0, forse nell\u2019area dell\u2019antica Caulonia, un\u2019altra colonia greca contemporanea a Locri. Alcuni dati storici fanno pensare che le miniere di Caulonia passarono sotto la giurisdizione di Locri alla met\ue0 del VI sec. a.C. e furono conquistate definitivamente agli inizi del IV sec. a.C., durante il periodo d\u2019uso dell\u2019officina metallurgica. Le parti del saggio dedicate alla storia e all\u2019archeologia sono state scritte da Marina Rubinich, direttore scientifico dello scavo a Maras\ue0 per conto dell\u2019Universit\ue0 di Udine

Study of 19th century inks from archives in the Palazzo Ducale (Venice, Italy) using various analytical techniques

Inks and paper are the main materials and components of library and archive collections. Since the Third century B.C. there has been a continual succession of ink recipes from all over the world, with varying levels of documentation of the recipes used; however, it was only in the 19th century that Europe became the main producer and industrial leader in the discovery of new products. The aim of this study is to find out more about this last historical period, when new inks were created that had never been known or used in the past. In this study we chose four inks: they belong to documents preserved in the archive of the Superintendence Beni architettonici per il paesaggio e per il patrimonio storico artistico ed etnoantropologico (BAPPDAD) of Venice Lagoon, held in the Palazzo Ducale (Ducal Palace) of Venice. Fourier Transform Infrared Spectroscopy (ATR), Scanning Electron Microscopy (SEM-EDS) and pyrolysis coupled to a gas chromatograph with mass spectrometric detector (Py–GC–MS) allowed us to obtain a qualitative characterization of organic and inorganic elements in three different ink typologies