Enabling 3D CT-scanning of cultural heritage objects using only in-house 2D X-ray equipment in museums.

Visualizing the internal structure of museum objects is a crucial step in acquiring knowledge about the origin, state, and composition of cultural heritage artifacts. Among the most powerful techniques for exposing the interior of museum objects is computed tomography (CT), a technique that computationally forms a 3D image using hundreds of radiographs acquired in a full circular range. However, the lack of affordable and versatile CT equipment in museums, combined with the challenge of transporting precious collection objects, currently keeps this technique out of reach for most cultural heritage applications. We propose an approach for creating accurate CT reconstructions using only standard 2D radiography equipment already available in most larger museums. Specifically, we demonstrate that a combination of basic X-ray imaging equipment, a tailored marker-based image acquisition protocol, and sophisticated data-processing algorithms, can achieve 3D imaging of collection objects without the need for a costly CT imaging system. We implemented this approach in the British Museum (London), the J. Paul Getty Museum (Los Angeles), and the Rijksmuseum (Amsterdam). Our work paves the way for broad facilitation and adoption of CT technology across museums worldwide.

Compositional and Micro-Morphological Characterisation of Red Colourants in Archaeological Textiles from Pharaonic Egypt.

When the imagination conjures up an image of an Egyptian mummy, it is normally one of a human body wrapped with undyed linen bandages. However, the reality was much more colourful, as shown by the set of red mummy shrouds and textile fragments from Pharaonic Egypt considered in this work. The textiles were subjected to scientific investigation with the main aim of shedding light on the sources of red colour and on the possible reasons for the different levels of colour fading. The red colourants were investigated using various non-invasive and micro-invasive approaches. The results pointed towards the presence of three sources of red colour, which, in increasing order of lightfastness, are safflower (Carthamus tinctorius), madder (Rubia spp.), and red ochre. Micro-morphological observations and elemental analyses also enabled some hypotheses to be formulated regarding the application of these colourants to the textiles. The results not only deepen our knowledge of dyeing technologies in ancient Egypt and shed new light on the function of red shrouds and textiles as part of the funerary practices of Pharaonic Egypt, but are also essential in planning the display and future preservation of these mummies and their associated textiles.

A multispectral imaging approach integrated into the study of Late Antique textiles from Egypt.

This work explores the use of multispectral imaging (MSI) techniques applied to the investigation of Late Antique (c. 250-800 AD) textiles found in Egypt. Although the use of these techniques is well-established in the study of polychrome surfaces, they have only been sparingly and often unsystematically applied to the investigation of textiles. The aim of this work is therefore to bridge this gap by showing how this non-invasive, relatively inexpensive and portable methodology can be used to map the photoluminescence and reflective characteristics of textiles under different wavelengths of light, and to provide qualitative and holistic insights into the chemical nature of the materials that compose them. Standardised acquisition and post-processing methods were applied to produce visible-reflected (VIS), ultraviolet-induced visible luminescence (UVL), infrared-reflected (IRR), infrared-reflected false colour (IRRFC), ultraviolet-reflected (UVR) and ultraviolet-reflected false colour (UVRFC) images that provided preliminary indications of the colourants used and their spatial distribution. This proved to be an important aid in planning more targeted and effective sampling strategies and facilitated comparisons between objects. Visible-induced visible luminescence (VIVL) and multiband-reflected (MBR) imaging were also explored for the first time with application to textiles, demonstrating their potential in mapping red and blue colourants respectively. The physical properties observed from all of these images were then related to the more detailed information provided by complementary non-invasive techniques, such as fibre optic reflectance spectroscopy (FORS), and micro-invasive approaches, such as high-performance liquid chromatography mass spectrometry (HPLC-MS). Guidelines towards the interpretation of complex MSI images and a discussion of the potential and limitations of relating multispectral data to chemical properties are presented. An important result of this work is the delineation of a protocol, which combines optical microscopy (OM), MSI, FORS and HPLC-MS and shows a high degree of potential, not only for the investigation of Late Antique textiles but for textiles in museum and historic collections generally.

The characterisation of shellac resin by flow injection and liquid chromatography coupled with electrospray ionisation and mass spectrometry.

A strategy based on electrospray ionisation (ESI) in negative mode coupled with quadrupole-time of flight (Q-ToF) detection techniques was adopted to characterise some samples of shellac resin. Flow injection analysis (FIA) was used to investigate the distribution of the components of the resin. Eight groups of compounds with increasing masses were detected and assigned to free acids, esters and polyesters with up to eight units. High pressure liquid chromatography (HPLC) enabled the compounds to be chromatographically separated. Accurate molecular masses and tandem mass (MS/MS) spectra interpretation were used to characterise the different compounds, assigning and/or suggesting molecular structures. In some cases, highly detailed information about the ester linkages was provided by the MS/MS spectra, enabling the different isomers to be distinguished. Oxidation products were also identified in the samples and differences were observed in terms of hydrolysis and oxidation. In addition to providing the first characterisation of shellac by HPLC-ESI-Q-ToF and an atlas of MS/MS spectra of shellac components, this work demonstrates the suitability of the proposed strategy for characterising the resin, and provides the identification of previously unknown degradation products and minor components. This represents a significant step forward in the chemical knowledge of this material.

Excited state dependent electron transfer of a rhenium-dipyridophenazine complex intercalated between the base pairs of DNA: a time-resolved UV-visible and IR absorption investigation into the photophysics of fac-[Re(CO)3(F2dppz)(py)]+ bound to either [poly(dA-dT)]2 or [poly(dG-dC)]2.

The transient species formed following excitation of fac-[Re(CO)(3)(F(2)dppz)(py)](+) (F(2)dppz = 11,12-difluorodipyrido[3,2-a:2',3'-c]phenazine) bound to double-stranded polynucleotides [poly(dA-dT)](2) or [poly(dG-dC)](2) have been studied by transient visible and infra-red spectroscopy in both the picosecond and nanosecond time domains. The latter technique has been used to monitor both the metal complex and the DNA by monitoring the regions 1900-2100 and 1500-1750 cm(-1) respectively. These data provide direct evidence for electron transfer from guanine to the excited state of the metal complex, which proceeds both on a sub-picosecond time scale and with a lifetime of 35 ps, possibly due to the involvement of two excited states. No electron transfer is found for the [poly(dA-dT)](2) complex, although characteristic changes are seen in the DNA-region TRIR consistent with changes in the binding of the bases in the intercalation site upon excitation of the dppz-complex.

A covalently linked phenanthridine-ruthenium(II) complex as a RNA probe.

A phenanthridine derivative covalently linked to a ruthenium complex yields an imaging probe whose fluorescence intensity and lifetime change substantially in the presence of RNA.

Photolysis of dibenzyl ketones sorbed on MFI zeolites in the presence of spectator molecules: cage effects, kinetics, and external surface sites characterization.

Over the past two decades, the photolytic reactions of dibenzyl ketones sorbed on zeolites have been investigated. The reported results are consistent with a supramolecular model that takes into account the physical and chemical nature of the structure of the zeolites and their effect on the reactive radical intermediates produced by photolysis of adsorbed molecules. The model incorporates various phenomena such as surface coverage, external and internal sorption, surface diffusion, radical sieving, and the resulting product distributions. This account reports direct evidence for the validation of the model through FT-IR spectroscopy and through a new method for "titrating" the binding sites via EPR spectroscopy. It is shown that it is possible to adjust and modulate the photolytic product distribution by varying the parameters of the system. The effects of co-adsorbed spectator molecules with different polarities, namely water, pyridine, and benzene, on the photolysis of o-methyldibenzyl ketone and dibenzyl ketone sorbed on MFI zeolites is examined. This study provides insights into a displacement mechanism caused by spectator molecules and further demonstrates how the product distribution of photolysis of sorbed ketones can be controlled. The kinetics of persistent radicals formed by photolysis of ketones sorbed on zeolites is directly monitored over time by EPR, providing a measure of the lifetime of these reactive organic intermediates. Finally, measurement of Langmuir isotherms was employed to provide classical evidence for the model.

Photooxidation of guanine by a ruthenium dipyridophenazine complex intercalated in a double-stranded polynucleotide monitored directly by picosecond visible and infrared transient absorption spectroscopy.

Transient species formed by photoexcitation (400 nm) of [Ru(dppz)(tap)2]2+ (1) (dppz = dipyrido[3,2-a:2',3'-c]phenazine; tap=1,4,5,8-tetraazaphenanthrene) in aqueous solution and when intercalated into a double-stranded synthetic polynucleotide, [poly(dG-dC)]2, have been observed on a picosecond timescale by both visible transient absorption (allowing monitoring of the metal complex intermediates) and transient infrared (IR) absorption spectroscopy (allowing direct study of the DNA nucleobases). By contrast with its behavior when free in aqueous solution, excitation of 1 when bound to [poly(dG-dC)]2 causes a strong increase in absorbance at 515 nm due to formation of the reduced complex [Ru(dppz)(tap)2]+ (rate constant=(2.0+/-0.2) x 10(9) s(-1)). The subsequent reformation of 1 proceeds with a rate constant of (1.1+/-0.2) x 10(8) s(-1). When the process is carried out in D2O, the rates of formation and removal of [Ru(dppz)(tap)2]+ are reduced (rate constants (1.5+/-0.3) x 10(9) and (0.7+/-0.2) x 10(8) s(-1) respectively) consistent with proton-coupled electron transfer processes. Picosecond transient IR measurements in the 1540-1720 cm(-1) region in D2O solution confirm that the reduction of 1 intercalated into [poly(dG-dC)]2 is accompanied by bleaching of IR ground-state bands of guanine (1690 cm(-1)) and cytosine (1656 cm(-1)), each with similar rate constants.

FRETView: a computer program to simplify the process of obtaining fluorescence resonance energy transfer parameters.

The process of modeling the fluorescence resonance energy transfer (FRET) process for a donor-acceptor pair can be rather challenging, yet few computer programs exist that allow such modeling to be done with relative ease. In order to address this, we have developed a Java-based program, FRETView, which allows numerous FRET parameters to be obtained with just a few mouse clicks. Being a Java-based program, it runs equally well on all the major operating systems such as Windows, Mac OS X, Linux, Solaris. The program allows the user to effortlessly input pertinent information about the donor-acceptor pair, including the absorption and/or emission spectra, and outputs the calculated FRET parameters in table format, as well as graphical plots.

Toward the development of prophylactic and therapeutic human papillomavirus type-16 lipopeptide vaccines.

Four lipid-core peptide systems were synthesized using stepwise solid-phase peptide synthesis, incorporating a sequence from the human papillomavirus type-16 (HPV-16) E7 protein (E744-62), for the purpose of developing vaccines against HPV-16 associated cervical cancer. d-Mannose was conjugated to the vaccine in order to investigate whether the targeting of dendritic cell mannose receptors would improve vaccine efficacy. The ability of the vaccines to clear or reduce the size of HPV-16 associated tumors was assessed in C57BL/6 (H-2b) mice using the TC-1 HPV-16 tumor model. Overall, significant reductions in the size of TC-1 tumors were observed in the mouse model, with the conjugation of mannose to these vaccines demonstrated to clear or reduce the size of TC-1 tumors to a greater extent than non-mannose-containing vaccines (37 out of 40 versus 21 out of 30 tumors cleared, respectively).

Solvent dependent photophysics of fac-[Re(CO)(3)(11,12-X(2)dppz)(py)](+)(X = H, F or Me).

The photophysical properties of [Re(CO)(3)(dppz)(py)](+) (dppz = dipyrido-[3,2-a:2',3'-c] phenazine) and its 11,12 substituted derivatives [Re(CO)(3)(dppzMe(2))(py)](+) and [Re(CO)(3)(dppzF(2))(py)](+) have been examined in organic and aqueous environments using phosphorescence and picosecond transient visible and infrared absorption spectroscopic methods. The roles of the intraligand IL(pi-pi*) and metal-to-ligand charge transfer MLCT(phz) excited states are evaluated and used to explain the major effect of difluoro-substitution, which is particularly remarkable in water, where the excited state of [Re(CO)(3)(dppzF(2))(py)](+) is strongly quenched.

Controlled diastereoselectivity at the alkene-geometry through selective encapsulation: E-Z photoisomerization of oxazolidinone-functionalized enecarbamates within hydrophobic nano-cavities.

Photoisomerization of encapsulated Z-enecarbamates within the hydrophobic chiral cavities of gamma-CD showed higher diastereoselectivities in the photoproducts than those obtained in solution. The selective encapsulation of the enecarbamates and the following isomerization process are both diastereoselectively controlled by gamma-CD.

Overcoming original antigenic sin to generate new CD8 T cell IFN-gamma responses in an antigen-experienced host.

The failure to mount effective immunity to virus variants in a previously virus-infected host is known as original antigenic sin. We have previously shown that prior immunity to a virus capsid protein inhibits induction by immunization of an IFN-gamma CD8+ T cell response to an epitope linked to the capsid protein. We now demonstrate that capsid protein-primed CD4+ T cells secrete IL-10 in response to capsid protein presented by dendritic cells, and deviate CD8+ T cells responding to a linked MHC class I-restricted epitope to reduce IFN-gamma production. Neutralizing IL-10 while delivering further linked epitope, either in vitro or in vivo, restores induction by immunization of an Ag-specific IFN-gamma response to the epitope. This finding demonstrates a strategy for overcoming inhibition of MHC class I epitopes upon immunization of a host already primed to Ag, which may facilitate immunotherapy for chronic viral infection or cancer.

Intranasal administration is an effective mucosal vaccine delivery route for self-adjuvanting lipid core peptides targeting the group A streptococcal M protein.

BACKGROUND: We investigated the lipid core peptide (LCP) system for mucosal vaccine delivery against infection with group A streptococcus (GAS)--the causative pathogen of rheumatic fever and rheumatic heart disease. METHODS: An LCP vaccine formulation containing 2 different peptide epitopes of the antiphagocytic M protein of GAS--a conformational epitope from the carboxyterminal conserved C-repeat region and an aminoterminal serotypic epitope--was intranasally administered to mice with cholera toxin B subunit or without additional adjuvant. RESULTS: Our data demonstrate that the LCP vaccine formulation induced the elicitation of antigen-specific systemic immunoglobulin G responses when administered with or without cholera toxin B subunit, whereas cholera toxin B subunit was required for the induction of antigen-specific mucosal immunoglobulin A responses. Immune serum samples from vaccinated mice were capable of opsonization of a homologous GAS strain, as well as opsonization of a heterologous GAS strain. Furthermore, mice were protected from GAS challenge following immunization with the LCP vaccine formulation, even in the absence of additional adjuvant. CONCLUSIONS: These data support the potential of the LCP system in the development of a self-adjuvanting, synthetic, peptide-based mucosal GAS vaccine for the prevention of diseases caused by GAS.

Phosphorylation state-responsive lanthanide peptide conjugates: a luminescence switch based on reversible complex reorganization.

[reaction: see text] A luminogenic probe for peptide dephosphorylation has been developed. It consists of a serine-/tyrosine-containing peptide modified on the N-terminus with a tryptophan residue and a DTPA chelate capable of binding Tb(3+). We propose a mechanistic model for the luminescence enhancement based on the interconversion of monomeric and dimeric lanthanide species, which is affected by the phosphorylation state of the serine or tyrosine residue. The optical switch reports effectively on phosphatase-catalyzed dephosphorylation in vitro.

Immunization with a tetraepitopic lipid core peptide vaccine construct induces broadly protective immune responses against group A streptococcus.

BACKGROUND: The development of a vaccine to prevent infection with group A streptococcus (GAS) is hampered by the widespread diversity of circulating GAS strains and M protein types, and it is widely believed that a multivalent vaccine would provide better protective immunity. METHODS: We investigated the efficacy of incorporating 3 M protein serotypic amino-terminal epitopes from GAS isolates that are common in Australian Aboriginal communities and a conformational epitope from the conserved carboxy-terminal C-repeat region into a single synthetic lipid core peptide (LCP) vaccine construct in inducing broadly protective immune responses against GAS after parenteral delivery to mice. RESULTS: Immunization with the tetraepitopic LCP vaccine construct led to high titers of systemic, antigen-specific IgG responses and the induction of broadly protective immune responses, as was demonstrated by the ability of immune serum to opsonize multiple GAS strains. Systemic challenge of mice with a lethal dose of GAS given 60 or 300 days after primary immunization showed that, compared with the control mice, the vaccinated mice were significantly protected against GAS infection, demonstrating that the vaccination stimulated long-lasting protective immunity. CONCLUSIONS: These data support the efficacy of the LCP vaccine delivery system in the development of a synthetic, multiepitopic vaccine for the prevention of GAS infection.

Two-photon induced uncaging of a reactive intermediate. Multiphoton in situ detection of a potentially valuable label for biological applications.

[reaction: see text] Two-photon induced Wolff rearrangement of a terphenyl diazoketone 1 was achieved by using focused laser pulses of 532 nm from a Q-switched Nd:YAG laser. The nonfluorescent terphenyl diazoketone 1 was transformed into a fluorescent ester derivative 4, which can be detected in situ using the focused laser pulses at 532 nm. Laser power dependence studies show that the Wolff rearrangement is induced by two-photon absorption of the terphenyl diazoketone 1, but suggests that more than two photons of 532 nm are involved (a multiphoton process) in excitation of the ester derivative 4.

Monitoring the effect of ultrafast deactivation of the electronic excited states of DNA bases and polynucleotides following 267 nm laser excitation using picosecond time-resolved infrared spectroscopy.

In this paper we demonstrate the use of picosecond time-resolved infrared spectroscopy (ps-TRIR) to monitor the early structural dynamics of DNA bases and polydeoxynucleotides following UV excitation in solution.

Development of a broadband picosecond infrared spectrometer and its incorporation into an existing ultrafast time-resolved resonance Raman, UV/visible, and fluorescence spectroscopic apparatus.

We have constructed a broadband ultrafast time-resolved infrared (TRIR) spectrometer and incorporated it into our existing time-resolved spectroscopy apparatus, thus creating a single instrument capable of performing the complementary techniques of femto-/picosecond time-resolved resonance Raman (TR3), fluorescence, and UV/visible/infrared transient absorption spectroscopy. The TRIR spectrometer employs broadband (150 fs, approximately 150 cm(-1) FWHM) mid-infrared probe and reference pulses (generated by difference frequency mixing of near-infrared pulses in type I AgGaS2), which are dispersed over two 64-element linear infrared array detectors (HgCdTe). These are coupled via custom-built data acquisition electronics to a personal computer for data processing. This data acquisition system performs signal handling on a shot-by-shot basis at the 1 kHz repetition rate of the pulsed laser system. The combination of real-time signal processing and the ability to normalize each probe and reference pulse has enabled us to achieve a high sensitivity on the order of deltaOD approximately 10(-4) - 10(-5) with 1 min of acquisition time. We present preliminary picosecond TRIR studies using this spectrometer and also demonstrate how a combination of TRIR and TR3 spectroscopy can provide key information for the full elucidation of a photochemical process.