To glove or not to glove? Investigations into the potential contamination from handling of paper-based cultural heritage through forensic fingerprinting approaches.

The handling of cultural heritage objects has become a highly debated topic in the last decade. The work and outcomes described in this paper are aimed to provide objective data to assist in making appropriate decisions as to whether or not wearing gloves is appropriate in a given situation. The forensic fingermark development techniques of 1,2-indandione and single metal deposition II were used to investigate the efficacy of handwashing and glove use to improve the information available when deciding whether to use gloves when handling paper objects. It was found that fingermarks did not permeate through polymer glove types but could through cotton gloves. It was also shown that the amounts of observable fingermark residues were greater 5 min after handwashing than if handwashing had not occurred, undermining previous arguments for not wearing gloves if hands could be washed before object handling.

Enhancing the evidential value of textile fibres Part 2: Application of a database-driven fibre comparison strategy to a cold-case investigation.

This paper describes the involvement of our laboratory in a Western Australian 'cold-case' investigation that spanned 24 years. The investigation was widely considered to be the largest in Australian history. During the investigative phase, our laboratory was tasked with the collection of trace evidence of all types in connection with a sexual assault and two homicides that were suspected to be related. Textile fibres represented the vast majority of trace evidence recovered. A much greater quantity of fibres (>10,800) was collected than would be typical for a routine case, as fibres of any colour or type were potentially of investigative value. The investigation was unprecedented in its scale, and presented numerous challenges in terms of evidence recovery, analysis, interpretation, reporting, and provision of testimony. A textile fibre microspectrophotometric (MSP) database was developed specifically for the interpretation of data in connection with the investigation. The database currently contains over 25,000 normalised and first derivative spectra of casework, validation and reference textile fibres. A fibre comparison strategy was devised, involving the identification of preliminary fibre groups on the basis of corresponding/similar MSP spectra, and verification of these groups via brightfield and fluorescence comparison microscopy. A potential link to an automotive source was identified for one of the homicide victims during the investigative phase. After identification of a suspect, a total of 98 fibres recovered from victims and from a seized motor vehicle were found to correspond in properties to six different fibre types from known textile sources in connection with the suspect. A highly publicised criminal trial was held, and textile fibre evidence provided a major contribution to the trial findings, in which the accused was found guilty of two homicides.

Chemometrics in forensic science: approaches and applications.

Forensic investigations are often reliant on physical evidence to reconstruct events surrounding a crime. However, there remains a need for more objective approaches to evidential interpretation, along with rigorously validated procedures for handling, storage and analysis. Chemometrics has been recognised as a powerful tool within forensic science for interpretation and optimisation of analytical procedures. However, careful consideration must be given to factors such as sampling, validation and underpinning study design. This tutorial review aims to provide an accessible overview of chemometric methods within the context of forensic science. The review begins with an overview of selected chemometric techniques, followed by a broad review of studies demonstrating the utility of chemometrics across various forensic disciplines. The tutorial review ends with the discussion of the challenges and emerging trends in this rapidly growing field.

Revealing the Elemental Distribution within Latent Fingermarks Using Synchrotron Sourced X-ray Fluorescence Microscopy.

Fingermarks are an important form of crime-scene trace evidence; however, their usefulness may be hampered by a variation in response or a lack of robustness in detection methods. Understanding the chemical composition and distribution within fingermarks may help explain variation in latent fingermark detection with existing methods and identify new strategies to increase detection capabilities. The majority of research in the literature describes investigation of organic components of fingermark residue, leaving the elemental distribution less well understood. The relative scarcity of information regarding the elemental distribution within fingermarks is in part due to previous unavailability of direct, micron resolution elemental mapping techniques. This capability is now provided at third generation synchrotron light sources, where X-ray fluorescence microscopy (XFM) provides micron or submicron spatial resolution and direct detection with sub-µM detection limits. XFM has been applied in this study to reveal the distribution of inorganic components within fingermark residue, including endogenous trace metals (Fe, Cu, Zn), diffusible ions (Cl-, K+, Ca2+), and exogeneous metals (Ni, Ti, Bi). This study incorporated a multimodal approach using XFM and infrared microspectroscopy analyses to demonstrate colocalization of endogenous metals within the hydrophilic organic components of fingermark residue. Additional experiments were then undertaken to investigate how sources of exogenous metals (e.g., coins and cosmetics) may be transferred to, and distributed within, latent fingermarks. Lastly, this study reports a preliminary assessment of how environmental factors such as exposure to aqueous environments may affect elemental distribution within fingermarks. Taken together, the results of this study advance our current understanding of fingermark composition and its spatial distribution of chemical components and may help explain detection variation observed during detection of fingermarks using standard forensic protocols.

Enhancing the evidential value of textile fibres: Part 1: Development of a spectral database and evaluative comparison strategy.

In numerous major crime cases undertaken at our laboratory the recovery of large numbers of fibres (>1000), particularly in cases with no known source, presented several challenges. These included the inability to effectively manage the data (i.e. combination of MSP spectra, microscopic characteristics, composition, etc.) and perform comparisons in an efficient manner. To address these challenges, and in response to a growing need for performing fibre comparisons, we developed a database of textile fibre microspectrophotometric (MSP) spectra. The database, designed to compare MSP spectra using a modified Pearson method of correlation, currently contains over 20,000 normalised and first derivative spectra of casework, validation and reference textile fibres. A comparison strategy for cases with a large number of questioned samples was devised, involving identification of critical fibres in the casework data set, development of preliminary fibre groups classified according to their corresponding/similar MSP spectra, and verification of the preliminary groups via brightfield and fluorescence comparison microscopy. The database has successfully been utilised for proficiency trials and casework with small questioned fibre sets. Furthermore, in a case involving a larger dataset (>4000 "unknown" fibres) the database assisted in the efficient classification of 156 distinct groups of interest, highlighting its utility in providing investigative leads for the identification of potential sources of the recovered fibres.

The effect of environmental degradation on the characterisation of automotive clear coats by infrared spectroscopy.

This paper presents a study into the chemical changes occurring within automotive clear coats as a result of environmental weathering, and their potential effect upon the characterisation of samples using infrared spectroscopy combined with chemometric modelling. Testing of three samples, collected from separate vehicles, exposed to the outside environment revealed no changes in model predictions over a 175 day period; however, incorrect predictions were observed following 435 days of exposure. Inspection of the corresponding infrared spectra revealed that these changes were likely due to the hydrolysis and photodegradation of polymer chains present in the clear coat, which were not observed in samples stored away from the outside environment over a one-year period. Analysis of previously weathered samples using synchrotron infrared microscopy found these changes occurred in a top-down fashion rather than within the bulk of the clear coat. This indicates that although weathering may affect the surface characterisation of clear coats over time, the targeting of deeper portions of the clear coat layer may still provide useful information as to the identity of the vehicle.

A new p-dimethylaminocinnamaldehyde reagent formulation for the photoluminescence detection of latent fingermarks on paper.

Dimethylaminocinnamaldehyde was re-evaluated as a wet contact reagent for the treatment of latent fingermarks on porous substrates. A new formulation (consisting of 0.028 g p-dimethylaminocinnamaldehyde, 0.84 mL glacial acetic acid, 6.2 mL ethyl acetate and 0.993 L 40-60 °C petroleum spirits) provides quick, sensitive and robust luminescent ridge detail within 3 h of treatment on both plain and thermal paper. Comparisons to existing formulations indicate improved visualisation and/or a more efficient process.

Synchrotron far-infrared spectroscopy of corroded steel surfaces using a variable angle of incidence.

Far-infrared spectroscopy, using a synchrotron source, has been used to study carbon steel corroded in CO2-saturated brine in the presence and absence of the corrosion inhibitor 2-mercaptopyrimidine (MPY), which allowed the steel surface roughness to be modified. The effect of the angle of incidence (θi, 30-80°) on the band intensity and observed bands of the spectra from these surfaces has been determined. For the MPY-treated steel (low surface roughness) the highest band intensity is observed at high θi (80°) and different bands were observed at different θi. In contrast, for the MPY-free steel (high surface roughness) the highest band intensity is observed at low θi (30°) and spectral content changes were not observed. The results are explained in terms of the roughness of the MPY-treated and MPY-free steels, and their effect on the level of diffusely reflected light of the incident infrared beam.

Synchrotron FTIR characterisation of automotive primer surfacer paint coatings for forensic purposes.

Synchrotron FTIR microspectroscopy was used to characterise and assess the chemical diversity of electrocoat primer, primer surfacer and basecoats of automotive paint samples from 75 vehicles representing a range of international car manufacturers. Significant diversity was found in the synchrotron FTIR data from the primer surfacer coats. Classification using principal component analysis revealed 14 discrete groups, which could be associated with the country of manufacture, the specific manufacturer and manufacturing plant, the year of manufacture of the vehicle and in some instances the number of layers in the paint system. The model generated from the primer surfacer was significantly more discriminating than a previous model generated from FTIR analysis of clear coats of the same vehicles. Analysis of the primer surfacer also avoids issues of possible environmental degradation and component migration seen with FTIR analysis of automotive clear coats. Providing such information concerning vehicle origin would aid in the development of investigative leads from the analysis of questioned automotive paint samples located at the scene of a crime or on the body/clothing of a hit and run victim.

Characterisation of chemical component migration in automotive paint by synchrotron infrared imaging.

Synchrotron infrared chemical imaging was employed to examine and assess the extent of interlayer component migration within multilayer automotive paint samples, with a particular emphasis on the cross-linking additive melamine. Two dimensional infrared chemical images revealed that melamine consistently diffuses in select paint samples from the underlying basecoat into the outermost clear coat layer. Pigments from the basecoat were also found to migrate into the adjoining layers. This is significant as the relative abundance of both melamine and pigments will vary greatly depending upon the region of the layer analysed. This component migration will undoubtedly impact the information gleaned from a questioned sample via library searching software or multivariate statistics. As a result, appropriate analytical protocols will need to be utilised to mitigate the effects of interlayer pigment and melamine diffusion, so as to afford a true representation of the composition of the coating for forensic identification purposes.

Chemical fingerprinting of adhesive tapes by GCMS detection of petroleum hydrocarbon products.

Pressure-sensitive adhesive tapes often represent key evidence of crimes such as assault, rape or homicide; thus, the development of analytical techniques able to contribute to a detailed characterization of these materials is of forensic importance. The gas chromatography-mass spectrometry (GCMS) analysis of the solvent extractable fractions of a suite of electrical and gaffer adhesive tapes spanning a range of colors and manufacturers identified a number of petroleum-derived hydrocarbons. Molecular and isotopic analyses of hydrocarbon constituents of complex materials have found wide analytical utility including the forensic investigation of oil spills and arson. Here, we investigate the utility of these techniques for characterizing the hydrocarbon composition of pressure-sensitive adhesive tapes for forensic correlation purposes. Subtle distinction of tape samples was evident in the GCMS distribution of several hydrocarbon groups including alkyl-naphthalenes, hopane and sterane biomarkers. Linear discriminant analysis of the abundances of these products provided high level differentiation of tape manufacturer. The distinction of different adhesive tape samples was further extended by measurement of their stable carbon isotopic values. The molecular and isotopic differences of the petroleum content of tapes are consistent with the use of different petroleum materials used in the manufacturing process and demonstrate the benefits of the combined use of complementary oil hydrocarbon characterization approaches. This study reveals the forensic potential of using established petroleum characterization methods for characterizing materials with a petroleum-derived hydrocarbon element.

Rapid nondestructive on-site screening of methylamphetamine seizures by attenuated total reflection fourier transform infrared spectroscopy.

The identification and quantification of illicit substances in the field is often desirable. Fourier transform infrared spectroscopy (FT-IR) has both qualitative and quantitative capabilities and field portable instruments are commercially available. Transmission infrared spectra of mixtures containing ephedrine hydrochloride, glucose, and caffeine and attenuated total reflection (ATR) infrared spectra of mixtures composed of methylamphetamine hydrochloride, glucose, and caffeine were used to develop principal component regression (PCR) calibration models. The root mean sum of errors of predictions (RMSEP) of all individual components in a mixture from a single measurement was <6% w/w, which reduced to approximately 3% w/w when triplicates were averaged. Sample mixing and grinding are essential to minimize the effect of heterogeneity, as deviations of up to 20% w/w were observed for single measurements of unground samples. Poor predictions of the components in a mixture occurred when samples were "contaminated" with substances not present in the calibration set, as would be expected. When only a single analyte (drug) was targeted, using a calibration set that contained both contaminated and uncontaminated samples, an RMSEP of approximately 4% w/w was achieved. The results demonstrate that ATR-FT-IR has the potential to quantify methylamphetamine samples, and possibly other licit or illicit substances, in at-seizure and on-site scenarios.

Intracrystalline urinary proteins facilitate degradation and dissolution of calcium oxalate crystals in cultured renal cells.

We have previously proposed that intracrystalline proteins would increase intracellular proteolytic disruption and dissolution of calcium oxalate (CaOx) crystals. Chauvet MC, Ryall RL. J Struct Biol 151: 12-17, 2005; Fleming DE, van Riessen A, Chauvet MC, Grover PK, Hunter B, van Bronswijk W, Ryall RL. J Bone Miner Res 18: 1282-1291, 2003; Ryall RL, Fleming DE, Doyle IR, Evans NA, Dean CJ, Marshall VR. J Struct Biol 134: 5-14, 2001. The aim of this investigation was to determine the effect of increasing concentrations of intracrystalline protein on the rate of CaOx crystal dissolution in Madin-Darby canine kidney (MDCKII) cells. Crystal matrix extract (CME) was isolated from urinary CaOx monohydrate (COM) crystals. Cold and [14C]oxalate-labeled COM crystals were precipitated from ultrafiltered urine containing 0-5 mg/l CME. Crystal surface area was estimated from scanning electron micrographs, and synchrotron X-ray diffraction was used to determine nonuniform strain and crystallite size. Radiolabeled crystals were added to MDCKII cells and crystal dissolution, expressed as radioactive label released into the medium, was measured. Increasing CME content did not significantly alter crystal surface area. However, nonuniform strain increased and crystallite size decreased in a dose-response manner, both reaching saturation at a CME concentration of 3 mg/ and demonstrating unequivocally the inclusion of increasing quantities of proteins in the crystals. This was confirmed by Western blotting. Crystal dissolution also followed saturation kinetics, increasing proportionally with final CME concentration and reaching a plateau at a concentration of approximately 2 mg/l. These findings were complemented by field emission scanning electron microscopy, which showed that crystal degradation also increased relative to CME concentration. Intracrystalline proteins enhance degradation and dissolution of CaOx crystals and thus may constitute a natural defense against urolithiasis. The findings have significant ramifications in biomineral metabolism and pathogenesis of renal stones.

The importance of a clean face: the effect of different washing procedures on the association of Tamm-Horsfall glycoprotein and other urinary proteins with calcium oxalate crystals.

This study was undertaken to determine whether the use of different washing procedures could explain dissident findings in published studies examining the role of urinary macromolecules in urolithiasis. Calcium oxalate monohydrate (COM) crystals were deposited from or added to the same sieved urine, washed with copious or limited amounts of distilled water, or with methanol, and examined by field emission scanning electron microscopy (FESEM). Demineralized extracts were analysed by SDS-PAGE and Western blotting for Tamm-Horsfall glycoprotein (THG), human serum albumin (HSA), osteopontin (OPN) and prothrombin fragment 1 (PTF1). Synchrotron X-ray diffraction (SXRD) with Rietveld whole-pattern peak fitting and profile analysis was used to determine non-uniform crystal strain and crystallite size in crystals generated from inorganic solutions in the presence of increasing concentrations of THG and prothrombin (PT). HSA and PTF1 were present in all demineralized crystal extracts, confirming their inclusion within COM. OPN was present in all extracts except those derived from pure inorganic COM crystals, because of its occlusion within small numbers of calcium oxalate dihydrate (COD) crystals contaminating the COM population. THG was absent from the demineralized extracts of all crystals washed copiously with water, but present in those washed with methanol or limited amounts of water. FESEM showed extraneous organic material associated only with crystals whose extracts contained THG, confirming that the protein does not bind permanently to the COM crystal surface and is not occluded within the mineral bulk. This was confirmed by SXRD, which showed that non-uniform strain and crystallite size remained unaltered in crystals grown in the presence of increasing THG concentrations. However, non-uniform strain increased and crystallite size decreased with increasing PT concentrations, demonstrating unambiguously that PT is included in COM crystals. It was concluded that scrupulous care must be taken to ensure the complete removal of extraneous THG adventitiously associated with CaOx crystals in order to avoid inaccurate analysis of crystal matrix protein content and possible misinterpretation of experimental data.

An in situ FTIR-ATR study of polyacrylate adsorbed onto hematite at high pH and high ionic strength.

FTIR-ATR was used to examine in situ the interaction of polyacrylate and hematite at pH 13. Static light scattering and mobility measurements were used to assess solution polyacrylate dimensions and hematite surface charge, respectively. Polyacrylate adsorption occurred only with the addition of electrolyte (e.g., NaCl), and it was found that excess cations, up to approximately 1 M, facilitated adsorption, above which the effect was found to plateau. At pH 13 and at low ionic strength, adsorption of polyacrylate onto hematite is facilitated by cations in solution shielding both the negative acrylate functionality of the polymer and the negative hematite surface. The shielding of the hematite surface continues to increase with increasing salt concentration up to a measured 3 M. Similarly, the shielding of the polymer increased with electrolyte concentration up to approximately 1 M salt, beyond which no further increase in shielding was observed. At this concentration the polymer assumes a finite minimum size in solution that ultimately limits the amount adsorbed. The dimension of the polymer in solution was found to be independent of monovalent cation type. Thus, at high pH and high ionic strength adsorption is determined by the degree of hematite surface charge reduction. The cation-hematite surface interaction was found to be specific, with lithium leading to greater polyacrylate adsorption than sodium, which was followed by cesium. The stronger affinity of lithium for the hematite surface over sodium and cesium is indicative of the inverse lyotropic adsorption series and has been rationalized in the past by the "structure-making-structure-breaking" model. These results provide a useful insight into the likely adsorption mechanism for polyacrylate flocculants at high pH and ionic strength onto residues in the Bayer processing of bauxite.

Intracrystalline proteins and urolithiasis: a synchrotron X-ray diffraction study of calcium oxalate monohydrate.

UNLABELLED: The existence of intracrystalline proteins and amino acids in calcium oxalate monohydrate was demonstrated by X-ray synchrotron diffraction studies. Their presence has implications for the destruction of calcium oxalate crystals formed in the urinary tract and the prevention of kidney stones. INTRODUCTION: Although proteins are present in human kidney stones, their role in stone pathogenesis remains unknown. This investigation aimed to characterize the nature of the relationship between the organic and mineral phases in calcium oxalate monohydrate (COM) crystals grown in human urine and in aqueous solutions of proteins and amino acids to clarify the function of proteins in urolithiasis. METHODS: COM crystals were grown in human urine and in aqueous solutions containing either human prothrombin (PT), Tamm-Horsfall glycoprotein (THG), aspartic acid (Asp), aspartic acid dimer (AspAsp), glutamic acid (Glu), glutamic acid dimer (GluGlu), or gamma-carboxyglutamic acid (Gla). Controls consisted of COM crystals precipitated from pure inorganic solutions or from human urine that had been ultrafiltered to remove macromolecules. Synchrotron X-ray diffraction with Rietveld whole-pattern peak fitting and profile analysis was used to determine nonuniform crystal strain and crystallite size in polycrystalline samples. RESULTS: Crystals precipitated from ultrafiltered urine had lower nonuniform strain than those grown in urine or in aqueous PT solution. Nonuniform strain was much lower in crystals grown in distilled water or in the presence of THG. For the amino acids, the highest nonuniform strain was exhibited by crystals grown in Gla solution, followed by Glu. Crystallite size was inversely related to nonuniform strain, with the effect being significantly less for amino acids than for macromolecules. CONCLUSIONS: Selected proteins and amino acids associated with COM crystals are intracrystalline. Although their incorporation into the mineral bulk would be expected to affect the rate of crystal growth, they also have the potential to influence the phagocytosis and intracellular destruction of any crystals nucleated and trapped within the renal collecting system. Crystals impregnated with protein would be more susceptible to digestion by cellular proteases, which would provide access to the crystal core, thereby facilitating further proteolytic degradation and mineral dissolution. We therefore propose that intracrystalline proteins may constitute a natural form of defense against renal stone formation.

Contribution of Raman spectroscopy to identification of biominerals present in teeth of Acanthopleura rehderi, Acanthopleura curtisiana, and Onithochiton quercinus.

Raman spectroscopic investigations of the major lateral teeth of the chitons Acanthopleura rehderi and Acanthopleura curtisiana indicate that, in addition to the magnetite of the cutting surface and a carbonated hydroxyapatite in the central tooth core, these species deposit limonite in place of the lepidocrocite reported for other members of the genus Acanthopleura. A comparison of the spectra from these species with those of Onithochiton quercinus, which also deposits limonite, indicates that the current assignment of these species to Acanthopleura may not be appropriate. Biomineralization of the major lateral teeth may be a useful parameter to include in the taxonomic classification of chiton species.

A new biomineral identified in the cores of teeth from the chiton Plaxiphora albida.

The hydrated iron(III) oxide limonite is reported for the first time as a biomineral. In situ laser Raman spectra of the tooth cores from major lateral teeth of the chiton Plaxiphora albida are compared with those of synthetic and mineral iron phosphates and iron oxides. Raman spectra measured on iron phosphate and iron oxide standard materials are shown to be easily distinguishable from one another. The central tooth cores of mature P. albida teeth do not show any evidence for the presence of a separate iron phosphate mineral. Rather, in each tooth a narrow band of the hydrated iron(III) oxide limonite is shown to separate the magnetite of the tooth surface from a central core region comprising both lepidocrocite and limonite. The high concentration of phosphorus in P. albida tooth cores, previously observed by energy dispersive spectroscopy, is not associated with a separate iron phosphate mineral, indicating that this element may be adsorbed onto the surface of the iron oxide minerals present. The failure to detect a separate iron(III) phosphate is discussed with reference to other chiton species that display high levels of iron and phosphorus in the cores of their mature major lateral teeth.

Alzheimer's beta-amyloid peptides compete for insulin binding to the insulin receptor.

The amyloid-beta (Abeta) peptide is neurotoxic and associated with the pathology of Alzheimer's disease (AD). We investigated the effect of Abeta peptides on insulin binding to the insulin receptor because it is known that (1) Abeta and insulin are both amyloidogenic peptides sharing a common sequence recognition motif, (2) Abeta and insulin are substrates for the same insulin degrading enzyme, and (3) impaired glucose metabolism is a characteristic event in the pathology of AD. We discovered that Abeta(1-40) and Abeta(1-42,) the main physiological forms, reduced insulin binding and receptor autophosphorylation. The reduction in binding was caused by a decrease in the affinity of insulin binding to the insulin receptor. This reduction was independent of the receptor concentration. The reverse, control peptide Abeta(40-1) did not reduce insulin binding or insulin receptor autophosphorylation. These results demonstrate that Abeta is a direct competitive inhibitor of insulin binding and action. We speculate that the increased levels of Abeta in Alzheimer's disease may be linked to the associated insulin resistance that has been observed previously in this disease.