Identifying diagonal cutter marks on thin wires using 3D imaging.

We present work on matching 2-mm-thick wires using optical 3D imaging methods. Marks on such small surfaces are difficult to match using a comparison microscope as this 2D imaging method does not provide height data about the sample surface. Moreover, these 2D microscopy images may be affected by illumination. Hence, the reference and investigated sample should be present at the same time. We employed scanning white light interferometry and confocal microscopy to provide quantitative 3D profiles for reliable comparison of samples that are unavailable for simultaneous analysis. We show that 3D profiling offers a solution by allowing illumination-independent sample comparison. We correctly identified 74 of 80 profiles using consecutive matching striae (CMS) criteria, and we were able to match samples based on profiles measured using different 3D imaging devices. The results suggest that the used methods allow matching cutter marks on thin wires, which has been difficult previously.

Statistical challenges in the quantification of gunshot residue evidence.

The discharging of a gun results in the formation of extremely small particles known as gunshot residues (GSR). These may be deposited on the skin and clothing of the shooter, on other persons present, and on nearby items or surfaces. Several factors and their complex interactions affect the number of detectable GSR particles, which can deeply influence the conclusions drawn from likelihood ratios or posterior probabilities for prosecution hypotheses of interest. We present Bayesian network models for casework examples and demonstrate that probabilistic quantification of GSR evidence can be very sensitive to the assumptions concerning the model structure, prior probabilities, and the likelihood components. This finding has considerable implications for the use of statistical quantification of GSR evidence in the legal process.

Bayesian adaptive approach to estimating sample sizes for seizures of illicit drugs.

A considerable amount of discussion can be found in the forensics literature about the issue of using statistical sampling to obtain for chemical analyses an appropriate subset of units from a police seizure suspected to contain illicit material. Use of the Bayesian paradigm has been suggested as the most suitable statistical approach to solving the question of how large a sample needs to be to ensure legally and practically acceptable purposes. Here, we introduce a hypergeometric sampling model combined with a specific prior distribution for the homogeneity of the seizure, where a parameter for the analyst's expectation of homogeneity (α) is included. Our results show how an adaptive approach to sampling can minimize the practical efforts needed in the laboratory analyses, as the model allows the scientist to decide sequentially how to proceed, while maintaining a sufficiently high confidence in the conclusions.

Statistical modelling of measurement errors in gas chromatographic analyses of blood alcohol content.

Headspace gas chromatographic measurements of ethanol content in blood specimens from suspect drunk drivers are routinely carried out in forensic laboratories. In the widely established standard statistical framework, measurement errors in such data are represented by Gaussian distributions for the population of blood specimens at any given level of ethanol content. It is known that the variance of measurement errors increases as a function of the level of ethanol content and the standard statistical approach addresses this issue by replacing the unknown population variances by estimates derived from large sample using a linear regression model. Appropriate statistical analysis of the systematic and random components in the measurement errors is necessary in order to guarantee legally sound security corrections reported to the police authority. Here we address this issue by developing a novel statistical approach that takes into account any potential non-linearity in the relationship between the level of ethanol content and the variability of measurement errors. Our method is based on standard non-parametric kernel techniques for density estimation using a large database of laboratory measurements for blood specimens. Furthermore, we address also the issue of systematic errors in the measurement process by a statistical model that incorporates the sign of the error term in the security correction calculations. Analysis of a set of certified reference materials (CRMs) blood samples demonstrates the importance of explicitly handling the direction of the systematic errors in establishing the statistical uncertainty about the true level of ethanol content. Use of our statistical framework to aid quality control in the laboratory is also discussed.

Cloning and expression of a Melanocarpus albomyces steryl esterase gene in Pichia pastoris and Trichoderma reesei.

The ste1 gene encoding a steryl esterase was isolated from the thermophilic fungus Melanocarpus albomyces. The gene has one intron, and it encodes a protein consisting of 576 amino acids. The deduced amino acid sequence of the steryl esterase was shown to be related to lipases and other esterases such as carboxylesterases. Formation of mature protein requires post-translational removal of a putative 18-amino-acid signal sequence and a 13-residue propeptide at the N-terminus. The intronless version of the Melanocarpus albomyces ste1 gene was expressed in Pichia pastoris under the inducible AOX1 promoter. The production level was low, and a large proportion of the total activity yield was found to be present intracellularly. However, the fact that steryl esterase activity was produced by P. pastoris cells carrying the expression cassette confirmed that the correct gene had been cloned. The ste1 gene was subsequently expressed in T. reesei under the inducible cbh1 promoter, and a clearly higher production level was obtained. About 60% of the total activity was bound to the fungal mycelium or to solid components of the culture medium, or existed as aggregates. Triton X-100 was successfully used to recover this activity. The heterologous production system in T. reesei provides a means of producing M. albomyces steryl esterase STE1 reliably in large scale for future studies.

Characterization of Melanocarpus albomyces steryl esterase produced in Trichoderma reesei and modification of fibre products with the enzyme.

Melanocarpus albomyces steryl esterase STE1 is considered to be an interesting tool for several industrial applications due to its broad substrate specificity. STE1 was produced in the filamentous fungus Trichoderma reesei in a laboratory bioreactor at an estimated production level of 280 mg l(-l). The properties of the purified recombinant enzyme (rSTE1), such as substrate specificity, molecular mass, pH optimum and stability and thermostability, were characterized and compared to the corresponding properties of the native enzyme. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis showed one band with a molecular weight of 60 kDa for rSTE1, whereas analytical gel filtration showed a dimeric structure with a molecular weight of 120 kDa. The rSTE1 was somewhat less stable under different conditions and had slightly lower activities on various substrates than the native STE1. The effects of rSTE1 on the properties of paper sheets and polyethylene terephthalate (PET) fabric were preliminarily evaluated. Due to the hydrolysis of triglycerides and steryl esters by the rSTE1 treatment, the tensile strength and hydrophilicity of the paper were increased. The rSTE1 treatment increased significantly the polarity of PET by hydrolysing the ester bonds in the polyester backbone. Dyeing of PET with methylene blue was also slightly improved after rSTE1 treatment.

The active site of cellobiohydrolase Cel6A from Trichoderma reesei: the roles of aspartic acids D221 and D175.

Trichoderma reesei cellobiohydrolase Cel6A is an inverting glycosidase. Structural studies have established that the tunnel-shaped active site of Cel6A contains two aspartic acids, D221 and D175, that are close to the glycosidic oxygen of the scissile bond and at hydrogen-bonding distance from each other. Here, site-directed mutagenesis, X-ray crystallography, and enzyme kinetic studies have been used to confirm the role of residue D221 as the catalytic acid. D175 is shown to affect protonation of D221 and to contribute to the electrostatic stabilization of the partial positive charge in the transition state. Structural and modeling studies suggest that the single-displacement mechanism of Cel6A may not directly involve a catalytic base. The value of (D2O)(V) of 1.16 +/- 0.14 for hydrolysis of cellotriose suggests that the large direct effect expected for proton transfer from the nucleophilic water through a water chain (Grotthus mechanism) is offset by an inverse effect arising from reversibly breaking the short, tight hydrogen bond between D221 and D175 before catalysis.