Genetic polymorphism of human alpha 2HS-glycoprotein: characterization and application to forensic hemogenetics

Human alpha 2HS-glycoprotein (AHSG) phenotypes were studied by isoelectric focusing of native and desialyzed samples followed by passive immunoblotting. At present a total of 15 different alleles could be distinguished. The usefulness of AHSG polymorphism in the field of forensic hemogenetics was demonstrated by markedly high variability at AHSG locus, reliability of phenotyping results and stability in blood stains.     

Recent and future trends in blood group typing

Blood group typing is the process of testing red blood cells to determine which antigens are present and which are absent. It is standard practice to test for A, B, and D (Rh) antigens and to perform tests for other antigens in selected cases. ABO blood group typing is confirmed by reverse grouping that detects expected isoagglutinins. Unexpected antibodies can be demonstrated by antibody screening tests. For transfusion, donor units compatible with the patient are selected. Prior to transfusion, a crossmatch is performed as a final check for incompatibility. This article describes the recent and future methods of blood group typing and testing of serological compatibility. In addition, methods for blood bank automation are presented.     

The use of library identification and common fragments for the identification of corticosteroids in forensic samples

The detection of corticosteroids and sex steroids in samples with no content indication, which are confiscated for forensic investigation, is a challenge in doping analysis. A screening method based on the identification of androgens, estrogens, gestagens, and their esters by means of a mass spectral library, along with a fast ultra-performance liquid chromatography–mass spectrometry (UPLC-MS) method, was recently developed in our lab for the analysis of dietary supplements. However, for forensic investigations, it is important to extend the scope of the method to corticosteroids in various matrices. Therefore, 36 corticosteroids were added to the mass spectral library, and the sample preparation step was modified so that androgens, gestagens, corticosteroids, and their esters could be analyzed with only one injection with the UPLC-MS method. A complementary tool to the existing library identification was found in the extraction of common fragment ions out of the full scan data obtained for the library search. The fragment ion with m/z 147 was found to be a good marker for the detection of steroids. Extra confirmation was obtained from the fragment ions with m/z 135 (for all steroids) and 237 (specific for corticosteroids) or from the fragment ions with m/z 77, 91, and 105. The effectiveness of this approach was evaluated on some samples previously screened for forensic investigation with thin-layer chromatography and confirmed with a targeted gas chromatography–mass spectrometry method. This study shows that the combination of the library identification and the common fragment ions approach can be a valuable tool in the detection of steroids without defining any target at the start of the analysis.     

The use of differential scanning calorimetry to identify methaqualone samples: forensic applications.

The d.s.c. curves of commercial samples of methaqualone—Qu$\text{aa}$lude (U.S.A.), Qu$\text{aa}$lude (Mexico), Sopar, Mandrax and Parest — are presented. Similar d.s.c. curves were found for Qu$\text{aa}$lude (U.S.A.), Qu$\text{aa}$lude (Mexico), and Mandrax, while Sopar and Parest exhibited different types of curves. New and old formulations of the Qu$\text{aa}$lude (U.S.A.) brand could be differentiated by differences in the tablet filler and binder. Unfortunately, d.s.c. could not be used to determine the country of origin of the drug, although it was useful to characterize it. The technique can be used to identify samples of 0.15 mg or less.     

DNA damage promotes mistyping in the allele specific oligonucleotide probing analysis of forensic samples

Five polymerase chain reaction (PCR) products which could not be reliably typed by allele-specific oligonucleotide (ASO) probing at the human leukocyte antigen (HLA) DQA1 locus were analyzed by polyacrylamide gel electrophoresis and direct sequencing. The first method revealed the preferential amplification of only one of the two alleles in two cases. Direct sequencing of PCR products allowed unambiguous genetic typing but a high number of artifacts was observed. Several of these artifacts occurred in the sequences recognized by the ASOs. This finding provides an explanation for the mistyping in the ASO probing procedure because Taq polymerase errors both created new genetic specificities and eliminated site-specific polymorphisms. Reversed-phase HPLC-MS of the five forensic templates showed a high degree of DNA damage. These data together indicate that the risk of mistyping when using the ASO probing procedure cannot be neglected in the forensic analysis of damaged DNA samples.     

The use of a rotating lead-disk electrode for electroanalytical purposes

The behavior of lead as an electrode material for electroanalytical purposes was studied in 1.0 M sulfuric acid. The useful negative range of potential was approximately ?0.57 to ?1.20 V vs. SCE for conventional voltammetry at a rotating disk electrode. For the sinusoidal hydrodynamic modulation technique, the negative limit was about ?1.25 V. Studies of the reduction of cadmium ion, thallium(I) ion, and p-nitroaniline showed that the behavior of a rotating lead-disk electrode was comparable to that expected on the basis of theory. Limits of detection are in the micromolar range.     

Extraction and concentration of vapors from fire debris for forensic purposes: Evaluation of the use of Radiello Passive Air Sampler

The Radiello Passive Air Sampler is one of the latest innovations developed for the sampling of pollutants in the air by passive headspace. It has been reported that its properties allow an enhanced sensitivity, reproducibility and adsorption capacity. It therefore appears to be of interest in the extraction of potential residues of ignitable liquids present in fire debris when arson is suspected.A theoretical approach and several laboratory tests have made it possible to precisely characterize in a forensic perspective the potential of the device in extracting and concentrating the vapors of ignitable liquids found in fire debris. Despite some advantages, the Radiello device appears to be less efficient than traditional axial symmetry samplers.     

Avoiding polyatomic interferences in measurements of lanthanides in uranium material for nuclear forensic purposes

Journal of Radioanalytical and Nuclear Chemistry - Measurements of the lanthanide series with ICP-SF-MS provide low detection limits but suffer from oxides of the lighter lanthanides interfering on...     

Subtyping of group specific component (GC) in human semen, blood and vaginal fluid by isoelectric focusing in immobilized pH gradients

The group specific component (GC) is stable and well suited for forensic casework. Isoelectric focusing of common GC variants from semen, seminal fluid, vaginal fluid and semen stains, on Immobiline DryPlates, pH 4.5-5.4, is of practical value in criminal investigations of sexual deliquencies. GC is present in normospermia and azoospermia seminal fluids and found in about 20% of the vaginal secretions. The GC patterns observed were similar and in accordance with the bands of the individual GC type in plasma/serum.     

Mass spectrometric detection of siRNA in plasma samples for doping control purposes

Small interfering ribonucleic acid (siRNA) molecules can effect the expression of any gene by inducing the degradation of mRNA. Therefore, these molecules can be of interest for illicit performance enhancement in sports by affecting different metabolic pathways. An example of an efficient performance-enhancing gene knockdown is the myostatin gene that regulates muscle growth. This study was carried out to provide a tool for the mass spectrometric detection of modified and unmodified siRNA from plasma samples. The oligonucleotides are purified by centrifugal filtration and the use of an miRNA purification kit, followed by flow-injection analysis using an Exactive mass spectrometer to yield the accurate masses of the sense and antisense strands. Although chromatography and sensitive mass spectrometric analysis of oligonucleotides are still challenging, a method was developed and validated that has adequate sensitivity (limit of detection 0.25–1 nmol mL⁻¹) and performance (precision 11–21%, recovery 23–67%) for typical antisense oligonucleotides currently used in clinical studies.     

Development and validation of a new 18 X-STR typing assay for forensic applications

With a unique inheritance pattern compared to autosomal short tandem repeats (A-STRs), X chromosomal STRs (X-STRs) have special usage in forensic relationship testing. In this study, we designed a multiplex amplification system (named TYPER-X19 multiplex assay) consisting of 18 STR loci spreading from 7.837 to 149.460 Mb on the X chromosomes (DXS9895, DXS8378, DXS9902, DXS6810, DXS7132, DXS10079, DXS6789, DXS7424, DXS101, DXS6797, DXS7133, DXS6804, GATA165B12, DXS10103, HPRTB, GATA31E08, DXS8377, and DXS7423), and the amelogenin. PCR primers were marked with four kinds of fluorophores including FAM, HEX, TAMRA, and ROX. The multiplex system was optimized and tested for precision, concordance, reproducibility, sensitivity, stability, DNA mixture, and species specificity according to the conventional validation guidelines. The results indicated that the system was accurate, reliable, and sensitive enough, and was suitable for common forensic case-type samples. In the population genetic study, a total of 148 alleles were detected at the 18 X-STR loci in 398 Southern Han Chinese. Relatively high combined power of discrimination in male (PD_m), power of discrimination in female (PD_f), mean paternity exclusion chance in trios (MEC_trio), and mean paternity exclusion chance in duos (MEC_Duo) by Desmarais were detected, and HPRTB-DXS10103 was in linkage disequilibrium. The results suggested that the TYPER-X19 multiplex assay was suitable for forensic applications.     

The use of solid-phase microextraction--gas chromatography in forensic analysis

A thorough review of the application of solid-phase microextraction (SPME) combined with gas chromatography for the analysis of forensic specimens is presented, including experimental results for several recent applications. The SPME applications covered in this comprehensive review include ignitable liquid residues (also referred to as accelerants), explosive traces, drugs and poisons from biological specimens, and other forensic applications. Recently developed SPME methods are also presented, including the analysis of ignitable liquid residues on human skin, odor signatures, and several drug applications such as free-fraction antipsychotic drug levels, blood alcohol casework, drink-tampering analysis, and gamma-hydroxybutyrate identification without the need for derivatization. SPME is shown to be an inexpensive, rapid, and sensitive method for the analysis of a variety of forensic specimens.     

The use of LA-SF-ICP-MS for nuclear forensics purposes: uranium isotope ratio analysis

This work describes the utilization of the laser ablation sector field inductively coupled plasma mass spectrometry (LA-SF-ICP-MS) technique for the determination of uranium isotopic composition in a highly enriched uranium sample. The measurements were performed on a continuous ablation with low energy density and defocusing, which demonstrated to be the optimum to reach the best signal stability. The measurements were improved by adjusting the following parameters: RF power, laser beam diameter, defocusing of laser beam, laser energy, laser energy density, auxiliary gas and sample gas. The ²³⁵U/²³⁸U isotope ratio with its respective uncertainty was 16.36 ± 0.15 and its precision was 1.12 % relative standard deviation. The uncertainties were estimated following the ISO GUM, with a confidence level of 95.45 % (k = 2.00). When compared the isotope abundances to the Round Robin Exercise Number 3’s average results a difference of 0.46 % has been found and when compared to supplier’s value, the difference was 0.41 %. The results presented by the measurements revealed that the LA-ICP-MS technique offers a rapid and accurate alternative to measure uranium isotope ratios without any sample preparation, since it allows carrying out the measurements straight on the sample. Moreover, it preserves the testimony—very important for safeguards and nuclear forensics purposes.     

Isoelectric focusing of group specific component in semen and vaginal stains on ultrathin immobilized pH gradient gels

Subtyping of the group specific component in secretions of semen and vaginal fluids is impossible with conventional detection systems. By means of a highly sensitive alkaline-phosphatase secondary antibody system the group specific component can reliably be detected in semen stains. Results with vaginal swabs were inconsistent and in saliva stains Gc activity could not be detected.     

Determination of metformin and other biguanides in forensic whole blood samples by hydrophilic interaction liquid chromatography-electrospray tandem mass spectrometry

Metformin is an anti-diabetic drug in the biguanide class which also includes phenformin and buformin. Because of the potential adverse effects of the biguanides, a reliable liquid chromatography-tandem mass spectrometry method using pneumatically assisted electrospray ionization was developed for the quantification of the drugs in both live and post-mortem human whole blood. The blood proteins were precipitated by the addition of a mixture of methanol and acetonitrile, and the extract was cleaned up by cation-exchange solid-phase extraction to eliminate ion suppression effects. The separation was performed by hydrophilic interaction liquid chromatography. Matrix-matched calibrants combined with isotope dilution of metformin were used for calibration. The detection limits were 0.01 mg/L for metformin and phenformin and the relative intra-laboratory reproducibility standard deviations were less than 6% at concentrations of 1-10 mg/L. The mean true recoveries were greater than 86%.     

The use of 2D diffractometry data for oriented samples in the choice of a unit cell

An approach for the substantiated choice of the indexing procedure of a diffraction pattern is described. The technique is based on a combined use of powder X-ray diffraction data in Bragg–Brentano schemes and 2D GIXD of mainly oriented polycrystalline samples.     

Visible and near-infrared chemical imaging methods for the analysis of selected forensic samples

This study investigated various chemical imaging methods for the forensic analysis of paints, tapes and adhesives, inks and firearm propellants (absorption and photoluminescence in the UV-vis-NIR regions). Results obtained using chemical imaging technology were compared with those obtained using traditional techniques. The results show that chemical imaging offers significant advantages in the forensic context, for example the ability to display visual and spectral results side by side and to reduce sample preparation, hence minimizing the risk of contamination. Chemical imaging produced a greater discriminating power than traditional techniques for most evidence types. Chemical imaging also eliminated different brands of ammunition based on the fluorescence characteristics of the propellant grains preserving the evidence for further analysis. It is expected that this technology will find broader forensic applications in the future.     

The group specific component/vitamin D binding protein (GC/DBP) system in the analysis of disputed paternities

The group-specific component (GC) was discovered in 1959, and in the same year a vitamin D binding protein (DBP) in human plasma was found; however, their identity was established as late as 1975. In the GC/DBP system three common alleles, GC*1F, GC*1S, and GC*2, determine six GC phenotypes: 1F, 1S, 2, 1F-1S, 2-1F and 2-1S, these common alleles having been found in all human populations studied. In addition, more than 120 GC variants have been discovered, with varying frequencies in different populations. The distribution of the common GC phenotypes and the presence of rare GC variant phenotypes render the GC/DBP system useful for the analysis of disputed paternities.