Rapid determination of furosemide by combined spot test/diffuse reflectance spectroscopy to detect doping in sport

This paper describes the development and application of a simple, cheap, and clean method for the quantification of furosemide in urine samples from athletes, to detect doping, using a combined spot test/diffuse reflectance spectroscopy procedure. The method is based on the complexation reaction of furosemide (5-(aminosulfonyl)-4-chloro-2-((furanylmethyl)amino)benzoic acid, dissolved in ethanol, with FeCl3 and the surfactant dodecyltrimethylammonium bromide (DTAB) in aqueous solution, yielding a colored compound on the surface of a filter paper. The reagent concentrations were optimized using a chemometric experimental design. The reflectometric measurements of the complex formed were carried out at 477nm. The linear range obtained was 1.65-9.00×10-3molL-1 of furosemide (R=0.997), and the detection and quantification limits were 4.9×10-4 and 1.62×10-3molL-1, respectively. The proposed method was successfully applied in the analysis of furosemide in spiked urine, demonstrating that it is a reliable alternative method for the detection of furosemide doping in sport. © 2012 Elsevier B.V..

Is characterizing the digital forensic facial reconstruction with hair necessary? A familiar assessors' analysis

Background: In the international scientific literature, there are few studies that emphasize the presence or absence of hair in forensic facial reconstructions. There are neither Brazilian studies concerning digital facial reconstructions without hair, nor research comparing recognition tests between digital facial reconstructions with hair and without hair. The miscegenation of Brazilian people is considerable. Brazilian people, and, in particular, Brazilian women, even if considered as Caucasoid, may present the hair in very different ways: curly, wavy or straight, blonde, red, brown or black, long or short, etc. For this reason, it is difficult to find a correct type of hair for facial reconstruction (unless, in real cases, some hair is recovered with the skeletal remains). Aims and methods: This study focuses on the performance of three different digital forensic facial reconstructions, without hair, of a Brazilian female subject (based on one international database and two Brazilian databases for soft facial-tissue thickness) and evaluates the digital forensic facial reconstructions comparing them to photographs of the target individual and nine other subjects, employing the recognition method. A total of 22 assessors participated in the recognition process; all of them were familiar with the 10 individuals who composed the face pool. Results and conclusions: The target subject was correctly recognized by 41% of the 22 examiners in the International Pattern, by 32% in the Brazilian Magnetic Resonance Pattern and by 32% in the Brazilian Fresh Cadavers Pattern. The facial reconstructions without hair were correctly recognized using the three databases of facial soft-tissue thickness. The observed results were higher than the results obtained using facial reconstructions with hair, from the same skull, which can indicate that it is better to not use hair, at least when there is no information concerning its characteristics. © 2013 Elsevier B.V. All rights reserved.

Wildlife forensic DNA and lowland tapir (Tapirus terrestris) poaching

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Genetic data of 15 autosomal STR loci: an analysis of the Araraquara population colonization (São Paulo, Brazil)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Latin-American Society Of Forensic Genetics (SLAGF) results of the Interlaboratory Quality Control Exercise 2010-2011

Latin-American Society of Forensic Genetics (SLAGF) Interlaboratory Quality Control Exercise (2010-2011) included the analysis of three bloodstain samples in FTA Classic Card (three persons, biologically unrelated) and one theoretical exercise. There were 56 participating laboratories from 13 Latin-American countries that belong to society, were reported 70 STRs, including autosomal and sex chromosome markers with consensus in 53 STRs with a rate in reporting errors of 2.3%. Fifty-six laboratories reported results in theoretical exercise with mistakes in calculation of IP for each marker. It is necessary to hold meetings to discuss the results of this exercise to reach conclusions and recommendations on all aspects of DNA forensics analysis and paternity test, to improve results and quality in the results of each laboratory. © 2011 Elsevier B.V.

Development and validation of a D-loop mtDNA SNP assay for the screening of specimens in forensic casework

Mitochondrial DNA (mtDNA) analysis is usually a last resort in routine forensic DNA casework. However, it has become a powerful tool for the analysis of highly degraded samples or samples containing too little or no nuclear DNA, such as old bones and hair shafts. The gold standard methodology still constitutes the direct sequencing of polymerase chain reaction (PCR) products or cloned amplicons from the HVS-1 and HVS-2 (hypervariable segment) control region segments. Identifications using mtDNA are time consuming, expensive and can be very complex, depending on the amount and nature of the material being tested. The main goal of this work is to develop a less labour-intensive and less expensive screening method for mtDNA analysis, in order to aid in the exclusion of non-matching samples and as a presumptive test prior to final confirmatory DNA sequencing. We have selected 14 highly discriminatory single nucleotide polymorphisms (SNPs) based on simulations performed by Salas and Amigo (2010) [1] to be typed using SNaPShotTM (Applied Biosystems, Foster City, CA, USA). The assay was validated by typing more than 100 HVS-1/HVS-2 sequenced samples. No differences were observed between the SNP typing and DNA sequencing when results were compared, with the exception of allelic dropouts observed in a few haplotypes. Haplotype diversity simulations were performed using 172 mtDNA sequences representative of the Brazilian population and a score of 0.9794 was obtained when the 14 SNPs were used, showing that the theoretical prediction approach for the selection of highly discriminatory SNPs suggested by Salas and Amigo (2010) [1] was confirmed in the population studied. As the main goal of the work is to develop a screening assay to skip the sequencing of all samples in a particular case, a pair-wise comparison of the sequences was done using the selected SNPs. When both HVS-1/HVS-2 SNPs were used for simulations, at least two differences were observed in 93.2% of the comparisons performed. The assay was validated with casework samples. Results show that the method is straightforward and can be used for exclusionary purposes, saving time and laboratory resources. The assay confirms the theoretic prediction suggested by Salas and Amigo (2010) [1]. All forensic advantages, such as high sensitivity and power of discrimination, as also the disadvantages, such as the occurrence of allele dropouts, are discussed throughout the article. © 2013 Elsevier B.V.

Dental fluorescence: Potential forensic use

In cases of identification of bones, skeletal segments or isolated bones, searching for biotypologic diagnostic data to estimate an individual's age enables comparing these data with those of missing individuals. Enamel, dentin and pulp undergo remarkable changes during an individual's life. The enamel becomes more mineralized, smoother and thinner, and deteriorates because of physiological and pathological factors. Dental pulp decreases in volume due to the deposition of secondary dentin; thus, the dentin becomes thicker with time. In natural teeth, the fluorescence phenomenon occurs in dentin and enamel and changes in those tissues may alter the expression of the natural tooth color. The aim of this study was to assess the correlation between age and teeth fluorescence for individuals from different age groups. The sample consisted of 66 randomly selected Brazilians of both genders aged 7-63 years old. They were divided into 6 groups: Group 1 - aged 7-12 years, Group 2 - aged 13-20 years, Group 3 - aged 21-30 years, Group 4 - aged 31-40 years, Group 5 - aged 41-50 years and Group 6 - aged between 51 and 63 years. Upper right or left central incisors were used for the study. Restored and aesthetic rehabilitated teeth were excluded from the sample. The measurement of tooth fluorescence was carried out via computer analysis of digital images using the software ScanWhite DMC/Darwin Systems - Brazil. It was observed that dental fluorescence decreases when comparing the age groups 21-30, 31-40, 41-50 and 51-63 years. The results also showed that there is a statistically significant difference between the groups 41-50 years and 21-30 years (p=. 0.005) and also among the group 51-63 years and all other groups (p< 0.005). It can be concluded that dental fluorescence is correlated with age and has a similar and stable behavior from 7 to 20 years of age. It reaches its maximum expected value at the age of 26.5 years and thereafter decreases. © 2013 Elsevier B.V.

Analysis of the procedures used to evaluate suicide crime scenes in Brazil: A statistical approach to interpret reports

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Forensic electrochemistry: indirect electrochemical sensing of the components of the new psychoactive substance Synthacaine

Synthacaine is a New Psychoactive Substance which is, due to its inherent psychoactive properties, reported to imitate the effects of cocaine and is therefore consequently branded as legal cocaine. The only analytical approach reported to date for the sensing of Synthacaine is mass spectrometry. In this paper, we explore and evaluate a range of potential analytical techniques for its quantification and potential use in the field screening Synthacaine using Raman spectroscopy, presumptive (colour) testing, High Performance Liquid Chromatography (HPLC) and electrochemistry. HPLC analysis of street samples reveals that Synthacaine comprises a mixture of methiopropamine (MPA) and 2-aminoindane (2-AI). Raman spectroscopy and presumptive (colour) tests, the Marquis, Mandelin, Simon's and Robadope test, are evaluated towards a potential in-the-field screening approach but are found to not be able to discriminate between the two when they are both present in the same sample, as is the case in the real street samples. We report for the first time a novel indirect electrochemical protocol for the sensing of MPA and 2-AI which is independently validated in street samples with HPLC. This novel electrochemical approach based upon one-shot disposable cost effective screen-printed graphite macroelectrodes holds potential for in-the-field screening for Synthacaine.