It has been repeatedly acknowledged that age-at-death estimation based on dentalcementum represents a partial and time-consuming method that hinders adoptionof this histological approach. User-friendly micrograph analysis represents a grow-ing request of cementochronology. This article evaluates the feasibility of using amodule to accurately quantify cementum deposits and compares the module's per-formance to that of a human expert. On a dental collection (n=200) of known-ageindividuals, precision and accuracy of estimates performed by a developed pro-gram (101 count/tooth;n=20,200 counts) were compared to counts performedmanually (5 counts/tooth;n=975 counts). Reliability of the software and agree-ment between the two approaches were assessed byintraclass correlation coeffi-cient and Bland Altman analysis. The automated module produced reliable andreproducible counts with a higher global precision than the human expert. Althoughthe software is slightly more precise, it shows higher sensitivity to taphonomic dam-ages and does not avoid the trajectory effect described for age-at-death estimationin adults. Likewise, for human counts, global accuracy is acceptable, but underestima-tions increase with age. The quantification of the agreement between the twoapproaches shows a minor bias, and 94% of individuals fall within the intervals ofagreement. Automation gives an impression of objectivity even though the region ofinterest, profile position and parameters are defined manually. The automated systemmay represent a time-saving module that can allow an increase in sample size, whichis particularly stimulating for population-based studies.

Arylcyclohexylamine (ACH) compounds represent a predominant faction within new psychoactive substances. Due to their powerful dissociative effects, they are used in recreational contexts but also in situations of drug-facilitated sexual assault, and therefore, they are a constant target of analysis by forensic experts. In recent years, their consumption has been notably high, especially the use of ketamine, presenting daily challenges for laboratories in the determination of this and other ACH analogues. This review comprises the recent strategies that forensic specialists use to identify and quantify ACH compounds in the laboratory with more traditional analytical techniques and technology, and on the point-of-care testing via sensor technology. The study focuses on analogues of phencyclidine (PCP), ketamine, and eticyclidine, highlighting the consistent need for higher sensitivity in the analysis of various samples collected from real cases and simulations of possible matrices. The review also emphasises the ongoing research to develop more sensitive, quicker, and more capable sensors.

The emergence of new psychoactive substances has earned a great deal of attention, and several reports of acute poisoning and deaths have been issued involving, for instance, synthetic opiates. In recent years, there have been profound alterations in the legislation concerning consumption, marketing, and synthesis of these compounds; rapid alert systems have also been subject to changes, and new substances and new markets, mainly through the internet, have appeared. Their effects and how they originate in consumers are still mostly unknown, primarily in what concerns chronic toxicity. This review intends to provide a detailed description of these substances from the point of view of consumption, toxicokinetics, and health consequences, including case reports on intoxications in order to help researchers and public health agents working daily in this area.

Allele frequencies and other relevant forensic parameters of 15 loci studied with Applied Biosystems AmpFlSTRs NGM Kit were calculated in a population of individuals residing in the South of Portugal. Blood stain samples were obtained from a total of 350 unrelated individuals involved in paternity testing casework. This new kit has five loci not present in any other kit used in our laboratory (Promega Powerplex 16 System and Applied Biosystems Identifiler Plus): D10S1248, D22S1045, D2S441, D1S1656 and D12S391. The NGM multiplex kit is used in our forensic casework as an auxiliary screening tool to solve deficient casework such as fatherless paternity testing, and to help in paternity investigations with one genetic incompatibility after the routine seventeen loci typing. Furthermore, this five loci included in the European Standard Set are also recommended by the European Network of Forensic Science Institutes (ENFSI) and the European DNA Profiling group (EDNAP). These studies are necessary to calculate statistical forensic parameters such as power of discrimination, power of exclusion or minimum allele frequencies. Statistical parameters such as heterozigoty, homozigoty or allele frequencies were determined with Arlequim V3.5 and results compared with the ones reported in previous similar studies. As final remark, is important to remember that this kind of study is highly important for the Forensic Laboratories accreditation achievement

Microextraction by packed sorbent (MEPS) efficiently combines extraction, pre-concentration, and cleanup in a single device comprising two parts: the MEPS syringe and the packed sorbent bed1. MEPS has been used for several bioanalytical applications, including extraction of endogenous metabolites, biomarkers, and drugs from biological samples. It is particularly useful in metabolomics and pharmacokinetic studies2,3. Regarding MEPS applicability to forensic toxicology, urine is the most used specimen, followed by oral fluid (despite of its relatively high viscosity). Protein precipitation followed by centrifugation and with, or without dilution of the supernatant is the most commonly reported approach. The most detected compounds in forensic settings using MEPS are drugs of abuse [opiates and opioids (26%), cocaine (13%) cathinones (11%), dissociative hallucinogens (11%), cannabinoids and amphetamines (9% each) and other drugs (10%)] and medicinal drugs [antidepressants (9%), benzodiazepines/Z drugs (4%)]. MEPS was also applied to a beverage for forensic purposes e.g. to evaluate its composition in drug-facilitated crimes. An important feature in MEPS is the miniaturization of the sorbent. A careful selection of the sorbent will allow working with complex matrices, separating the target analytes from interferences and improve recoveries. The most widely selected sorbent was the silica based C18 that is a popular reversed-phase material (41%). Starting in the 2000s, new modifications of sorbents appeared. Overall, what the future holds for MEPS applications in forensic toxicology is promising, and ongoing research and technological advancements are likely to enhance the capabilities of MEPS approaches, making this technique an increasingly valuable tool for toxicological investigations.

The biological individualization of samples is studied through DNA STR loci analysis and is the principal goal of Forensic Genetics. STR loci study was initiated in the 90s on the majority of Forensic Laboratories. vWA is one of the most studied STR loci all over the world. It is included in any STR set proposed by Forensic Groups as the 13 core loci of the CODIS System used in routine cases by most Forensic Genetic Laboratories. When performing paternity investigation and identification cases with STR loci, once in a while we came through rare or new alleles. Sequencing is essential to confirm the structure of these alleles. Also in mutation cases, especially in primer binding mutation cases, allele sequence should be performed. The high primer binding mutation observed in this locus and the interest in structure analysis of rare alleles observed in certain population groups were the main reasons for performing vWA sequence. More than one hundred vWA samples have been sequenced with DNA Sequencing Kit Big Dye Terminator Cycle Sequencing with forward and reverse primers (Kimpton et al, 1992 GenBank M25858). Sequencing was performed in an ABIPRISM 377 DNA Sequencer and analyzed by Sequencing Analysis. In this study we will present the structural analysis of vWA alleles from allele 11 to allele 22 studied in two main populations – a Portuguese population and an African population mainly from Cabo Verde Islands and Angola

Amphetamines consumption is still an important public health issue, namely in terms of compounds variability and disposition to consumers. However, some of them, still classic substances, keep standing in the illicit market, with remarkable and continuous success. That is the case of MDMA. Nevertheless, there is always new information and data on MDMA intoxication, both in vivo as in post-mortem context. The authors report an intoxication case with MDMA in an 18 years old male, who was considered a potential organ donor, after a cardiac arrest. Whole blood samples were collected in vivo, at the Emergency Room (ER), in different moments, and post-mortem, at the Forensic Pathology Service of the National Institute of Legal Medicine and Forensic Science. After a general screening procedure, samples were extracted by an SPE procedure (OASIS® MCX) followed by a GC-MS single quad analysis. The post-mortem whole blood sample was positive for Lidocaine (< 500 ng/mL), compatible with ER intervention, and positive for MDMA (2278 ng/mL) and MDA (49 ng/mL), while the whole blood samples collected in vivo (during the maintenance of the individual under advanced life support), were positive for MDMA (between 504 ng/mL and 1918 ng/mL) and MDA (between 20 ng/mL and 89 ng/mL). The samples were negative for other substances, namely ethanol, other drugs of abuse and medicines. Interpretation of these results is pivotal to understand the behaviour of the substance. So, in this case, MDMA post-mortem behaviour should be carefully evaluated, considering as possible influencers, in the specific context of the case, the time lapse between the death verification, the maintenance of the advanced life support and the body manipulation for organ collection purposes. Also referred and discussed is the ante-mortem/post-mortem ratio of MDMA obtained values, compared with literature references. No doubt that death was due to MDMA intoxication, but information from the in vivo samples analysis suggests that this type of sample should also be considered, in a complementary role, whenever possible.

Acetic acid treatment for an accurate differentiation between ancient and recent human bones was assessed using Raman and FTIR-ATR spectroscopies. Each set of skeletal samples was analysed by these techniques, prior and after chemical washing, in order to determine the variations in bone´s chemical composition and crystallinity. Bone samples were collected from several independent sources: recent bones burned under controlled experimental conditions or cremated, and archaeological (XVII century and Iron Age). The effect of acetic acid, expected to impact mostly on carbonates, was clearly evidenced in the spectra of all samples, particularly in FTIR-ATR, mainly through the bands typical of A- and B-carbonates. Furthermore, as seen for crematoria and archaeological samples, acetic acid was found to remove contaminants such as calcium hydroxide. Overall, acetic acid treatment can be an effective method for removing carbonates (exogenous but possibly also endogenous) and external contaminants from bone. However, these effects are dependent on the skeletal conditions (e.g. post-mortem interval and burning settings). In addition, this chemical washing was shown to be insufficient for an unequivocal discrimination between recent and archaeological skeletal remains. Based on the measured IR indexes, only cremated bones could be clearly distinguished.