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- Forensic genetic analysis of South Portuguese population with the six dye Powerplex® Fusion 6CPublication . Vieira Da Silva, Cláudia; Afonso Costa, Heloísa; Porto, Maria João; Cunha, E; Corte Real, F.; Amorim, AntónioAs an improvement in efficiency and in Human Discrimination Power, the new six dye multiplex kit PowerPlex® Fusion 6C System, by Promega, available for human identification can co-amplify 27 loci, in a single reaction, have been introduced in the last years with great success. This kit allows the amplification and detection of autosomal loci included in the expanded Combined DNA Index System CODIS, plus the loci Penta D, PENTA E and SE33 as well as Amelogenin for gender determination. Furthermore, this kit includes three Y –STRs (DYS391, DYS576 and DYS570), allowing allelic attribution in a total of 27 loci. This genetic markers extension satisfies not only CODIS but also European Standard Set recommendations. Thinking about continuous human migration movements, especially in a very cosmopolitan region like Lisbon and south of Portugal, and also, in keeping population studies and actualized STR databases we decided to update our previous studies. Our sample is composed of 600 unrelated individuals, from paternity testing with laboratory identity anonymised. DNA was extracted by Prep-n-go BufferTM(Thermo-Fisher Scientific). PCR amplification was performed with PowerPlex® Fusion 6C System, according to manufacturer’s guidelines. Fragment analysis was carried out in an Applied Biosystems® 3500 Genetic Analyser. Electrophoresis results were analysed with GeneMapper® ID-X v1.4. Allele frequencies and population statistics, including Hardy-Weinberg equilibrium p-values from exact test probabilities and forensic parameters were calculated with adequate software. In conclusion, our population information was updated in order to apply most recent data in our casework weight of evidence.
- Investigação de parentesco biológico: a importância de marcadores adicionais em casos de especial complexidadePublication . Rodrigues, Diogo; Vieira Da Silva, Cláudia; Carvalho, Mónica; Afonso Costa, Heloísa; Sampaio, Lisa; Cunha, E; Corte Real, F.; Amorim, AntónioA grande maioria das perícias de investigação de parentesco biológico realizadas pelo Instituto Nacional de Medicina Legal e Ciências Forenses (INMLCF) têm inicio com a ordem do Tribunal para realização da mesma. O mais frequente é o Tribunal dar ordem para nos serem presentes, como intervenientes, um trio constituído por um suposto pai, uma mãe e um(a) filho(a), havendo, no entanto, variações quanto ao número ou tipo de intervenientes, o que pode resultar em maior dificuldade em apresentar resultados com a robustez desejada. Em qualquer dos casos, genericamente, as conclusões possíveis de um estudo de parentesco biológico e mais concretamente de um estudo de paternidade são a exclusão ou não exclusão de paternidade relativamente ao suposto pai em estudo. A conclusão pela não exclusão é sempre acompanhada pela valorização estatística dos resultados, designadamente através do cálculo e apresentação do Índice de Parentesco (IP) e da Probabilidade de Parentesco (W). No caso de uma perícia de investigação de parentesco em que nos seja presente unicamente um filho biológico do suposto pai e um suposto filho biológico, sem possibilidade de estudo das amostras das respetivas mães biológicas nem da amostra biológica do suposto pai, e nos é pedido o estudo sobre a possibilidade de ambos serem filhos biológicos do mesmo pai, a impossibilidade de exclusão da paternidade pode estar associada a valores calculados de IP que podem ser pouco robustos. O ensaio Investigator® HDplex permite o estudo de marcadores genéticos adicionais aos habitualmente estudados na rotina pericial do INMLCF.
- QUANTIFILER®TRIO DNA method performance in a collection of ancient samplesPublication . Vieira- Silva, C.; Lopes, J.; Afonso Costa, H.; Ribeiro, T.; Porto, M.J.; Dias, M; Cunha, E; Amorim, A,During the past few years significant progress has been made in solving technical challenges associated with STR profiling including the ability to analyze degraded DNA and low amounts of DNA. The result of these changes is that useful STR profiles can now be obtained from previously untypeable forensic DNA samples. Analysis of DNA from ancient material represents an important role in molecular anthropology, although there are many limitations concerning low DNA quantity and/or degraded DNA, and/or PCR inhibitors. These factors can make it difficult to decide whether to continue with STR analysis, which STR panel to use and how much DNA to add to PCR reaction. With all these constraints, DNA quantification represents an important tool to decide which method will follow in order to improve workflow and have good results in less time-consuming. The Quantifiler® Trio DNA method provides a quality index (QI) to detect the presence of degraded DNA along with PCR inhibitors.This guide allows the selection of the optimal short tandem repeat (STR) analysis chemistry (autosomal, or miniSTR) and streamlines the workflow while increasing downstream analysis success rates. In order to compare DNA quality from different extraction methods, samples from 46 exhumed Middle Ages individuals were extracted with modified phenol-chloroform method and also PrepFiler Express BTA™ method. DNA was quantified with Quantifiler® Trio DNA Quantification in an Applied Biosystems® 7500 Real-Time PCR System. Results were analyzed and allow us to point Quantifiler® Trio method as an important tool in pre-STR typing methods in ancient samples
- Technical Note: The Forensic Anthropology Society of Europe (FASE) Map of Identified Osteological CollectionsPublication . Petaros, Anja; Caplova, Zuzana; Verna, Emeline; Adalian, Pascal; Baccino, Eric; de Boer, Hans H.; Cunha, E; Ekizoglu, Oguzhan; Ferreira, Maria Teresa; Fracasso, Tony; Kranioti, Elena F.; Lefevre, Philippe; Lynnerup, Niels; Ross, Ann; Steyn, Maryna; Obertova, Zuzana; Cattaneo, CristinaIdentified (documented) osteological collections represent an important resource in the development of forensic anthropology standards and methods as well as a precious tool for learning and training of practitioners. Even though the number of papers presenting identified collections worldwide increases, many of the collections have still not been divulged to the scientific community in sufficient detail to ascertain their exact number. The Forensic Anthropology Society of Europe (FASE) therefore developed a tool that goes beyond sporadic publications: the FASE Map of Identified Osteological Collections, which is freely accessible and continuously updated and revised. The online map is available at http://forensicanthropology.eu/osteological-collections/. The map of skeletal collections was created in 2017 and currently displays information on 153 identified osteological collections (43 of them categorized as contemporary) located in 41 different countries. This article offers a short analysis of the type, geographical location and content of the collections included in the map. The aim of this article and the map as such is to provide a useful resource to facilitate research planning and teaching in forensic anthropology and related disciplines.
- Challenges and (Un)Certainties for DNAm Age Estimation in FuturePublication . Correia Dias, Helena; Cunha, E; Corte Real, F.; Manco, LicínioAge estimation is a paramount issue in criminal, anthropological, and forensic research. Because of this, several areas of research have focused on the establishment of new approaches for age prediction, including bimolecular and anthropological methods. In recent years, DNA methylation (DNAm) has arisen as one of the hottest topics in the field. Many studies have developed age- prediction models (APMs) based on evaluation of DNAm levels of many genes in different tissue types and using different methodological approaches. However, several challenges and confounder factors should be considered before using methylation levels for age estimation in forensic contexts. To provide in-depth knowledge about DNAm age estimation (DNAm age) and to understand why it is not yet a current tool in forensic laboratories, this review encompasses the literature for the most relevant scientific works published from 2015 to 2021 to address the challenges and future directions in the field. More than 60 papers were considered focusing essentially on studies that developed models for age prediction in several sample types
- El papel de las Ciencias Forenses en la identificación de restos óseos - informe de un caso (poster)Publication . Costa Lopes, Miguel; Abreu, Ana; Amorim, António; Ribeiro, Teresa; Cunha, Eugénia; Eiras, Luísa
- Automated age‐at‐death estimation by cementochronology: Essential application or additional complication?Publication . Bertrand, Benoit; Vercauteren, Martine; Cunha, E; Bécart, Anne; Gosset, Didier; Hédouin, ValeryIt 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.
- The unidentified skeletal collection of Capuchos Cemetery (Santarém) housed at the University of CoimbraPublication . José, Andreia; Tomé, Laura; Coelho, Catarina; Cunha, E; Umbelino, Cláudia; Ferreira, Maria TeresaThe aim of this article is to present to the scientific and academic community a new osteological collection housed at the Department of Life Sciences from the University of Coimbra (Portugal), the Unidentified Skeletal Collection of the Capuchos Cemetery. The skeletons were collected from the same cemetery as those of the 21st Century Identified Skeletal Collection, however their identity is unknown. The collection is comprised of 73 individuals, of which 68 are adults of both sexes (34 females, 33 males, and one individual of unknown sex) and five are non-adult individuals. It is estimated that the majority of adult individuals have a European ancestry (n = 52; 76.47%). The skeletons are reasonably preserved, although several are incomplete (n = 8; 10.96%) as result of both taphonomic changes occurred during the burial, and also due to experimental researches performed (some of which destructive). Regarding the osteopathology, it was observed that the majority of adult individuals have pathological changes (n = 67; 98.53%), with degenerative pathology being the most frequent in the sample. In addition, 13 individuals (17.81%) exhibit medical devices and/or signs of surgical procedures.The Unidentified Skeletal Collection of the Capuchos Cemetery is an osteological collection that, although not containing individual biographical data, has contributed to teaching, research and development of new methods for Biological and Forensic Anthropology in subjects such as osteology, morphology, biological profile, paleopathology, and cremains
- Pesquisa de SARS-CoV-2 em cadáveres: experiência da Delegação do Sul do Instituto Nacional de Medicina Legal e Ciências ForensesPublication . Logrado, Diana; Inácio, Ana Rita; Amorim, António; Santos, Carlos dos; Cunha, E