INMLCF - Artigos Científicos
Permanent URI for this collection
Browse
Recent Submissions
- Sudden Cardiac Death: The Role of Molecular Autopsy with Next-Generation SequencingPublication . Santos, Agostinho; Amorim, António; Cainé, Laura; Fadoni, JenniferMolecular autopsy is a term employed to describe the investigation of the cause of death through the analysis of genetic information using biological samples collected post-mortem. Its utility becomes evident in situations where conventional medico-legal autopsy methods are not able to identify the cause of death, i.e., in sudden cardiac death (SCD) cases in young individuals, where deaths are commonly due to genetic cardiac conditions, such as cardiomyopathies and channelopathies. The recent advancement in high-throughput sequencing techniques, such as next-generation sequencing (NGS), has allowed the investigation of a high number of genomic regions in a more cost-effective and faster approach. Unlike traditional sequencing methods, which can only sequence one DNA fragment at a time, NGS can sequence millions of short polynucleotide fragments simultaneously. This parallel approach reduces both the time and cost required to generate large-scale genomic data, making it a useful tool for applications ranging from basic research to molecular autopsy. In the forensic context, by enabling the examination of multiple genes or entire exomes and genomes, NGS enhances the accuracy and depth of genetic investigations, contributing to a better understanding of complex inherited diseases. However, challenges remain, such as the interpretation of variants of unknown significance (VUS), the need for standardized protocols, and the high demand for specialized bioinformatics expertise. Despite these challenges, NGS continues to offer significant promise for enhancing the precision of molecular autopsies. The goal of this review is to assess the effectiveness of contemporary advancements in molecular autopsy methodologies when applied to cases of SCD in young individuals and to present an overview of the steps involved in the analysis of NGS data and the interpretation of genetic variants.
- X-chromosomal STRs: Metapopulations and mutation ratesPublication . Gusmão, L.; Antão-Sousa, S.; Faustino, M.; Abovich, M.A.; Aguirre, D.; Alghafri, R.; Alves, C.; Amorim, A.; Arévalo, C.; Baldassarri, L.; Barletta-Carrillo, C.; Berardi, G.; Bobillo, C.; Borjas, L.; Braganholi, D.F.; Brehm, A.; Builes, J.J.; Cainé, Laura; Carvalho, E.F.; Carvalho, Mónica; Catelli, L.; Cicarelli, R.M.B.; Contreras, A.; Corach, D.; Di Marco, F.G.; Diederiche, M.V.; Domingues, P.; Espinoza, M.; Fernandéz, J.M.; García, M.G.; García, O.; Gaviria, A.; Gomes, I.; Grattapaglia, D.; Henao, J.; Hernandez, A.; Ibarra, A.A.; Lima, G.; Manterola, I.M.; Marrero, C.; Martins, J.A.; Mendoza, L.; Mosquera, A.; Nascimento, E.C.; Onofri, V.; Pancorbo, M.M.; Pestano, J.J.; Plaza, G.; Porto, M.J.; Posada, Y.C.; Rebelo, M.L.; Riego, E.; Rodenbusch, R.; Rodríguez, A.; Rodríguez, A.; Sanchez-Diz, P.; Santos, S.; Simão, F.; Siza Fuentes, L.M.; Sumita, D.; Tomas, C.; Toscanini, U.; Trindade-Filho, A.; Turchi, C.; Vullo, C.; Yurrebaso, I.; Pereira, V.; Pinto, N.The analysis of STRs located on the X chromosome has been one of the strategies used to address complex kinship cases. Its usefulness is, however, limited by the low availability of population haplotype frequency data and lack of knowledge on the probability of mutations. Due to the large amount of data required to obtain reliable estimates, it is important to investigate the possibility of grouping data from populations with similar profiles when calculating these parameters. To better understand the partition of genetic diversity among human populations for the X-STRs most used in forensics, an analysis was carried out based on data available in the literature and new data (23,949 haplotypes in total; from these 10,445 new) obtained through collaborative exercises within the Spanish and Portuguese Working Group of the International Society for Forensic Genetics. Based on the available population data, a similarity in X-STR profiles was found in European populations, and in East Asian populations, except for some isolates. A greater complexity was found for African, South American, and South and Southeast Asian populations, preventing their grouping into large metapopulations. New segregation data on 2273 father/mother/daughter trios were also obtained, aiming for a more thorough analysis of X-STR mutation rates. After combining our data with published information on father/mother/daughter trios, no mutations were detected in 13 out of 37 loci analyzed. For the remaining loci, mutation rates varied between 2.68 × 10−4 (DXS7133) and 1.07x10−2 (DXS10135), being 5.2 times higher in the male (4.16 ×10−3) than in the female (8.01 ×10−4) germline.
- Phytocanabinoids and synthetic cannabinoids: from recreational consumption to potential therapeutic use – a reviewPublication . Teixeira, Helena M.Cannabinoids are part of the most popular group of illicit substances in the Western world. The word “cannabinoid” refers to any chemical substance, regardless of structure or origin, that binds to the body’s cannabinoid receptors and that has effects similar to those produced by the Cannabis plant. Regarding their origin, cannabinoids can be classified into endocannabinoids, phytocannabinoids and synthetic cannabinoids. The behavioral and physiological effects of cannabinoids have received particular attention over the last few decades, including sensations of euphoria, relaxation and loss of concentration, with their repeated use being associated with short and long-term side effects, including respiratory and cardiovascular disorders, cognitive changes, psychoses, schizophrenia and mood disorders. On the other hand, recent investigations have proposed a promising therapeutic potential of cannabinoid-based drugs for a wide range of medical situations, including neurological and psychiatric disorders, among other indications. The growing popularity in the use of cannabinoid-based compounds, both for recreational and therapeutic purposes, has been accompanied by an equally continuous and growing evolution of knowledge regarding their potential harmful and beneficial effects. However, there are several open questions and challenges to be answered, which require more and better investigations. This article’s main objectives are: i) to understand the importance of the action of cannabinoids in humans; ii) identify the different types of cannabinoids that exist and understand the differences in their action; iii) distinguish the legislative framework for cannabinoid consumption; iv) identify the possible adverse effects of cannabinoid consumption, as well as their potential benefits; v) know the existing medical-scientific evidence in terms of therapeutic potential, particularly in relation to aspects of safety and efficacy; vi) encourage critical thinking about the recreational consumption and therapeutic use of cannabinoids, based both on currently available evidence and gaps in knowledge.
- Determination of Arylcyclohexylamines in Biological Specimens: Sensors and Sample Pre-Treatment ApproachesPublication . Pelixo, Rodrigo; Barroso, Mário; Gallardo, Eugenia; Rosado, TiagoArylcyclohexylamine (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.
- Volumetric Absorptive Microsampling in ToxicologyPublication . Pires, Bruno; Catarro, Gonçalo; Soares, Sofia; Gonçalves, Joana; Rosado, Tiago; Barroso, Mário; Araujo, André R. T. S.; Gallardo, EugeniaVolumetric absorptive microsampling (VAMS) is an emerging technique in clinical and forensic toxicology. It is recognized as a promising alternative to traditional sampling methods, offering an accurate and minimally invasive means of collecting small volumes of biological samples, such as blood, urine, and saliva. Unlike conventional methods, VAMS provides advantages in terms of sample stability, storage, and transportation, as it enables samples to be collected outside laboratory environments without requiring refrigeration. This review explores several VAMS methodologies, with a particular focus on its application for the quantification of drugs and other substances in clinical and forensic toxicology. It compares VAMS to other microsampling techniques, such as dried blood spots (DBSs), highlighting VAMS's superiority in addressing issues related to sample volume consistency and environmental impact. Despite its advantages, VAMS also presents certain limitations, including higher costs and difficulties in detecting underfilled samples. Overall, VAMS stands out as a microsampling technique with the potential to enhance patient compliance and operational efficiency, positioning itself as a viable tool for toxicological analysis in both clinical and forensic contexts.
- Determination of Antiepileptics in Biological Samples—A ReviewPublication . Martinho, João; Simão, Ana Y.; Barroso, Mário; Gallardo, Eugenia; Rosado, TiagoEpilepsy remains a disease that affects many people around the world. With the development of new drugs to treat this condition, the importance of therapeutic drug monitoring continues to rise and remains a challenge for the medical community. This review article explores recent advances in the detection of antiepileptic drugs across various sample types commonly used for drug monitoring, with a focus on their applications and impact. Some of these new methods have proven to be simpler, greener, and faster, making them easier to apply in the context of therapeutic drug monitoring. Additionally, besides the classic use of blood and its derivatives, there has been significant research into the application of alternative matrices due to their ease of sample collection and capacity to reflect drug behavior in blood. These advances have contributed to increasing the efficacy of therapeutic drug monitoring while enhancing its accessibility to the population.
- α-Pyrrolidinohexanophenone (α-PHP) and α-Pyrrolidinoisohexanophenone (α-PiHP): A ReviewPublication . Dinis, Pedro; Franco, João; Margalho, CláudiaNew Psychoactive Substances are currently a serious and growing problem affecting public health worldwide. By 2022, 1184 of these substances had been identified over a period of 16 years. Within these, α-pyrrolidinohexanophenone (α-PHP) and α-pyrrolidinoisohexanophenone (α-PiHP) have emerged, two synthetic cathinones from the pyrovalerone derivates subgroup that are positional isomers of each other. Alpha-PHP appeared on the Japanese illicit drug market in 2014 and, two years later, α-PiHP was identified for the first time in China. They were placed in schedule II on the list of Psychotropic Substances under International Control in 2020 and in March 2023, respectively. Both cathinones have no therapeutic potential for medical use and therefore are abused for recreational habits, which can lead to fatalities. The most frequent adverse effects reported are cardiac, psychiatric, and neurologic, and fatal intoxications have already been described. In Portugal, their consumption and consequent seizures are more prevalent on the archipelagos, which has been aggravating the health situation. In conclusion, these types of substances are a challenge for forensic toxicology since they are easily synthesized, modified, and placed on the market. Therefore, more studies to develop analytical methods to detect them and more comprehensive legislation should be applied. Thus, this review aimed to address the legislative, physicochemical, toxicological, and analytical aspects of both substances.
- Human identification by medical findings in a forensic anthropology contextPublication . Lemos, Yara Vieira; Furtado, Alexandre Neves; Lima, Adriana Zatti; Dionísio, Alexander Santos; Araújo, Ricardo Moreira; Cunha, EugéniaThis article presents a series of three complex forensic cases that posed significant challenges for identifying human remains. These include a mass dam disaster, burnt human remains, and extensively decomposed human remains. Positive identification was achieved using a shadow positioning technique with imaging comparisons of medical findings. After establishing the biological profile, medical data were evaluated with digital radiography and computed tomography examinations the human remains. These aimed to replicate the original (intravitam) traits in the same angulation to examine postsurgical characteristics, as well as the anatomical, pathological, and morphological features, which were sufficient to establish a positive scientific identification. Technological advancements tend to reveal additional skeletal details, making medical data comparisons significantly more effective in the context of anthropological identification. These cases demonstrate that the possibility of identification should never be ignored, even in situations with advanced decomposition.
- Solving cold cases: The importance of new identification techniques in old cases of forensic anthropologyPublication . Santos, Gabriela Araujo dos; Castro Espicalsky, Talita Lima de; Custódio, Leticia Regina Alfaia; Calmon Silva, Melina; Cunha, EugéniaScientific and technological developments experienced by forensic sciences have increased the possibility of human identification. Forensic anthropology has developed worldwide and has made significant progress in Brazil in the last 10 years. This context has allowed forensic medicine institutes to use new techniques to resolve cold cases of human identification. This work aimed to report the methodology, development, and partial results of the LAFOL Identifies Operation, carried out at the Forensic Anthropology and Forensic Dentistry Laboratory (LAFOL) of the Medico-Legal Institute of Porto Velho. We carried out a multidisciplinary expert approach on human skeletal remains archived over the last 20 years to identify these individuals using current scientific methods. Eighty-one cold cases of skeletonized and/or mummified human skeletal remains were reanalyzed; they were archived in the LAFOL Laboratory collection as unidentified. Anthropological analyses showed that the collection is predominantly composed of adult individuals (86.4%), between 21 and 39 years old (55.6%), and males (62.9%). In the human skeletal remains identified, forensic anthropology played a key role in the case screening process. We expected that the results presented in this work will inspire other Medico-Legal Institutes in Brazil and worldwide to establish task forces for the reanalysis of cold cases in their institutes, promoting new forensic dentistry and anthropological comparisons and collecting biological samples to feed the database of genetic profiles of missing persons.
- Cranial surgical approaches in the 21st Century Identified Skeletal CollectionPublication . Ferreira, Maria Teresa; Coelho, Catarina; Gaspar, Rosa Ramos; de Matos, Daniela P.; Pereira, Ricardo; Cunha, Eugénia; Oliveira-Santos, InêsIn Forensic Anthropology, identification is comparing information from a person whose fate and whereabouts are unknown with data from a person who is present but whose identity is unknown. The rarer and more distinctive the characteristics observed during the anthropological investigation, the greater the probability of reaching an identification. Taking this into account, cranial surgeries can be considered good individualization factors. The cases of skulls with lesions compatible with surgical procedures from the 21st Century Identified Skeletal Collection are presented, and their identification potential is discussed. This collection comprises 302 adult individuals, of which four females (aged 55 to 94 years) and four males (aged 67 to 92 years) present cranial lesions of surgical origin. The results of the macroscopic and imaging analyses (conventional x-rays and CTs) are presented, along with a detailed description and location of each lesion. In five cases there is clear evidence of bone remodelling. This research, which combines anthropological analysis with imaging and neurosurgical insights, demonstrates the importance of correctly interpreting these lesions, illustrating how important is a complete and interdisciplinary analysis for forensic expertise