Ciências e Técnicas Laboratoriais Forenses

Palavras d´El Rei ? Uma Investigação Forense Multidisciplinar do Enigmático Memorando de 1919

Sun, 01 Jan 2023 00:00:00 GMT

Palavras d´El Rei ? Uma Investigação Forense Multidisciplinar do Enigmático Memorando de 1919 Ribeiro, Rogério Eduardo Cerqueira Pereira Na sequência da implantação da República, entre 1910 e 1926, Portugal viveu um período politicamente conturbado. Com D. Manuel II exilado em Inglaterra, alguns monárquicos em Portugal, ansiando pela restauração da Monarquia, solicitaram o aval do Rei exilado para dar início a um movimento revolucionário militar. Foi então elaborado um “Memorando”, que após alegada consulta ao Rei, por parte do seu Conselheiro Ayres de Ornellas, passou a ostentar a enigmática expressão “Go On! Palavras d'El Rei”, indicando o suposto consentimento. Conhecendo-se a oposição de D. Manuel à restauração da monarquia pela via revolucionária, preferindo a via política, pacífica e ordenada, face ao documento no qual, alegadamente, anuiu com a manobra revolucionária, emerge uma contradição. O presente trabalho pretende contribuir para o esclarecimento da mesma, pelo relacionamento de diversos factos históricos e pela avaliação da veracidade/autoria dos manuscritos do Memorando. Para tal, foram pesquisados diferentes arquivos bibliográficos e analisados diversos documentos e obras que permitiram aprofundar o enquadramento histórico e encontrar, na verdade, dois Memorandos: i) “Memorando Manuscrito”, detentor de um corpo de texto supostamente manuscrito por Hipólito Raposo (político monárquico) e de manuscritos laterais de suposta autoria de Ayres de Ornellas (supostamente assinados pelo mesmo), e ii) “Memorando Dactilografado”, detentor de um corpo de texto dactilografado e de manuscritos laterais de suposta autoria de Ayres de Ornellas (supostamente assinados pelo mesmo). Desta forma, foram também recolhidos manuscritos da autoria inequívoca de Hipólito Raposo e de Ayres de Ornellas para tornar possível a investigação forense. Assim, a Escrita Manual e a Linguística constante dos manuscritos de autoria inequívoca, foram então comparadas, com a Escrita Manual e a Linguística constante dos manuscritos presentes nos dois Memorandos. Foram ainda utilizados Métodos Computacionais para coadjuvar a análise da Escrita Manual realizada. A análise comparativa de Escrita Manual realizada permitiu afirmar que as várias similitudes e as reduzidas diferenças encontradas, entre os manuscritos de autoria inequívoca e os constantes nos dois Memorandos, suportam, moderadamente, as seguintes preposições: i) Hipólito Raposo é o autor da escrita manuscrita do corpo do texto presente no Memorando Manuscrito, ii) Ayres de Ornellas é o autor dos manuscritos laterais presentes no Memorando Manuscrito, iii) Ayres de Ornellas é o autor da escrita da assinatura presente no Memorando Manuscrito, iv) Ayres de Ornellas é o autor dos manuscritos laterais presentes no Memorando Dactilografado, e v) Ayres de Ornellas é o autor da escrita da assinatura presente no Memorando Dactilografado. Quanto à análise computacional da Escrita Manual, as correspondências da distribuição dos ângulos dos contornos das letras não permitiram atribuir uma maior probabilidade de autoria a Hipólito Raposo ou a Ayres de Ornellas nos diversos documentos comparados, devido a diferenças insuficientes entre autores e, por vezes, cumulativamente, resultados inconsistentes. Desta forma os Métodos Computacionais deram um resultado inconclusivo. Por último, através da análise de Linguística Forense, após a análise dos corpora de textos e marcadores de estilo, foi difícil realizar a atribuição de uma autoria inequívoca aos corpora de texto analisados. Contudo, foi possível identificar, embora em número insuficiente, diversas características linguísticas e marcadores linguísticos comuns, no sentido de identificar Hipólito Raposo como possível autor do corpo do texto e Ayres de Ornellas como possível autor dos manuscritos laterais presentes no Memorando Manuscrito. Pese embora os resultados da análise computacional da Escrita Manual tenham sido inconclusivos e os resultados da análise de Linguística Forense não tenham permitido chegar a conclusões assertivas de autoria, os mesmos também não foram contraditórios com os resultados obtidos na análise de Escrita Manual. Desta forma, podemos assim concluir, com moderada certeza, que Hipólito Raposo e Ayres de Ornellas são os autores dos dizeres manuscritos e assinaturas atribuídas aos mesmos, conforme anteriormente descrito, constantes dos Memorandos. Tal facto, apoia a teoria de que o Rei alegadamente consentiu a tentativa da restauração da Monarquia pela via revolucionária. Palavras-Chave: Rei D. Manuel II; Hipólito Raposo; Ayres de Ornellas; Análise de Escrita Manual; Linguística Forense.; Following the implementation of the Republic, between 1910 and 1926, Portugal lived a politically unstable period. With D. Manuel II exiled in England, some monarchists in Portugal, anxious for the restoration of the Monarchy, requested the endorsement of the exiled King to begin a revolutionary military movement. A “Memorando” was then written, which, upon alleged consultation to the King by his Counselor Ayres de Ornellas, appeared presenting the enigmatic handwritten expression “Go On! Palavras d'El Rei”, indicating the purported consent. Knowing D. Manuel´s opposition to the restoration of the Monarchy through revolutionary means, preferring the peaceful, political, and orderly way, facing the document in which, allegedly, granted the revolutionary manoeuvre, a contradiction emerges. This present work aims to contribute to the clarification of the same, relating several historical facts and evaluating the authenticity/authorship of the Memorando manuscripts. For that purpose, different bibliographical archives were researched and several documents and works were analysed, allowing to deepen the historical framework and find, in fact, two Memorandos: i) "Manuscript Memorando", holder of a body text supposedly handwritten by Hipólito Raposo (monarchical politician) and side manuscripts of supposed authorship of Ayres de Ornellas (supposedly signed by the same), and ii) "Typewritten Memorando", holder of a typewritten body text and side manuscripts of supposed authorship of Ayres de Ornellas (supposedly signed by the same). In this way, manuscripts of unequivocal authorship of Hipólito Raposo and Ayres de Ornellas were also collected, making the forensic investigation possible. Therefore, the Handwriting and Linguistic of the manuscripts of unequivocal authorship were then compared with the Handwriting and Linguistic of the manuscripts present on both Memorandos. Computational Methods were additionally used to assist the carried-out Handwriting analysis. The performed comparative analysis of Handwriting allowed to affirm that the various similarities and reduced differences that were found, between the manuscripts of unequivocal authorship and the ones on both Memorandos, support, moderately, the following prepositions: i) Hipólito Raposo is the author of the handwriting of the body text present in the Manuscript Memorando, ii) Ayres de Ornellas is the author of the side manuscripts present in the Manuscript Memorando, iii) Ayres de Ornellas is the author of the signature handwriting present in the Manuscript Memorando, iv) Ayres de Ornellas is the author of the side manuscripts present in the Typewritten Memorando, and v) Ayres de Ornellas is the author of the signature handwriting present in the Typewritten Memorando. As for the computational analysis of the handwriting, the correspondences of the distribution of the angles of the contours of the letters did not allow assigning a higher probability of authorship to Ayres de Ornellas or to Hipólito Raposo in the several compared documents, due to insufficient differences between authors and, sometimes, cumulatively, inconsistent results. Therefore, the Computational Methods gave an inconclusive result. Finally, through the Forensic Linguistic analysis, after analysing the corpora of texts and style markers, it was difficult to attribute an unequivocal authorship to the corpora of texts analysed. However, it was possible to identify, although in insufficient number, several common linguistic characteristics and style markers, in order to identify Hipólito Raposo as a possible author of the body text and Ayres de Ornellas as a possible author of the side manuscripts present in the Manuscript Memorando. Although the results of the computational analysis of Handwriting had been inconclusive and the results of Forensic Linguistics have not allowed to reach assertive conclusions, the same were also not contradictory to the results obtained in the Handwriting analysis. Therefore, we can conclude, with moderated certainty, that Hipólito Raposo and Ayres de Ornellas are the authors of the handwritten sayings and signatures attributed to them, as previously described, in the Memorandos. This fact supports the theory that the King allegedly consented the attempt to restore the Monarchy through revolutionary means.

Toxicity of the 3,4- methylenedioxymethamphetamine (MDMA) and its enantiomers to Daphnia magna, after isolation by semipreparative chromatography

Sun, 01 Jan 2023 00:00:00 GMT

Toxicity of the 3,4- methylenedioxymethamphetamine (MDMA) and its enantiomers to Daphnia magna, after isolation by semipreparative chromatography Costa, Ana Rita Fernandes Miranda da Psychoactive substances (PAS) have been frequently documented in aquatic systems causing oncern for their potential to interfere with biochemical, cellular, physiological,and behavioural mechanisms of non-target organisms. 3,4-Methylenedioxymethamphetamine (MDMA) is among the most consumed PAS in theworld. However, the United States Food and Drugs Administration (USFDA) recentlyapproved a trial to assess its pharmacological potential in patients with post-traumaticstress disorder. MDMA is a chiral substance, sold on the illicit market exclusively as a racemate (R,SMDMA).After consumption, human metabolism is enantioselective, S(+)-MDMA undergo preferential metabolism over R(−)-MDMA, which leads to enrichment of the R(−)-enantiomer in excretions. Its occurrence in the environment arises from directdisposal of sewage, clandestine laboratories or ischarges of effluents from Wastewater Treatment Plants (WWTPs) due to the ineficciency of WWTP to complete eliminatedrugs residues. Studies on the toxicity of this compound in non-target organisms are scarce and lack of information on enantioselectivity. Thus, the objective of this work was to evaluate the enantioselective potential on MDMA toxicity using Daphnia magna as a freshwater animal model. For this, the MDMA enantiomers were separated by semi-preparativechromatography using a semi-preparative column with amylose tris-3,5-dimethylphenylcarbamate adsorbed on minopropyl silica (APS-Nucleosil - 500 Å, 7 μm; 20% g/ g). Enantiomers were obtained with an enantiomeric purity > 97% and used inecotoxicity assay. The sub-chronic assay was initiated with neonates (< 24 h, day 0) through day 8, using three concentrations for the racemate, 0.1, 1.0 and 10.0 μg/L, two concentrations for the enantiomers (0.1 and 1.0 μg/L) and a control group. Each experimental unit consisted of a group of 15 organisms and 5 replicates for each concentration or control and morphophysiological, behavioural, reproductive and biochemical parameters were determined at different stages of the organism'sdevelopment. Changes were observed for some of the analyzed parameters as well as enantioselectivity. For example, an increase in body size was observed in organisms exposed to (R,S)-MDMA at day 8 (adults) and an enantioselective effect with significantly reduced body growth in organisms xposed to the S(+)-enantiomer also at day 8 (adults). Changes in ii swimming behaviour were observed with increasing swimming speed and total distance travelled in organisms exposed to (R,S)-MDMA at all concentrations. On the contrary, a decrease in the total distance travelled was observed in organisms exposed to the enantiomers but enantioselective effects were not observed. No reproductive or biochemical changes were observed in either racemate or enantiomer exposure except for acetylcholinesterase and catalase activity, whose activity decreased in organisms exposed to the highest concentration of (R,S)-MDMA (10 μg/L). This study demonstrated that MDMA can affect the development and swimming behaviour of daphnia including at environmental concentrations and that these effects may be enantioselective, but no reproductive and biochemical changes were observed for the majority of the parameters analysed. However, it is essential to carry out additional studies to complement the resultsobtained, for an accurate assessment of the potential environmental risks of thissubstance.

Characterisation of organic and inorganic constituents in cemetery soils and its application as forensic evidence

Sat, 01 Jan 2022 00:00:00 GMT

Characterisation of organic and inorganic constituents in cemetery soils and its application as forensic evidence Queirós, Sara Filipa de Sousa O solo é uma matriz complexa, composta por matéria inorgânica e orgânica, utilizada em múltiplos contextos, incluindo na investigação forense. A sua elevada capacidade de retenção permite estudar compostos provenientes de um corpo em decomposição sobre/dentro do solo, associá-los às diferentes fases de decomposição cadavérica, e estimar há quanto tempo o corpo está naquele local. A translação de um corpo humano também pode ser comprovada na eventualidade de se identificarem compostos provenientes da sua decomposição, sem que este esteja no local em análise. A decomposição cadavérica é afetada por fatores extrínsecos e intrínsecos ao corpo. Ao estudar a decomposição cadavérica, devem considerar-se todas as variáveis envolvidas, para que melhor se entenda a relação entre o processo de decomposição e os compostos orgânicos encontrados no solo. Muitos estudos foram desenvolvidos ao longo dos anos, sendo que, por questões éticas, a maioria foi realizada em animais (principalmente porcos). Contudo, existem diferenças que devem ser consideradas, uma vez que podem influenciar as conclusões dos estudos. Assim, torna-se pertinente a realização de novos estudos com humanos, objetivando-se a ampliação do conhecimento científico nesta área. Posto isto, neste trabalho foram recolhidas 69 amostras de solo de 5 sepulturas humanas em 2 cemitérios portugueses (Monte D’Arcos e Castelo de Mértola). Caracterizou-se inorganicamente o solo das campas (pH, cor e humidade) e determinou-se o seu teor de matéria orgânica. Posteriormente, as amostras recolhidas no interior da sepultura foram analisadas por Cromatografia Gasosa Acoplada a Espectroscopia de Massa para identificar ácidos gordos e esteroides. Os resultados obtidos mostraram que as amostras recolhidas a norte do país têm maior conteúdo de humidade, e a sul verificaram-se valores mais altos de pH e matéria orgânica; a análise da cor não revelou grandes diferenças entre os cemitérios. Relativamente à caracterização orgânica, não foram definidos nenhuns marcadores orgânicos que se relacionassem com a fase específica da decomposição cadavérica, nem com a parte do corpo proveniente. No entanto, identificou-se a presença de ácidos gordos característicos da decomposição humana (ácido mirístico, palmítico, oleico e esteárico), comprovando a importância do estudo destes lípidos em casos forenses. Adicionalmente, foi possível verificar que solos mais ácidos e menos ricos em humidade e matéria orgânica, apresentam menos ácidos gordos. Por fim, não foram encontrados esteroides biomarcadores da degradação humana, refletindo a necessidade de uma futura otimização do protocolo aplicado. Futuramente, deverá também alargar-se o estudo a mais cemitérios com diferentes tipologias de solo.; Soil is a complex matrix, composed of inorganic and organic matter, used in multiple contexts, including forensic investigation. Its high retention capacity makes it possible to study compounds from a decomposing body on/within the soil, associate them with the different phases of cadaveric decomposition, and estimate how long the body has been there. The translation of a human body can also be proven if compounds from its decomposition are identified, without the body being at the site under analysis. Cadaveric decomposition is affected by both extrinsic and intrinsic factors to the body. When studying cadaveric decomposition, all the variables involved must be considered to better understand the relationship between the decomposition process and the organic compounds found in the soil. Many studies have been developed over the years, most of which, for ethical reasons, have been carried out on animals (mainly pigs). However, there are differences that should be considered since they may influence the conclusions of the studies. Thus, it becomes pertinent to carry out new studies with humans, aiming expanding scientific knowledge in this area. Therefore, in this study 69 soil samples were collected from 5 human graves in 2 Portuguese cemeteries (Monte D'Arcos and Mértola Castle). The soil of the graves was inorganically characterised (pH, colour and moisture) and its organic matter content was determined. Subsequently, the samples collected inside the grave were analysed by Gas Chromatography Coupled with Mass Spectroscopy to identify fatty acids and steroids. The results obtained showed that the samples collected in the north of the country have higher moisture content, and in the south, there were higher values of pH and organic matter; the colour analysis did not reveal great differences between the cemeteries. Regarding the organic characterisation, no organic markers were defined that could be related with the specific phase of the cadaveric decomposition, nor with the part of the body provenance. However, the presence of fatty acids characteristic of human decomposition (myristic, palmitic, oleic and stearic acids) was identified, proving the importance of studying these lipids in forensic cases. Additionally, it was possible to verify that soils thar are more acidic and less rich in humidity and organic matter, present less fatty acids. Finally, no biomarker steroids of human degradation were found, reflecting the need for a future optimisation of the applied protocol. In the future, the study should also be extended to more cemeteries with different soil typologies.

Enantioselective ecotoxicity of psychoactive substances in Daphnia magna

Sat, 01 Jan 2022 00:00:00 GMT

Enantioselective ecotoxicity of psychoactive substances in Daphnia magna Gomes, Cristiano Manuel Araújo Psychoactive substances (PAS) are emergent contaminants frequently detected in aquatic ecosystems that may pose environmental risks in aquatic organisms even at low concentrations (ng to µg/L). Many PAS are chiral substances commercialized as racemate. Amphetamine (AMP) is a central nervous system stimulant used in the treatment of attention deficit hyperactivity disorder narcolepsy, and obesity. AMP is a chiral substance that exhibits enantioselective in its pharmacological activity being the (S)-AMP more potent and clinically effective than (R)-AMP. On the other hand, AMP is frequently used as a recreative drug. Due to its high consume and low biodegradability AMP has been detected in wastewaters and surface waters and can occur as enantiomeric mixtures or pure enantiomers. In this context, the evaluation of its enantioselectivity in eco-toxicity is crucial for a better understanding of AMP environmental risk on non-target organisms in freshwater ecosystems. Therefore, the present study aimed to investigate the enantioselectivity of AMP in toxicity on the aquatic invertebrate daphnia (Daphnia magna), used as a model organism to assess different biomarkers of toxicity. For that, neonates (less than 24h old) were exposed to 0.1; 1, and 10 µg/L of the racemate (rac-AMP) and to 0.1 and 1 µg/L of pure enantiomers, (R)-AMP and (S)-AMP, for 8 days. At selected 3, 5, and 8-days of exposure, different parameters were determined as morphophysiology (on days 3 and 8, as body size and heart rate, area, and length); swimming behaviour (on day 5, as swimming speed, active time, and total distance); reproduction (on day 8, number of eggs per daphnia, number of daphnia with eggs and number of neonates) and biochemical parameters (on day 8, like oxidative stress, catalase (CAT) and acetylcholinesterase (AChE) enzymatic activities). Data showed a significant decrease in body size found for organisms exposed to (S)-AMP comparatively to (R)-enantiomer and to the racemate. Effects on the development and functioning of heart were observed with a significant decrease in heart rate for both racemate and enantiomers though a decrease was observed in the organisms exposed to the racemate at days 3 and 8, while for the enantiomers a decrease was observed at day 3 with an enantioselective effect at 0.1 ug/L (lower decrease for (R)-AMP) but on day 8 no differences were found. A significant decrease in the heart area was observed for both enantiomers. Regarding swimming behaviour, different results were observed between racemate and enantiomers. (rac)-AMP caused a reduction of the total travelled distance while for both enantiomers an increase in total travelled distance was noted though no enantioselective effects were observed. No changes were observed in active time in the organism exposed to (rac)-AMP while a reduction of the active time was observed for both enantiomers. No changes in the number of eggs per daphnia or the number of daphnia with eggs were observed for the racemate, however, a tendency to increase of the number of neonates at 0.1 and 1 μg/L was observed while a significant increase was found at 10 μg/L. Regarding enantiomers, a significant difference was found between enantiomers with a decrease in the organisms exposed to (S)-AMP in contrast to the increase in the organisms exposed to (R)-AMP at 1 μg/L. Though no significant differences were observed for the number of daphnia with eggs, a tendency to increase was observed in the organisms exposed to (R)-AMP. These results show that AMP affects reproductive performance of daphnia, and these effects are enantiomer dependent. Changes in biochemical parameters were also observed with a significant decrease in reactive oxygen species (ROS) and CAT activity for enantiomers. These results showed that AMP can interfere with different biomarkers of toxicity and these effects can be enantioselective demonstrating the relevance and providing evidence for the need for this kind of study for an accurate environmental risk assessment. Additionally, some of the effects were observed at environmental reported concentrations (0.1 and 1 μg/L) AMP both racemate and enantiomers can cause adverse effects on D. magna reinforcing the concern of invertebrate medium- and long-term exposure to AMP.