This research paper discusses the role of virtual environments in digital forensic investigations. With virtual environments becoming more prevalent as an analysis tool in digital forensic investigations, it’s becoming more important for digital forensic investigators to understand the limitation and strengths of virtual machines. The study aims to expose limitations within commercial closed source virtual machines and open source virtual machines. The study provides a brief overview of history digital forensic investigations and virtual environments, and concludes with an experiment with four common open and closed source virtual machines; the effects of the virtual machines on the host machine as well as the performance of the virtual machine itself. My findings discovered that while the open source tools provided more control and freedom to the operator, the closed source tools were more stable and consistent in their operation. The significance of these findings can be further researched by applying them in the context of exemplifying reliability of forensic techniques when presented as analysis tool used in litigation.

Biological material (fluids or tissues) whether from the victim or suspect is often collected as forensic evidence, and methods to obtain and analyze the DNA found in that material have been well established. The type of body fluid (i.e. blood, saliva, semen, vaginal secretions, and menstrual blood) from which the DNA originated is also of interest, and messenger RNA typing provides a specific and sensitive means of body fluid identification. In order for mRNA profiling to be utilized in routine forensic casework, RNA of sufficient quantity and quality must be obtained from biological fluid stains and the methods used for RNA analysis must be fully compatible with current DNA analysis methodologies. Several DNA/RNA co-extraction methods were evaluated based on the quantity and quality of DNA and RNA recovered and were also compared to standard non-co-extraction methods. The two most promising methods, the in-house developed NCFS co-extraction and the commercially available AllPrep DNA/RNA Mini kit, were then optimized by improving nucleic acid recovery and consistency of CE (capillary electrophoresis) detection results. The sensitivity of the two methods was also evaluated, and DNA and RNA profiles could be obtained for the lowest amount of blood (0.2 µL) and saliva and semen (1 µL) tested. Both extraction methods were found to be acceptable for use with forensic samples, and the ability to obtain full DNA profiles was not hindered by the co-extraction of RNA. It is generally believed that RNA is less stable than DNA which may prevent its use in forensic casework. However, the degradation rates of DNA and RNA in the same biological fluid stain have not been directly compared. To determine the relative stability of DNA and RNA, the optimized NCFS co-extraction protocol was used to isolate DNA and RNA from iv environmentally compromised stains. Dried blood, saliva, and semen stains and vaginal secretions swabs were incubated at set temperatures and outside for up to 1 year. Even at 56°C, DNA and RNA were both stable out to 1 year in the blood and semen stains, out to 3 months (DNA) and 1 year (RNA) in the saliva stains, and out to 6 months (DNA) and 3 months (RNA) in the vaginal secretions swabs. The recoverability of both nucleic acids was reduced when the samples were exposed to increased humidity, sunlight, and rain. In general, DNA and RNA stability was found to be similar with a loss in ability to obtain a DNA or RNA profile occurring at the same time point; however, there were instances where RNA body fluid markers were detected when a poor/no DNA profile was obtained, indicating that RNA in dried stains is sufficiently stable for mRNA body fluid typing to be used in forensic casework.

No / Forensic archaeology is mostly defined as the use of archaeological methods and principles within a legal context. However, such a definition only covers one aspect of forensic archaeology and misses the full potential this discipline has to offer. This volume is unique in that it contains 57 chapters from experienced forensic archaeological practitioners working in different countries, intergovernmental organisations or NGO’s. It shows that the practice of forensic archaeology varies worldwide as a result of diverse historical, educational, legal and judicial backgrounds. The chapters in this volume will be an invaluable reference to (forensic) archaeologists, forensic anthropologists, humanitarian and human rights workers, forensic scientists, police officers, professionals working in criminal justice systems and all other individuals who are interested in the potential forensic archaeology has to offer at scenes of crime or places of incident. This volume promotes the development of forensic archaeology worldwide. In addition, it proposes an interpretative framework that is grounded in archaeological theory and methodology, integrating affiliated behavioural and forensic sciences.

This project constitutes an instructional unit for honors biology that involves the use of science in the field of criminal investigation and forensics. Before beginning the unit, the learners should have mastered basic laboratory skills, including use of the microscope. They should also have an understanding of the basic structure and function of DNA and its role in heredity and protein synthesis. The standard time frame is 24 days with 70-minute periods, but can be easily adjusted to meet classroom needs. Several instructional strategies enhance student learning and make science fun. The unit is inquiry-driven and activity-based. Students are surprised by the crime, gather and analyze evidence, and work towards proposing an explanation. This real world problem involves the use of cooperative learning and a variety of assessment techniques.

D.Phil.

Esta tese é composta por três capítulos. O primeiro capítulo teve por objetivo analisar quais as técnicas de reconstrução nasal mais consistentes em contexto forense através de uma revisão sistemática da literatura. No total, existem 15 métodos de reconstrução nasal descritos na literatura. Para localizar a ponta do nariz e estimar a projeção nasal em indivíduos brasileiros, o método que se demonstrou, em meta-análise, o mais apropriado foi o que considera que o ângulo (ponto Pronasale) formado pelas retas que partem dos pontos Rhinion e Prosthion possui valor aproximado de 90º. O capítulo II teve como propósito testar, mediante análise de imagens tomográficas, o método desenvolvido em indivíduos brasileiros para estimar a localização da ponta do nariz em casos de reconstrução facial forense. De acordo com a técnica, o ângulo (ponta do nariz) formado pela união de retas traçadas a partir dos pontos Rhinion e Prosthion corresponde a 90º. O programa Horos® foi utilizado para analisar as imagens dos exames de tomografia computadorizada de feixe cônico. Os critérios de seleção da amostra foram semelhantes aos utilizados no trabalho que propõe o método, assim como as análises realizadas. Os resultados mostraram que o ângulo de interesse correspondeu em média a 96.5º. Esta diferença resultou em uma inacurácia de aproximadamente 3 mm da localização estimada da ponta do nariz em relação à sua localização real. Em termos práticos, esta diferença não impede o reconhecimento facial. O terceiro capítulo teve o propósito de testar o método proposto para estimar a largura nasal de indivíduos brasileiros em reconstrução facial forense, além de determinar parâmetros em tecido duro para estimar a largura nasal e verificar relações entre o tipo facial e a morfologia do perfil nasal. A amostra foi composta por 246 imagens de tomografia computadorizada de feixe cônico (feminino: 183; masculino: 63). O programa Horos® foi utilizado. As análises em tecido duro foram realizadas na visualização de reconstrução multiplanar com projeção de máxima intensidade igual a zero, e a análise do perfil nasal foi realizada em tecido mole na ferramenta de visualização de superfícies. Os resultados mostram que o método proposto para estimar a largura nasal em brasileiros apresentou uma inacurácia de aproximadamente 1 mm, fato que não interfere no reconhecimento facial. Não houve relação de proporção direta entre a largura do nariz e a abertura piriforme, nem entre a largura do nariz e a distância intercanina, mesmo quando foram consideradas variáveis como sexo, idade e tipo facial. O biótipo facial longo apresentou relação moderada com o perfil nasal reto (r=0.037). / This thesis contains three chapters. The first chapter aimed to conduct a systematic review to analyze what are the most consistent techniques of nasal reconstruction in a forensic context. There are fifteen nasal reconstruction methods described in the literature. To predict the Pronasale point and the nasal projection, the method that considers that the confluence between the lines traced from Rhinion and from Prosthion is an angle (Pronasale point) with a value of 90º presented good results when meta-analysis was performed. The aim of the second chapter was to test, trough CT images, the method developed on a sample of Brazilian subjects to predict the Pronasale point in forensic facial reconstruction. According to this technique, the union of the Rhinion and the Prosthion lines creates an 90º angle (Pronasale point). The software Horos® was used in order to perform the analysis on the cone-beam tomography images. The inclusion criteria of the sample were the same used by the primary study as well as the performed analysis. The results showed a mean value of 96.5º from the projection Pronasale and Rhinion. This difference results in an inaccuracy of approximate 3 mm of the Pronasale point and it is not impeditive for facial recognition of a familiar face. The third chapter aimed to test the method proposed to predict nasal length in Brazilian subjects, to determine hard tissue parameters to predict nasal length and to observe if there was any relation between facial pattern and nasal profile. The sample was composed by 246 cone beam computed tomography (183 female, 63 male). The software Horos® was used. The hard tissue analysis were performed on 3D multiplanar reconstruction with maximum intensity projection equal to zero, and the 3D surface rendering was used to verify the nasal profile in soft tissue. The proposed method to predict nasal length presented an inaccuracy of approximate 1 mm and it certainly does not influence facial recognition of familiar faces. There was no relation between nor nasal length and piriform aperture nor nasal length and intercanine distance even when sex, age and facial type were considered. The long facial type showed moderate relation with strength nasal profile (r=0.037).

O presente trabalho analisa diferentes aspectos da perícia médico legal, visando buscar formas efetivas de otimizar seus laudos e aferir melhores resultados na busca da verdade dos fatos na Justiça brasileira. Para tanto, é feita uma análise etimológica e uma breve retrospectiva da história da perícia, como forma de melhor entender e contextualizar suas bases e origens. Após, é analisada a realidade brasileira nesta área, em especial no Estado de São Paulo, mediante a verificação da estrutura dos locais de perícia, bem como da qualidade das intervenções dos poderes legislativo e executivo nesta área. É discutida, ainda, a questão da desvinculação da perícia à Segurança Pública no Brasil, bem como é realizado um breve estudo de direito comparado, em que são focadas as diferenças no tratamento legal da perícia entre o ordenamento jurídico brasileiro e os ordenamentos estrangeiros. / This study analyzes several aspects of forensic expertise, aiming to seek effective ways to optimize the experts reports, consequently benchmarking best results in the search for the truth in investigations carried out in Brazil. Therefore, it was performed an etymological analysis and a brief retrospective of the history of forensic sciences as a way to understand and contextualize its origins and bases. In sequence it was analyzed the Brazilian reality in this field, especially in the State of São Paulo, by means of checking the structure of the locations where the States official examinations are conducted, as well as the quality of the interventions of the government and the parliament in this field. This work also discusses the issue of untying the States official examinations from the Citizen Security authorities in Brazil, as well as performs a brief study of comparative Law, which is focused on the differences related to the legal treatment of forensic sciences on foreign Legal systems compared to the Brazilian Legal system.

A necropsia é essencial para a identificação da causa de morte e dos processos que culminaram no óbito do animal. O presente estudo visou à pesquisa da real ocorrência de morte súbita e inesperada em cães submetidos à necropsia, e à determinação da discrepância entre as suspeitas dos proprietários em relação ao estado de saúde prévio e à causa de morte de seus cães, quando comparadas às conclusões necroscópicas. Este trabalho também focou nas alterações morfológicas e funcionais nas glândulas adrenais de cães necropsiados, a fim de se estudar a aplicação de possíveis marcadores da resposta adrenal ao estresse sofrido pelo animal na iminência do óbito. Foram utilizados os dados de 82 cães necropsiados na FMVZ-USP para análise da ocorrência real de morte súbita nestes animais. As alterações morfológicas nas adrenais de 46 cães necropsiados foram avaliadas através de análises morfométricas e histopatológicas. Também foram avaliados os índices de proliferação e apoptose nas células do córtex adrenal em relação à causa de morte do animal, através da marcação imunoistoquímica para o antígeno nuclear de proliferação celular (PCNA) e para BAX e Bcl-2, proteínas envolvidas na regulação da apoptose. A análise das alterações funcionais sofridas pelas adrenais focou na quantificação das concentrações das catecolaminas adrenalina e noradrenalina na medula adrenal dos cães por cromatografia líquida de alta eficiência com detecção eletroquímica (CLAE-DE). Dentre os resultados encontrados, a suspeita do proprietário de que seu cão sofreu uma morte súbita e inesperada é muito maior do que a real ocorrência deste tipo de morte em cães, sendo o óbito por decorrência de complicações de doenças muito mais frequente. As características morfométricas das adrenais dos cães apresentaram maior influência pelo peso corpóreo do animal e pela presença ou ausência de hiperplasia cortical do que pela associação com doenças crônicas ou condições agudas. Cães que vieram a óbito em decorrência de complicações de doenças crônicas exibiram fibrose em região corticomedular e focos de infiltrado inflamatório, ausentes nos animais com morte súbita ou doenças agudas, além de maior ocorrência de hiperplasia adrenocortical. Cães que sofriam de alterações cardíacas crônicas apresentaram alterações histopatológicas significativas mais marcantes em suas adrenais, como necrose, fibrose e depleção vacuolar cortical. Por outro lado, a congestão severa nas adrenais foi um achado mais frequente nos animais previamente saudáveis que sofreram morte súbita ou que vieram a óbito por doenças agudas. A avaliação dos índices de proliferação celular e apoptose no córtex das adrenais através da marcação imunoistoquímica para PCNA e BAX e Bcl-2, respectivamente, não apresentou potencial relevante para o estudo dos efeitos do estresse por doenças crônicas sobre as adrenais de cães. As concentrações de adrenalina e noradrenalina na medula adrenal se mostraram muito maiores em cães machos quando comparados às fêmeas. Os achados deste estudo podem auxiliar nas conclusões da necropsia, sendo especialmente relevantes em casos médico-legais, nos quais todos os achados possíveis devem ser relatados e analisados a fim de se prover um diagnóstico preciso, seguro e incontroverso / The necropsy is essential to identify the cause of death and the processes that culminated in the death of the animal. The present study aimed to research the actual occurrence of sudden and unexpected death in dogs subjected to necropsy, and to determine the discrepancy between the suspicions of owners regarding the previous state of health or cause of death of their dogs when compared to the gross findings. This work also focused on the morphological and functional changes in the adrenal glands of dogs submitted to necropsy in order to study the application of possible markers of adrenal stress response experienced by the animal on the verge of death. The data of 82 dogs necropsied in FMVZ-USP was used for analysis of the actual occurrence of sudden death in these animals. The morphological changes in the adrenal glands of 46 necropsied dogs were evaluated by morphometric and histopathological analysis. The proliferation rates and apoptosis in cells of the adrenal cortex regarding the cause of death of the animal were also evaluated by immunohistochemical staining for the nuclear antigen of cellular proliferation (PCNA) and BAX and Bcl-2, proteins involved in apoptosis regulation. The analysis of functional changes in the adrenal gland focused on quantifying the concentrations of the catecholamines adrenaline and noradrenaline in the adrenal medulla of dogs by high-performance liquid chromatography with electrochemical detection (HPLC-DE). Regarding the results of this study, the owners’ suspicion that their dog suffered a sudden and unexpected death is much greater than the actual occurrence of this type of death in dogs, while death due to complications from diseases is far more frequent. The morphometric characteristics of the adrenal glands of dogs showed greater influence by the body weight of the animal and the presence or absence of cortical hyperplasia than by association with chronic or acute conditions. Dogs that died due to chronic disease complications exhibited fibrosis in the corticomedullary region and foci of inflammatory infiltrate, absent in animals with sudden death or acute diseases, as well as higher incidence of adrenocortical hyperplasia. Dogs suffering from chronic cardiac conditions showed more marked significant histopathological changes in their adrenal glands, such as necrosis, fibrosis and cortical vacuolar depletion. On the other hand, severe congestion in the adrenal glands was a frequent finding in previously healthy animals who suffered sudden death or that died from acute illnesses. The evaluation of cell proliferation rates and apoptosis in the cortex of the adrenal by immunohistochemical staining for PCNA and BAX and Bcl-2, respectively, did not show significant potential for studying the effects of stress by chronic diseases in the adrenal glands of dogs. The adrenaline and noradrenaline concentrations in the adrenal medulla were much higher in male dogs when compared to females. This study’s findings may help the necropsy conclusions, which are particularly relevant in forensic cases where all possible findings should be reported and analyzed in order to provide an accurate, confident and uncontroversial diagnosis

A necropsia é essencial para a identificação da causa de morte e dos processos que culminaram no óbito do animal. O presente estudo visou à pesquisa da real ocorrência de morte súbita e inesperada em cães submetidos à necropsia, e à determinação da discrepância entre as suspeitas dos proprietários em relação ao estado de saúde prévio e à causa de morte de seus cães, quando comparadas às conclusões necroscópicas. Este trabalho também focou nas alterações morfológicas e funcionais nas glândulas adrenais de cães necropsiados, a fim de se estudar a aplicação de possíveis marcadores da resposta adrenal ao estresse sofrido pelo animal na iminência do óbito. Foram utilizados os dados de 82 cães necropsiados na FMVZ-USP para análise da ocorrência real de morte súbita nestes animais. As alterações morfológicas nas adrenais de 46 cães necropsiados foram avaliadas através de análises morfométricas e histopatológicas. Também foram avaliados os índices de proliferação e apoptose nas células do córtex adrenal em relação à causa de morte do animal, através da marcação imunoistoquímica para o antígeno nuclear de proliferação celular (PCNA) e para BAX e Bcl-2, proteínas envolvidas na regulação da apoptose. A análise das alterações funcionais sofridas pelas adrenais focou na quantificação das concentrações das catecolaminas adrenalina e noradrenalina na medula adrenal dos cães por cromatografia líquida de alta eficiência com detecção eletroquímica (CLAE-DE). Dentre os resultados encontrados, a suspeita do proprietário de que seu cão sofreu uma morte súbita e inesperada é muito maior do que a real ocorrência deste tipo de morte em cães, sendo o óbito por decorrência de complicações de doenças muito mais frequente. As características morfométricas das adrenais dos cães apresentaram maior influência pelo peso corpóreo do animal e pela presença ou ausência de hiperplasia cortical do que pela associação com doenças crônicas ou condições agudas. Cães que vieram a óbito em decorrência de complicações de doenças crônicas exibiram fibrose em região corticomedular e focos de infiltrado inflamatório, ausentes nos animais com morte súbita ou doenças agudas, além de maior ocorrência de hiperplasia adrenocortical. Cães que sofriam de alterações cardíacas crônicas apresentaram alterações histopatológicas significativas mais marcantes em suas adrenais, como necrose, fibrose e depleção vacuolar cortical. Por outro lado, a congestão severa nas adrenais foi um achado mais frequente nos animais previamente saudáveis que sofreram morte súbita ou que vieram a óbito por doenças agudas. A avaliação dos índices de proliferação celular e apoptose no córtex das adrenais através da marcação imunoistoquímica para PCNA e BAX e Bcl-2, respectivamente, não apresentou potencial relevante para o estudo dos efeitos do estresse por doenças crônicas sobre as adrenais de cães. As concentrações de adrenalina e noradrenalina na medula adrenal se mostraram muito maiores em cães machos quando comparados às fêmeas. Os achados deste estudo podem auxiliar nas conclusões da necropsia, sendo especialmente relevantes em casos médico-legais, nos quais todos os achados possíveis devem ser relatados e analisados a fim de se prover um diagnóstico preciso, seguro e incontroverso / The necropsy is essential to identify the cause of death and the processes that culminated in the death of the animal. The present study aimed to research the actual occurrence of sudden and unexpected death in dogs subjected to necropsy, and to determine the discrepancy between the suspicions of owners regarding the previous state of health or cause of death of their dogs when compared to the gross findings. This work also focused on the morphological and functional changes in the adrenal glands of dogs submitted to necropsy in order to study the application of possible markers of adrenal stress response experienced by the animal on the verge of death. The data of 82 dogs necropsied in FMVZ-USP was used for analysis of the actual occurrence of sudden death in these animals. The morphological changes in the adrenal glands of 46 necropsied dogs were evaluated by morphometric and histopathological analysis. The proliferation rates and apoptosis in cells of the adrenal cortex regarding the cause of death of the animal were also evaluated by immunohistochemical staining for the nuclear antigen of cellular proliferation (PCNA) and BAX and Bcl-2, proteins involved in apoptosis regulation. The analysis of functional changes in the adrenal gland focused on quantifying the concentrations of the catecholamines adrenaline and noradrenaline in the adrenal medulla of dogs by high-performance liquid chromatography with electrochemical detection (HPLC-DE). Regarding the results of this study, the owners’ suspicion that their dog suffered a sudden and unexpected death is much greater than the actual occurrence of this type of death in dogs, while death due to complications from diseases is far more frequent. The morphometric characteristics of the adrenal glands of dogs showed greater influence by the body weight of the animal and the presence or absence of cortical hyperplasia than by association with chronic or acute conditions. Dogs that died due to chronic disease complications exhibited fibrosis in the corticomedullary region and foci of inflammatory infiltrate, absent in animals with sudden death or acute diseases, as well as higher incidence of adrenocortical hyperplasia. Dogs suffering from chronic cardiac conditions showed more marked significant histopathological changes in their adrenal glands, such as necrosis, fibrosis and cortical vacuolar depletion. On the other hand, severe congestion in the adrenal glands was a frequent finding in previously healthy animals who suffered sudden death or that died from acute illnesses. The evaluation of cell proliferation rates and apoptosis in the cortex of the adrenal by immunohistochemical staining for PCNA and BAX and Bcl-2, respectively, did not show significant potential for studying the effects of stress by chronic diseases in the adrenal glands of dogs. The adrenaline and noradrenaline concentrations in the adrenal medulla were much higher in male dogs when compared to females. This study’s findings may help the necropsy conclusions, which are particularly relevant in forensic cases where all possible findings should be reported and analyzed in order to provide an accurate, confident and uncontroversial diagnosis

[Truncated abstract] The collection of buccal cells is common practise in the epidemiological and forensic science. Unlike venipuncture collection of blood; it is a safer, non-invasive method for collection of biological material. The methods by which these cells are collected from the inner cheek of an individual and stored are the key elements in preserving DNA. Typically, forensic samples require long term storage. Samples are commonly collected on cotton swabs and stored moist at low to ultra-low temperatures (less than -20oC). Although this is the method of choice in most forensic facilities, there are drawbacks. The samples are inherently contaminated with microflora within the oral cavity and the moisture allows a plethora of microorganisms to grow. As the time frame that has elapsed from collection to storage increases, there is an exponential increase in bacterial cells. Storage of containers containing swabs coated with cells at temperatures below 20oC is also costly due to requirements for large freezers which are running and monitored over 24 hours. In the pass 10 to 15 years, researchers have focussed on alternative ways to store buccal cells. The FTA card system by Whatman is one such development. The FTA card is unique in that it provides a means for the collection of buccal cells for storage at room temperature. DNA profiling from samples stored in this way for 11 years has been successfully achieved. The filter paper matrix of the FTA card binds and subsequently lyses cells. ... (2) The second component of this thesis describes a study which subjected cells on buccal swabs to various conditions of increased temperature over periods of time to establish if DNA could be amplified. The aim was to mimic exposure to the vigours of field conditions, particularly in the extreme local environments that prevail in the United Arab Emirates. a. Initially, buccal cells stored at -20oC over 360 days were used to mimic standard archiving procedures. The cells were subsequently transferred to FTA cards, amplified and profiled by using ABI AmpFLSTR Identifiler PCR Amplification Kit (Applied Biosystems, Foster City, CA). Complete STR profiles were successfully recovered from the archived swabs. In most cases 100% of alleles were recovered, suggesting that it is feasible to transfer DNA from properly archived buccal swabs to FTA cards. b. The second phase involved the storage of fresh swabs that had been artificially aged by using incubation temperatures ranging from 40oC to 100oC. Partial profiles resulted from artificially aged samples, indicating that the prevailing conditions prior to low temperature storage of the swabs plays an important role in ensuring cellular integrity and thus, DNA quality. Results from this study suggest that it is possible for biological samples stored under correct conditions to be transferred from swabs to FTA card. In combination, the two chapters presented in this study show that it is feasible to transfer achieved forensic biology samples from swabs to the FTA card system. However, it is necessary to ensure that the samples are treated in the correct manner so as to minimise contamination from external sources and to maintain the correct environmental state to maintain intact cells and usable DNA.