In the field of postmortem toxicology, principles from pharmacology and toxicology are combined in order to determine if exogenous substances contributed to ones death. In order to make this determination postmortem and (whenever available) antemortem blood samples may be analyzed. This project focused on evaluating the relationship between postmortem and antemortem blood drug levels, in order to better define an interpretive framework for postmortem toxicology. To do this, it was imperative to evaluate the differences in antemortem and postmortem drug concentrations, determine the role microbial activity and evaluate drug stability. Microbial studies determined that the bacteria Escherichia coli and Pseudomonas aeruginosa could use the carbon structures of drugs as a source of food. This would suggest prior to sample collection, microbial activity could potentially affect drug levels. This process however would stop before toxicologic evaluation, as at autopsy blood samples are stored in tubes containing the antimicrobial agent sodium fluoride. Analysis of preserved blood determined that under the current storage conditions sodium fluoride effectively inhibited microbial growth. Nonetheless, in many instances inconsistent drug concentrations were identified. When comparing antemortem to postmortem results, diphenhydramine, morphine, codeine and methadone, all showed significantly increased postmortem drug levels. In many instances, increased postmortem concentrations correlated with extended postmortem intervals. Other drugs, such as alprazolam, were likely to have concentration discrepancies when short antemortem to death intervals were coupled with extended postmortem intervals. While still others, such as midazolam followed the expected pattern of metabolism and elimination, which often resulted in decreased postmortem concentrations. The importance of drug stability was displayed when reviewing the clonazepam/ 7-aminoclonazepam data, as the parent drug commonly converted to its metabolite even when stored in the presence of a preservative. In instances of decreasing postmortem drug concentrations the effect of refrigerated storage could not be ruled out. A stability experiment, which contained codeine, produced data that indicated concentrations could continue to decline under the current storage conditions. The cumulative data gathered for this experiment was used to identify concentration trends, which subsequently aided in the development of interpretive considerations for the specific analytes examined in the study.

Decomposition chemistry refers to the biochemical degradation processes which occur in soft tissue as decomposition proceeds. This study aims to investigate the relationship between the release of decomposition fluids into contrasting soil environments and their potential correlation with the presence of a decomposing carcass. Soil from two different carcass decomposition trials was utilized to determine if seasonal and soil variation altered the soils reaction to a carcass. The reaction was determined by investigating the soil available phosphorus, extractable lipid-phosphate, pH, moisture and fatty acid content. A significant increase in the relative concentration of extractable lipid-phosphate, soil available phosphorus, and fatty acid content was identified, confirming the flux in the microbial biomass in the soil. Contrary to these nutrients, there were no notable changes in the soil pH and moisture content. The findings of this study were able to highlight the future forensic potential of these techniques and demonstrate a need for further research.

Postmortem microbial communities are being extensively studied for their utility in forensic science investigations. Microbial communities associated with decomposition, necrobiome, have been shown to react in a predictable manner to the postmortem interval or time since death. These communities are affected by environmental factors such as temperature and humidity which can cause variabilities in the community structural and functional turnover. However, the transmigration patterns across organs and functional activity as decomposition progresses is still relatively unknown in a highly controlled system. This study aims to describe the community structural changes that take place during a highly controlled decomposition in mice along with one of the first representations of visualizing transmigration and detecting functional pathway differences between postmortem times. Although, postmortem microbial communities have been viewed under the scope of forensics, there are also extended uses of early postmortem microbiome communities that represent the antemortem microbiome for health research. Additionally, in this study we aim to provide evidence for the use of the early postmortem microbiome as a public health surveillance tool by detecting antibiotic resistance determinants with their corresponding bacterial genera in human autopsies. These results have provided important baseline microbial community structure and function data for forensic research in murine models and have identified antibiotic resistance determinants of high public health concern in human autopsy samples.

Violence is a dreadful phenomenon spread throughout the world, resulting in unfortunate events that can ultimately cause death. It is known that some countries play a much worrying role in this scenario than others. Brazil is one of them. The present study has focused on identifying the use of cocaine within 105 postmortem cases arriving at the Institute of Legal Medicine of São Paulo (IML-SP) through analytic toxicological methods and latter applying genetic testing to see whether the presence of certain single nucleotide polymorphisms (SNPs) is more predominant within users rather than non-users, which would help to better understand one\'s susceptibility to abuse the drug. Both blood and hair samples have been analysed through ultra-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) in order to distinguish between recent or chronic cocaine use among violent individuals whose violence has ultimately leaded to their death. Two dilute-and-shoot methods have been validated and used for this purpose, and the final residue was analysed through the UPLC-ESIMS/ MS system. From the 105 postmortem cases, a rather high proportion of cocaine and its metabolites was found. A chronic use of the drug was denoted in 53% of the cases, which were positive for cocaine and benzoylecgonine, followed by 43% for norcocaine, 40% for cocaethylene and 13% for anhydroecgonine methyl ester, in hair. As for blood, reflecting the use of cocaine prior to death, 51% of the cases have shown to be positive for benzoylecgonine, followed by 41% for cocaine, 23% for cocaethylene and 20% for norcocaine. These findings suggest a probable association between the use of the drug and risky/violent behaviours. Genetic wise, a significant difference has been observed for SNP rs4263329 from the BCHE gene in its dominant model, with higher frequencies of the genotypes A/G and G/G seen in cocaine users rather than non-users (OR=8.91; 95%CI=1.58-50.21; ρ=0.01). Likewise, also SNP rs6280 from the DRD3> gene presented a significant association in both its additive and dominant model, suggesting that the C allele may be playing a role in cocaine use as both genotypes T/C and C/C were significantly more frequent in users than non-users. This association was not lost when adjusted for covariants using logistic regression (OR=4.96; 95%CI=1.07; ρ=0.04). Finally, a statistically significant association (ρ = 0.003) was also encountered among individuals with both A/G and G/G genotypes within SNP rs4263329 and the use of cocaine HCl (f(A/G+G/G)=44.7%) versus crack-cocaine (f(A/G+G/G)=7.7%) and nonusers (f(A/G+G/G)=16.2%). In conclusion, this study has found significant associations within two SNPs related to cocaine use, however, due to several inherent limitations, these must be confirmed by further studies with a higher number of subjects and within a more controlled setting. Definite assumptions may not be made at this point and future researches are to be conducted. / A violência é um fenômeno aterrador espalhado por todo o mundo, resultando em eventos que podem, em última instância, causar a morte. Sabe-se que, em alguns países esse cenário é mais preocupante que em outros. O Brasil é um deles. O presente estudo teve como objetivo identificar o uso de cocaína em 105 casos postmortem provenientes do Instituto de Medicina Legal de São Paulo (IML-SP) por meio de métodos toxicológicos analíticos e posterior aplicação de testes genéticos para verificar se a presença de determinados polimorfismos de nucleotídeo único (SNPs) é mais predominante dentro dos usuários do que dos não usuários, o que explicaria uma possível suscetibilidade de um indivíduo ao abuso da droga. Amostras de sangue e cabelo foram analisadas através de cromatografia líquida de ultra-eficiência acoplada a espectrometria de massas e ionização por electrospray (UPLC-ESI-MS/MS) para distinguir entre uso recente ou crônico de cocaína entre indivíduos violentos cuja violência levou à sua morte. Para tal, dois métodos de extração baseados na técnica de \"dilute-and-shoot\" foram validados e utilizados para esse fim, e o resíduo final foi analisado através de um sistema UPLC-ESI-MS/MS. Dos 105 casos postmortem, foi encontrada uma proporção significativa de cocaína e seus produtos de biotransformação. O uso crônico da droga foi denotado em 53% dos casos, sendo estes positivos para cocaína e benzoilecgonina, seguidos de 43% para norcocaína, 40% para cocaetileno e 13% para anidroecgonina metil éster, no cabelo. Quanto ao sangue, refletindo o uso de cocaína antes da morte, 51% dos casos mostraram-se positivos para benzoilecgonina, seguido de 41% para cocaína, 23% para cocaetileno e 20% para norcocaína. Esses dados corroboram a hipótese provável da relação entre o uso da droga e comportamentos de risco/violentos. Quanto à genética, uma diferença significativa foi observada para o SNP rs4263329 do gene BCHE em seu modelo dominante, com maiores frequências dos genótipos A/G e G/G vistos em usuários de cocaína ao contrário de não usuários (OR=8,91; 95%IC=1,58-50,21; ρ=0,01). Da mesma forma, também o SNP rs6280 do gene DRD3 apresentou uma associação significativa tanto no seu modelo aditivo quanto dominante, sugerindo que o alelo C pode estar desempenhando um papel no uso de cocaína, pois ambos os genótipos T/C e C/C foram significativamente mais frequentes nos usuários do que não usuários. Essa associação não foi perdida quando ajustada para co-variáveis usando regressão logística (OR=4,96; 95%IC=1,07; ρ=0,04). Finalmente, uma associação estatisticamente significativa (ρ=0,003) também foi encontrada entre indivíduos com ambos os genótipos A/G e G/G dentro do SNP rs4263329 e o uso de cocaína HCl (f(A/G + G/G)=44,7%) versus crack (f(A/G + G/G)=7,7%) e não usuários (f(A/G + G/G)=16,2%). Em conclusão, este estudo encontrou associações significativas em dois SNPs relacionados ao uso de cocaína, no entanto, devido a várias limitações inerentes, estas devem ser confirmadas por mais estudos com um maior número de indivíduos e dentro de um cenário mais controlado. Hipóteses definitivas não poderão ser feitas neste momento e futuras pesquisas devem ser conduzidas.

Violence is a dreadful phenomenon spread throughout the world, resulting in unfortunate events that can ultimately cause death. It is known that some countries play a much worrying role in this scenario than others. Brazil is one of them. The present study has focused on identifying the use of cocaine within 105 postmortem cases arriving at the Institute of Legal Medicine of São Paulo (IML-SP) through analytic toxicological methods and latter applying genetic testing to see whether the presence of certain single nucleotide polymorphisms (SNPs) is more predominant within users rather than non-users, which would help to better understand one\'s susceptibility to abuse the drug. Both blood and hair samples have been analysed through ultra-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) in order to distinguish between recent or chronic cocaine use among violent individuals whose violence has ultimately leaded to their death. Two dilute-and-shoot methods have been validated and used for this purpose, and the final residue was analysed through the UPLC-ESIMS/ MS system. From the 105 postmortem cases, a rather high proportion of cocaine and its metabolites was found. A chronic use of the drug was denoted in 53% of the cases, which were positive for cocaine and benzoylecgonine, followed by 43% for norcocaine, 40% for cocaethylene and 13% for anhydroecgonine methyl ester, in hair. As for blood, reflecting the use of cocaine prior to death, 51% of the cases have shown to be positive for benzoylecgonine, followed by 41% for cocaine, 23% for cocaethylene and 20% for norcocaine. These findings suggest a probable association between the use of the drug and risky/violent behaviours. Genetic wise, a significant difference has been observed for SNP rs4263329 from the BCHE gene in its dominant model, with higher frequencies of the genotypes A/G and G/G seen in cocaine users rather than non-users (OR=8.91; 95%CI=1.58-50.21; ρ=0.01). Likewise, also SNP rs6280 from the DRD3> gene presented a significant association in both its additive and dominant model, suggesting that the C allele may be playing a role in cocaine use as both genotypes T/C and C/C were significantly more frequent in users than non-users. This association was not lost when adjusted for covariants using logistic regression (OR=4.96; 95%CI=1.07; ρ=0.04). Finally, a statistically significant association (ρ = 0.003) was also encountered among individuals with both A/G and G/G genotypes within SNP rs4263329 and the use of cocaine HCl (f(A/G+G/G)=44.7%) versus crack-cocaine (f(A/G+G/G)=7.7%) and nonusers (f(A/G+G/G)=16.2%). In conclusion, this study has found significant associations within two SNPs related to cocaine use, however, due to several inherent limitations, these must be confirmed by further studies with a higher number of subjects and within a more controlled setting. Definite assumptions may not be made at this point and future researches are to be conducted. / A violência é um fenômeno aterrador espalhado por todo o mundo, resultando em eventos que podem, em última instância, causar a morte. Sabe-se que, em alguns países esse cenário é mais preocupante que em outros. O Brasil é um deles. O presente estudo teve como objetivo identificar o uso de cocaína em 105 casos postmortem provenientes do Instituto de Medicina Legal de São Paulo (IML-SP) por meio de métodos toxicológicos analíticos e posterior aplicação de testes genéticos para verificar se a presença de determinados polimorfismos de nucleotídeo único (SNPs) é mais predominante dentro dos usuários do que dos não usuários, o que explicaria uma possível suscetibilidade de um indivíduo ao abuso da droga. Amostras de sangue e cabelo foram analisadas através de cromatografia líquida de ultra-eficiência acoplada a espectrometria de massas e ionização por electrospray (UPLC-ESI-MS/MS) para distinguir entre uso recente ou crônico de cocaína entre indivíduos violentos cuja violência levou à sua morte. Para tal, dois métodos de extração baseados na técnica de \"dilute-and-shoot\" foram validados e utilizados para esse fim, e o resíduo final foi analisado através de um sistema UPLC-ESI-MS/MS. Dos 105 casos postmortem, foi encontrada uma proporção significativa de cocaína e seus produtos de biotransformação. O uso crônico da droga foi denotado em 53% dos casos, sendo estes positivos para cocaína e benzoilecgonina, seguidos de 43% para norcocaína, 40% para cocaetileno e 13% para anidroecgonina metil éster, no cabelo. Quanto ao sangue, refletindo o uso de cocaína antes da morte, 51% dos casos mostraram-se positivos para benzoilecgonina, seguido de 41% para cocaína, 23% para cocaetileno e 20% para norcocaína. Esses dados corroboram a hipótese provável da relação entre o uso da droga e comportamentos de risco/violentos. Quanto à genética, uma diferença significativa foi observada para o SNP rs4263329 do gene BCHE em seu modelo dominante, com maiores frequências dos genótipos A/G e G/G vistos em usuários de cocaína ao contrário de não usuários (OR=8,91; 95%IC=1,58-50,21; ρ=0,01). Da mesma forma, também o SNP rs6280 do gene DRD3 apresentou uma associação significativa tanto no seu modelo aditivo quanto dominante, sugerindo que o alelo C pode estar desempenhando um papel no uso de cocaína, pois ambos os genótipos T/C e C/C foram significativamente mais frequentes nos usuários do que não usuários. Essa associação não foi perdida quando ajustada para co-variáveis usando regressão logística (OR=4,96; 95%IC=1,07; ρ=0,04). Finalmente, uma associação estatisticamente significativa (ρ=0,003) também foi encontrada entre indivíduos com ambos os genótipos A/G e G/G dentro do SNP rs4263329 e o uso de cocaína HCl (f(A/G + G/G)=44,7%) versus crack (f(A/G + G/G)=7,7%) e não usuários (f(A/G + G/G)=16,2%). Em conclusão, este estudo encontrou associações significativas em dois SNPs relacionados ao uso de cocaína, no entanto, devido a várias limitações inerentes, estas devem ser confirmadas por mais estudos com um maior número de indivíduos e dentro de um cenário mais controlado. Hipóteses definitivas não poderão ser feitas neste momento e futuras pesquisas devem ser conduzidas.

Thesis (M.A.)--Texas State University--San Marcos, 2010. / Vita. Appendices: leaves 59-98. Includes bibliographical references (leaves 99-104).

Limited experimentation has been conducted on fractography of fresh versus dry bones. The present project examined the presence of select fractographic features on wet and dry bone specimens over a time interval of 15 months. The experimental remains consisted of a total of 81 white-tailed deer (Odocoileus virginianus) long bones as a proxy for human long bones. A subsample of 15 long bones that were defleshed of most external soft tissue was subjected to blunt force trauma every 30 days for a total of three months. After these three months passed, a subsample of 15 bones was subjected to blunt force trauma every 90 days for the remaining 12 months. Following fracturing, the long bones were macerated and the fractures on the long bones were inspected, and observations were recorded and photographed. The author hypothesized that the presence (or absence) of fractographic features including hackle patterns, bone mirror, cantilever curls, and arrest ridges, on the fractured long bones would differ on fresh versus dry bones. Therefore, the difference in fractographic features found on the fresh versus dry bones would allow greater separation of perimortem from postmortem fractures. Other fracture characteristics such as fracture angle, fracture surface texture, fracture jaggedness, number of fragments produced, and type of fracture produced were also observed as part of the data collected in this research to potentially confirm the findings and results of previous studies conducted on differentiating between perimortem and postmortem trauma on bone. This study disproved the hypothesis by concluding that the presence (or absence) of fractographic features is not greatly affected by time exposure and therefore, does not aid in distinguishing between fresh bone and dry bone fractures. Fractographic features were present and absent on bone specimens during all postmortem intervals. The only statistically significant difference discovered was that bone hackle patterns are more commonly observed than cantilever curls on bones with a later PMI. Other general trends observed were that the number of bones showing bone hackle patterns increased over time and the number of bones showing bone mirror decreased over time. In addition, the results of the study revealed that the only fracture characteristic that showed a slightly significant difference with time of exposure was the fracture surface texture produced. The probability of a bone showing intermediate fracture surfaces is statistically significantly higher than a bone showing rough fracture surfaces when the represented PMI is fresh. The probability of a bone showing intermediate fracture surfaces is statistically significantly higher than a bone showing smooth fracture surfaces when the represented PMI is dry. The present study showed that the fracture characteristics including fracture angle, fracture type, number of fragments produced, and fracture jaggedness were not greatly influenced by exposure of time but, certain patterns and trends were recognized. The number of bones showing sharp fracture angles increased over time, while the number of bones showing intermediate fracture angles stayed stagnant. Bones showing comminuted fractures also increased with the progression of drying time. The average number of fragments produced were high during both fresh and dry PMIs and low throughout the transitional postmortem intervals. Bones showing jagged, intermediate, and not jagged broken ends increased with the progression of time however, not jagged broken ends only began to appear in the sample starting at a PMI of 90 days.

No / The susceptibility of RNA to enzymatic degradation has been considered as a tool to estimate time-since-death in forensic samples, and it has previously been demonstrated that the choice of tissue is an important factor. In this study we have extracted RNA from decaying bone and bone marrow under the hypothesis that the delayed onset of putrefaction may render them a useful source in this context. In a preliminary study, total RNA was extracted from bone and bone marrow that had been sampled from six skeletally mature rabbits at time points between zero and 31 days after death. The levels of three specific RNA transcripts could be quantified using real-time polymerase chain reaction. Bioanalyzer results show rRNA bands in bone marrow samples up to 21 days postmortem. We hereby propose bone marrow as a potential source for postmortem RNA in forensic studies.

In homicide cases, suitcases provide concealment and ease of transport of a body and reduce the likelihood of detection. When estimating a minimum postmortem interval (mPMI), it is necessary to understand the taphonomic processes that occur when a body is concealed within a suitcase. In this study, the experimental carcasses consisted of pig (Sus scrofa Linnaeus) heads concealed within either hard shell or fabric suitcases whereas control pig heads were decomposed naturally. All control pigs heads attracted blowflies (Calliphoridae) immediately and mummified within days, whereas the experimental pigs heads had an oviposition delay of approximately 72 hours and decomposed resulting in skeletonization. Differences in the arthropod species present varied between the experimental and control pigs heads. Temperature comparisons revealed significant (p <0.001) differences between the experimental and control pigs heads. How a corpse decomposes when a suitcase is used for body disposal must be considered when estimating the mPMI.

The development and improvement of methods used for the estimation of the postmortem interval (PMI) is a common area of research in forensic science. This research was conducted to physically and chemically analyze pig carcass decomposition on a soil surface using conventional and newly developed methods for the potential use in estimating the PMI. Photographs of pig carcasses decomposing on forested and open land were scored using a decomposition scoring system and decomposition scores were related to accumulated degree days (ADD). Overall, the ADD values were significantly different for the two groups of carcasses; however, the ADD values for the onset of each score demonstrated more similarity between groups. Decomposition scoring results also indicated that refinements must be made to the calculation of ADD to allow for a meaningful comparison of pig and human decomposition. The decomposition of pig carcasses altered the water content, pH and fatty acid content of soil. The fatty acids, myristic, palmitic, palmitoleic, stearic and oleic acids were successfully extracted and analyzed from decomposition soil. Palmitic, stearic and oleic acids were the most abundant fatty acids detected whilst the levels of myristic and palmitoleic acids were negligible in comparison. A three peak fatty acid cycle was also observed for each fatty acid. Variations in soil pH and fatty acid content of decomposition soil have the potential to indicate the presence of a decomposition site. Furthermore, a nonlinear diffusion model was developed to predict the development of the cadaver decomposition island (CDI) in soil over time. The simulation of the model indicated that the diffusion model has the potential to generate PMI estimations for early stages of decomposition by corresponding the effective radius of the CDI to a particular time point. The general findings of this research indicate that more accurate methods for PMI estimations can potentially be developed with further research. / UOIT