The question of post-mortem interval (PMI) or time since death is often the most sought after piece of information associated with a medical death investigation. Based on the observation that DNA degradation disproportionately affects the analysis of larger genetic loci, it was proposed that DNA degradation, as a result of autolysis or putrefaction, could prove suitable as a potential rate-of-change indicator of PMI. Nine randomly amplified polymorphic DNA (RAPD) analysis primers and three sets of directed amplification primers were evaluated to determine their suitability for use in assessing the degree of DNA fragmentation in tissue samples. They were assessed for amplicon specificity, total DNA target sensitivity, allele monomorphism and the observance of degradation-based profile changes. Markers meeting the requisite criteria were then used to assess a range samples degraded under controlled and uncontrolled conditions. Tissue samples collected from seven domestic pigs (Sus scrofa) were incubated under controlled laboratory or uncontrolled field conditions to produce samples simulating those potentially collected in a forensic case. DNA samples isolated from these specimens were then analyzed at those loci which had been determined to meet the requisite criteria. Collectively, data generated from these analyses indicate that genetic profiles generated by this approach can provide information useful for estimating the post-mortem interval, with the locus and amplicons used being most useful during the first 72 hours after death.

The objective of this study was to compare prediction of meat tenderness by means of gene technologies (markers) with established physical estimates of meat tenderness. Weaned, young bulls (n = 60) were selected on phenotype from various commercial producers to represent a Brahman (Bos indicus; n = 20), Simmental (continental Bos Taurus; n = 20) and Nguni (Sanga; n = 20) crossbred group. After being raised under intensive feedlot conditions the animals were slaughtered according to normal South African slaughter procedures at an A-age (10 - 12 months) with a fatness class of two or three (lean-medium fatness). At slaughter the carcasses were not electrical stimulated because electrical stimulation influences the processes of meat tenderness, and the emphasis was on the expression of the inherent tenderness characteristics without external post mortem influences. Carcasses were halved, chilled at 4 ˚C within 2 hours post mortem. The M. longissimus thoracis et lumboram (LT and LL) of the right and left sides were removed from the third last rib to the last lumbar vertebra and sub sampled for shear force evaluations, SDS-PAGE, Western-blotting, myofibril fragmentation (MFL), sarcomere length (SL), calpain, calpastatin, total collagen, % collagen solubility and marker analysis. The position of sampling for each test was consistent and the different samples were either frozen immediately at -20 ˚C or -80 ˚C or vacuum packed and aged (2 ± 2 ˚C) for 7 or 14 days post mortem. Two single nucleotide polymorphism (SNP) markers were employed in this study for the bovine CAPN1 gene, which is found or situated on bovine chromosome 29, namely a SNP marker which is situated on exon nine (CAPN1-316) and the other on intron 17 (CAPN1-4751). The inhibitor, calpastatin (CAST) found on chromosome seven was also analysed in this study. Both the CAST markers (CAST and CAST-Brahman) lie in the three prime untranslated regions (3' UTR) of the CAST gene. Genotype data of two- markers were used to determine the two-marker haplotypes. The results of the study showed that differences exist in meat quality of the different crossbreds. Brahman- and Nguni-crosses had lower shear force values (more tender) than that of the Simmentaler-crosses under these specific experimental conditions. The pH decline did not differ significantly (p > 0.001) between the crossbreds. The carcass temperature and temperature decline rate although similar between the Brahman- and the Simmentaler-crosses for three hours and up to eight hours post mortem differed significantly from that of the Nguni-crosses (p < 0.006). The sarcomere lengths were mostly under 1.7 µm, which indicate that shortening (caused by rapid chilling) can not be eliminated. No significant differences were found between the different crossbreds for the calpastatin levels but significant differences were found for the µ-calpain activity and µ-calpain / calpastatin activity ratios. The Brahman-crosses had longer myofibril fragment lengths on average, indicating lower proteolysis / myofibrillar fragmentation compared to the other crossbreds. Myofibrillar protein degradation (titin, nebulin, desmin) and myofibrillar protein formation (30 kDa) during post mortem ageing was examined as a confirmation for the myofibril fragment length results. Significant differences between the crossbreds were found for titin degradation. Nguni-cross animals had significantly (p < 0.033) more titin present than the other crossbreds. Nebulin degradation showed a significant (p < 0.038) breed effect at 7 days post mortem and a significant degradation rate difference for breed types between 1 day and 7 days post mortem (p < 0.03) and 1 day and 14 days post mortem (p < 0.034). Desmin degradation evaluated with SDS-PAGE and Western-blotting indicated that the Simmentaler-cross animals had significantly (p < 0.018; p < 0.024, respectively) lower desmin levels compared to the other crossbreds. For the 30 kDa proteins there were no significant (p > 0.001) differences in data evaluated at 1 day, 7 and 14 days post mortem as well as for the formation rate. The results indicates that Brahman-crosses had the highest frequency for haplotypes that are associated with increased shear force, and thus tougher meat compared to the Simmentaler-crosses that had the highest frequency for haplotyes that are associated with lower shear force, and thus more tender meat, while Nguni-crosses were intermediate. A multiplex marker system incorporating both markers (316 and 4751) and indexes for the markers at CASTand CAPN1 genes were evaluated in this study. Considering the average index for the interactions between the CAST and CAPN1 genes, it can be concluded that the Nguni-cross was overall the breed with the highest potential for inherently tender meat. In general, the animals in this study had the tendency for tougher meat. The genetic markers (CAST and CAPN1) showed no association with Warner-Bratzler shear force (WBSF) (p > 0.05), which suggest that various mechanisms and environmental factors may be involved and give another outcome compared to the genetic make up. Simple correlation coefficients were generated between the different characteristics measured. If the group of animals in this study is indeed a typical representation of South African feedlot finished crossbred animals, the relatively high WBSF values emphasise the challenge to manipulate their intrinsic tenderness potential by making use of various pre- and post-slaughter techniques and procedures. / Dissertation (MSc(Agric) (Animal Science))--University of Pretoria, 2008. / Animal and Wildlife Sciences / unrestricted

O objetivo deste trabalho foi investigar a incidência de carnes DFD (dark, firm, dry) em um frigorífico comercial de frango e determinar as suas propriedades funcionais comparando com as do PSE (pale, soft, exudative) e Normal em filés de peito, Pectoralis major. Para a avaliação da ocorrência desses tipos de cores, as aves das linhagens Cobb e Ross receberam banho calmante, com aspersão de água em temperatura ambiente por aproximadamente 10 minutos, associado à ventilação, imediatamente antes do seu abate. Em seguida, foram sacrificadas e desossadas e os filés foram analisados ao tempo de 90min e 24h post mortem, para coleta de valores de pH e L* e classificados em carnes PSE, Normal e DFD, com base nos resultados de 24h post mortem (n=329). O resultado da incidência foi de 24,92, 73,55 e 1,52% para amostras PSE, Normal e DFD, respectivamente. O valor de L* ao tempo de 90 min post mortem foi de 51,57 (&#177;1,91), 48,55 (&#177;1,94) e 41,67 (&#177;1,80), respectivamente, enquanto para o pH foi de 6,10 (&#177;0,48), 6,31 (&#177;0,59) e 7,15 (&#177;0,43) para carnes PSE, Normal e DFD, respectivamente. Após 24h post-mortem os resultados foram para L* 54,32 (&#177;1,09), 50,56 (&#177;1,64), 42,23 (&#177;1,80) e para o pH 5,69 (&#177;0,14), 5,78 (&#177;0,14), 6,05 (&#177;0,43) respectivamente para amostras PSE, Normal e DFD, significativamente diferente (P<0,05). Para o estudo das propriedades funcionais, amostras (n=374) foram selecionadas nas linhas de produção, classificadas em carnes PSE, Normal e DFD de acordo com o L* e pH 24h post mortem. Foram realizadas as medidas do exsudado formado, perda de umidade durante o cozimento e estabilidade da emulsão. Os resultados encontrados na medida do exsudado foram 4,73 (&#177;2,61)%, ,55 (&#177;2,16)%, 1,11 (&#177;1,01)% para PSE, Normal e DFD respectivamente, significativamente diferentes (P<0,05). A medida da perda de umidade durante o cozimento para as amostras PSE foi de 26,03 (&#177;2,93)%, para o grupo Normal a quebra foi de 25,38 (&#177;3,96)% e para o grupo DFD 22,89 (&#177;2,65)%. Entre as amostras dos grupos PSE e Normal não ocorreu diferença significativa (P<0,05), apesar da leve tendência para menor quebra de peso para grupo considerado Normal, enquanto que a quebra de peso para o grupo Normal e DFD apresentou resultados significativamente diferentes (P<0,05) com diferença significativa de 2,50% para DFD. Finalmente, para a medida de instabilidade da emulsão os resultados foram 1,04 (&#177;0,32)%, 0,78 (&#177;0,20)%, 0,70 (&#177;0,12)%, respectivamente para PSE, Normal e DFD, significativamente diferente (P<0,05). Concluindo, as matérias-primas que apresentam características DFD tendem a conferir melhor qualidade funcional com melhor capacidade de absorção, retenção de umidade, baixo exsudado e baixa perda durante o cozimento. A constatação da diferença nas cores do filé na linha de processamento pode padronizar os processos contribuindo para a distribuição adequada das amostras de carne PSE, Normal e DFD especificamente para diferentes tipos de processamento trazendo benefícios econômicos às industrias e qualidade sensorial aos consumidores. / The objective of this work was to investigate the incidence of DFD (dark, firm, dry) meat, in a commercial chicken plant and to determine its functional properties in comparison to PSE (pale, soft, exudative) and Normal breast fillet, Pectoralis major. In order to evaluate the occurrence of different color types, commercial Cobb and Ross chickens were submitted to water shower and ventilation treatments during 10 minutes immediately before slaughtering. Birds (n=329) were sacrificed, deboned and analyzed after 90 minutes and 24h post mortem for pH and L* values evaluation and submitted to classification as PSE, Normal, DFD meat based upon 24h post mortem samples. The occurrence was 24.92, 73.55 and 1,52% for samples PSE, Normal and DFD meats, respectively. L* values for 90 minutes post mortem were 51.57 (&#177;1.91), 48.55 (&#177;1.94) and 41.67 (&#177;1.80), respectively, while for pH values were 6.10 (&#177;0.48), 6.31 (&#177;0.59) and 7.15 (&#177;0.43) for PSE, Normal e DFD meats, respectively. After 24h post-mortem, L* values results were 54.32 (&#177;1.09), 50.56 (&#177;1.64), 42.23 (&#177;1.80) for PSE, Normal and DFD meats, respectively, and pH values were 5.69 (&#177;0.14), 5.78 (&#177;0.14), 6.05 (&#177;0.43) respectively for PSE, Normal and DFD meats. All results were significantly different (P<0.05). For functional properties evaluation, samples (n=374) were selected from production lines and ranked as PSE, Normal e DFD meats according to L* and pH 24h post mortem. Measurement for formed exudate, cook drip loss and emulsion stability was carried out. Results for formed exudate were 4.73 (&#177;2.61), 2.55 (&#177;2.16), 1.11 (&#177;1.01)% for PSE, Normal e DFD meats, respectively and for cook drip loss for PSE 26.03 (&#177;2.93), for Normal 25.38 (&#177;3.96) and for DFD 22.89 (&#177;2.65)%, respectively. No significantly difference (P<0,05) was found between PSE and Normal samples although there was a tendency for lower weight loss for normal group while between normal and DFD samples a significantly difference (P<0.05) was found. Finally, for emulsion estability results were 1.04 (&#177;0.32), 0.78 (&#177;0.20), 0.70 (&#177;0.12)%, respectively for PSE, Normal and DFD samples, significantly different (P<0,05). In conclusion, broiler breast fillet meat which presents DFD characteristics tends to have better functional properties for water holding capacity, lower exudate formation and lower cook drip loss. The standardization of colour differences detection in broiler fillet meat in the processing line could direct these samples specifically to PSE, normal and DFD meats for processing meat products accordingly in order to have economical benefits to the poultry industries and particularly sensorial meat products improvement for consumers.

Tesis (Magister en Derecho con y sin Menciones) / Actividad Formativa Equivalente a Tesis (AFET)

This research focused on the evaluation of iris recognition on post-mortem subjects. It was to determine if iris image captures were suitable from post-mortem subjects and if the captures contained the features required to be used in recognition scenarios. One commercially available iris camera was used, the IriShield USB MK2120U. In order to complete this research, it was first necessary to obtain images from subjects that contain the proper features, including sharpness, pupil size, and image quality. The images were captured during three different conditions that would be possible to find under real-world circumstances. The first condition was as the decedent came into the coroner’s office before the vitreous fluid was sampled from the eyes. The second condition was after the vitreous fluid was sampled from the deceased. Sampling vitreous fluid is a common autopsy procedure. This second condition would also be similar to a subject with a punctured eye. The third condition was after replacing the volume of vitreous fluid with saline solution. Replacing the vitreous with saline restored the round shape to the eye. This study found that high quality images can be captured from a post-mortem eye and that matching images across conditions results in positive identification.<br>

High Performance Liguid Chromatography was used to measure degradation of nucleotides in human cadavers for the purpose of prediction of post mortem interval. Endogenous nucleotides were extracted from integumentary tissue of six(6) human cadavers using six percent(6%) tricholoacetic acid. Linear regression statistical techniques were used to determine linearity of degradation of various nucleotide pools.

No description available.

A interpretação dos achados laboratoriais no estabelecimento da causa mortis consiste na integração dos conhecimentos sobre a toxicocinética e toxicodinâmica do agente, conhecimentos de sua redistribuição post mortem (RPM) e achados necroscópicos que possibilitem o nexo causal entre o toxicante e o efeito letal. Neste sentido, é importante considerar que somente as concentrações de cocaína (COC) e seus metabólitos podem não ser determinantes na interpretação da causa de morte, podendo ser útil o cotejamento com outros parâmetros, como os níveis de neurotrasmissores que representem o mecanismo de ação do fármaco. Assim, este trabalho teve por objetivo investigar a RPM da COC e seu metabólito benzoilecgonina (BE) em três segmentos do tecido encefálico (TE), no humor vítreo (HV) e sangue (SG), bem como determinar as concentrações de catecolaminas e indolaminas no encéfalo para avaliar a aplicação da neuroquímica post mortem (NPM) na toxicologia forense. No estudo de RPM foram quantificados os níveis de COC e BE em três segmentos do TE (córtex frontal, núcleos da base e cerebelo), no HV e no SG através de método por cromatografia líquida de alta eficiência (HPLC) acoplada ao detector de arranjo de diodos. Os estudos de neuroquímica foram realizados empregando-se HPLC acoplada ao detector eletroquímico. Os resultados indicaram que as concentrações médias de COC foram maiores no TE, seguido por SG e HV (3,09, 2,92 e 1,71 &#181;g/mL, respectivamente), enquanto para BE foram maiores em SG, seguido por HV e TE (6,12, 1,39 e 0,87 &#181;g/mL, respectivamente). As concentrações de COC se apresentaram distribuídas uniformemente nos três segmentos de TE e apresentaram alta correlação com o HV. Adicionalmente, a média de concentrações de dopamina total foi maior no grupo de indivíduos com amostras positivas para COC, sendo verificado diferença significativa entre este grupo e o de indivíduos com amostras negativas para o fármaco de interesse. Os resultados demostraram que o estudo de RPM e da NPM constituem ferramentas aplicáveis na interpretação da causa e maneira de morte. / In case of intoxication, the interpretation of analytical results to assess the cause and process of death requires knowledge about toxicokinetics, toxicodynamic, postmortem redistribution, and autopsy elements. Cocaine-related deaths occur mainly after prolonged drugs use and the presence of cocaine (COC) in fluids or tissues does not prove that death was due to COC consumption, and the interpretation of postmortem concentrations is even more complicated than attempts at making such correlations in the living. The objectives of this study were to investigate the post mortem redistribution (PMR) of COC and its metabolite benzoylecgonine (BE) in three segments of brain (frontal cortex, base nucleous, and cerebellum), vitreous humor, and blood. In additional, catecholamines and indolamines were quantified in brain in order to evaluate the usefulness of post mortem neurochemistry (PMN) in forensic toxicology. In PMR studies were quantified the COC and BE levels in three brain (BR) segments, in vitreous humor (VH), and blood (BL) by High Performance Liquid Chromatography (HPLC) with diode array detection, and for neurochemistry studies the neurotransmitters were quantified by HPLC with electrochemical detection. A homogenous distribution of COC and BE within frontal cortex, base nucleous, and cerebellum was found. The COC media concentrations were 3.09, 2.92 e 1.71&#181;g/mL in BR, BL and VH, respectively, and the BE media concentrations were 6.12, 1.39 e 0.87 &#181;g/mL in BL, VH, and BR, respectively. The COC concentrations in VH show high correlation with brain. The media total dopamine concentration was significant higher in COC positive group. These findings suggest that the studies of PMR and PMN by neurotransmitters levels may be useful to assess the cause and process of death.

Esta dissertação de Mestrado tem por objetivo explicitar a relação entre senhores e escravos, analisando as alforrias no contexto da transmissão de bens e partilha de heranças, entre 1850 e 1888, nas freguesias da Sé, Brás, Santa Efigênia, Penha, Conceição de Guarulhos, Juquery, Nossa Senhora do Ó, São Bernardo e Cotia, que eram os Distritos de Paz da cidade de São Paulo. Pretende-se, partindo das manumissões registradas nos testamentos, principalmente as condicionadas à morte do proprietário de cativos, verificar a obtenção da liberdade jurídica, entendida como conquista do status jurídico de forro pelo ex-cativo e seus possíveis significados. Para tal enfoque, serão analisados os testamentos, inventários post-mortem e as cartas de liberdade registradas na Capital. Desse modo, será possível confrontar a vontade dos testadores à forma como esta pôde ser cumprida, após a morte, levando em consideração o patrimônio amealhado, as atitudes dos herdeiros nas sucessões e a legislação exposta nas Ordenações Filipinas, na Constituição de 1824, nos decretos e leis excepcionais, no Direito Romano e no direito consuetudinário, dispositivos que influenciavam nos assuntos relativos à escravidão. / This Masters Degree Dissertation has as subject to demonstrate the relationship between masters and slaves, analysing the alforrias in the context of transmition of goods and sharing of heritages, from 1850 to 1888, in the Freguesias of Sé, Brás, Santa Ifigênia, Penha, Conceição de Guarulhos, Juquery, Nossa Senhora do Ó, São Bernardo and Cotia, which were the Distritos de Paz of São Paulo City. We aim to verify the obtention of legal freedom, understood as a conquest of legal status as forro by the ex-captive and the possibles meanings of it, starting from manumitions registered on wills, mainly the conditional manumitions linked to the death of captive owners. Through this perspective, the wills, inventories post-mortem and freedom letters registered on the city capital will be analysed. By this way, it will be possible to confront the wishes of the slave owners with the mode those ones could be accomplished after the owners death, taking in account the patrimony amassed, the atitudes of the heirs concerning the succession, and the laws exposed in the Ordenações Filipinas, in the Constitution of 1824, in the decrees and exceptional laws, in Roman and Common Law, in all these legal documents that had influence on the themes related to slavery.

A interpretação dos achados laboratoriais no estabelecimento da causa mortis consiste na integração dos conhecimentos sobre a toxicocinética e toxicodinâmica do agente, conhecimentos de sua redistribuição post mortem (RPM) e achados necroscópicos que possibilitem o nexo causal entre o toxicante e o efeito letal. Neste sentido, é importante considerar que somente as concentrações de cocaína (COC) e seus metabólitos podem não ser determinantes na interpretação da causa de morte, podendo ser útil o cotejamento com outros parâmetros, como os níveis de neurotrasmissores que representem o mecanismo de ação do fármaco. Assim, este trabalho teve por objetivo investigar a RPM da COC e seu metabólito benzoilecgonina (BE) em três segmentos do tecido encefálico (TE), no humor vítreo (HV) e sangue (SG), bem como determinar as concentrações de catecolaminas e indolaminas no encéfalo para avaliar a aplicação da neuroquímica post mortem (NPM) na toxicologia forense. No estudo de RPM foram quantificados os níveis de COC e BE em três segmentos do TE (córtex frontal, núcleos da base e cerebelo), no HV e no SG através de método por cromatografia líquida de alta eficiência (HPLC) acoplada ao detector de arranjo de diodos. Os estudos de neuroquímica foram realizados empregando-se HPLC acoplada ao detector eletroquímico. Os resultados indicaram que as concentrações médias de COC foram maiores no TE, seguido por SG e HV (3,09, 2,92 e 1,71 &#181;g/mL, respectivamente), enquanto para BE foram maiores em SG, seguido por HV e TE (6,12, 1,39 e 0,87 &#181;g/mL, respectivamente). As concentrações de COC se apresentaram distribuídas uniformemente nos três segmentos de TE e apresentaram alta correlação com o HV. Adicionalmente, a média de concentrações de dopamina total foi maior no grupo de indivíduos com amostras positivas para COC, sendo verificado diferença significativa entre este grupo e o de indivíduos com amostras negativas para o fármaco de interesse. Os resultados demostraram que o estudo de RPM e da NPM constituem ferramentas aplicáveis na interpretação da causa e maneira de morte. / In case of intoxication, the interpretation of analytical results to assess the cause and process of death requires knowledge about toxicokinetics, toxicodynamic, postmortem redistribution, and autopsy elements. Cocaine-related deaths occur mainly after prolonged drugs use and the presence of cocaine (COC) in fluids or tissues does not prove that death was due to COC consumption, and the interpretation of postmortem concentrations is even more complicated than attempts at making such correlations in the living. The objectives of this study were to investigate the post mortem redistribution (PMR) of COC and its metabolite benzoylecgonine (BE) in three segments of brain (frontal cortex, base nucleous, and cerebellum), vitreous humor, and blood. In additional, catecholamines and indolamines were quantified in brain in order to evaluate the usefulness of post mortem neurochemistry (PMN) in forensic toxicology. In PMR studies were quantified the COC and BE levels in three brain (BR) segments, in vitreous humor (VH), and blood (BL) by High Performance Liquid Chromatography (HPLC) with diode array detection, and for neurochemistry studies the neurotransmitters were quantified by HPLC with electrochemical detection. A homogenous distribution of COC and BE within frontal cortex, base nucleous, and cerebellum was found. The COC media concentrations were 3.09, 2.92 e 1.71&#181;g/mL in BR, BL and VH, respectively, and the BE media concentrations were 6.12, 1.39 e 0.87 &#181;g/mL in BL, VH, and BR, respectively. The COC concentrations in VH show high correlation with brain. The media total dopamine concentration was significant higher in COC positive group. These findings suggest that the studies of PMR and PMN by neurotransmitters levels may be useful to assess the cause and process of death.