Thesis (M. Sc.)--University of Canterbury, 2006. / Typescript (photocopy). Includes bibliographical references (leaves (184-185). Also available via the World Wide Web.

Thesis (Ph. D.)--West Virginia University, 2004. / Title from document title page. Document formatted into pages; contains ix, 174 p. : ill. (some col.). Vita. Includes abstract. Includes bibliographical references (p. 166-170).

Thesis (Ph. D.)--University of Rhode Island, 2004. / Typescript. Includes bibliographical references (leaves 138-141).

Thesis (M.S.)--West Virginia University, 2006. / Title from document title page. Document formatted into pages; contains vi, 56 p. : ill. (some col.). Includes abstract. Includes bibliographical references (p. 55-56).

This thesis is divided into two parts, each of which examines aspects of bloodstain analysis where gravity is the main force applied to blood. Part I is a preliminary study on the dynamics of blood flow on various inclined surfaces and examines the use of blood analogs for easy test replication. The flow of uncoagulated human blood at different volumes and temperatures was examined on wood at a set angle of 1.5°, and on glass at varying incline angles. Glycerol solutions of 59% and 42% were used to represent blood at 23°C and 37°C respectively. Glycerol flow trials of similar volumes were conducted on wood, PVC and glass. Fluid flow plots of distance versus time exhibited double exponential curve behaviour, although a power-law relationship derived by H. E. Huppert's (1982) flow expression was obtained for blood flowing on inclined wood. Blood flow exhibited several observable characteristics; a decreasing width of the leading edge over time, and streaking and component separation of the leading region at very low speeds. On a glass surface, the width of the initial flow region decreased and initial speed increased with increasing angles. The glycerol analogs used in this study did not represent their blood counterparts well due to differences in physical properties of the fluids. Part II of this study focuses on the forensic value of passive bloodstains on three fabrics; 100% cotton drill, 65/35 polyester cotton, and 100% Shantung silk. 26 µL drops of 37°C human blood were deposited onto the three fabrics and paper from a height of 14 cm at various impact angles. The stains were photographed and analysed qualitatively and quantitatively using computational methods. 100% cotton drill, 65/35 polyester cotton and ironed 100% Shantung silk provided useful forensic values such as direction of travel and angle of impact. Overall, this study has provided useful preliminary data for further research work.

Being able to detect if drugs were used in the commission of a crime, and if so what drugs, is of great importance. For many cases these tests can be carried out on an intimate blood or urine sample (one recovered directly from the subject in question), however this may not always be the case. In cases where a dried bloodstain is the only source of biological material, identifying the presence of drugs affecting an individual at the time of stain deposition has not been well studied. Towards this goal, two methods of detection of drugs of abuse in dried bloodstains were evaluated: lateral flow immunoassay test cards and gas chromatography-mass spectrometry. Stains were created using certified drug-free blood spiked with analytes of interest, and were then extracted and introduced into each testing method. Both methods proved effective for the detection of one or two of the five chosen analytes (amphetamine, cocaine, morphine 3-ß-D-glucuronide, phencyclidine and 11-nor-Δ9-tetrahydracannabinol), even after 24 hours of drying at room temperature. With further method optimization and more thorough method development, these methods may, in the future, be effectively used for drug detection in forensic stains. However, neither method evaluated in this study was able to detect all of the drugs tested.

Made available in DSpace on 2016-08-10T10:38:45Z (GMT). No. of bitstreams: 1 LUCIANA MAIA ESCHER DOS SANTOS.pdf: 3201320 bytes, checksum: f5c8c078f60bbba17912e8c39742ed0a (MD5) Previous issue date: 2013-06-24 / Molecular Biology shown to be an effective tool in forensic laboratories for the ability to identify an individual from minute amounts of biological samples such as blood, bones, semen, hair, teeth, nails, spittle, urine and other biological fluids recovered from the crime scene. One of the main biological evidence found at a crime scene are traces of bloodstains. This study aimed to analyze the molecular profiles of biological samples exposed to the chemiluminescent reagent luminol based on different storage times (48 hours and 30 days). With the measurement of all samples can be inferred that in the first 48 hours of storage, there was obtained DNA and varying concentrations after application of the chemiluminescent reagent. The samples were amplified by PCR using a multiplex system AmpFlSTR ® Select NGM and capillary electrophoresis. Molecular profiles were obtained complete and incomplete denoting specificity as the quality and quantity of the sample analyzed. The selection of molecular markers mini-STRs greatly contributed to the success of these profiles, allowing the study and analysis of degraded material. Our data suggest that the degradation of the DNA molecule exposed to the chemiluminescent reagent was higher in the samples compared to those containing diluted whole blood impregnation. Therefore, analyzing the bloodstains samples exposed to luminol in shorter storage provide molecular profiles compatible to clash samples. / A Biologia Molecular mostrou-se uma ferramenta efetiva nos laboratórios forenses pela capacidade de identificar um indivíduo a partir de quantidades ínfimas de amostras biológicas como sangue, ossos, sêmen, cabelo, dentes, unhas, saliva, urina, entre outros fluidos biológicos recuperados no local do crime. Uma das principais evidências biológicas encontradas em local de crime são vestígios de substâncias hematóides. Esta pesquisa visou a análise de perfis moleculares de amostras biológicas expostas ao reagente quimioluminescente a base de luminol em diferentes tempos de armazenamento (48 horas e 30 dias). Com a quantificação de todas as amostras pode-se inferir que nas primeiras 48 horas de armazenamento, obtiveram-se concentrações variáveis de DNA e após a aplicação do reagente quimioluminescente. As amostras foram amplificadas por PCR utilizando um sistema multiplex AmpFlSTR® NGM SElect e eletroforese capilar. Foram obtidos perfis moleculares completos e incompletos denotando inespecificidade conforme a qualidade e quantidade da amostra analisada. A seleção dos 17 marcadores moleculares mini-STRs em muito contribuiu para o sucesso desses perfis, permitindo estudo e análise de material degradado. Os dados indicam que a degradação da molécula de DNA exposta ao reagente quimioluminescente foi maior nas amostras diluídas em relação àquelas contendo impregnação com sangue total. A análise estatística das amostras com e sem exposição ao reagente quimioluminescente comparado ao grupo controle e em diferentes concentrações, mostrou não haver diferença estatísticamente significativa entre os valores quantificados obtidos. Ainda assim, analisar as amostras hematóides expostas ao luminol em menor tempo de armazenamento proporcionarão perfis moleculares compatíveis ao confronto de amostras.

With current scientific technologies, a significant amount of genetic information can be obtained from biological evidence found at a crime scene. Not only is it possible to identify the donor of the evidence through routine DNA profiling techniques, but new RNA based methods are being developed to determine the tissue type as well as the physical characteristics of the donor. Despite the information that can be obtained, the ability to determine the age or time the biological material was deposited at the crime scene has eluded the forensic community thus far. Timing is critically important as it could help police determine when the crime was committed. In this body of work an investigation was conducted into whether the degradation rates of nucleic acid macromolecules could serve as molecular clocks for age estimations. An attempt was made to gain a better understanding of the degradation products produced from an internal urban environment and to develop an optimal assay accordingly. A number of different RNA based techniques for ageing both hair and blood samples were also examined. Degradation assays have been traditionally designed around amplicon size however, it was established that testing loci stability is an essential requirement in the optimisation process. The results presented in this thesis suggest the reliability of the data can be increased when the two competing target species are selected from the same loci, which minimised the effect of loci susceptibility to degradation. It was determined that blood stains aged up to 60 days in an internal urban environment were best distinguished (in terms of age estimations) by using targets that differed in size by 170 to 240 base pairs, with one of the targets being between 200 and 300 base pairs in length. Despite using a robust TH01 qPCR assay it was established that an internal “urban” environment was not as stable as predicted and that seasonal temperature variation had a large effect on degradation rates. Interpretation of the results was therefore limited suggesting these optimised target sizes may only be relevant to the winter months. Using a carefully designed hermetically sealed dry swab we were able to remove moisture and inhibit the growth of DNA consuming micro-organisms. It was determined that bacteria alone can cause a 2-fold increase in the degradation rate of a sample aged at room temperature. In terms of integrity, storing samples at room temperature in a moisture free environment was equivalent to storing standard samples (exposed to normal humidity levels) in refrigerated conditions. It was also determined that the effect of bacterial degradation can be halved by lowering the storage temperature from room temperature to 4°C. RNA was examined in an attempt to reduce the large variations that had inhibited previous DNA methodologies. IL-6 and TNF-α were initially selected due to their rapid post-extraction change in expression levels. However, their levels were highly variable, unpredictable and therefore not suitable for this type of analysis even on samples that had been aged for only ten days. It is thought that their dynamic roles in a number of haemopoietic processes could be responsible for the poor results. A new RNA methodology, as described by Nolan et al (2008) was used to analyse samples that had been aged over 80 days. Four targets, AMICA1, MNDA, CASP1 and GAPDH were chosen based on their cell lineage as it was hypothesised that inter-donor variation could be reduced by using targets confined to the granulocytic cell lineage. Using the novel 3’/5’ assay, AMICA1, MNDA and CASP1 all performed poorly and no correlation could be determined between the 3’/5’ ratio and sample age. GAPDH showed some encouraging results with a correlation of 0.912 (age to 3’/5’ ratio) although initial stability over the first 20 days and the inter-donor variation were still limiting factors. It was also thought that the various mRNA degradation processes, in particular the 5’/3’ exonuclease activity, contributed to the poor results generally. A large inter-donor variation was a common aspect to all the blood based methodologies trialled. This meant that none of the methods had any practical value. As a result, an alternative RNA method was used to determine if it was possible to age another forensically important type of biological evidence; hair. Using a Reverse Transcription Quantitative PCR (RT-qPCR) assay, we monitored the Relative Expression Ratio (RER) of two different RNA species (18S rRNA and B-actin mRNA) in hair samples that were aged naturally over a period of three months. Overall the results presented here suggest that the age of hair samples containing follicular tags can be approximated using a second order polynomial (Age = 3.31RER2 - 2.85RER – 0.54), although with limitations.

The success or failure of any criminal investigation often depends on the recognition of physical evidence left at a crime scene and the proper analysis of that evidence. Crime scenes that involve bloodshed often contain a wealth of information in the form of blood patterns, the location, and its cause. Any criminal investigation has specific tasks, from the time when the crime is reported to the reconstruction of crime scenes. A lot of work needs to be done. Once the investigation starts at the crime scene, BPA needs to be done at the crime scene and the investigating officer must identify this evidential tool. The investigating officer should not necessarily have specialised training in blood pattern analysis, but rather know when to use these experts at their bloody crime scenes. With the interviews and docket analysis done, the researcher found that this was a problem as the investigating officers, either had no knowledge on the subject of BPA or very little knowledge on this research. The purpose of this study was to determine the use of BPA to CSR, and for the investigating officer to realise that it is not just a bloody crime scene, but also contains a wealth of evidence. The researcher had two research questions. Once the investigating officer follows the objectives of criminal investigation, they should be able to have a strong case against the perpetrators. How could BPA be used in the reconstructing of a crime scene? The researcher wanted to bring it to the investigating officers’ attention that it is not just a bloody crime scene, but rather that it contains a wealth of evidence, which can give them a perspective of the movement of both the victim and perpetrator during the commencement of the crime. Regardless of the lack of knowledge of BPA, it is proposed that investigating officers are to be informed, either through station lectures or by yearly refresher workshops and courses of the evidential tool of BPA. When the bloody crime scene is reconstructed with the use of BPA, an insight of what transpired at the crime scene will help them to finalise their cases. For recommendations, it is proposed that investigating officers are to be trained in more in depth courses in criminal investigation as well as crime scene reconstruction and evidence collection using FSL. / Criminology and Security Science / M.Tech. (Forensic Investigation)