In forensic casework analysis it is necessary to obtain genetic profiles from increasingly smaller amounts of biological material left behind by perpetrators of crime. The ability to obtain profiles from trace biological evidence is demonstrated with so-called ‘touch DNA evidence’ which is perceived to be the result of DNA obtained from shed skin cells transferred from donor to an object or person during physical contact. However, the current method of recovery of trace DNA involves cotton swabs or adhesive tape to sample an area of interest. This "blindswabbing" approach may result in the recovery of biological material from different individuals resulting in admixed DNA profiles which are often difficult to interpret. Profiles recovered from these samples are reported to be from shed skin cells with no biological basis for that determination. A specialized approach for the isolation of single or few cells from ‘touch DNA evidence’ is necessary to improve the analysis and interpretation of recovered profiles. Here we describe the development of optimized and robust micro volume PCR reactions (1-5 μL) to improve the sensitivity and efficiency of ‘touch DNA’ analysis. These methods will permit not only the recovery of the genetic profile of the donor of the biological material, but permit an identification of the tissue source of origin using mRNA profiling. Results showed that the 3.5 uL amplification volume, a fraction of the standard 25 uL amplification volume, was the most ideal volume for the DNA assay, as it had very minimal evaporation with a 50% profile recovery rate at a single cell equivalent input (~5 pg) with reducing amplification volume alone. Findings for RNA showed that by reducing both amplification steps, reverse transcriptase PCR (20 uL) and body fluid multiplex PCR (25 uL), to iv 5 uL, ideal results were obtained with an increase in sensitivity and detection of six different body fluids down to 50 pg. Once optimized at the trace level, the assays were applied to the collection of single and few cells. DNA findings showed that about 40% of a full profile could be recovered from a single buccal cell, with nearly 80% of a full profile recovered from only two cells. RNA findings from collected skin particles of "touched" surfaces showed accurate skin detection down to 25 particles and detection in one clump of particles. The profiles recovered were of high quality and similar results were able to be replicated through subsequent experiments. More studies are currently underway to optimize these developed assays to increase profile recovery at the single cell level. Methods of doing so include comparing different locations on touched surfaces for highest bio-particle recovery and the development of physical characteristics of bio-particles that would provide the most ideal results

Background: Accidental exposure to patients’ body fluids (BFs) is an occupational hazard among health care workers (HCWs). The study aimed at describing the patterns of exposure to patients’ BFs among HCWs at a university hospital in Ethiopia. Methods: A contextual descriptive cross-sectional design was used for this study. Self-administered questionnaires were used to collect data. Results: The one year and professional life prevalence of occupational exposure to patients’ BFs among HCWs was 33.5% and 66.5% respectively. Circumstances that led to participants’ exposures to patients’ BFs include needle stick injuries to fingers and splashes to the eyes (82.4%); conducting procedures included blood withdrawal (10.8%) and inserting intravenous infusions (8.1%) and recapping of used needles (12.2%). Conclusion: Findings of this study generally indicated that occupational exposures to patients’ BF of different types and circumstances were common among all categories of HCWs in the study site. This high finding of BF exposure should not be over looked. HCWs should follow the universal precaution protocol and PEP need to be strengthened / Health Studies / M.A. (Public Health)

South African forestry workers are predisposed to dehydration due to the heavy physical activity they perform in impermeable regulation safety clothing in hot and often humid environments where the availability of a variety of suitable fluids at reasonable temperatures is limited. As dehydration reduces both physical and mental capacity the potential consequences include decreased productivity and an increased risk for injury. The aim of this cross sectional observational study was to determine the prevalence and severity of dehydration in rural forestry workers in both winter (minimum and maximum daily temperatures 3-22°C) and autumn (minimum and maximum daily temperatures 14-27°C). The convenience sample included 103 workers in autumn (Nelspruit, n=64 males, n=39 females, mean age 37.32 years, mean BMI 22.3 kg/m2) and 79 in winter (Richmond, n=68 males, n=11 females, mean age 25.85 years, mean BMI 22.2 kg/m2). The sample included chainsaw operators, chainsaw operator assistants, debarkers and stackers. The risk of heat illness was moderate in Nelspruit (average daily temperature 21.1°C 67% rh) and low in Richmond (average daily temperature 17.0°C 39% rh). The prevalence of dehydration was determined by urine specific gravity (USG) measurements. Percent loss of body weight in the course of the shift was used to determine the severity of dehydration. In Nelspruit 43% (n=43) and in Richmond 47% (n=37) of the forestry workers arrived at work dehydrated (USG>1.020 g/ml). Pre break this had increased to 49% (n=49) in Nelspruit and 55% (n=33) in Richmond. By the end of shift the number of dehydrated forestry workers had significantly increased to 64% (n=64, p≤0.001) in Nelspruit and 63% (n=42, p=0.043) in Richmond. A minimum of 21% (n=2) in Nelspruit and 23% (n=15) in Richmond of the forestry workers had lost more than 2% of their body weight which could significantly decrease work capacity and work output as well as mental and cognitive ability. Dehydration was not related to season (winter/autumn), gender or job category. In Nelspruit 23% (n=23) and in Richmond 13% (n=10) arrived at work overhydrated (USG<1.013 g/ml). Pre break this had decreased to 14% (n=14) in Nelspruit and 10% (n=6) in Richmond. By the end of shift 4% (n=4) in Nelspruit and 2% (n=1) in Richmond had remained overhydrated and without correcting for fluid and food intake, 5% (n=5) had gained over 2% of their body weight in Nelspruit while none had gained weight in Richmond. Overhydration was not related to season (winter/autumn), gender or job category. Physical symptoms at the end of shift included tiredness (24%), toothache (13%) and headaches (10%) although these did not correlate to end of shift USG readings (p=0.221). The fluid requirements for male workers (n=8) who did not eat or drink across the shift was 439 ml per hour. The contractors were unaware of how much fluid should be supplied to workers and how much fluid they actually supplied. The only fluid provided by the contractors was water at the ambient air temperature which was the main source of fluid for the majority. Some forestry workers brought a limited variety of other fluids including amahewu, tea and cold drinks to work. At least 40% of the work force investigated, started their shift already compromised to work to capacity (USG>1.020 g/ml). The prevalence of dehydration had increased by the break emphasizing the need to begin drinking early on in the shift. The majority of forestry workers were dehydrated at the end of the shift. A significant proportion was dehydrated to the extent (>2%) that both work capacity and mental ability would be significantly compromised. A select group of forestry workers were drinking excessive amounts of fluid and were therefore susceptible to potentially fatal dilutional hyponatremia especially as water was the primary source of fluid. Dehydration in both autumn and winter was identified as being a significant but preventable risk. As a consequence of overhydration, a small group of forestry workers may be susceptible to dilutional hyponatremia. Fluid intake guidelines for males of 450 ml per hour appeared to be safe and were within the recommendations of the American College of Sports Medicine. Fluid guidelines for females need investigation. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2008.

Background: Accidental exposure to patients’ body fluids (BFs) is an occupational hazard among health care workers (HCWs). The study aimed at describing the patterns of exposure to patients’ BFs among HCWs at a university hospital in Ethiopia. Methods: A contextual descriptive cross-sectional design was used for this study. Self-administered questionnaires were used to collect data. Results: The one year and professional life prevalence of occupational exposure to patients’ BFs among HCWs was 33.5% and 66.5% respectively. Circumstances that led to participants’ exposures to patients’ BFs include needle stick injuries to fingers and splashes to the eyes (82.4%); conducting procedures included blood withdrawal (10.8%) and inserting intravenous infusions (8.1%) and recapping of used needles (12.2%). Conclusion: Findings of this study generally indicated that occupational exposures to patients’ BF of different types and circumstances were common among all categories of HCWs in the study site. This high finding of BF exposure should not be over looked. HCWs should follow the universal precaution protocol and PEP need to be strengthened / Health Studies / M.A. (Public Health)

Se sintetizaron nanocompuestos híbridos en bloque de poli(etil metacrilato-co-hidroxietil acrilato) 70/30 wt%/sílice, P(EMA-co-HEA)/SiO2, con distintas proporciones de sílice hasta el 30 wt%. El procedimiento de síntesis consistió en la copolimerización de los monómeros orgánicos durante la polimerización sol-gel simultánea de tetraetoxisilano, TEOS como precursor de sílice. El TEOS se hidroliza eficientemente y condensa dando lugar a sílice, y presenta una distribución homogénea en forma de agregados inconexos de nanopartículas de sílice elementales en los híbridos con bajos contenidos de sílice (<10 wt%) o redes continuas interpenetradas con la red orgánica tras la coalescencia de los agregados de sílice (>10 wt%). La red polimérica orgánica se forma en los poros producidos en el interior de las nanopartículas elementales de sílice, y también en los poros formados entre los agregados de nanopartículas. Los nanohíbridos con contenidos de sílice intermedios (10-20 wt%) exhibieron las propiedades más equilibradas e interesantes: i) refuerzo mecánico de la matriz orgánica conseguida gracias a redes de sílice continuas e interpenetradas, ii) buena capacidad de hinchado debida a la expansión de la red orgánica no impedida todavía por un esqueleto de sílice rígido, y a un número alto de grupos silanol terminales hidrófilos (concentraciones inorgánicas en los alrededores de la coalescencia), y iii) mayor reactividad superficial debido a un contenido relativo bastante elevado de grupos polares silanol terminales disponibles en las superficies. La 'bioactividad' o capacidad de los materiales en bloque de formar hidroxiapatita (HAp) sobre sus superficies fue estudiada in vitro sumergiéndolos en fluido biológico simulado (simulated body fluid, SBF). La formación de la capa de HAp viene controlada por el mecanismo y el tiempo de inducción a la nucleación de la misma, que dependen a su vez de la estructura de la sílice. / Vallés Lluch, A. (2008). P(EMA-co-HEA)/SiO2 hybrid nanocomposites for guided dentin tissue regeneration: structure, characterization and bioactivity [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/3795

Indiana University-Purdue University Indianapolis (IUPUI) / In recent years, magnesium alloys, due to their high strength and biocompatibility, have attracted significant interest in medical applications, such as cardiovascular stents, orthopedic implants, and devices. To overcome the high corrosion rate of magnesium alloys, coatings have been developed on the alloy surface. Most coating methods, such as anodic oxidation, polymer coating and chemical conversion coating, cannot produce satisfactory coating to be used in human body environment. Recent studies demonstrate that micro-arc oxidation (MAO) technique can produce hard, dense, wear-resistant and well-adherent oxide coatings for light metals such as aluminum, magnesium, and titanium. Though there are many previous studies, the understanding of processing conditions on coating performance remains elusive. Moreover, previous tests were done in simulated body fluid. No test has been done in a cell culture medium, which is much closer to human body environment than simulated body fluid. In this study, the effect of MAO processing time (1 minute, 5 minutes, 15 minutes, and 20 minutes) on the electrochemical behaviors of the coating in both conventional simulated body fluid and a cell culture medium has been investigated. Additionally a new electrolyte (12 g/L Na2SiO3, 4 g/L NaF and 4 ml/L C3H8O3) has been used in the MAO coating process. Electrochemical behaviors were measured by performing potentiodynamic polarization and electrochemical impedance spectroscopy tests. In addition to the tests in simulated body fluid, the MAO-coated and uncoated samples were immersed in a cell culture medium to investigate the corrosion behaviors and compare the difference in these two kinds of media. The results show that in the immersion tests in conventional simulated body fluid, the 20-minute MAO coated sample has the best resistance to corrosion due to the largest coating thickness. In contrast, in the cell culture medium, all MAO coated samples demonstrate a similar high corrosion resistance behavior, independent of MAO processing time. This is probably due to the organic passive layers formed on the coating surfaces. Additionally, a preliminary finite element model has been developed to simulate the immersion test of magnesium alloy in simulated body fluid. Comparison between the predicted corrosion current density and experimental data is discussed.

Ces travaux de thèse s'articulent autour de deux projets : les études structure-fonction de l'aminopeptidase A, d'une part, et celles du récepteur de l'apéline, d'autre part. I/ L'aminopeptidase A (APA, EC 3.4.11.7) est une aminopeptidase monozinc membranaire qui, dans le cerveau, produit l'angiotensine (Ang) III à partir de l'Ang II. L'Ang III est l'un des principaux peptides effecteurs du système rénine-angiotensine cérébral qui exerce un effet stimulateur tonique sur le contrôle central de la pression artérielle chez le rat hypertendu. Ainsi le blocage de l'APA par un inhibiteur spécifique et sélectif, l'EC33 ou sa prodrogue, le RB150, normalise la pression artérielle dans deux modèles expérimentaux d'hypertension artérielle (HTA). L'APA constitue une cible thérapeutique potentielle pour le traitement de l'HTA qui justifie le développement de nouveaux inhibiteurs de cette enzyme plus puissants et plus sélectifs que l'EC33 et avec un profil pharmacodynamique et pharmacocinétique amélioré par rapport au RB150. Pour cela, nous avons construit un modèle tridimensionnel (3D) de l'APA sur la base de la structure cristallographique de l'APA humaine récemment publiée. Nous avons ensuite validé ce modèle par des études structure-fonction par modélisation moléculaire et mutagénèse dirigée en démontrant l'implication, d'un résidu du sous-site S1 dans la spécificité de substrat acide de l'APA et de deux résidus formant le sous-site S2' interagissant avec le résidu P2' acide d'inhibiteurs tripeptidiques précédemment développés dans le laboratoire.II/ L'apéline est le ligand naturel du récepteur orphelin humain APJ (ApélineR), un récepteur à sept domaines transmembranaires couplé aux protéines G. L'apéline et son récepteur sont impliqués dans le maintien de l'équilibre hydrique et des fonctions cardiovasculaires. L'ApélineR constitue une cible thérapeutique potentielle dans le traitement de l'insuffisance cardiaque et des rétentions hydriques. Etant donné que la demi-vie de l'apéline dans la circulation sanguine est de l'ordre de la minute, l'objectif est de développer des analogues de l'apéline métaboliquement stables. Pour développer de tels composés, nous avons entrepris de comprendre comment l'apéline se lie à son récepteur et comment elle l'active. Dans ce but, nous avons construit un modèle 3D de l'ApélineR basé sur la structure cristallographique du récepteur aux chimiokines, CXCR4. Nous avons validé ce modèle par des études structure-fonction par modélisation moléculaire et mutagénèse dirigée. Nous avons identifié à la surface du récepteur, les résidus acides des boucles extracellulaires qui interagissent avec les résidus basiques de l'apéline. Nous avons ensuite développé des analogues de l'apéline-17 (K17F) métaboliquement stables par deux stratégies différentes. Premièrement, nous avons substitué chacun des résidus de l'apéline par son énantiomère de la série D ou par un acide aminé synthétique. Deuxièmement, nous avons ajouté une chaîne fluoroalkyle à l'extrémité N-terminale de l'apéline. Ces deux stratégies ont permis d'obtenir plusieurs composés dont les plus actifs sont le P92 et le LIT01-196 qui conservent des propriétés pharmacologiques identiques à celles de K17F et qui présentent une demi-vie plasmatique largement supérieure à celle du peptide endogène. Ces deux analogues se sont révélés particulièrement actifs in vivo avec une capacité à diminuer la pression artérielle et à réduire la sécrétion de vasopressine dans le sang conduisant à une augmentation de la diurèse aqueuse. Les modèles 3D validés de l'APA et de l'ApélineR seront utilisés pour des campagnes de criblage in silico de chimiothèques virtuelles afin de découvrir de nouveaux inhibiteurs de l'APA et des agonistes de l'ApélineR qui pourraient conduire à terme à de nouveaux candidats-médicaments. Ces composés pourraient être utiles pour le traitement de l'HTA et de l'insuffisance cardiaque. / The doctoral work was divided in two parts, one on the structure-function studies of aminopeptidase A, and the second one, on those of the apelin receptor. I/ Aminopeptidase A (APA) is a membrane bound monozinc aminopeptidase which generates, in the brain, angiotensin (Ang) III from Ang II. Ang III is one of the main effector peptides of the brain renin-angiotensin system, which exerts a tonic stimulatory action on the control of blood pressure in hypertensive rats. Thus, the blockade of brain APA by a specific and selective inhibitor, EC33 or its prodrug, RB150, normalizes blood pressure in two animal models of arterial hypertension (HTA). APA constitutes a potential therapeutic target for the treatment of HTA that justifies the development of more potent and selective APA inhibitors than EC33, with enhanced pharmacodynamic and pharmacokinetic profiles when compared to RB150. With this aim, we built a three dimensional (3D) model of APA based on the recently published crystal structure of human APA. We validated this model by structure-function studies combining molecular modeling and site-directed mutagenesis demonstrating the crucial role of one residue in the S1 subsite responsible for substrate specificity of APA for N-terminal acidic amino-acid residues and two other residues constituting the S2' subsite of APA involved in the binding of the P2' acidic residue of tripeptidic inhibitors, previously developed in the laboratory. II/ Apelin is the endogenous ligand of the human orphan receptor named APJ (ApelinR), a G protein-coupled receptor. Apelin and ApelinR are involved in the control of body fluid homeostasis and cardiovascular functions. ApelinR constitutes a potential therapeutic target for the treatment of heart failure and water retentions. Given that apelin half-life in the blood circulation is in the minute range, we aimed to develop potent metabolically stable apelin analogs.. In this context, it is necessary to understand how apelin binds to ApelinR and how it is activated. To do so, we build a 3D model of ApelinR based on the crystal structure of the chemokine receptor, CXCR4. We validated this model by structure-function studies by molecular modeling and site-directed mutagenesis. We showed that apelin interacts with the receptor through interactions between the basic residues of the peptide and the acidic residues of the ApelinR, located in the extracellular loops. ,We then developed metabolically stable apelin-17 (K17F) analogs following two different strategies. First, we substituted each residue of K17F by its D-isomer or a synthetic amino-acid. Secondly, we added a fluoroalkyl chain at the N-terminal part of K17F. These two strategies allowed to significantly improve plasma half-life of the modified peptides for several hours without modifying their pharmacological properties as compared to K17F. Two apelin metabolically stable analogs, P92 and LIT01-196, were found to have significantly higher in vivo activity than K17F with a strong capacity to decrease blood pressure and to inhibit vasopressin release in the blood stream inducing an increased aqueous diuresis. These new validated 3D models will be now used to perform in silico screening of virtual chemical libraries to discover new APA inhibitors and ApelinR agonists that could ultimately lead to new drug candidates. These compounds could be useful for the treatment of HTA and heart failure.

Serious and violent crime in South Africa (SA) has been on the rise in the last few years. The SAPS has been stretched to the maximum in its efforts of trying to stabilise the situation, and has applied various tactics and strategies to eradicate crime. This included, among other things, changes to the basic training programme and reintroduction of specialised units. The success or failure of any criminal investigation will still often depend on the detection and analysis of physical evidence found on the crime scene. Crimes such as rape will always leave behind physical evidence in the form of body-fluids. The detectives need not be experts in order to detect or analyse this physical evidence, but should be experienced enough to know which experts or investigative aids must be used to ensure maximum recovery of the evidence. The purpose of this study was to determine how the Biological Body-fluid Dog (BBFD) can assist detectives in the investigation of rape cases. The researcher has chosen a unique investigative aid in the form of man’s best friend, namely “The Police K9” (canine). The literature shows that trained police dogs have achieved outstanding success in numerous fields around the world, i.e. from narcotic busts, detection of explosives, to countering terrorist threats, to the most chilling search-and-rescue operations. The SAPS has sent its search-and-rescue dogs to many countries abroad to assist in natural disasters. Dogs have been trained by various police agencies for various purposes, but the BBFD dog is unique to the SAPS, and is trained to detect only human blood and semen. The use of K9s in the complex forensic science environment can never be doubted or overlooked. The main problem facing the Booysens SAPS was the low arrest and conviction rate in rape cases, due to a lack of evidence. The researcher hopes to broaden the detectives’ knowledge and skills regarding the objectives of crime investigation, with special focus on detection of physical evidence at rape crime scenes. The BBFD dog is trained to detect minute amounts of body fluid on any type or size of surface, including veld, bush areas, vehicles, carpets, grass, bedrooms, etc. / Police Practice / M.A. (Criminal Justice)