Poly Brominated Diphenyl Ethers (PBDEs) are known flame retardants with highly persistent and lipophilic in nature. The continued usage of PBDE in various products amplifies the human burden of PBDEs. It is therefore, important to study the potential toxicological and/or biological effects of PBDE exposure in human. In this study we investigated the mode of action of PBDE induced toxicity in human liver by exposing human hepatocarcinoma cells in a time (24-72h) and dose (0-100μM) dependent manner. The highest test dose caused an inhibition in cell viability up to 50% after 72h, whereas lower doses (<50μM) showed slight increase in cell viability. Likewise, higher doses caused significant accumulation of intracellular ROS over time. Further, increase in caspase-3 enzyme levels and DNA fragmentation showed that, lower brominated PBDEs induce liver toxicity through accumulation of toxic metabolites and reactive oxygen species over time leading to caspase-mediated apoptotic cell death.

HIV/AIDS is the most threatening and challenging infectious diseases of our time, with the highest increase of newly infected cases reported. This infectious disease was discovered in the early eighties under homosexual men and was later to be discovered in heterosexuals. HIV is a systemic immunosuppressive disorder which causes a depletion of CD4+ T cells and develops into the acquired immunodeficiency syndrome - AIDS. Africa is the continent most affected by HIV/AIDS with the southern parts of Africa having the highest prevalence rates compared to the rest of Africa. Statistics indicate that AIDS is responsible for 3% of deaths in children worldwide - one in seven people dying of an HIV-related illness is a child under the age of 15 years. It was stated by the WHO that countries should develop improved antiretrovirals regimes for the prevention of mother-to-child transmission. Difficulties in administering antiretrovirals (ARVs) to patients (especially children) are the strict dosage regimes and the severe adverse reactions. These factors complicate patient adherence. The list of problems in treating patients is endless and includes the distribution, stability as well as the low efficacy of these drugs. Most of the above mentioned problems and obstacles related to ARVs and ARV treatment could be minimized or eliminated by the use of a stable and effective drug delivery system. Enhancing ARV treatment may be accomplished by the use of the Pheroid™ drug delivery system. Pheroids™ consists mainly of fatty acids and sterile nitrous oxide gassed water. Pharmacological active substances are entrapped into submicron and micron sized structures called Pheroids™. Research showed promising results and advantages in delivering drugs through oral and transdermal routes using Pheroid™ technology. The focus of this study was to test the possible enhancement of the efficacy of antiretrovirals using Pheroid™ technology. The assays used to study this possible enhancement were a modified neutral red and a modified 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay. These assays confirmed and illustrated the toxic and protective properties of the tested ARVs (stavudine, lamivudine and nevirapine). An MT-2 cell line was used and infected with an HIV-1 strain, SW7-TCL. Applying Pheroid™ technology in these assays resulted in massive cell death, due to increased ARV toxic levels within the cells. Viability tests proved that Pheroids™ had no effect on the viability of cells at the concentration typically used. This confirmed the enhancing properties of Pheroids™ in the delivery of drugs into the cells. The MTT assay was further adapted from a seven day incubation period to a three day incubation period. By using a low concentration series and a three day incubation period the loss of cells through toxicity was partially overcome. One of the problems that arose form this study was the non-reproducibility of the results. Absorbance levels fluctuated at specific concentrations of the same ARV, which cause difficulties in comparing results. This result was repeatedly confirmed in this syncytium forming infection model. In conclusion, Pheroid™ technology enhanced the delivery of ARVs into the cells although it resulted in cell death. Both the neutral red and MTT assays were found to be inaccurate but further development, research and assay optimization could result in improved in vitro studies. The article format was used for this thesis, as described in the general academic rules in section A.13.7.3 of the North West University. Chapter 1 deals with HIV/AIDS related problems, statistics and treatment obstacles. Chapter 2 is a summary of the cell viability assays used in this study. Pheroid™ technology and its application to ARV treatment are dealt with in chapter 3. The proposed article for submission in the journal Cell Death and Differentiation has been included in chapter 4. Some of the results from the study are reported in the article and annexures, whilst other results are shown and discussed in Chapter 5. Chapter 6 gives a conclusion and final summary of this study. All other experimental methods and results are enclosed in the annexures, as is the "Guide for authors" for the article. / Thesis (M.Sc. (Pharmaceutics))--North-West University, Potchefstroom Campus, 2008.

Syftet med studien var att undersöka svenska hamnars möjlighet att investera i land­strömsteknik och sedan betala tillbaka investeringen genom försäljning av elektricitet till ankommande fartyg. Studien är en systematisk litteraturöversikt baserad på en sys­tematisk litteratursökning som sedan presenteras i en teoretisk fallstudie. Litteraturöversikten utgår från fem studier som beskriver kostnader gällande utbyggnad av landströmsteknik.   Första delen av resultatet består av beräkningar för vad investeringskostnaderna för utbyggnad av landströmsteknik till en enstaka kaj kan uppskattas till. Därefter beräk­nades kostnader för att generera en viss mängd elektricitet ombord och jämfördes med kostnaden för att köpa den från marknätet. Sedermera beräknades en vinstmarginal om elektriciteten säljs för samma pris som den kostat om den genererats ombord i dieselgenerator. Slutligen beräknades vilket prispåslag på elförsäljning en hamn kan göra för att få igen investe­ringskostnaderna på fem år samt hur detta pris står sig i jämförelse med kostnaden att generera samma elektricitet ombord.   Studien visar att det finns goda möjligheter för den teoretiska standardkajen att investera i landströmsteknik och få sina investeringar betalda på fem år, under förutsättningen att ankommande far­tyg är utrustade med möjligheten att ta emot elektricitet från land. / The purpose of the study was to investigate Swedish ports possibility to invest in onshore power supply equipment and pay back the initial investment with income from selling electricity to arriving ships. The study is a systematic review based on a sys­tematic search and presented as a theoretical case study. The review was based on five studies that describe costs of providing a port with onshore power supply.   The first part of the findings is calculations of the investment cost for onshore power supply for one single berth. Next the calculations of the costs to generate a certain amount of electricity on board was compared with the costs of buying said amount of electricity from shore providers. Then a profit margin if the electricity was sold for the same price as it would have cost to generate onboard in diesel generators was calculated. Finally the in­crease to the electricity sale price a port could add and still repay the initial investment over five years was calculated. This electricity sale price was then compared to what it would have cost to generate onboard.   The findings shows that there is a good possibility for the theoretical standard quay to refund their investment in onshore power supply over five years, under circumstances where ships visiting the port are equipped to receive the electricity.

Food retail with large supermarkets consumes significant amounts of energy. The environmental impact is also significant because of the indirect effect from CO2 emissions at the power stations and due to the direct effect arising from refrigerant leakage to the atmosphere. The application of trigeneration (local combined heat, power and refrigeration) can provide substantial improvements in the overall energy efficiency over the conventional supermarket energy approach of separate provision of electrical power and thermal energy. The use of natural refrigerants such as CO2 offers the opportunity to reduce the direct impacts of refrigeration compared to conventional systems employing HFC refrigerants that possess high global warming potential. One approach through which the overall energy efficiency can be increased and the environmental impacts reduced, is through the integration of trigeneration and CO2 refrigeration systems where the cooling generated by the trigeneration system is used to condense the CO2 refrigerant in a cascade arrangement. This research project investigates experimentally and theoretically, through mathematical modelling and simulation, such a system and its potential application to supermarkets. A small size CO2 refrigeration system for low and medium food temperature applications was designed and constructed to enable it to be integrated with an existing trigeneration system in the refrigeration laboratory at Brunel University to form an integrated trigeneration and CO2 refrigeration test facility. Prior to the construction, the design of the system was investigated using mathematical models developed for this purpose. The simulations included the CO2 refrigeration system, CO2 evaporator coils and the integration of the trigeneration and CO2 refrigeration systems. The physical size of the design and component arrangement was also optimised in a 3D AutoCAD model. A series of experimental tests were carried out and the results showed that the medium temperature system could achieve a very good COP, ranging from 32 to 60 due to the low pumping power requirement of the liquid refrigerant. The low temperature system performed with average steady state COP of 4, giving an overall refrigeration system COP in the range between 5.5 and 6. Mathematical models were also developed to investigate the application of the integrated trigeneration and CO2 refrigeration system in a case study supermarket. The models were validated against test results in the laboratory and manufacturers’ data. The fuel utilisation efficiency and environmental impacts of different trigeneration and CO2 refrigeration arrangements were also evaluated. The results indicated that a system comprising of a sub-critical CO2 refrigeration system integrated with a trigeneration system consisting of a micro-turbine based Combined Heat and Power (CHP) unit and ammonia-water absorption refrigeration system could provide energy savings of the order of 15% and CO2 emission savings of the order of 30% compared to conventional supermarket energy systems. Employing a trigeneration system with a natural gas engine based CHP and Lithium Bromide-Water sorption refrigeration system, could offer energy savings of 30% and CO2 emission savings of 43% over a conventional energy system arrangement. Economic analysis of the system has shown a promising payback period of just over 3 years compared to conventional systems.

In this thesis the effects of individual and multiple environmental stressors (physical and chemical) are examined in inbred and outbred zebrafish (Danio rerio, Hamilton), a model species used in ecotoxicology and environmental risk assessment (ERA). The central question addressed is, are inbred laboratory animals representative and protective of wild populations? That is, are inbred fish equally or more sensitive to chemicals and other stressors compared with more outbred (wild) fish? A combination of tools and approaches incorporating traditional (eco)toxicology and population genetics have been employed, together with more contemporary molecular genetics and population modelling, to compare and contrast a range of responses in inbred and outbred zebrafish exposed to the endocrine disrupting chemical clotrimazole and/or temperature elevation in the laboratory. The choice of test species was based on our broad understanding of its basic biology, extending from the molecular level to the population level, and its wide use as a model organism in (eco)toxicology. Selection of the test chemical clotrimazole and temperature was based on a shared mode of action, aromatase inhibition, and therefore their ability to block oestrogen production, impair reproduction, promote male development and skew population sex ratios in zebrafish. A cascade of responses were compared in inbred and outbred zebrafish, including changes in the levels of expression of genes for gonadal aromatase and other steroidogenic enzymes, circulating sex steroid hormones, gonadal sex differentiation and development (via gonadal histopathology) and reproductive fitness (female fecundity, paternity and viability of embryos). Amongst the most striking results were directional skews in sex ratio towards males in response to clotrimazole (Chapter 5) and elevated temperature exposure (Chapter 7). Inbred fish were generally more responsive compared to outbreds, which showed evidence of physiological and developmental compensation, resulting in lower male-sex skews and superior fitness in terms of male reproductive success (paternity and viability of embryos). The greater effects observed in inbred fish were attributed to inbreeding×environment interactions and the amplification of inbreeding depression. Although no empirical genetic evidence of this mechanism is presented (loss of heterozygosity at quantitative trait loci and concomitant loss of heterosis and/or the expression of recessive, deleterious alleles in homozygotes), supporting evidence was provided by increased phenotypic variance in some apical endpoints in inbred fish, including specific growth rate and fecundity. This increased variance also has the potential to counteract the higher levels of response observed in inbreds, because the power to detect statistically significant changes in responses is reduced. This trade-off was demonstrated for specific growth rate. Crucially, significant male-sex ratio skews (>80%) were induced at substantially lower clotrimazole exposure concentrations (1.7 µg l-1) in combination with elevated temperature (33°C), compared with exposure concentrations (43.7 µg l-1) generating similar sex ratio skews at the standard test temperature of 28°C. These temperatures represent current and predicted 2100 (elevated) mean temperatures in the zebrafish’s native India and Bangladesh. Although the lowest observed effect concentration was an order of magnitude above the predicted environmental concentration for clotrimazole, it is conceivable that combined environmental exposures to similarly acting chemicals (e.g. other azole compounds used in crop protection, veterinary and human medicine) could produce similar effects to those we observed. The consequent effects of sex ratios skews and reduced fitness (fecundity and embryo viability) on per capita population growth rate (r) and extinction probability were predicted in inbred versus outbred zebrafish populations using stochastic population viability analysis. The results showed that the observed male-skews >80% threaten small zebrafish populations with fewer than 100 breeding adults (<20 adult females). However, small reductions of 2-3% in embryo-juvenile (age 0+) survivorship (including simulated inbreeding depression) were more influential on r and extinction probability than large sex ratio skews and/or reduced female fecundity. The results presented in this thesis support the contention that chemical effects may be exacerbated by other environmental stressors, but also illustrate the importance of considering biological (genetic), as well as physical and chemical interactions in cumulative ERA. Greater sensitivity of inbred versus outbred organisms to the effects of environmental stressors on sexual differentiation and reproductive fitness offers a margin of safety to ERA and the protection of wildlife populations (excluding those that are severely inbred and critically endangered). This is because, as originally stated, laboratory organisms used in ERA are generally more inbred than their wild counterparts. Nevertheless, more attention should be paid to the origin, breeding history and genetics of laboratory strains. This will help to ensure consistency between studies and testing laboratories and provide more confidence in extrapolating the results to wild populations.

L’objectif de ce travail est de caractériser l’effet de l’érythromycine sur l’activité bactérienne et l’inhibition de l’épuration dans les réacteurs de traitement des eaux usées urbaines. L’étude a montré l’importance du temps d’exposition à l’érythromycine. L’inhibition de la nitrification et de l’épuration de la DCO a été mesurée sur une période de 4 h pour des concentrations supérieures à 1 mg/L d’érythromycine alors que l’inhibition de l’épuration pour une concentration d’érythromycine de 4 µg/L a été mesurée suite à 20 h d’exposition. L’origine de la liqueur mixte est également déterminante : l’érythromycine a inhibé la nitrification avec les boues de Nancy mais n’a pas inhibé la nitrification avec les boues et Épinal. Par contre, l’inhibition de l’épuration de la DCO a été mesurée pour les deux boues. Des analyses microscopiques ont montré qu’en présence d’érythromycine les flocs bactériens se fractionnaient suite aux lyses bactériennes. La microscopie en épifluorescence et la microscopie confocale combinées à des marqueurs de viabilité fluorescents ont permis de déterminer les vitesses de mortalité bactérienne. Ces études ont mis en évidence un temps de latence qui précède la mortalité des bactéries. Ce temps de latence pourrait être lié aux mécanismes d’adsorption de l’antibiotique et de diffusion dans les flocs, ainsi qu’à la vitesse biologique de mortalité. Enfin, un marquage simultané Gram et viabilité (microscopie confocale 3D au cours du temps) a montré que l’érythromycine pourrait modifier la composition bactérienne des boues en sélectionnant les bactéries les moins sensibles / This study examines the effect of erythromycin on activated sludge bacteria and the inhibition of the pollution removal in batch reactors treating urban wastewater. Results showed the importance of exposure time to erythromycin. Inhibition of nitrification and COD removal was measured during a 4 h period for erythromytcin concentrations higher than 1 mg/L. A 4 µg/L erythromycin concentration inhibited COD removal during a 20 h exposure time. The effect of erythromycin on nitrification was variable depending on the sludge origin. Erythromycin inhibited the specific nitrification rate with sludge from Nancy WWTP, but increased the nitrification rate at the other facility (Epinal WWTP). The cell lysis resulted in destruction of activated sludge flocs. Microscopic techniques (epifluorescence and confocal laser scanning microscopy (CLSM)), combined with a fluorescent viability indicator, allowed us to study erythromycin time-kill activity.Viability staining results showed a latency time before the lower antibiotic concentrations began to kill bacteria. This latency time could be related to antibiotic adsorption and diffusion into activated sludge flocs as well as the rate of bateria death. The effect of erythromycin according to the bacterial Gram type was investigated with 3-dimensional Confocal Laser Scanning Microscopy (CLSM) time-lapse imaging combined with a Gram and Viability staining. Gram+ bacteria had a higher mortality rate than the Gram- bacteria. This result suggests that antibiotic in wastewater could change the activated sludge bacteria composition according to their Gram type by selecting bacteria the less sensitive to the antibiotics

This study is aimed to evaluate the degradation characteristics, cell viability and host tissue responses of PDLLA/PRGD/beta-TCP (PRT) composite nerve scaffold, which was fabricated by poly(D, L-lactic acid) (PDLLA), RGD peptide(Gly-Arg-Gly-Asp-Tyr, GRGDY, abbreviated as RGD) modified poly-{(lactic acid)-co-[(glycolic acid)-alt-(L-lysine)]}(PRGD) and beta-tricalcium phosphate (beta-TCP). The scaffolds' in vitro degradation behaviors were investigated in detail by analysing changes in weight loss, pH and morphology. Then, the 3-(4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2 -H-tetrazolium bromide (MTT) assay and cell live/dead assay were carried out to assess their cell viability. Moreover, in vivo degradation patterns and host inflammation responses were monitored by subcutaneous implantation of PRT scaffold in rats. Our data showed that, among the tested scaffolds, the PRT scaffold had the best buffering capacity (pH - 6.1-6.3) and fastest degradation rate (12.4%, 8 weeks) during in vitro study, which was contributed by the incorporation of beta-TCP nanoparticles. After in vitro and in vivo degradation, the high porosity structure of PRT could be observed using scanning electron microscopy. Meanwhile, the PRT scaffold could significantly promote cell survival. In the PRT scaffold implantation region, less inflammatory cells (especially for neutrophil and lymphocyte) could be detected. These results indicated that the PRT composite scaffold had a good biodegradable property; it could improve cells survival and reduced the adverse host tissue inflammation responses.

Hospital-acquired infections are to date a major challenge in the patient safety. The proliferation of pathogens such as Staphylococcus aureus, Escherichia coli and Klebsiella pneumoniae is often reported in connection with textiles, which represent a significant source of transmission. This leads most often to the contamination and cross-contamination of the hospitalized patient and the hospital staff. A promising approach and the immediate objective of this research is the application of probiotics to a textile fabric. They provide preliminary evidence in being able to inhibit pathogenic bacteria growth through their competitive mechanism. During this study, screen-printing was used as a method to apply probiotics on a polyester fabric. The viability of probiotics on the fabric was evaluated in the agar plate test method. Samples that exhibited a growth of grown out colonies were further tested regarding their efficacy towards the abovementioned pathogens. This was determined in a competition test, that included the individually inoculation of the samples with the different bacteria strains. Contact-angle measurements and abrasion resistance as well as the durability were tested in order to investigate the applicability of the fabric and scanning electron microscope images were taken to detect probiotics and to evaluate the quality of the print on the fabric. Major findings included that probiotics were able to survive on the polyester fabric and that these viable probiotics are further successful in the growth inhibition of pathogenic bacteria. Thereby their efficacy against pathogens is related to the viability the probiotics. As neither the probiotics used in this study, nor the other ingredients are considered as hazardous, this process is environment-friendly. This work increases the understanding of probiotics mechanism and their survival and competition behaviour on a textile material. It opens a way in the reduction of hospital-acquired infections through the beneficial effects of probiotics.

Master of Science / Department of Animal Sciences and Industry / Barry J. Bradford / Wet corn gluten feed (WCGF) is commonly included in lactation rations for dairy cattle. Research at Kansas State University has shown that increasing WCGF inclusion decreased ruminal pH. Lack of adequate particle size may result in this decrease and can impact efficiencies, animal health and longevity. A study was conducted to look at the effects of feeding WCGF while maintaining > 10% of particles > 19 mm across diets. We hypothesized that as WCGF increased, DMI and milk yield would increase while ruminal pH would be maintained. Seven ruminally-cannulated, lactating Holstein cows were used in an incomplete 4 × 4 Latin square design with treatments of 0, 12.4, 24.5 or 35.1% WCGF across 4 periods of 21 d. Alfalfa hay was used to maintain particle size. All diets met particle size goals; however, as WCGF increased, the proportion of particles > 19 mm decreased (P = 0.01) and cows changed their sorting behavior in favor of particles > 19 mm (P = 0.03) and against particles on the bottom screen (P < 0.01) and pan (P = 0.01). As WCGF increased, ruminal pH and ECM/DMI were not affected, yet DMI (P = 0.02) and milk yield (P = 0.02) increased quadratically. Milk protein, lactose and fat concentrations were not affected; however, milk protein (P = 0.004; linear) and lactose (P = 0.02; quadratic) yields increased. In a separate study, active dry yeast (ADY) products, commonly used in the dairy industry to support ruminal health, were evaluated for product guarantees and effects of storage and storage medium. Few products received through normal distribution met product guarantees (1 of 6; experiment 1) and after 3 mo of storage cell viability dropped significantly (P < 0.01). In the second experiment, products were stored in ground corn or in a vitamin-trace mineral mix (VTM). Depressions in viability caused by high-temperature storage were partially mitigated when ADY products were stored with a VTM. Although both mediums resulted in lower cell viabilities after storage at 40ºC, VTM cell viabilities were significantly (P = 0.02) higher than ground corn.

>Magister Scientiae - MSc / Introduction The blood-testis barrier (BTB) is formed by tight junctions between adjacent Sertoli cells. The barrier formed by these tight junction helps to create a specialized environment for spermatogenesis and provide an immunological barrier to protect developing germ cells. Methamphetamine (Meth) is known as neurotoxin however, its effects on the male reproductive system, especially on Sertoli cells and, the BTB are not well established. Therefore, this study aimed to determine the effects of Meth on the TM4 mouse testis Sertoli cell line and on the integrity of the BTB permeability. Materials and Methods This study investigated the effect of selected concentrations of Meth (0.1 μM, 1 μM, 10 μM, 20 μM and 100 μM) on TM4 mouse testis Sertoli cell line for 24 until 96 hours, using two treatments: an “acute” study (24 hrs exposure) and a “chronic” study, where treatment occurred on a daily basis over 96 hrs. The following parameters were investigated: viability, cell proliferation, mitochondrial activity, monolayer permeability.