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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
41

Assessing Usability of Products in the Low Vision Field

Wing, Craig Jason Tam 15 February 2007 (has links)
Student Number : 9804058J - MSc dissertation - School of Information and Electrical Engineering - Faculty of Engineering and the Built Environment / This paper presents the implementation of usability engineering into a device to meet the requirements of a Visually Impaired Person (VIP). Users of such a device may suffer from conditions such as Macular Degeneration, Diabetes and HIV/AID’s related disorders. Since these disorders affect a person’s vision, the device enlarges the desired text to reduce the effects of loss of vision. Other functionality may include image manipulation and colour modification. A usability engineering framework is incorporated into the design as well as accommodating user requirements in the design process. Usability principles are implemented, hence meeting the aims of effectiveness, efficiency, learnability, satisfaction and context of use. The device is examined via heuristic evaluation and usability testing from specialists and end users, with comments, ratings and times recorded. Research indicates that this device successfully implements usability engineering techniques and provides a cost effective, highly functional device for the VIP.
42

Analysis of metatranscriptomes from an acidophilic electricity generating community treating acidic mining wastewaters

Palma, Daniela January 2018 (has links)
Human ́s constant need for metals requires unsustainable mining and refining of metalore. As a result, highly contaminated wastewaters are discharged in the environmentcompromising the nearby habitat together with all its life forms. Microbial fuel cells arebioelectrochemical systems (BES’s) that use microorganisms to convert organic andinorganic matter, producing electricity as the final product. This technology has shownto have great potential for application for bioremediation of wastewaters. This thesisdescribes the gene expression and the taxonomical abundance of an acidophilic,electricity generating community that was used to treat mining wastewaters in amicrobial fuel cell. A complete metatranscriptomics analysis has been performed onduplicate MFC anode acidophilic microbial community generating electricity frominorganic sulfur compounds (ISC) oxidation at extremely low pH. The analysis showsthat the most expressed genus is Ferroplasma-like, the genus Acidithiobacillus-like isfollowing along with Sulfobacillus-like and Thermoplasma-like. Some of the generaexpressed show behaviours never described before suggesting that potentially, newspecies have been selected. The reactions of the sulfur pathway are regulated mostly bytwo genera: Acidithiobacillus-like during the disproportionation of tetrathionate, andFerroplasma-like by expressing the hdr gene that catalyses the reaction from elementalsulfur to sulfite, the sulfite is then converted to sulfate. The hdr gene has not previouslybeen found in F. acidarmanus-like suggesting that the specie might have been selectedfor this trait. Acidithiobacillus-like genus has a bigger role for the energy conservationand the electron transport in the sample, however the data are not sufficient to point outwhich gene has the major role in the process. The CO2 fixation in the chamber wasconsiderably low as a result from a significant carboxysomes production, bacterialcompartiments involved in the carbon dioxide fixation. The transcripts abundanceregarding the metal resistance genes have shown low expression suggesting that thecells were not under stress. This result is indicated by the synthesis of a transcriptionalrepressor protein that had prevented a significant production of metal resistanceenzymes. Likewise, the pH homeostasis plot does not show vast transcripts abundances,indicating that the cells were thriving under conditions not far from the optimum.
43

Modelo multicritério de apoio à decisão para classificação de risco em barragens

Silva, Julierme Siriano da 19 June 2017 (has links)
O objetivo deste estudo foi comparar modelos importantes de classificações de risco em barragens (Índice Global de Risco Modificado, USACE, COGERH e Resolução CNRH n° 143), empregados por instituições no Brasil e no Mundo, na forma padrão e utilizando adaptações para o método multicritério ELECTRE TRI. Propôs-se também um método que atendesse às especificidades do ELECTRE TRI com apoio de um decisor com conhecimento técnico em segurança de barragens. Essas classificações foram aplicadas nas barragens dos projetos de irrigação Rio Formoso e Manuel Alves sob gestão da SEPLAN - TO. Levantaram-se informações técnicas por meio de projetos executivos e visitas técnicas nas construções para a realização do enquadramento das barragens nas classificações, tanto para os métodos já existentes, como para o método proposto pelo estudo que utilizou especificamente o ELECTRE TRI. Para a barragem do projeto Rio Manuel Alves a classificação foi de Baixo Risco. As barragens do Projeto Rio Formoso apresentaram classificações de Alto Risco e Risco Extremo. / The objective of this study was to compare important models of risk classifications in dams (Global Modified Risk Index, USACE, COGERH and Resolution CNRH n ° 143), employed by institutions in Brazil and in the world, in the standard form and using adaptations to the method Multicriteria ELECTRE TRI. It was also proposed a method that would meet the specificities of ELECTRE TRI with the support of a decision maker with technical knowledge on dam safety. These classifications were applied to the dams of the Rio Formoso and Manuel Alves irrigation projects under the management of SEPLAN - TO. Technical information was obtained through executive projects and technical visits to the constructions for the implementation of the classification of dams in the classifications, both for the existing methods and for the method proposed by the study that specifically used ELECTRE TRI. For the Rio Manuel Alves dam, the classification was Low Risk. The Rio Formoso Project dams presented High Risk and Extreme Risk ratings.
44

Bioassay assessment of mine pit lake water for aquaculture and biodiversity conservation

Neil, Luke L January 2007 (has links)
For decades, researchers have been working to better understand the effects of acid and metal toxicity to aquatic organisms and to develop ways to effectively mitigate these detrimental effects. Acid mine drainage (AMD) causes environmental pollution that affects many countries having historic or current mining industries. Both surface and underground mining have the potential to create AMD and therefore adversely affect the local and adjacent environments. Through malignant mining and farming practices, damming rivers and the changing climate, surface freshwater resources are rapidly being reduced in Australia. Mine pit lakes offer a large freshwater resource that if managed correctly may sustain significant environmental, social and economic benefits from selected end uses for the future. However, assessment of mine pit lakes needs to be achieved to validate end use options. The Collie Basin located in the southwest of Western Australia is a coal mining area with many abandoned open cast mines (OCM). Some of these OCM have filled with water forming mine pit lakes that are affected by AMD. The result is large freshwater bodies with moderate to high concentrations of metals and a low pH. High concentration of metals combined with low pH is of environmental concern to both the aquatic and terrestrial ecosystem. Nevertheless, remediation techniques for AMD are available and applicable to these acidic lakes. However, the lakes in the Collie Basin are low in sulphatic compounds, therefore, restricting the use of the most common remediation treatment of bioremediation with sulphate reducing bacteria. Three remediation treatments were assessed for there efficacy in toxicity amelioration to three mine pit lakes. The treatments assessed were Limestone addition, phosphorus addition and a Rapid Catalytic Oxidation (RCO) treatment. / Toxicity assessment was achieved by means of comparing physico-chemical data to guideline trigger values and biological assessment. Biological assessment was achieved with three commonly used but ecologically and geographically relevant species. The bioassay species were the alga Chlorella protothecoides, the crustacean Ceriodaphnia cf dubia and the protozoa Tetrahymena thermophila. Aquaculture is being trialled adjacent to one of the pit lakes in limestone treated mine pit lake water. Therefore, the aquaculture fish species Bidyanus bidyanus early life stages were also used to assess residual toxicity from treated mine pit lake water. Limestone remediation of pit lake water demonstrated good reduction of dissolved metal concentrations with the exception of Zn and the ability to increase pH to circum-neutral. Biological assessment of limestone treated pit lake water showed that toxicity was removed to the three bioassay species and to the aquaculture species B. bidyanus early life stages. A larger field- scale mesocosm experiment with limestone treatment in situ is recommended. Assessment of the mesocosm experiment with biological and chemical analysis will confirm the efficacy of this treatment for full scale use.
45

Genexpression und Wirkung von Faktoren der Blutgerinnungskaskade und des Komlementsystems in humanen retinalen Pigmentepithel (RPE)-Zellen

Dott, Britta 28 March 2012 (has links) (PDF)
Eine lokale Aktivierung des Komplementsystems im RPE ist ein pathogener Faktor der AMD. Neben der Wirkung von angiogenen Faktoren wie VEGF könnte eine Aktivierung des Blutgerinnungssystems im RPE dazu beitragen, dass sich aus einer trockenen eine feuchte AMD entwickelt. Dies könnte auf mehreren Ebenen geschehen: Gerinnungsfaktoren könnten die Expression der Komplementfaktoren und der angiogenen Faktoren regulieren sowie Wirkungen auf die Proliferation und Migration der RPE-Zellen besitzen. Eine Stimulierung der Proliferation und Migration der RPE-Zellen trägt zur Ausbildung von CNV-Membranen bei. Es ist aber bis jetzt nichts darüber bekannt, ob RPE-Zellen Faktoren des Blutgerinnungssystems exprimieren und ob z.B. Thrombin (als zentrale Protease des Blutgerinnungssystems) die Genexpression von Komplementfaktoren und von VEGF im RPE beeinflusst. Die Ziele der vorliegenden Dissertation waren daher: ● Nachweis der mRNA-Expression von Blutgerinnungs- und Komplementfaktoren im RPE; ● Nachweis der Wirkung von Thrombin auf die Expression von VEGF und von Komplementfaktoren, sowie auf die Proliferation und Migration der RPE-Zellen; und ● Nachweis der Wirkung der Komplementfaktoren C5a und C9 auf die Sekretion von VEGF und die Proliferation und Migration der RPE-Zellen.
46

Structure and function of food webs in acid mine drainage streams

Hogsden, Kristy Lynn January 2013 (has links)
Acid mine drainage (AMD) is a significant environmental issue worldwide, which often causes severe contamination and marked species losses in receiving streams. However, little is known about how this stress alters food webs and ecosystem function. I conducted a literature review, which revealed that AMD-impacted streams generally had depauperate benthic communities dominated by a few tolerant species and impaired ecosystem processes. Next, using survey and experimental-based approaches, I investigated food web structure and energy flow in these highly stressed streams, which typically have low pH (< 3), high concentrations of dissolved metals (Al, Fe), and substrata coated with metal hydroxide precipitates, on the South Island, New Zealand. Inputs of AMD caused substantial loss of consumers and reduced the overall number of links between species generating small and simplified food webs, with few invertebrates and no fish. Comparative analysis of food webs from a survey of 20 streams with either anthropogenic or natural sources of acidity and metals, indicated that anthropogenic sources had a stronger negative effect on food web properties (size, food chain length, number of links); an effect driven primarily by differences in consumer diversity and diet. However, the presence of fewer trophic levels and reduced trophic diversity (detected using isotopic metrics), were common structural attributes in AMD-impacted webs along a pH gradient, regardless of impact level. Furthermore, complementary dietary analyses of consumer gut contents and stable isotope signatures (δ13C and 15N) confirmed that primary consumers fed generally on basal resources and that there were few predatory interactions, which reflected low densities of small-bodied chironomids. This suggests that food quantity was unlikely to limit primary consumers but that reduced prey availability may be an additional stressor for predators. In these radically re-structured food webs, trophic bottlenecks were generated at the primary consumer level and energy flow to higher consumers was disrupted. However, streams still retained some limited function, including slow leaf litter breakdown, which provided detrital resources and supported the small food webs. Overall, my findings have furthered our understanding of these highly stressed stream ecosystems by providing new insights into interactions among species and trophic levels that structure food webs and enable function.
47

Diatom communities across a gradient of acid mine drainage on the West Coast, South Island, New Zealand

Schowe, Kate January 2012 (has links)
Acid mine drainage (AMD) is a major environmental issue worldwide. On the West Coast of the South Island, New Zealand, numerous catchments receive AMD, with significant negative impacts on in-stream flora and fauna. Diatoms are commonly regarded as powerful biological indicators and may be found in high abundance in AMD-contaminated streams; however, relatively little work has been done on diatoms in mining environments in New Zealand. Initially, I conducted a survey of epiphytic diatom communities in 39 streams ranging from non-impacted reference streams to those severely impacted by AMD. Streams were assigned to one of four classes along an AMD gradient: circum-neutral reference, naturally acidic reference, moderately impacted, and severely impacted. There was a wide range in diatom taxonomic richness in reference and moderately impacted streams (8 – 33 taxa). Taxonomic richness was greatly reduced in severely impacted streams (1 – 5 taxa) at a threshold of pH 3.4 and was dominated by Pinnularia cf. acidophila (69 – 100% relative abundance). Community composition differed between circum-neutral reference, moderately, and severely impacted streams; however, naturally acidic and moderately impacted streams had similar diatom communities primarily composed of acid-tolerant Eunotia and Frustulia species. This indicated that diatoms are strongly structured by pH and able to tolerate moderate conductivity and metal concentrations without a corresponding shift in community composition. Survey data were then used to develop two diatom-based indices for streams impacted by AMD: a single Biotic Index and a Multimetric Index. While neither index was able to distinguish naturally acidic from moderately impacted streams, both indices successfully categorised streams as circum-neutral reference, moderately or severely impacted by AMD. These indices may be useful in assessing AMD impact on circum-neutral streams or in identifying when a stream has crossed a threshold from moderately to severely impacted by AMD. Diatoms would be especially useful as bioindicators of AMD if they respond rapidly to a change in mine discharge. To test this, mature algal biofilms were reciprocally transferred between circum-neutral reference streams and streams of varying degrees of AMD over a period of 13 days. Diatom mortality increased rapidly in the reciprocal transfer between reference and severely impacted streams. Reference communities resembled the ambient diatom community of severely impacted streams 13 days post-transfer. However, in the reverse transfer, a change in community composition was slow to occur. Diatoms respond faster to an increase in pollution than to pollution amelioration. Overall, results indicated that diatom communities may be a useful tool for monitoring the presence and magnitude of AMD in New Zealand streams.
48

Sulphate removal from industrial effluents through barium sulphate precipitation / Swanepoel H.

Swanepoel, Hulde. January 2011 (has links)
The pollution of South Africa’s water resources puts a strain on an already stressed natural resource. One of the main pollution sources is industrial effluents such as acid mine drainage (AMD) and other mining effluents. These effluents usually contain high levels of acidity, heavy metals and sulphate. A popular method to treat these effluents before they are released into the environment is lime neutralisation. Although this method is very effective to raise the pH of the effluent as well as to precipitate the heavy metals, it can only partially remove the sulphate. Further treatment is required to reduce the sulphate level further to render the water suitable for discharge into the environment. A number of sulphate removal methods are available and used in industry. These methods can be divided into physical (membrane filtration, adsorption/ion exchange), chemical (chemical precipitation) and biological sulphate reduction processes. A literature study was conducted in order to compare these different methods. The ABC (Alkali – Barium – Calcium) Desalination process uses barium carbonate to lower the final sulphate concentration to an acceptable level. Not only can the sulphate removal be controlled due to the low solubility of barium sulphate, but it can also produce potable water and allows valuable by–products such as sulphur to be recovered from the sludge. The toxic barium is recycled within the process and should therefore not cause additional problems. In this study the sulphate removal process, using barium carbonate as reactant, was investigated. Several parameters have been investigated and studied by other authors. These parameters include different barium salts, different barium carbonate types, reaction kinetics, co–precipitation of calcium carbonate, barium–to–sulphate molar ratios, the effect of temperature and pH. The sulphate removal process was tested and verified on three different industrial effluents. The results and conclusions from these publications were used to guide the experimental work. A number of parameters were examined under laboratory conditions in order to find the optimum conditions for the precipitation reaction to take place. This included mixing rotational speed, barium–to–sulphate molar ratio, initial sulphate concentration, the effect of temperature and the influence of different barium carbonate particle structures. It was found that the reaction temperature and the particle structure of barium carbonate influenced the process significantly. The mixing rotational speed, barium–to–sulphate dosing ratios and the initial sulphate concentration influenced the removal process, but not to such a great extent as the two previously mentioned parameters. The results of these experiments were then tested and verified on AMD from a coal mine. The results from the literature analysis were compared to the experiments conducted in the laboratory. It was found that the results reported in the literature and the laboratory results correlated well with each other. Though, in order to optimise this sulphate removal process, one has to understand the sulphate precipitation reaction. Therefore it is recommended that a detailed reaction kinetic study should be conducted to establish the driving force of the kinetics of the precipitation reactions. In order to upgrade this process to pilot–scale and then to a full–scale plant, continuous reactor configurations should also be investigated. The sulphate removal stage in the ABC Desalination Process is the final treatment step. The effluent was measured against the SANS Class II potable water standard and was found that the final water met all the criteria and could be safely discharged into the environment. / Thesis (M.Ing. (Chemical Engineering))--North-West University, Potchefstroom Campus, 2012.
49

Sulphate removal from industrial effluents through barium sulphate precipitation / Swanepoel H.

Swanepoel, Hulde. January 2011 (has links)
The pollution of South Africa’s water resources puts a strain on an already stressed natural resource. One of the main pollution sources is industrial effluents such as acid mine drainage (AMD) and other mining effluents. These effluents usually contain high levels of acidity, heavy metals and sulphate. A popular method to treat these effluents before they are released into the environment is lime neutralisation. Although this method is very effective to raise the pH of the effluent as well as to precipitate the heavy metals, it can only partially remove the sulphate. Further treatment is required to reduce the sulphate level further to render the water suitable for discharge into the environment. A number of sulphate removal methods are available and used in industry. These methods can be divided into physical (membrane filtration, adsorption/ion exchange), chemical (chemical precipitation) and biological sulphate reduction processes. A literature study was conducted in order to compare these different methods. The ABC (Alkali – Barium – Calcium) Desalination process uses barium carbonate to lower the final sulphate concentration to an acceptable level. Not only can the sulphate removal be controlled due to the low solubility of barium sulphate, but it can also produce potable water and allows valuable by–products such as sulphur to be recovered from the sludge. The toxic barium is recycled within the process and should therefore not cause additional problems. In this study the sulphate removal process, using barium carbonate as reactant, was investigated. Several parameters have been investigated and studied by other authors. These parameters include different barium salts, different barium carbonate types, reaction kinetics, co–precipitation of calcium carbonate, barium–to–sulphate molar ratios, the effect of temperature and pH. The sulphate removal process was tested and verified on three different industrial effluents. The results and conclusions from these publications were used to guide the experimental work. A number of parameters were examined under laboratory conditions in order to find the optimum conditions for the precipitation reaction to take place. This included mixing rotational speed, barium–to–sulphate molar ratio, initial sulphate concentration, the effect of temperature and the influence of different barium carbonate particle structures. It was found that the reaction temperature and the particle structure of barium carbonate influenced the process significantly. The mixing rotational speed, barium–to–sulphate dosing ratios and the initial sulphate concentration influenced the removal process, but not to such a great extent as the two previously mentioned parameters. The results of these experiments were then tested and verified on AMD from a coal mine. The results from the literature analysis were compared to the experiments conducted in the laboratory. It was found that the results reported in the literature and the laboratory results correlated well with each other. Though, in order to optimise this sulphate removal process, one has to understand the sulphate precipitation reaction. Therefore it is recommended that a detailed reaction kinetic study should be conducted to establish the driving force of the kinetics of the precipitation reactions. In order to upgrade this process to pilot–scale and then to a full–scale plant, continuous reactor configurations should also be investigated. The sulphate removal stage in the ABC Desalination Process is the final treatment step. The effluent was measured against the SANS Class II potable water standard and was found that the final water met all the criteria and could be safely discharged into the environment. / Thesis (M.Ing. (Chemical Engineering))--North-West University, Potchefstroom Campus, 2012.
50

Untersuchungen zur Expression von Interleukin-10 nach Transfektion humaner retinaler Pigmentepithelzellen und dessen Einfluss auf die Proliferation von T-Lymphozyten in vitro

Poschinger, Katharina 28 November 2004 (has links) (PDF)
Bei der Altersabhängigen Makuladegeneration (AMD) handelt es sich um eine Erkrankung des Auges, die die Macula lutea, die Stelle des schärfsten Sehens betrifft. Sie ist verbunden mit der Degeneration von RPE-Zellen, die zur Dystrophie von Photorezeptoren und damit zum Verlust des zentralen Sehvermögens führt. Eine ähnliche Pathophysiologie ist bei der sogenannten Retinalen Pigmentepitheldystrophie (RPED) des Hundes zu beobachten. Die Transplantation von gesunden RPE-Zellen in das betroffene Gebiet stellt eine vielversprechende Therapiemöglichkeit dar. Die Transplantatabstoßung als Kom-plikation schränkt die klinische Anwendung ein. Eine beim Patienten nach Transplantation lebenslang durchgeführte systemische Immunsuppression ist mit erheblichen Nebenwirkungen verbunden. Deshalb bietet die Gentherapie unter Einbezug immunsuppressiver Zytokine wie beispielsweise des Interleukin-10 (IL-10) eine Lösung. In der vorliegenden Arbeit wurde ein selbst konstruierter IL-10-Expressionsvektor (Plasmid pCIneoIL-10) mittels Gentransfer in humane RPE-Zellen in vitro eingebracht. Untersucht wurde die Wirkung des sezernierten IL-10 auf die Proliferation von allogenen T-Lymphozyten mit und ohne allogene Makrophagen als professionelle antigenpräsentierende Zellen (APC). Neben humanen Spender RPE-Zellen (Spender-hRPE-Zellen) wurde eine immortalisierte Permanent-Zelllinie (hTERT-RPE1-Zellen) eingesetzt, deren Hauptvorteil in einer gleichbleibend hohen Wachstumsrate lag. Als transientes Transfektions-system für den Transfer von IL-10-DNA in hRPE-Zellen wurden kationische Lipide gewählt. Drei verschiedene Lipidformulierungen wurden miteinander verglichen und das optimale Transfektionsreagenz:DNA-Verhältnis, mit dem die höchste Transfektionseffizienz erreicht werden konnte, evaluiert. Eine Transfektionseffizienz von 23,3 ± 9,0 % (hTERT-RPE1-Zellen) beziehungsweise 10,3 ± 4,5 % (Spender-hRPE-Zellen) konnte erreicht werden. Die Transfektion hatte weder einen negativen Einfluss auf die Vitalität der hRPE-Zellen, noch wurde der natürliche Zelltod, die Apoptose, erhöht. Die IL-10-mRNA-Expression wurde mittels RT-PCR nachgewiesen. Lediglich bei den transfizierten hRPE-Zellen konnte IL-10-mRNA gefunden werden. Mittels ELISA konnte das IL-10-Protein gemessen werden. Die Sekretion des IL-10 in den Kulturüberstand von transfizierten hRPE-Zellen wurde dafür über einen Zeitraum von 7 Tagen untersucht. Es konnte festgestellt werden, dass die maximale IL-10-Proteinkonzentration bei beiden Zelllinien am Tag 3 mit Werten von 10,3 ± 0,8 ng/ml (hTERT-RPE1-Zellen) und 3,1 ng/ml (Spender-hRPE-Zellen) lag. Es bestand überdies eine positive Korrelation zwischen Transfektionseffizienz und synthetisiertem IL-10. Es wurde außerdem gezeigt, dass durch Stimulation mit dem immunmodulatorischen Zytokin Interferon-gamma (IFN-g) hRPE-Zellen MHC Klasse II-Moleküle vermehrt exprimierten. Damit sind sie ebenso wie die Makrophagen zur Antigenpräsentation fähig. Die Wirkung des von den transfizierten hRPE-Zellen sezernierten IL-10 auf die Proliferation von T-Lymphozyten wurde zwischen Tag 2 und Tag 6 (hTERT-RPE1-Zellen) beziehungsweise zwischen Tag 2 und Tag 4 (Spender-hRPE-Zellen) photometrisch untersucht. Die Proliferation allogener T-Lymphozyten mit beziehungsweise ohne Makrophagen konnte durch das sezernierte IL-10 supprimiert werden. Bei den hTERT-RPE1-Zellen lag ohne die Anwesenheit von professionellen APC am Tag 6 eine signifikante Reduktion der T-Lymphozytenproliferation vor, während bei Kokultivierung mit Makrophagen Signifikanzen am Tag 5 und Tag 6 erkennbar waren. Die immunsuppressive Wirkung von IL-10 konnte mittels Anti-IL-10-Antikörper neutralisiert werden. Damit wurde bewiesen, dass die proliferations-supprimierende Wirkung auf IL-10 zurückzuführen war. Diese Ergebnisse könnten demnach neue Möglichkeiten zur Verhinderung einer Abstoßungsreaktion nach RPE-Zelltransplantation bei Patienten mit AMD eröffnen / Age-related macular degeneration (AMD) is a disease of eyes affecting the macula lutea, the area of the retina with the highest density of retinal pigment epithelial cells (RPE cells). The disease is characterized by degeneration of RPE cells resulting in dystrophy of photoreceptors and finally loss of central vision. Transplantation of healthy RPE cells is a promising possibility for therapy but rejection of the allotransplant limits clinical application. One way to avoid this complications is a systemic immunosuppression of the recipient but this is combined with many side effects. In this thesis a self-constructed IL-10 expression vector (plasmid pCIneoIL-10) has been transferred into human RPE cells in vitro by gene transfer. In addition to human donor RPE cells a permanent RPE cell line (hTERT-RPE1 cells) was employed. Kationic lipids were used as transient transfection system for transfer of pCIneoIL-10 into hRPE cells. Three different lipid formulations and various ratios of transfection reagent:DNA were evaluated for highest transfection efficacy. With the optimized protocols a transfection efficacy of 23,3 ± 9,0 % (hTERT-RPE1 cells) and 10,3 ± 4,5 % (donor hRPE cells) was achieved. A negative influence on the viability of the hRPE cells after transfection was not observed. The IL-10 mRNA expression was analysed by reverse transcription-polymerase chain reaction (RT-PCR). Only in transfected hRPE cells the IL-10 mRNA-amplicon with 383 bp in size was found. Secretion of IL-10 protein in the cell culture supernatants of transfected hRPE cells was investigated using an enzyme-linked immunosorbent assay (ELISA) daily for 7 days. The IL-10 protein concentrations peaked at day 3 with 10,3 ± 0,8 ng/ml (hTERT-RPE1 cells) and 3,1 ng/ml (donor hRPE cells). The amount of secreted IL-10 positively correlated with transfection efficacy. After stimulation with the immunmodulatory cytokine interferon-gamma (IFN-g) the expression of MHC class II molecules on hRPE cells is increasing. Therefore they are able to present antigens similar to macrophages. Hence, the effects of recombinantly expressed IL-10 on the proliferation of allogeneic T lymphocytes were investigated both with and without allogeneic macrophages as professional antigen presenting cells (APC). Proliferation of T lymphocytes has been investigated colorimetrically between day 2 and day 6 (hTERT-RPE1 cells) and day 2 and day 4 (donor hRPE cells) respectively. The proliferation of allogeneic T lymphocytes with and without macrophages could be suppressed by the secreted IL-10. Signifikant reduction of proliferation was observed at day 6 in absence of professional APC (14,1 ± 1,1 % to 100% of untransfected control) and between day 5 (44,1 ± 4,9 %) and day 6 (37,4 ± 6,3%) in the presence of macrophages. It was possible to neutralize the immunosuppressive effect of IL-10 with anti-IL-10 antibodies. Proving that the suppressive effect of T lymphocyte proliferation was caused by IL-10. Thus, the specific IL-10 gene transfer into hRPE cells prior to transplantation may prevent rejection process and could prove a reliable method to help prevent loss of central vision due to AMD.

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