>Magister Scientiae - MSc / Saccharomyces cerevisiae is an important host in industrial biotechnology. This yeast is the host of choice for the first and second-generation biofuels for ethanol production. Genome modification in S. cerevisiae has been extremely successful largely due to this yeast’s highly efficient homology-directed DNA repair machinery. The advent of CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) genome editing technology has made multi-gene editing in yeast more accessible. In this study, we aimed at targeting the Cas9 to multiple genomic positions for integrating multiple genes at different sites. We have developed two CRISPR-Cas9 systems, based on published one- and two-plasmid systems, for application in S. cerevisiae strains. In this study, these CRISPR-Cas9 systems were used to transform fungal heterologous genes into yeast using the electroporation transformation method. We first utilized the CRISPR systems for targeting the T.r.eg2 gene to single locus chromosomal sites for single copy integration. Subsequently, we then targeted the same gene to repeated sequences in the genome, namely the delta sites, for multi-copy integration. The procedure was repeated with a different gene, T.e.cbh1, integrated into the same sites to ascertain reporter gene specific effects. High integration efficiency was achieved, since all the strains successfully integrated the genes. However, we discovered significant differences in enzyme activities between the two genes when targeted to different loci, as well as varying copy numbers as determined by qPCR. The T.e.cbh1 gene was highly expressed by yeast transformants targeted at the repeated delta sequences used for multi-copy integration, reaching maximum levels of 248 mU/gDCW. The T.r.eg2 gene was highly expressed in yeast transformants targeted to the single locus site on chromosome 12, reaching a maximum of 160U/gDCW, though it was shown that off-target integration likely occurred. We then used the information from these observations to construct a CBP yeast strain containing three cellulase genes: T.r.eg2, T.e.cbh1, and S.f.BGL1. Significant differences in enzyme activities were observed between the three genes, and it was shown that the S.f.BGL1 gene was poorly expressed by the CBP yeast strain, whereas the T.r.eg2 gene was highly expressed. Notably, due to the fact that marker containing plasmids could be cured from these strains, many additional genetic changes can still be made. Overall, our two CRISPR-Cas9 systems were efficient at engineering strains that produce recombinant proteins and can be used in future studies for a variety of applications, including metabolic engineering in S. cerevisiae

Today there are no requirements on burglary protection in the building regulation “Boverketsbyggregler”. Instead SSF Stöldskyddsföreningen has developed norms for physical security.SSF 200:5 is a norm that treats burglary protection in buildings and business premises, and isused as a basis for the insurance companies own description of security classification.The main purpose of the report is to get an increased understanding of what requirementsregarding burglary protection there are, and who is demanding them. The report also aims toexplore how people in the industry deal with security related work in the design stage.The questions at issue are all related to burglary protection. To answer these questions threemethods have been used. Interviews/surveys, physical tests, and a comparison between SSF200:5 and the insurance companies description of security classification.The interviews show that the burglary protection can vary depending on who the claimant is.It shines a light on a problem that occurs in the construction process, that is, discussingsecurity related questions too late. This can lead to compromised burglar security andincreased construction costs.The comparison shows that SSF 200:5 is not fully comparable with the insurance companiesdescription of security classification. Easy measures like door and window locks can beequated, meanwhile more complicated measures like wall constructions, can not.Using the test results, alternative wall constructions that satisfy the norm have been presented.Also improvements of SSF 200:5 have been discussed in favor of the whole industry.

Thesis (MScEng)--Stellenbosch University, 2012 / ENGLISH ABSTRACT: Through many years of research, a number of production schemes have been developed for converting lignocellulosic biomass into transport fuels. These technologies have been assessed through a number of techno-economic studies for application in a particular context in terms of the technical and economic feasibility. However, previous studies using these methods have tended to lack vigour in various aspects. Either the energy efficiency of the processes were not maximised through adequate heat integration, or a competing technology which existed was not considered. From an economic perspective, the financial models would often lack the vigour to account for the risk and uncertainty that is inherent in the market prices of the commodities. This phenomenon is especially relevant for the biofuel industry that faces the full fledge of uncertainties experienced by the agricultural sector and the energy sector. Furthermore, from an environmental perspective, the techno-economic studies had often ignored the environmental impacts that are associated with biofuel production. Thus, a comparative study could have favoured an option due to its economic feasibility, while it could have had serious environmental consequences. The aim of this study was to address these issues in a South African context, where biofuels could be produced from sugarcane bagasse. The first step would be to modify an existing simulation model for a bioethanol scenario that operates with a Separate Hydrolysis and Fermentation (SHF process) configuration into a second processing scenario that operates with a Simultaneous Saccharification and Fermentation (SSF process) configuration using reliable experimental data. The second step was to ensure that the maximum energy efficiency of each scenario was realised by carrying out pinch point analysis as a heat integration step. In contrast to these biological models is the thermochemical model that converts bagasse to gasoline and diesel via gasification, Fischer-Tropsch synthesis and refining (GFT process). While there were no significant advances in technology concerning this type of process, the energy efficiency was to be maximised with pinch point analysis. The GFT process obtained the highest energy efficiency of 50.6%. Without the affects of pinch point technology, the efficiency dropped to 46%, which thus emphasises the importance of heat integration. The SSF had an efficiency of 42.8%, which was superior to that of the SHF at 39.3%. This resulted from a higher conversion of biomass to ethanol in the SSF scenario. Comparing the SHF model to an identical model found in literature that did not have pinch point retrofits, this study showed lower efficiency. This arose because the previous study did not account for the energy demands of the cold utility systems such as the cooling tower operation, which has been shown in this study to account for 40% of the electrical energy needs. The economic viability of all three processes was assessed with Monte Carlo Simulations to account for the risks that the fluctuations in commodity prices and financial indices pose. This was accomplished by projecting the fluctuations of these parameters from samples of a historical database that has been transformed into a probability distribution function. The consequences were measured in terms of the Net Present Value (NPV) and Internal Rate of Return (IRR) for a large number of simulations. The results of these variables were aggregated and were then assessed by testing the probability that the NPV<0, and that the IRR recedes below the interest rate of 12.64%. The investment was thus deemed unfeasible if these probabilities were greater than 20%. Both biological models were deemed profitable in terms of this standard. The probabilities were 13% for the SSF and 14% for the SHF. The GFT process however was deemed completely unfeasible because the probability that the NPV<0 was 78%. Given that the GFT process had the highest energy efficiency, this result arises mainly because the capital investment of 140,000USD/MWHHV of biomass energy input is to enormous for any payback to be expected. The environmental footprint of each process was measured using Life Cycle Assessments (LCAs). LCAs are a scientifically intricate way of quantifying and qualifying the effects of a product or process within a specified boundary. The impacts are assessed on a range of environmental issues, such as Global Warming, Acidification, Eutrophication and Human toxicity. Furthermore, if the project under concern has multiple output products, then the impacts are distributed between the output products in proportion to the revenue that each generates. The impacts were either relative to the flow of feedstock, which was 600MW of bagasse, or to the functional unit, which was the amount of fuel required to power a standard vehicle for a distance of 1 kilometre. In either case, the GFT scenario was the least burdening on the environmental. This was expected because the GFT process had the highest energy efficiency and the process itself lacked the use of processing chemicals. Relative to the feedstock flow, the SSF was the most environmentally burdening scenario due to the intensive use of processing chemicals. Relative to the functional unit, the SHF was the most severe due to its low energy efficiency. Thus, the following conclusions were drawn from the study:  The GFT is the most energy and environmentally efficient process, but it showed no sign of economic feasibility. iv  There is no significant difference in the economic and environmental evaluation of the SSF and SHF process, even though the SSF is considered to be a newer and more efficient process. The major cause of this is because the setup of the SSF model was not optimised. / AFRIKAANSE OPSOMMING: Deur baie jare van navorsing is ‘n aantal produksie-skemas vir die omskakeling van lignosellulose biomassa na vloeibarebrandstof ontwikkel. Hierdie tegnologië is geassesseer ten opsigte van die tegniese en ekonomiese haalbaarheid deur middel van tegno-ekonomiese studies in bepaalde tekste. Tog het hierdie vorige studies besliste beperkings gehad. Of die energie-doeltreffendheid van die proses is nie gemaksimeer deur voldoende hitte-integrasie nie, of 'n mededingende tegnologie wat bestaan is nie oorweeg nie. Vanuit 'n ekonomiese perspektief, was die finansiële modelle dikwels nie die omvattend genoeg om rekening te hou met die risiko en onsekerheid wat inherent is in die markpryse van die kommoditeite nie. Hierdie verskynsel is veral relevant vir die biobrandstof bedryf wat die volle omvang van onsekerhede ervaar waaraan die landbousektor en die energiesektoronderhewig is. Verder het die tegno-ekonomiese studies dikwels die omgewingsimpakte wat verband hou met biobrandstofproduksie geïgnoreer. Dus kon ‘n opsie deur die ekonomiese haalbaarheid bevoordeel word, ten spyte van die ernstige omgewingsimpakte wat dit kon inhou. Die doel van hierdie studie was om hierdie kwessies aan te spreek in 'n Suid-Afrikaanse konteks, waar biobrandstof uit suikerriet bagasse geproduseer kan word. Die eerste stap was om 'n bestaande simulasiemodel vir 'n bio-scenario wat met Afsonderlike Hidroliese en Fermentasie (SHF proses) stappe werk, te modifiseer vir 'n tweede verwerking scenario wat met 'n gelyktydige Versuikering en Fermentasie (SSF proses) konfigurasie werk. Die verandering is gedoen deur die gebruik van betroubare eksperimentele data. Die tweede stap was om te verseker dat elke scenario die maksimum energie-doeltreffendheid het, deur 'n hitte-integrasie stap, wat gebruik maak van “pinch-point” analise. In teenstelling met hierdie biologiese modelle, is daar die thermochemiese roete waar petrol en diesel van bagasse vervaardig word via vergassing, Fischer-Tropsch-sintese en rafinering (GFT proses). Daar was geen betekenisvolle vooruitgang in tegnologie vir hierdie proses nie, maar die energie-doeltreffendheid is gemaksimeer word deur energie-integrasie. Die GFT proses toon die hoogste energie-doeltreffendheid van 50,6%. Sonder die invloed van energie-integrasie het die doeltreffendheid gedaal tot 46%, wat dus die belangrikheid van hitte-integrasie beklemtoon. Die SSF het 'n effektiwiteit van 42,8% gehad, wat beter was as dié 39,3% van die SHF opsie. Hierdie hoër effektiwiteit wasas gevolg van die hoër omskakeling van biomassa na etanol in die SSF scenario. Die energie doeltreffendheid vir die SHF-model was laer as met 'n identiese model (sonder energie-integrasie) wat in die literatuur gevind wat is. Dit het ontstaan omdat die vorige studie nie 'n volledig voorsiening gemaak het met die energie-eise van die verkillingstelselsnie, wat tot 40% van die elektriese energie behoeftes kan uitmaak. Die ekonomiese lewensvatbaarheid van al drie prosesse is bepaal met Monte Carlo simulasies om die risiko's wat die fluktuasies in kommoditeitspryse en finansiële indekse inhou, in berekening te bring. Hierdie is bereik deur die projeksie van die fluktuasies van hierdie parameters aan die hand van 'n historiese databasis wat omskep is in 'n waarskynlikheid verspreiding funksie. Die gevolge is gemeet in terme van die netto huidige waarde (NHW) en Interne Opbrengskoers (IOK) vir 'n groot aantal simulasies. Die resultate van hierdie veranderlikes is saamgevoeg en daarna, deur die toets van die waarskynlikheid dat die NPV <0, en dat die IRR laer as die rentekoers van 12,64% daal, beoordeel. Die belegging is dus nie realiseerbaar geag as die waarskynlikhede meer as 20% was nie. Beide biologieseprosesse kan as winsgewend beskou word in terme van bostaande norme. Die waarskynlikhede was 13% vir die SSF en 14% vir die SHF. Aangesien die NHW van die GFT-proses onder 0 met ‘n waarskynlikheid van 78% is, is die opsie as nie-winsgewend beskou. Gegewe dat die GFT-proses die hoogste energie-doeltreffendheid het, is die resultaat hoofsaaklik omdat die kapitale belegging van 140,000 USD / MWHHV-biomassa energie-inset te groot is, om enige terugbetaling te verwag. Die omgewingsvoetspoor van elke proses is bepaal deur die gebruik van Lewens Siklus Analises (“Life Cycle Assessments”) (LCAS). LCAS is 'n wetenskaplike metodeom die effek van ‘n produk of proses binne bepaalde grense beide kwalitatief en kwantitatief te bepaal. Die impakte word beoordeel vir 'n verskeidenheid van omgewingskwessies, soos aardverwarming, versuring, eutrofikasie en menslike toksisiteit. Voorts, indien die projek onder die saak verskeie afvoer produkte het, word die impakte tussen die afvoer produkte verdeel, in verhouding tot die inkomste wat elkeen genereer. Die impak was met of relatief tot die vloei van roumateriaal (600MW van bagasse), of tot die funksionele eenheid, wat die hoeveelheid van brandstof is om 'n standaard voertuig aan te dryf oor 'n afstand van 1 kilometer. In al die gevalle het die GFT scenario die laagste belading op die omgewing geplaas. Hierdie is te verwagte omdat die GFT proses die hoogste energie-doeltreffendheid het en die proses self nie enige addisionele chemikalieë vereis nie. Relatief tot die roumateriaal vloei, het die SSF die grootse belading op die omgewing geplaas as gevolg van die intensiewe gebruik van verwerkte chemikalieë. Relatief tot die funksionele eenheid, was die SHF die swakste as gevolg van sy lae energie-doeltreffendheid.

<p>Detta arbete, inom ämnet Kristendomens historia, presenterar en svenskkyrklig orden, ”Franciskus Tredje Orden inom Svenska kyrkan” (F.T.O.), som tillhör den ”Tredje Orden” (TSSF) inom ”The Society of Saint Francis” (SSF), en ordensgemenskap inom den Anglikanska kyrkan, Church of England. Orden är öppen för kvinnor och män, vigda (biskopar, präster och diakoner) och lekfolk som lever vanliga liv i familj och samhälle, med en vilja att leva sina liv utifrån en tydlig inspiration av den helige Franciskus och hans liv.</p><p>Syftet med arbetet är att undersöka och redogöra för de primärkällor som finns från tiden av F.T.O.: s grundande för att svara på frågan om huruvida Ordens uppkomst var förenlig eller ej med den Svenska kyrkan, dess tro, lära och bekännelse.</p><p>Undersökningen inleds med en deskriptiv del gällande bakgrunden till frågeställningen där jag redogör något för reformationen och vad detta medförde för Svenska kyrkan gällande klosterliv. Sedan följer en redogörelse och en hermeneutiskt och dogmatisk analys av de, för tiden av Ordens grundande, aktuella källorna.</p><p>En sammanfattning av resultatet som framkommit är det att grundandet av F.T.O. i början av 1970-talet var förenligt med den Svenska kyrkan. Vi kan se i källorna att Ordens syfte och annat typiskt för Orden, korresponderar väl med vad som uttrycks gällande tro, lära och bekännelse i Kyrkolagen 1686 och de, förutom Bibeln, samlade bekännelseskrifterna för den Svenska kyrkan. Genom detta arbete konstaterar vi att ett ordensliv likt F.T.O. kan tillämpas inom ett evangeliskt-lutherskt kyrkosamfund och att det på intet sätt är synonymt med den gärningsfromhet inom klosterväsendet som reformatorerna så tydligt vände sig mot under medeltiden.</p> / <p>This study, in the subject the history of Christianity, present an order in the Church of Sweden, “Saint Francis Third Order within the Church of Sweden” (F.T.O.), which is a part of the “Third Order” (TSSF) within “The Society of Saint Francis” (SSF), a community within the Anglican Church, Church of England. The order is open for women and men, ordained (bishops, priests and deacons) and lay people who live ordinary lives in family and society, with a desire to live their lives inspired by Saint Francis and his life.</p><p>My purpose with this study is to examine and describe the prime sources from the time of the foundation of the F.T.O. to answer the question about whether the foundation of the Order was compatible or not with the Church of Sweden, their faith, doctrine and confession.</p><p>The examination starts with a descriptive part about the background to the study problem where I give some facts about the reformation and what that brings for the Church of Sweden in relation to monastery life. Then comes a description and a hermeneutics and a dogmatic analysis of the, for the time of the foundation of the Order, current sources.</p><p>A summery of the upcoming results of this study is that at the foundation of F.T.O. in 1970ths was compatible with the Church of Sweden. In the sources we can see that the Orders purpose and other typical for the Order corresponds well with what they say about faith, doctrine and confession in the church law from 1686 and the, except the holy Bible, confession scripts for the Church of Sweden. Through this work we establish the fact that an order life like that we see in F.T.O. could practices in an evangelical-Lutheran church and it is not in any way synonymic with that phenomena called “action piety” within the monastery life that the men’s of the reformation so clearly said no to under the middle age.</p>

Detta arbete, inom ämnet Kristendomens historia, presenterar en svenskkyrklig orden, ”Franciskus Tredje Orden inom Svenska kyrkan” (F.T.O.), som tillhör den ”Tredje Orden” (TSSF) inom ”The Society of Saint Francis” (SSF), en ordensgemenskap inom den Anglikanska kyrkan, Church of England. Orden är öppen för kvinnor och män, vigda (biskopar, präster och diakoner) och lekfolk som lever vanliga liv i familj och samhälle, med en vilja att leva sina liv utifrån en tydlig inspiration av den helige Franciskus och hans liv. Syftet med arbetet är att undersöka och redogöra för de primärkällor som finns från tiden av F.T.O.: s grundande för att svara på frågan om huruvida Ordens uppkomst var förenlig eller ej med den Svenska kyrkan, dess tro, lära och bekännelse. Undersökningen inleds med en deskriptiv del gällande bakgrunden till frågeställningen där jag redogör något för reformationen och vad detta medförde för Svenska kyrkan gällande klosterliv. Sedan följer en redogörelse och en hermeneutiskt och dogmatisk analys av de, för tiden av Ordens grundande, aktuella källorna. En sammanfattning av resultatet som framkommit är det att grundandet av F.T.O. i början av 1970-talet var förenligt med den Svenska kyrkan. Vi kan se i källorna att Ordens syfte och annat typiskt för Orden, korresponderar väl med vad som uttrycks gällande tro, lära och bekännelse i Kyrkolagen 1686 och de, förutom Bibeln, samlade bekännelseskrifterna för den Svenska kyrkan. Genom detta arbete konstaterar vi att ett ordensliv likt F.T.O. kan tillämpas inom ett evangeliskt-lutherskt kyrkosamfund och att det på intet sätt är synonymt med den gärningsfromhet inom klosterväsendet som reformatorerna så tydligt vände sig mot under medeltiden. / This study, in the subject the history of Christianity, present an order in the Church of Sweden, “Saint Francis Third Order within the Church of Sweden” (F.T.O.), which is a part of the “Third Order” (TSSF) within “The Society of Saint Francis” (SSF), a community within the Anglican Church, Church of England. The order is open for women and men, ordained (bishops, priests and deacons) and lay people who live ordinary lives in family and society, with a desire to live their lives inspired by Saint Francis and his life. My purpose with this study is to examine and describe the prime sources from the time of the foundation of the F.T.O. to answer the question about whether the foundation of the Order was compatible or not with the Church of Sweden, their faith, doctrine and confession. The examination starts with a descriptive part about the background to the study problem where I give some facts about the reformation and what that brings for the Church of Sweden in relation to monastery life. Then comes a description and a hermeneutics and a dogmatic analysis of the, for the time of the foundation of the Order, current sources. A summery of the upcoming results of this study is that at the foundation of F.T.O. in 1970ths was compatible with the Church of Sweden. In the sources we can see that the Orders purpose and other typical for the Order corresponds well with what they say about faith, doctrine and confession in the church law from 1686 and the, except the holy Bible, confession scripts for the Church of Sweden. Through this work we establish the fact that an order life like that we see in F.T.O. could practices in an evangelical-Lutheran church and it is not in any way synonymic with that phenomena called “action piety” within the monastery life that the men’s of the reformation so clearly said no to under the middle age.

In most casting applications, dendritic microstructure morphology is not desired because it leads to poor mechanical properties. Forced convection causing sufficient shearing in the mushy zone of the partially solidified melt is one of the means to suppress this dendritic growth. The dendrites formed at the solid-liquid interface are detached and carried away due to strong fluid flow to form slurry. This slurry, consisting of rosette or globular particles, provides less resistance to flow even at a high solid fraction and can easily fill the die-cavity. The stated principle is the basis of a new manufacturing technology called “semi-solid forming” (SSF), in which metal alloys are cast in the semi-solid state. This technique has numerous advantages over other existing commercial casting processes, such as reduction of macrosegregation, reduction of porosity and low forming efforts. Among all currently available methods available for large scale production of semisolid slurry, the cooling slope is considered to be a simple but effective method because of its simple design and easy control of process parameters, low equipment and running costs, high production efficiency and reduced inhomogeneity. With this perspective, the primary objective of the present research is to investigate, both experimentally and numerically, convective heat transfer and solidification on a cooling slope, in addition to the study of final microstructure of the cast billets. Some key process parameters are identified, namely pouring temperature, slope angle, slope length, and slope cooling rate. A systematic scaling analysis is performed in order to understand the relative importance of the parameters in influencing the final properties of the slurry and microstructure after solidification. A major part of the present work deals with the development of an experimental set up with careful consideration of the range of process parameters involved by treating the cooling slope as a heat exchanger. Subsequently, a comprehensive numerical model is developed to predict the flow, heat transfer, species concentration solid fraction distribution of aluminum alloy melt while flowing down the cooling slope. The model uses a variable viscosity relation for slurry. The metal-air interface at the top during the melt flow is tracked using a volume of fluid (VOF) method. Solidification is modeled using an enthalpy based approach and a volume averaged technique. The mushy region is modeled as a multi-layered porous medium consisting of fixed columnar dendrites and mobile equiaxed or fragmented grains. In addition, the solidification model also incorporates a fragmentation criterion and solid phase movement. The effects of key process parameters on flow behavior involving velocity distribution, temperature distribution, solid fractions at the slope exit, and macrosegregation, are studied numerically and experimentally for aluminium alloy A356. The resulting microstructures of the cast billets obtained from the experiments are studied and characterized. Finally the experimental results are linked to the model predictions for establishing the relations involving interdependence of the stated key process parameters in determining the quality of the final cast products. This study is aimed towards providing the necessary guidelines for designing a cooling slope and optimizing the process parameters for desirable quality of the solidified product.