Abstract   This exam project is an effort to derivatize one side of magnetic beads with one kind of molecule  , and another one on the opposite side. First the surface of the sphere is loaded with a suitable linker with, e.g. amino or hydroxyl groups. In the second step, these groups are derivatized with a photosensitive protecting group such as Nitroveratryloxycarbonyl. In the third step, the particles are placed on a surface and then irradiated with UltraViolet light (320 nm) from above, which will cleave off the Nitroveratryloxycarbonyl on the upper half, while leaving in place the ones at the lower half. The linker groups of the upper half can now be derivatized by other reagents of choice. The remaining Nitroveratryloxycarbonyl groups can be removed by suspending the particles in a solvent and then exposing them to UltraViolet light. Finally the linker groups on this half of the particles can be derivatized by a second reagent. Magnetic particles were marked with FITC, two different kinds of magnetic particles were selected, sikastar-NH2 function and sikastar-COOH function. Five different solvents were used to wash the magnetic particles and remove the bounded FITC, solvents are Acetone, 1-butanol, DMSO, 4-propanol, and Urea. Magnetic particles sikastar-NH2 and sikastar-COOH were washed with Tween 20 and SDS to remove non-specific binding of FITC. Sikastar particles were treated with IgG*FITC in constant presence of the following solvents: PBS*10, Pluronic-F127, Tween 20. Pegylation of sikastar particles got done to reduce non-specific binding. Derivatisation of Nitroveratryloxycarbonyl got done and specific bindning of IgG*FITC to micromer particles got done by protein thiolation. When a different concentration of FITC was tested to control specific and non-specific binding to sikastar functions, we observed that we had a specific binding to sikastar-NH2 in the lowest concentration. In choice of magnetic particles we had specific binding with sikastar-NH2. Using a different solvents Acetone, 1-butanol, 4-propanol, and Urea  to remove bounded FITC, sikastar-NH2 showed stronger fluoresence than sikastar-COOH after washing because of specific binding and it was difficult to remove FITC with Acetone, 1-butanol,  4-propanol,and Urea, on the other hand DMSO could remove bounded FITC from sikastar particles. When we washed magnetic particles sikastar-NH2 and sikastar-COOH with Tween 20 and SDS to remove non-specific binding of FITC, we could see that magnetic particles showed fluoresence in both functions due to non-specific binding. When sikastar particles got treated with IgG*FITC in constant presence of solvents PBS*10, Pluronic-F127, and Tween 20, we had a specific binding between sikastar particles and IgG*FITC in a presence of pluronic-F127. Pegylation of sikastar particles with a different kind of a PEG was possibl to reduce non-specific bindning. The derivatisation of Nitroveratryloxycarbonyl could be done in a N2 environment, and Nitroveratryloxycarbonyl-sikastar-NH2 could be radiated with UltraViolet light to remove Nitroveratryloxycarbonyl. Also thiolation method could be used to perform specific binding of IgG*FITC to micromer particles.

På uppdrag av Hifab Byggprojektledaren AB:s (Hifab AB) filial i Karlstad kommer riskerna för spridning av miljöstörande avfall såsom asbest, dioxin, kvicksilver samt PCB att inventeras och utvärderas vid rivning av kemisk industrienhet vid Akzo Nobels anläggning i Skoghall. Inventeringen begränsas till att omfatta den gamla kloralkalifabriken byggd 1938-1946 innehållande cellsalar, likriktarutrymmen samt personaldelar. Riktad provtagning bygger på tidigare erfarenheter och kunskaper av liknande objekt. Byggnationsår kan ge indikationer på vilka ämnen som kan misstänkas finnas i ev. byggnad då vissa material användes under olika epoker. Metoden är ekonomiskt fördelaktig men ger inte en heltäckande bild över hur föroreningen är spridd. Vid riskanalysen är den så kallade MIFO-modellen använd, där bedömning sker från tillgänglig data från den riktade provtagningen. Efter genomgången gasskyddsutbildning gavs möjlighet att okulärbesikta gamla cellsalarna med tillhörande personalutrymmen, förråd samt transformator- och likriktarutrymmen. Prover togs för senare analys av asbest och övrig provtagning gällande kvicksilver på platsen är daterade 1999 av Envipro. Analysresultaten påvisade ingen asbest i proverna. Enligt uppgift från ansvarig personal på Akzo Nobel har tidigare PCB-prover visat sig vara negativa. Dock har stora mängder kvicksilver påträffats i objektet. Slutsatsen är att i cellsalen har halter som överstiger Naturvårdsverkets gränsvärden gällande mindre känslig markanvändning av kvicksilver uppmätts. Detta är ett stort miljöproblem som måste åtgärdas så fort som möjligt.

Sammanfattning Syftet med den här studien var att utveckla och optimera FISH (Fluorescense In Situ Hybridisation) tekniken som en snabb och ganska billig metod för detektion av laktobaciller. Det vill säga att kunna på objektsglas använda FISH tekniken för att identifiera laktobaciller på artnivå med fluorescensmärkta prober mot 16S och 23S RNA. FISH är en allmän och användbar metod för att detektera och lokalisera mikroorganismer eller en specifik grupp av mikroorganismer i provet (1). Metoden detekterar DNA- eller RNA- sekvenser med hjälp av fluorescensmärkta prober som hybridiseras specifikt med komplementära målsekvenser i intakta celler (2). Detta innebär att man behåller cellmorfologin och tillför en lättdetekterad fluorescerande färg. I början av studien utvaldes två grupper av bakterier, gramnegativa bakterier som har tunnare cellvägg, vilket underlättar hybridiseringen och grampositiva laktobacillus med en tjockare cellvägg. Enligt tidigare undersökningar gav FISH tekniken bra resultat om rätt probe användes för rätt organism. Som anvisning till den här studien användes en tidigare studie på E.coli K12.(6). I början av den här studien användes samma prober som i studien med E.coli K12. Bakterierna i grupp 1 valdes utifrån homologi mellan probernas sekvens och målsekvensen hos bakterierna. De hade 100 % homologi med probe 1 och hög homologi, 83-100 %, med probe 2 ,vilket väl överensstämmer med resultatet för E. coli. Bakterierna odlades i lämpliga medier och prov togs från log-fas. Bakterierna behandlades med hybridiseringsbuffert och studerades under fluorescensmikroskop. Stark fluorescens iakttogs i flertalet av bakterierna i de fall som homologi mellan probe och målsekvens var hög. Effekten av tiden för förvaring i kyl efter skörd på fluorescensförmågan studerades också. Fluorescensen efter en dag jämfördes med fluorescensen efter 15 dagar och 30 dagar hos samma bakterie. Bakterierna fotograferades i fluorescensmikroskop samt ljusmikroskop och resultaten presenteras i respektive tabeller och/eller bilder. Resultaten visade en sänkning i fluorescensstyrka och i antal bakterier som lyste redan efter 15 dagar. Efter 30 dagar hade nästan alla bakterier upphört att reagera med proben, oavsett vilken art som studerades. Slutligen studerades kvaliteten på tvättningar. Fluorescensbilder togs av bakterierna efter en tvätt och jämfördes med bilder efter 2 och tre tvättar. Ingen stor skillnad observerades efter flera tvättningar jämförd med en tvätt. Studien visade att den teknik som utvecklades är användbar för att detektera specifika sekvenser både i grampositiva och gramnegativa bakterier.

Abstract This degree thesis was made in cooperation with Chematur Engineering in Karlskoga. Many of the environmental issues of today are a result of the motor traffic. Consumption of fossil fuels harms our environmental through formation of carbon dioxide and other greenhouse gases. To get under control with our environmental issues it’s important to find substitutes for oil that are both cheap and environmentally friendly. Ethanol has a long history as a motor fuel and is both enviromentally friendly has a high efficiency. According to the EU commissions directives for motor fuels, 5, 75 % of all the fuels in Sweden should be renewable in 2010. Ethanol is international the most widely used and imported renewable fuel. 70 millions litres of ethanol is manufactured in Sweden yearly and the majority comes from the factory in Norrköping owned by Lantmännen. In order to make the Swedish ethanol more competitive the import duty has been increased in 2007. The ethanol produced in Sweden mainly is made from wheat and grain, while the production in USA mainly is corn based. Ethanol can be manufactured by fermentation from all materials containing suger or some form of chemical composition that can be transformed into suger. The raw material used for ethanol production can be divided into three groups: suger, starch and cellulose. Biostil is a process developed in order to make ethanol production as cheap and environmentally friendly as possible. This method is used to manufacture both fuel and technichal ethanol. The raw material used is all kinds of starch containing material. Chematur Engineering has developed their own concept within biostil called BiostilÒ 2000. Biostil is a recycling concept that gives a high ethanol yield and lower water consumption. To be able to develop the biostil furher, Chematur Engineering aims to build a pilot plant on Kilsta industrial estate. / Sammanfattning Detta examensarbete gjordes i samarbete med Chematur Engineering i Karlskoga. Många av dagens miljöproblem kommer från biltrafiken. Förbrukning av fossila bränslen som exempelvis olja skadar vår miljö genom de utsläpp som bildas i form av koldioxid och andra växthusgaser. För att kunna få bukt med våra miljöproblem krävs det att substitut till olja hittas som är både kostnadseffektiva och miljövänliga. Etanol har länge använts som drivmedel för fordon, det är ett mer miljövänligt alternativ samtidig som det också har hög effektivitet. 5,75 % av de drivmedel som används i Sverige år 2010 ska vara förnyelsebara enligt EUs direktiv. Etanol är det förnyelsebara drivmedel som används mest internationellt och importeras till högsta grad. I Sverige idag tillverkas ca 70 miljoner liter om året och Lantmännens anläggning i Norrköping står för den största delen. Etanolen som produceras i Sverige kommer främst från spannmål som vete och korn, medan etanolproduktionen i ex. USA mestadels är majsbaserad. Etanol kan framställas med fermentering från alla material innehållande socker eller någon form av kemisk sammansättning som kan omvandlas till socker. De råvaror som används vid etanolframställning via fermentering delas in i tre typer: socker, stärkelse och cellulosamaterial. Biostil är en process utvecklad för att göra etanolproduktion så billig och miljövänlig som möjligt. Processen används för att tillverka både bränsleetanol och teknisk sprit och man använder sig av socker eller stärkelseinnehållande råvaror. Chematur Engineering har utvecklat ett eget koncept inom Biostil kallat BiostilÒ 2000. Biostil går i kort ut på att få ut så mycket etanol som möjligt ur biomassa genom recirkulation och andra åtgärder för att minska exempelvis vattenanvändningen. Tanken är att Chematur Engineering ska bygga en biostil pilotanläggning på Kilsta industriområde för att kunna utveckla processen ytterligare.

SAMMANFATTNING De textilindustrier som finns i u-länderna är oftast tekniskt enkla i sin utformning, bl.a. med avseende på det färgade avloppsvatten som släpps ut utan rening. Detta ger upphov till miljöförstöring pga. att vattnet innehåller höga salthalter, alkalinitet och stark färg. Det ger även upphov till hälsoproblem då flera av de färger som används vid textilfärgning kan blir enzymatiskt nedbrutna i människans matsmältningssystem och bilda cancerogena substanser. Syftet med denna studie var att undersöka om skogsflis, fetvadd och vitrötesvamp kan användas för att bryta ner färger som används vid textilfärgning i synnerhet i fattigare länder där problemen med utsläppen från textilfärgningsfabriker är stort. Huvudtanken med projektet är att med små medel förbättra miljö och hälsoförhållandena där textilindustrin befinner sig. I denna studie användes tre olika textilfärger tillsammans med skogsflis, fetvadd och vitrötesvamp. I försöken med skogsflis har avfärgningen av textilfärgerna varit mycket effektiv. Detta har kunnat bekräftas genom analys med spektofotometer och genom visuella iakttagelser. Försök har även gjorts för att se om skogsflis kan återanvändas. Det konstaterades att den kunde återanvändas. Försöket gav bättre resultat än om man använder ny skogflis. Detta konstaterades genom att avfärgningen gick snabbare med använd skogsflis än med ny skogsflis. Försök gjordes med vitrötesvamp (Bjerkandera sp. BOL-13) på maltagarplattor innehållande textilfärg. Även vitrötesvamp var effektiv vid avfärgning av textilfärgerna. Slutsatser som vi kan dra av vår studie är att användning av skogsflis och vitrötesvamp är effektiv vid avfärgning av textilvatten. Denna metod har stor potential för vidare utveckling, för att sedan kunna användas där billiga och enkla metoder behövs.

Prostate cancer is the most detected cancer in males and one of the leading causes of cancer mortality in the western society. Prostate Specific Antigen (PSA) is since the 1980´s widely used as a serum marker but can not distinguish between prostate cancer and benign prostate hyperplasia. PSA is a glycoprotein with a molecular weight of 30 kDa and consists of an active 237 amino acid residues polypeptide with five disulphide bonds and approximately 7-12 % carbohydrates. The aim of the project was to develop an immunochromatographic test for measuring the total concentration of PSA in urine and to verify, by using size exclusion chromatography, if PSA in urine was free or complexed with other proteins. However, the first issue was to deal with the variable urine composition and the occurrence of precipitate in urine which can bind proteins. Six different immunochromatographic test systems were developed and the concentration of PSA was measured in urine specimens from normal individuals and from patients with prostate cancer by using two selected systems. The most sensitive immunochromatographic system showed a detection limit of 1.2 ng/L which is 130 times more sensitive than presently available commercially enzyme immunoassays. The precipitates in urine were dissolved by pH-adjustment and addition of chelator and detergent. Only 0.5% PSA was detected in the supernatant of centrifuged urine in comparison to when the precipitates where dissolved and these findings confirm that PSA in urine is in precipitate. The median concentration in normal male urine was 106µg/L. The highest value of 991µg/L was obtained in urine from a patient with prostate cancer but several of the urine specimens from patients showed non-detectable values. Four normal and patient urine specimens were separated by size exclusion chromatography and the fractions were measured by two immunochromatographic test systems. For all specimens a single peak was eluted in the same position which was consistent with a 30kDa molecular weight protein and no larger complex of PSA could be found. However, it was found that measurable PSA during storage in the fraction tubes could disappear rapidly.

At Rottneros pulp mill mechanical pulp is bleached with hydrogen peroxide. The changes of bleaching chemicals that are used for different grades are taken from bleaching tables; however these tables are old and need to be updated. The purpose of this thesis was to be the basis of new documentation. In this thesis three pulps where studied with the aim to find the optimal bleaching chemical changes (total NaOH/VP-quote) for each pulp. The pulps were taken from the filter before the bleaching tower and the pulps were bleached in the lab. The most important pulp property in this study was the brightness and how that was influenced by the chemical dosages. A fiberline inspection was also done to examine the brightness gain during the process. Finally a study was carried out to examine how freeness affected the brightness for low- and high freeness pulps. The studies showed that reduced chemical consumption was possible to achieve for all pulps studied. For pulp CA 683 a total NaOH/VP-ratio of 0,8 was suggested, while the ratio were 1,1 for CA 970 and 0,7 for CS 770. Lower freeness increased the brightening of the pulp at the same chemical consumption.

Iron overload is a serious clinical condition caused by excessive iron in the body, which can be largely prevented by the use of iron-specific chelating agents. At the moment there are only a few chelators in clinical use for the treatment of Fe overload. One of them, and so far the best working one, is desferrioxamine (DFO). This iron chelator has a major disadvantage of being orally inactive and is given by long and frequent subcutaneous infusions (12-24 hours/5-6 days/week) to patients. 1 Consequently, the design of an orally active, nontoxic, selective iron chelator has become a high priority. To design an iron chelator for clinical use, the important factors to consider are metal selectivity and affinity, ligand-metal complex stability, bioavailability and toxicity. The best iron chelator should be highly selective for iron(III) in order to minimize chelation of other biological essential metal ions which could lead to deficiency with prolonged usage. Favouring the Fe (III) oxidation state avoids Fenton chemistry and the production of toxic free radicals. This report deals with complexes of aroylpicolylhydrazines. Chemical and biological testing has established that these molecules function extremely well as iron chelators in vivo. The most effective candidates are more efficient at promoting iron efflux (release of iron) from cells than the existing drug DFO. Although the ligands discussed in this report, are selective for iron, there is no record of how they interact with other essential metal ions (Mn, Co, Ni, Cu and Zn) in our body and it is this that is the topic of this report. It was found that the complexes with H2PPH (N,N’-bis(α-picolinoyl)hydrazine) probably built polymeric complexes and therefore they were almost totally insoluble in all common solvents. The complexes of H2BPH (N-(benzoyl)-N’-(picolinoyl)hydrazine)) were also problematic in terms of insolubility. Another problem was decomposition of the complexes in solution, with resultant precipitation of the ligand and that happened with all of the ligands. The ligand H2TPH formed complexes with Ni and Co, [MII(HL)2] and [MIII(L)2]-, and crystals were grown suitable for x-ray structure studies. The cobalt complex is the first one reported of its kind. There was also crystals grown suitable for x-ray work of a protonated picolylhydrazide and that is also the first crystal structure reported.

<p>Electrowinning is an energy demanding process, in which the electrolysis accounts for the major part of the energy consumption. The anodic reaction in the electrolysis of zinc from sulphate based electrolytes is oxygen evolution, which has a standard potential of 1.23 V at 25 ºC. The lead anodes used in zinc electrowinning today have a high overpotential for oxygen evolution and the operating anode potential is as high as 2 V vs NHE. Since the lead anodes are not stable some of the lead can dissolve and incorporate in the zinc metal on the cathode. The lead anodes are also undesired because of health concerns. By introducing formic acid or methanol oxidation as anode reaction, the anode potential could be lowered significantly. This would reduce the energy consumption and also enable the use of new types of electrodes.</p><p>The general aim of this thesis was to investigate how depolarised anodes for methanol and formic acid oxidation can be used in electrowinning processes.</p><p>Several electrodes were shown to be suitable as anodes in zinc electrowinning. A high activity was obtained with porous electrodes, whereas a platinum coated titanium was the most stable electrode. The choice of electrode will however always be a compromise between activity, stability and costs. All electrodes were deactivated with time and the best method for reactivation tested in this study, was periodic current reversal (PCR).</p><p>An operating procedure was established for methanol and formic acid oxidation on high surface area electrodes, Pt-TySAR^®. With the use of PCR at process current density, a potential lower than 0.7 V vs NHE could be maintained for 36 hour in synthetic electrolyte. The use of formic acid oxidation resulted in lower potentials than the use of methanol oxidation.</p><p>The activities for methanol and formic acid oxidation in industrial electrolyte were very low and chloride impurities were shown to cause the major part of the activity reduction. Even small amounts of chloride (10^-6-10^-5M) affected the activity of the electrodes. The chloride impurities are considered as the main problem to overcome in order to introduce the oxidation of formic acid or methanol in the zinc electrowinning process.</p> / QC 20101004

<p>The thesis deals with various ways of utilization of biomass. Chapter 1 compares three biomass types: birch wood Betula sp., marine brown alga Fucus vesiculosus, and terrestrial moss Pleurozium schreberi, as precursors for preparation of biosorbents for removal of copper ions from diluted water solutions. Small sample doses (0.5 g/100ml) of the biosorbents prepared from alga and moss enabled more than 90 % removal of Cu (II) ions from diluted water solutions (5-20 mg/l). The sample from birch wood was less effective. The maximum sorption capacities (Xm) determined from the experimental equilibrium isotherms by applying the Langmuir model showed that the alga had the best copper-binding ability (Xm = 23.4 mg/g), followed by the moss (Xm = 11.1 mg/g), and the sawdust (Xm = 4.9 mg/g). The performance of the biosorbent prepared from birch was not satisfactory. The regeneration of the sorbents from alga and moss was performed using diluted HCl as eluent. No visible damages or performance losses were detected after five sorptiondesorption cycles.</p><p> Chapter 2 deals with MnOx-Pd/Alumina-La catalysts for abatement of the emissions from wood combustion. Of primary interest is the calcination temperature used in preparation of the catalysts. Several catalysts are prepared using various calcination temperatures, 500, 600, 700 and 800 oC for 4 h in air and their activities and stabilities are compared. The activity tests were performed using gaseous mixtures containing combustibles representative for the flue gases from wood combustion, carbon monoxide (2500 ppm), methane (200 ppm) and naphthalene (50 ppm). The catalytic oxidation tests were performed in presence of 10 % O2, 12 % H20, 12 % CO2 and N2 (balance). The concentrations of the components in the gaseous mixture and the total flow of the mixture correspond to those in the flue gases from combustion (gas flow 2.5 l/min corresponding to a space velocity of approximately 20000 h-1). In presence of the catalysts carbon monoxide (CO) and naphthalene (C10H8) ignite almost simultaneously in the interval 150-200 ºC and are totally converted at temperatures a little above 200 ºC. The light-off temperatures of methane (CH4) are in the interval 600-650 ºC, and total conversion is reached at around 700 ºC. The most suitable calcination temperature for the catalysts prepared here is 700 ºC. Lower temperatures, 500 and 600 ºC, seem to result in formation of less stable catalysts. The catalysts calcined at higher temperature, 800 ºC, have stable performance in repeated tests, but lower activity.</p><p> Chapter 3 presents results from literature study on corrosion and deposit formation in combustion of biofuels. Contributing to understanding the reasons for corrosion and the methods for its abatement are the primary goals. The scope is limited to deeper insight of the role of chlorine and alkali in combustion of biomass and the possibilities for hampering their corrosive effects. The role of additives decreasing the corrosion and deposit formation as well as the effect of water and the prospective for availability of low-chlorine biofuels have also been examined.</p>