In this thesis, the main concern has been to improve the reliability of different parts of the analytical workflow (Paper I, II, IV &V). Additionally, one of the resulting optimized methods was used in a real application (Paper III). Paper I-II concerned the extraction of acrylamide (AA) from foods. In Paper I different parameters such as sample particle size, extraction solvent, extraction time and extraction temperature were optimized, leading to a method that showed good agreement with the assigned AA levels of several proficiency test samples. Later, after the publication of the paper, the method showed good performance in a collaborative trial validation, in terms of trueness, repeatabil­ity and reproducibility figures. It was labeled “undoubtedly fit for the purpose”. In Paper II, it was shown that the ‘extra’ amounts of AA obtained during extraction of foods with an alkaline aqueous solution was not due to improved extractability of AA from the food matrix. Strongly alkaline conditions seemed to rather induce net formation of AA from water-soluble precursors formed during thermolysis. This phenomenon should therefore be regarded as an extraction artifact. Paper III was an application of the optimized method from Paper I, where it was used to study the reduction of AA in potato chips (crisps) by using pre-treatments and frying at reduced pressure. There were significant reductions in AA, down to below the limit of quantification (5 µg/kg) for the method. Paper IV-V concerned analysis of anthocyanins (AC) in red onion. In Paper IV, a new separation method using capillary electrophoresis was developed, and its rapidness combined with an acidic background electrolyte helped in preventing AC degradation. Furthermore, its alternative separation mechanism is a complement to that of the more commonly used liquid chromatography technique. In Paper V, simultaneous extraction and degradation of anthocyanins from red onion was studied in a static batch reactor at 110ºC. The extraction and degradation kinetics were successfully separated, and an ideal theoretical extraction curve was constructed by compensating mathematically for degradation effects, showing that more anthocyanins, 21 to 36% depending on different species, could be extracted if no degradation occurred. The results give important information about the different kinetics competing during an extraction procedure, and also show that quantitative extraction is not to recommend in the batch system used in the study.

Biologically inspired materials are being developed with the aim of improving the integration of medical implants and minimizing non-desirable host reactions. A promising strategy is the design of topographically patterned surfaces that resemble those found in the extracellular environment. Nanoporous alumina has been recognized as a potential biomaterial and as an important template for the fabrication of nanostructures. In this thesis in vitro studies were done to elucidate the role of alumina nanoporosity on the inflammatory response. Specifically, by comparing alumina membranes with two pore sizes (20 and 200 nm in diameter). Complement and platelet activation were evaluated as well as monocyte/macrophage behaviour. Whole blood was incubated with the alumina membranes and thereafter the biomaterial surfaces were evaluated in terms of protein and platelet adhesion as well as procoagulant properties. The fluid phase was analyzed for complement activation products and platelet activation markers. Besides, human mononuclear cells were cultured on the alumina membranes and cell adhesion, viability, morphology and release of pro-inflammatory cytokines were evaluated. The results indicated that nanoporous alumina with 200 nm pores promotes higher complement activation than alumina with 20 nm pores. In addition, platelet response to nanoporous alumina was found to be highly dependent on the material porosity, as reflected by differences in adhesion, PMP generation and procoagulant characteristics. A clear difference in monocyte/macrophage adhesion and activation was found between the two pore size alumina membranes. Few but highly activated cells adhered to the 200 nm membrane in contrast to many but less activated monocytes/macrophages on the 20 nm surface. The outcome of this work emphasizes that nanotopography plays an important role in the host response to biomaterials. Better understanding of molecular interactions on nano-level will undoubtedly play a significant role in biomaterial implant development and will contribute to design strategies for controlling specific biological events.

Inter-molecular interactions have significant influences on linear and nonlinear optical properties of molecules including one- and two-photon absorptions, emissions, and various high order nonlinear polarizations. The related investigation has become an active and challenging research area. The theoretical structure-to-property relationship obtained from quantum chemical calculations of single organic conjugated molecules often can not be directly applied to real materials in condensed phases. One has to consider the effect of environment, that is, inter-molecular interactions, where the model systems experience in real experiments or applications. The change of molecular conformations under all kinds of interactions and its effects on linear and nonlinear optical properties are the central issue of this thesis. Special attentions have been paid to symmetrical diamino substituted distyrylbenzene chromophores with different torsional angles, two dipolar merocyanine dyes of various orientations, two isomers of a V-shaped 2-hydroxypyrimidine derivative and their various dimers, and the structural fluctuations of interacting polar chromophores in solutions. Quantum chemical methods in combination with molecular dynamics simulations have been employed to study molecular conformations and optical properties in solutions, in particular the solvent and aggregation effects on one- and two-photon absorption. More specifically, time-dependent density functional theory has been used for all electronic calculations, while the polarizable continuum model and supermolecule approach have also been employed to take into account solvent effects. Moreover, the propagation of an ultrashort laser pulse through a one-dimensional asymmetric organic molecular medium which possesses large permanent dipole moments has been simulated by solving full Maxwell-Bloch equations using predictor-corrector finite-difference time-domain method. We have focused on the supercontinuum generation of spectra and the formation of attosecond pulses. / QC20100630

Two small bordering catchments in Bispgården, Central Sweden, wereinvestigated in regard to soil solution and stream water chemistry during the frostfree seasons of 2003-2007. Both catchments were drained by first order streams,Fanbergsbäcken and Gråbergsbäcken, and in Fanbergsbäckens catchment anextensive investigation of the soil and soil solution chemistry was conducted bylysimeter and centrifugation sampling. The area of intensive soil solutioninvestigation was situated in a slope towards a stream incorporating a rechargearea, with podzolic soil, and a discharge area close to the stream with an arenosolsoil. Samples were continuously taken in both the recharge- and the discharge areaof the slope, and stream water was sampled in the streams of both catchments. Themain variables of interest of the study were the interactions, the influence ofdissolved organic carbon and the effects of wood ash application to soil solutionand stream water.The natural variations and the interactions between soil solution and streamwater were monitored during 2003-2004. In soil solution, most of the investigatedsubstances tended to increase during the growing season, due to weathering andmicrobial degradation of biota. Ca, Mg, Al and Fe were highly associated todissolved organic carbon (DOC) throughout the catchment. The low molecularfraction of DOC seemed to have a higher impact on the soil processes in therecharge area, while high molecular DOC was more important for transport ofcations in the discharge area and the stream water.The concentration of different substances in the two streams differedsignificantly, even though the catchments were similar in size, shape andforestation. The seasonal patterns of most of the substances measured weresignificantly correlated between the streams, however. Cations and pH correlatedwell with DOC and flow. The flow pattern driven by precipitation seems to be thedriver of the stream water chemistry.Wood ash was applied at a dosage of 3 ton/ha to one of the catchments in theautumn of 2004, to investigate the initial effects on the soil solution- and streamwater chemistry. WAA is recommended by the Swedish Forest Agency tocounteract acidification in soil and runoff that may be caused by an intensivebiomass harvesting. The impact of the WAA was studied during 2005-2006.Compared to the control temporarily higher concentrations of K, Ca and SO4 wereobserved in the soil solution of the ashed area. In the stream water the effects of theWAA were easier to distinguish due to higher sampling frequency. The strongesteffect was seen for K, but increases in the stream water were also noted for DOC,Ca, Mg, Si, Cl and malonate. No increase in pH could be statistically verifiedhowever, and overall the initial effects of the WAA seem mild. / Två angränsande avrinningsområden i Bispgården i centrala Sverigeundersöktes under den snöfria säsongen 2003-2006. Båda avrinningsområdenadränerades av första ordningens bäckar och i det ena, Fanbergsbäckensavrinningsområde, gjordes en omfattande undersökning av mark och markvattenkemin.Markvatten provtogs genom centrifugering och med lysimetrar.Provtagningen gjordes i inströmningsområdet där jordmånen var en typiskpodzol samt närmare bäcken, i utströmningsområdet, där jordmånen var enarenosol. Bäckvatten provtogs i båda avrinningsområdena. Interaktioner mellanmark, markvatten och bäckvatten undersöktes med särskilt fokus på inverkan avorganiskt kol samt påverkan av askåterföring.De naturliga variationerna i markkemin samt interaktionerna mellan mark- ochbäck vatten undersöktes 2003-2004. I markvatten ökade merparten av de studeradeämnena under provtagningssäsongen beroende av ökad vittring och recirkulationav biota och fallförna. Ca, Al och Fe var i stor utsträckning associerade till löstorganiskt kol (DOC). Den lågmolekylära fraktionen av DOC hade en störreinverkan på markprocesserna i inströmningsområdet, medan den högmolekyläradelen var viktigare för transport av katjoner i utströmningsområdet samt ibäckvattnet.Koncentrationsnivåerna av olika substanser i bäckarna uppvisade skillnadertrots avrinningsområdenas yttre likheter med avseende på storlek, form ochbeskogning. Trots de kvantitativa skillnaderna erhölls emellertid liknandesäsongsvariationer i koncentrationerna, vilket indikerade att kemin i huvudsakstyrdes av mängden DOC som i sin tur berodde av avrinningen som drevs avnederbörden. Höga halter av Ca, Mg, Al och Fe återfanns associerade tillhögmolekylärt DOC i bäckvattnet i en utsträckning som inte rapporterats tidigare.På hösten 2004 spreds 3 ton aska/ha till Fanbergsbäckens avrinningsområde föratt undersöka de initiala effekterna på mark- och bäckvatten kemin. Askåterföringbör ske minst en gång per omloppstid vid helträdsavverkning, i enlighet medSkogsstyrelsens rekommendationer, främst för att motverka försurning i mark ochavrinnande vatten som antas uppstå vid intensivt uttag av biomassa. De initialaeffekterna av askåterföringen på mark- och bäckvattnets kemi studerades under2005-2006 och Gråbergsbäcken kunde användas som obehandlad kontroll vidundersökning av vattenkemin i Fanbergsbäcken. I undersökningen av markvattenviåterfanns stora säsongsvariationer, vilket gjorde det svårt att urskilja eventuellaeffekter av askåterföringen. Tillfälligt högre värden av K, Ca och SO4 återfannsdock i det askade området i jämförelse med kontrollområdet. På grund av högreprovtagningsfrekvens i bäckvattnet var det lättare att påvisa förändringar ibäckvatten kemin. Framförallt märktes en signifikant ökning av K jämfört medkontrollbäcken. Sådana ökningar, om än inte lika markanta återfanns också förDOC, Ca, Mg, Si, Cl och malonat. En tendens till ökat pH kunde observeras, mendenna kunde inte verifieras statistiskt. De initiala effekterna av askåterföringen varsåledes till synes milda, och den eftertraktade pH-effekten erhölls ej i dennaundersökning.

Background: Most errors in the venous blood testing process are preanalytical, i.e. they occur before the sample reaches the laboratory. Unlike the laboratory analysis, the preanalytical phase involves several error-prone manual tasks not easily avoided with technological solutions. Despite the importance of the preanalytical phase for a correct test result, little is known about how blood samples are collected in hospitals. Aim: The aim of this thesis was to survey preanalytical procedures in hospitals to identify sources of error. Methods: The first part of this thesis was a questionnaire survey. After a pilot study (Paper I), a questionnaire addressing clinical chemistry testing was completed by venous blood sampling staff (n=314, response rate 94%) in hospital wards and hospital laboratories (Papers II–IV). The second part of this thesis was an experimental study. Haematology, coagulation, platelet function and global coagulation parameters were compared between pneumatic tube-transported samples and samples that had not been transported (Paper V). Results: The results of the questionnaire survey indicate that the desirable procedure for the collection and handling of venous blood samples were not always followed in the wards (Papers II–III). For example, as few as 2.4% of the ward staff reported to always label the test tube immediately before sample collection. Only 22% of the ward staff reported to always use wristbands for patient identification, while 18% reported to always use online laboratory manuals, the only source of updated information. However, a substantial part of the ward staff showed considerable interest in re-education (45%) and willingness to improve routines (44%) for venous blood sampling. Compared to the ward staff, the laboratory staff reported significantly higher proportions of desirable practices regarding test request management, test tube labelling, test information search procedures, and the collection and handling of venous blood samples, but not regarding patient identification. Of the ward staff, only 5.5% had ever filed an error report regarding venous blood sampling, compared to 28% of the laboratory staff (Paper IV). In the experimental study (Paper V), no significant preanalytical effect of pneumatic tube transport was found for most haematology, coagulation and platelet function parameters. However, time-to-clot formation was significantly shorter (16%) in the pneumatic tube-transported samples, indicating an in vitro activation of global coagulation. Conclusions. The questionnaire study of the rated experiences of venous blood sampling ward staff is the first of its kind to survey manual tasks in the preanalytical phase. The results suggest a clinically important risk of preanalytical errors in the surveyed wards. Computerised test request management will eliminate some, but not all, of the identified risks. The better performance reported by the laboratory staff may reflect successful quality improvement initiatives in the laboratories. The current error reporting system needs to be functionally implemented. The experimental study indicates that pneumatic tube transport does not introduce preanalytical errors for regular tests, but manual transport is recommended for analysis with thromboelastographic technique. This thesis underscores the importance of quality improvement in the preanalytical phase of venous blood testing in hospitals.

Traditionally, rolling mills use emulsions based on a mixture of oil and water for lubrication. Since two years ago SAPA has been using (instead of oil) a synthetic lubricant so called conditional emulsion for hot-rolling of aluminum. This lubricant is water based and homogenous at ambient temperature, but switches to a two-phase system at heating above the cloud point. This project aims to validate and if necessary modify an existing HPLC method for quantifying two out of three key components (A, B and C) in the conditional emulsion. Attempts to develop a method to quantify the pH adjusting components, X and Y were also made. These two methods are required to optimize the lubricant. Due to the complexity of the components, it has been difficult to present a method for quantification, and HPLC with ELS detection was chosen after a long series of trials. Due to a few uncontrollable parameters the proposed analysis method has tendencies to be unstable. The column used is sensitive to changes in equilibrium and ELSD is also less sensitive and less reproducible than the commonly used UV-detector. While the proposed assay method shows somewhat large relative standard deviations the method has been shown to produce sufficiently precise and accurate data for the intended purpose. Development of a method for the pH-adjusting components X and Y was more difficult than expected. For some reason their difference in chemical properties does not show satisfying impact in the chromatograms. This method is still in its cradle and needs further development.

Övergången från grundskolan till gymnasieskolan är en stor händelse för många elever. Detställs högre och fler krav på eleverna när de börjar gymnasiet och deras förkunskaper harbetydelse för hur väl de klarar av utbildningen. I denna undersökning fick elever i årskurs ettpå det naturvetenskapliga programmet svara på frågor om de anser att de hade tillräckligakunskaper i de naturvetenskapliga ämnena från högstadiet. Undersökningen visar att de flestatycker att de hade tillräckliga kunskaper i biologi, kemi och matematik men inte i fysik.Kursplanen för fysik på gymnasiet skiljer sig från de andra kursplanerna, genom att det intestår lika tydligt att undervisningen ska bygga på elevernas kunskaper i grundskolan.Respondenterna i undersökningen fick även svara på frågor om vilket ämne som var svårt,roligt och om de läste i NO-block eller i separata ämnen på högstadiet. Undersökningen visaratt det finns en skillnad mellan flickor och pojkar när det gäller deras attityder och intresse.

Immobilization of different bio- and organic molecules on solid supports is fundamental within many areas of science. Sometimes, it is desirable to obtain a directed orientation of the molecule in the immobilized state. In this thesis, the copper (I) catalyzed Huisgen 1,3-dipolar cycloaddition, referred to as a “click chemistry” reaction, was explored as a means to perform directed immobilization of small molecule ligands on gold surfaces. The aim was to synthesize alkyne- and azide-terminated alkanethiols that would form well-organized self assembled monolayers (SAMs) on gold from the commercially available substances orthoethylene glycol and bromo alkanoic acid. N-(23-azido-3,6,9,12,15,18,21-heptaoxatricosyl)-n-mercaptododekanamide/hexadecaneamide (n = 12, 16) were successfully synthesized and allowed to form SAMs of different compositions to study how the differences in density of the functional groups on the surface would influence the structure of the monolayer and the click chemistry reaction. The surfaces were characterized by different optical methods: ellipsometry, contact angle goniometry and infrared reflection-absorption spectroscopy (IRAS). The click reaction was found to proceed at very high yields on all investigated surfaces. Finally, the biomolecular interaction between a ligand immobilized by click chemistry on the gold surfaces and a model protein (bovine carbonic anhydrase) was demonstrated by surface plasmon resonance using a Biacore system.

The insulin-like 3 peptide is autocrine/paracrine insulin-related hormone with a size of approximately 6kDa [1]. It mediates through a leucine richG-coupled receptor named LGR8. INSL3 is mainly expressed in human Leydig cells and is directly responsible for migration of the testis during the pre-natal period in maledevelopment. [2] INSL3 mRNA has recently been verified in human platelets whereas no mRNA has been detected for LGR8 (by Sanofi-Aventis GmbH in Frankfurt,Germany), indicating that INSL3 might be released through paracrine functions at sites of platelet adhesion and aggregation upon a vascular injury.Furthermore, has activated platelets been shown to translate essential proteins upon activation, in a term called “signal-dependent protein synthesis”.The B-Cell lymphoma-3 protein (BCL-3) is an example of such a protein [3], and there is a possibility that INSL3 might be also. In this thesis we wanted to detect the relaxin- like peptide 3 hormone (INSL3). (Its function, location and the timeframe of its release, when/if it issecreated in stimulated platelets).The source of platelet-derived INSL3 can be found with Western blotting and Enzyme immunoassay. Detection of the insulin-like 3 peptide in human platelets turned out to be a difficult challenge due to the small amount of INSL3 secretion uponplatelet activation; hence the total amount of INSL3 produced might be below detection limit.

Flowcytometric technology has been widely used for measurement of the absolute numbers of T-lymphocytes subsets in Human Immunodeficiency virus (HIV), defining the disease state, monitoring antiviral treatment, and identifying any risk for opportunistic infections. A manual preparing of the samples is required. More recently an automated and enclosed blood cell counting, Celldyn Sapphire has been introduced. In this study the performance of the Flow cytometer EPICS XL as a reference method for analysis of CD3+, CD4+ and CD8 T-lymphocytes was evaluated with blood from 40 individual’s samples. EPICS XL was also compared with Celldyn Sapphire in the analysis of T-lymphocyte subsets in 39 blood samples from patients with low, high and normal lymphocyte counts. The result showed that the precision was high for both EPICS XL (2.5%) and Celldyn (10%). The method was linear over a wide range. Comparisons of CD3+, CD4+, and CD8+ T-lymphocytes analysis showed high coefficients of correlation (r0.9) and agreement (y>0.9x) between two instruments. A lower degree of agreement was observed at low concentration of CD3+ and CD4+ T-lymphocytes (0.757, 0.739). This means that cell counts obtained by Celldyn were 30% lower than those obtained with EPICS XL. This study shows that both EPICS XL and Celldyn Sapphire were suitable for CD4+ and CD8+ T cell counts. It is however preferable to use Flowcytometry for counting of low concentration of CD4+ T-lymphocytes (<200 cells/µL).