Helices in proteins have substantial permanent dipole moments arising from the nearly perfect alignment of the individual dipole moments of each peptide bond. Interaction with this helix "macrodipole" is thought to perturb the pKa values of basic or acidic residues at the helix termini. The goal of this project is to investigate the effect of the helix confonnation on the proton affinities ofbasic amino acids placed at theN- or Ctenninus of helical model peptides in the gas phase. Several series of model peptides having a basic residue, lysine (K) or 2,3- diaminopropionic acid (Dap ), located at either terminus were synthesized by solid phase peptide synthesis using conventional techniques or the amino acid fluoride approach. Proton affinities were determined for several basic amino acids and peptides using mass spectrometry by applying the extended Cooks' kinetic method. Favorable conformations and theoretical proton affinities were probed using computational chemistry. The proton affinities determined for Na-acetyl-(L)-lysine, Ac-AK, Ac-KA, and Ac-KAA are 236.8 ± 1.9 kcal mol-1 , 249.4 ± 2.0 kcal mol-1 , 241.5 ± 1.9 kcal mol-1 , and 244.4 ± 2.0 kcal mol-1 respectively. The large negative entropy changes for each of the peptides upon protonation ( -11.2 to - 21.7 cal mol-1 K- 1 ) are consistent with globular confmmations adopted by the protonated peptides due to extensive intramolecular hydrogen bonding. The measured proton affinities of the peptides increased with the size of the peptide as expected. However, the measured proton affinity of the peptide with C-terminal lysine, Ac-AK, is substantially higher than that of the con·esponding peptide with N-terrninal lysine, Ac-KA, contrary to expectations. Proton affinities determined for these compounds using computational chemistry are in reasonable agreement with experimental results. Additionally, proton affinities calculated for helical polyalanine and Aib (aaminoisobutytic acid) modified polyalanine peptides with C-terminal basic residues (Ac AnK and Ac-(AibA)n-Dap) are much larger than proton affinities calculated for the corresponding peptides with N-terminal basic residues. These results indicate that the helix dipole has a substantial effect on the basicity of residues at the helix termini.

Potassium is an essential element, which is important in many plant processes. It functions as a major osmotic and is involved in the regulation of turgor during cell growth or stomatal movements. It is also important for maintaining membrane potencial. In plants, potassium transporters from the KT/HAK/KUP family are involved in the transport of K+ . Some of them are important in the uptake of K+ from the enviroment (HAK5, KUP7), others in regulation of cell turgor (KUP2, KUP6, KUP8). In Arabidopsis thaliana, less characterized KT/HAK/KUP transporters include KUP5 and KUP9, which I studied in this diploma thesis. In this diploma thesis, I analyzed the growth phenotype of kup5 mutant plants. The results show that kup5 mutant plants are not more sensitive to K+ deficiency than wild-type plants, therefore KUP5 is probably not involved in the K+ uptake from the enviroment. Kup5 mutant plants were larger than wild-type plants, had larger root and hypocotyl cells as well as longer root meristematic zone. This growth phenotype suggests that KUP5 is involved in the regulation of cell growth, probably through turgor regulation. Using the pKUP5::KUP5-GFP construct, the KUP5 protein was localized in the ER, but this localization needs further verification. Using the pKUP5:GUS construct, KUP5 expression was...

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第20281号 / 医博第4240号 / 新制||医||1021(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 髙折 晃史, 教授 岩田 想, 教授 萩原 正敏 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

京都大学 / 0048 / 新制・課程博士 / 博士(ｴﾈﾙｷﾞｰ科学) / 甲第20775号 / エネ博第360号 / 新制||エネ||71(附属図書館) / 京都大学大学院エネルギー科学研究科エネルギー基礎科学専攻 / (主査)教授 片平 正人, 教授 森井 孝, 教授 木下 正弘 / 学位規則第4条第1項該当 / Doctor of Energy Science / Kyoto University / DGAM

No description available.

Föreliggande uppsats syfte är att synliggöra teaterpedagogers syn på sin erfarenhet av lajv i relation till teaterundervisningen. Genom kvalitativa intervjuer har tre teaterpedagoger intervjuats som alla ser att deras teaterundervisning har påverkats av deras lajverfarenhet. Övergripande resultat i studien är att flera lajvpraktiker kunnat infogas i teaterundervisningen och understödja kvalitéer och processer kring kollaborativt skapande, rollarbete och reflektionsarbete. Fyra praktiker lyfts. Mini-lajvande är en praktik pedagogerna använder där eleverna lämnar teatermanuset och utforskar nya situationer med sina karaktärer. Den dramaturgiska principen ”play to lose” används för att eleverna ska skapa intressant dramaturgi och öppna situationen för andra elever att kunna delta. Ett slags transparensarbete beskrivs inspireras av lajvets form där eleverna får insyn i pedagogens planering och medkonstruerar sina kunskaper och lektioner. Pedagogerna arbetar även kring elevernas känslor för att omhänderta starka reaktioner i läromomenten och för att eleverna ska lära sig skilja mellan känsloreflektioner och gestaltningsreflektioner. Teaterklassrummet har även analyserats som en läromiljö med informella och formella kvalitéer, vilket blir en möjlig förklaring för varför lajvpraktikerna stundvis kan infogas sömlöst i teaterundervisningen, och varför stora hinder ibland uppstår. Resultaten sätts i relation till tidigare forskning i lajv- och teaterundervisningsfälten, och vidare diskuteras resultatet. För det första diskuteras huruvida lajvet introducerar helt nya praktiker, eller om praktikerna snarare sätter form för redan befintliga praktiker. För det andra diskuteras varför vissa väletablerade lajvpraktiker inte återfinns i informanternas beskrivningar. För det tredje lyfts huruvida teaterläromiljön ska anta flest formella eller informella kvalitéer, och om läromiljön har en unik möjlighet att vara dynamisk.

Antibodies, as forerunners in the field of biological drugs, are originally an organism’s answer to the invasion of different pathogens. Today, antibodies are a common treatment for many chronic diseases such as the immune-mediated inflammatory diseases rheumatoid arthritis or psoriasis. It is suspected that the cytokines interleukin 17a (IL17a) and interleukin 17c (IL17c) are involved in those diseases and are commonly treated with antibodies that inhibit the cytokines. Even though antibodies have been a huge success as biological drugs they also have downsides when it comes to their production, size and stability. In quest of finding alternatives to antibodies in diagnostics and therapy, a novel class of biologics has been developed. So-called alternative scaffold proteins are small polypeptide chains that can be engineered to show affinity towards different biomarkers. ABD-Derived Affinity ProTeins or ADAPTs are one example of these alternative scaffolds that can be modified to bind a biomarker as target and keep their affinity to Human Serum Albumin (HSA) at the same time, making them bispecific. In this project, twenty-four previously selected ADAPT binder candidates that have shown good prospects towards IL17a and IL17c in previous experiments were cloned, produced, purified and characterized to determine if they show potential as tools in diagnostics or therapy of autoimmune diseases. The proteins were produced in E. coli, purified by affinity chromatography and characterized using Surface Plasmon Resonance (SPR), Circular Dichroism (CD) and Size Exclusion Chromatography (SEC). All candidates were successfully cloned into E. coli and out of these, 10 could be produced and 5 showed affinity towards their target using SPR. Examination by SEC and CD showed that the protein variants did not seem to be structurally stable and hints of impurities in the samples could be detected. This and a low yield could be further confirmed via SDS-PAGE. In conclusion, binders were produced that could theoretically be promising candidates as tools in diagnostics or therapy of chronic diseases were IL17a and/or IL17c are important. Nevertheless, in order to support these claims further investigations and developments are necessary. / Antikroppar, som föregångare inom området biologiska läkemedel, är ursprungligen en organisms svar på invasionen av olika patogen. Idag är antikroppar en vanlig behandling för många kroniska sjukdomar, såsom de immunmedierade inflammatoriska sjukdomarna reumatoid artrit eller psoriasis. Cytokinerna interleukin 17a (IL17a) och interleukin 17c (IL17c) tros vara involverade i dessa sjukdomar och behandlas vanligtvis med antikroppar som hämmar cytokinerna. Trots att antikroppar har varit en stor framgång som biologiska läkemedel har de också nackdelar när det gäller deras produktion, storlek och stabilitet. För att hitta alternativ till antikroppar inom diagnostik och terapi har en ny klass av biologiska läkemedel utvecklats. Så kallade alternative scaffold proteins är små polypeptidkedjor som kan manipuleras för att visa affinitet gentemot olika biomarkörer. ABD-Derived Affinity ProTeins eller ADAPTs är ett exempel på dessa alternative scaffolds som kan modifieras för att binda en biomarkör som mål utan att påverka affiniteten till Humant Serum Albumin (HSA), vilket gör dem bispecifika. I detta projekt klonades, producerades, renades och karakteriserades tjugofyra tidigare utvalda ADAPT-bindarkandidater som har visat goda förutsättningar gentemot IL17a och IL17c i tidigare experiment. Proteinerna producerades i E. coli, renades genom affinitetskromatografi och karakteriserades med användning av Surface Plasmon Resonance (SPR), Circular Dichroism (CD) och Size Exclusion Chromatography (SEC). Alla kandidater klonades framgångsrikt i E. coli och av dessa kunde 10 produceras. Fem bindare visade affinitet till deras mål med SPR. Undersökning med SEC och CD visade dock att proteinvarianterna inte var strukturellt stabila och antydan till föroreningar kunde detekteras i proverna. Detta och ett lågt utbyte kunde ytterligare bekräftas via SDS-PAGE. Sammanfattningsvis kunde bindare producerades och dessa kan teoretiskt vara lovande kandidater till diagnostik eller terapi av kroniska sjukdomar där IL17a och/eller IL17c är viktiga. För att stödja dessa påståenden krävs dock ytterligare experiment och utveckling av bindarna.

京都大学 / 新制・課程博士 / 博士(工学) / 甲第23513号 / 工博第4925号 / 新制||工||1769(附属図書館) / 京都大学大学院工学研究科材料化学専攻 / (主査)教授 大塚 浩二, 教授 松原 誠二郎, 教授 秋吉 一成 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DFAM

No description available.

Frontal sinus variation has been used in forensic anthropology to aid in positive identification since the 1920s. As radiographic technology has evolved, so has the quality and quantity of data that radiologists and anthropologists can collect during an individual’s lifetime. This has led to new methodology when comparing antemortem and postmortem radiographic images. The current study aims to look at frontal sinus morphology and dimensional variations on computed tomography (CT) scans, as these are currently the most commonly collected images showing the frontal sinus in its entirety used in clinics in the United States. This study assessed 307 individuals for the morphological analysis and 325 individuals in the dimensional analysis. These individuals represented females and males from Asian, African, European, and Latin American derived groups. It is hypothesized that frontal sinus shape variations will cluster based on assigned sex and population affinities. Similarly, it is hypothesized that dimensional variation, specifically the maximum height, maximum width, and maximum depth, will show statistically significant clustering based on assigned sex and population affinities. The frontal sinus outlines from the CT images were transferred into SHAPE v1.3 in order to run an elliptical Fourier analysis. The dimensional data was measured directly from the images using a MicroDicom viewer. All of the statistical analyses, including Pearson’s Chi-squared and ANOVA tests, were run in R studio. Results indicated that morphologically there is no statistically significant clustering based on assigned sex or population affinity. However, there was statistically significant clustering dimensionally when tested against both assigned sex and population affinity using an ANOVA, indicating that the interactive effects of sexual dimorphism and population affinity influence the dimensions but not the shape of the frontal sinus. These results add to the foundational knowledge that practitioners have surrounding frontal sinus indicating that assigned sex and population affinity have impact on the approximate dimensions of this structure while these variables do not have a statistically significant effect on morphological variation. The results also speak to the idiosyncratic nature of the frontal sinus and bolster confidence using morphological variations as a mean to individuate.