京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第22504号 / 農博第2408号 / 新制||農||1077(附属図書館) / 学位論文||R2||N5284(農学部図書室) / 京都大学大学院農学研究科農学専攻 / (主査)教授 白岩 立彦, 教授 丸山 伸之, 准教授 田中 朋之 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DGAM

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Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

In dieser Arbeit wird die Erzeugung der Charmonium-Zustände J/psi und psi(2S) in Proton-Kern-Reaktionen bei Schwerpunktsenergien von 42 GeV untersucht. Die Daten wurden mit dem HERA-B-Experiment aufgenommen, das in HERA den Protonstrahl nutzt, um Protonen an den Kernen verschiedener Drahttargets zu streuen. Mehr als 150 000 J/psi und 2400 psi(2S)-Zerfälle in Myon-Paare wurden beobachtet. Die HERA-B-Daten decken den kinematischen Bereich in Feynman-x (x_F) zwischen -0.36 und 0.10 und im Transversalimpuls (p_T) bis 4.5 GeV/c ab. Die x_F - und p_T-Verteilungen für die Produktion von J/psi und psi(2S) wurden für Kohlenstoff- und Wolfram-Targets gemessen. Die x_F-Verteilung stimmt gut mit nicht-relativistischen QCD-Rechnungen überein. Bei der p_T-Verteilung findet man einen klaren Anstieg des mittleren p_T für schwerere Kerne: Kohlenstoff = 1.244 +- 0.003 +- 0.034 GeV/c und Wolfram = 1.336 +- 0.004 +- 0.041 GeV/c. Der Vergleich der Erzeugungsraten von psi(2S) und J/psi ergibt: (B''_{muon muon} sigma(psi(2S)) ) / (B_{muon muon} sigma(J/psi)) = 165 +- 9 +- 4 ( x 0.0001) für Kohlenstoff und 154 +- 16 +- 8 ( x 0.0001) für Wolfram. Diese Verältnisse wurden auch für die x_F - und p_T-Verteilungen bestimmt. / In this thesis the production of the charmonium states J/psi and psi(2S) in proton-nucleus collisions at a center-of-mass energy of 42 GeV is studied. The data have been taken by the HERA-B experiment which uses the HERA proton beam to scatter protons off the nuclei of different wire targets. Over 150 x 10^3 J/psi and 2400 psi(2S) decaying to muon pairs are observed. The HERA-B data cover the kinematical range in Feynman x (x_F) between -0.36 and 0.10 and in transverse momentum (p_T) up to 4.5 GeV/c. The x_F and p_T differential distributions for the production of J/psi and psi(2S) mesons are measured for carbon and tungsten target materials. The J/psi x_F distribution agrees well with non-relativistic QCD calculations. From the p_T distributions we find a clear increase of the average p_T for heavier nuclei: Carbon = 1.244 +- 0.003 +- 0.034 GeV/c and Tungsten = 1.336 +- 0.004 +- 0.041 GeV/c. We compared the production rates of psi(2S) to J/psi mesons: (B''_{muon muon} sigma(psi(2S)) ) / (B_{muon muon} sigma(J/psi)) = 165 +- 9 +- 4 ( x 0.0001) for carbon and 154 +- 16 +- 8 ( x 0.0001) for tungsten. These ratios have also been determined for the x_F and p_T distributions.

Les allergies alimentaires à l'arachide et au lait posent un problème majeur de santé publique, en particulier chez les enfants. Nous avons développé un modèle cellulaire de dégranulation, afin d'analyser si la liaison d'un allergène à ses IgE spécifiques, mesurée in vitro, a bien la capacité de déclencher la phase effectrice de la réaction allergique. Ce modèle repose sur l'utilisation de la lignée cellulaire RBL SX-38 développée par JP Kinet, mastocytes de rat immortalisés et modifiés pour exprimer le récepteur de haute affinité aux IgE humaines. Le développement de ce modèle pour son utilisation en microplaques de 96 puits a consisté notamment en l'optimisation de différents paramètres critiques pour la réalisation des 2 phases du test : la phase de sensibilisation des cellules par les IgE de patients allergiques et la phase de déclenchement par les allergènes auxquels le patient est sensibilisé. Ce modèle a permis de montrer l'importance des caséines et des albumines 2S (Ara h 2 et Ara h 6) dans cette étape clé de la réaction allergique au lait et à l'arachide, confirmant les observations sérologiques et cliniques disponibles. L'effet de traitements thermiques sur l'allergénicité d'un allergène majeur de l'arachide (Ara h 1), a été évalué dans le cadre du programme européen EuroPrevall. Les résultats obtenus montrent la diminution de la réactivité d'Ara h 1 après chauffage de la protéine isolée en solution. Par contre, la même protéine préparée et purifiée à partir d'arachide grillée présente une forte réactivité. Ces résultats suggèrent qu'un traitement à température élevée de la graine entraine une augmentation de l'allergénicité de Ara h 1 du fait de ses interactions avec les autres constituants de la graine. Le modèle cellulaire de dégranulation développé apparaît être un outil pertinent pour l'étude de la fonctionnalité biologique de l'interaction allergène-IgE, permettant une meilleure compréhension de la relation entre la structure d'une protéine alimentaire et son allergénicité, voire une évaluation du risque allergique des aliments ne nécessitant pas d'essais cliniques systématiques.

This thesis presents the results of a comprehensive multi-wavelength study of the high mass X-ray binary 2S 0114+650. This enigmatic source has previously been proposed to be the first in a new class of super-slow X-ray pulsars, containing a neutron star revolving once every 2.7 h. The 11.6 d orbital period of this system has been well established in both X-ray and optical wavelengths. During the initial stages of the research presented in this thesis we discovered an additional 30.7 d ???super-orbital??? modulation in the X-ray emission from this source. While similar periodicities seen in other X-ray binaries are commonly attributed to the precession of a warped accretion disc, the observational evidence suggests the absence of such a disc in 2S 0114+650. The purpose of this project is thus to determine the nature of the super-orbital modulation and to better constrain the astrophysical parameters of this system. To investigate the long-term variability we analysed ~8.5 yr of archived data from the Rossi X-ray Timing Explorer space telescope. The problem of the spurious ~24 h periods in this data was solved as a by-product of these studies. Follow-up pointed observations were obtained with this satellite in order to examine the spectral and temporal behaviour over the spin, orbital and super-orbital timescales. Independent confirmation of the super-orbital modulation was performed using ~2 yr of data from the INTEGRAL satellite obtained during a long-term monitoring campaign of the Cassiopeia region. The evolution of the spin, orbital and super-orbital periods over ~10 yr was examined using archived data from the Rossi X-ray Timing Explorer satellite. Radio observations were performed with the Giant Meterwave Radio Telescope to search for any radio emission associated with this source and to determine whether it is variable over the known periodicities. Near infrared observations were performed with the Mt Abu telescope to determine whether a Be star nature can be ruled out for the optical component. Finally, a statistical analysis of the properties of the confirmed super-orbital X-ray binaries was performed in order to search for commonalities between these systems.

In the first part of the thesis, protein-ligand interactions were investigated using the chaperone LcrH, from Yersinia as target protein. The structure of a peptide encompassing the amphipathic domain (residue 278-300) of the protein YopD from Yersinia was determined by NMR in 40% TFE. The structure of YopD278-300 is a well defined α-helix with a β-turn at the C-terminus of the helix capping the structure. This turn is crucial for the structure as peptides lacking the residues involved in the turn are unstructured. NMR relaxation indicates that the peptide is not monomeric. This is supported by intermolecular NOEs found from residue Phe280 to Ile288 and Val292 indicative of a multimeric structure with the helical structures oriented in an antiparallel manner with hydrophobic residues forming the oligomer. The interaction with the chaperone LcrH was confirmed by 1H relaxation experiments and induced chemical shift changes in the peptide Protein-ligand interactions were investigated further in the second paper using a different approach. A wide range of substances were used in screening for affinity against the chaperones PapD and FimC from uropathogenic Escherichia coli using 1H relaxation NMR experiments, surface plasmon resonance and 19F NMR. Fluorine NMR proved to be advantageous as compared to proton NMR as it is straight forward to identify binding ligands due to the well resolved 19F NMR spectra. Several compounds were found to interact with PapD and FimC through induced line-broadening and chemical shift changes for the ligands. Data corroborate well with surface plasmon resonance and proton NMR experiments. However, our results indicate the substances used in this study to have poor specificity for PapD and FimC as the induced chemical shift is minor and hardly no competitive binding is observed. Paper III and IV is an investigation of the structural features of the allergenic 2S albumin Ber e 1 from Brazil nut. Ber e 1 is a 2S albumin previously identified as the major allergen of Brazil nut. Recent studies have demonstrated that endogenous Brazil nut lipids are required for an immune response to occur in vivo. The structure was obtained from 3D heteronuclear NMR experiments followed by simulated annealing using the software ARIA. Interestingly, the common fold of the 2S albumin family, described as a right-handed super helix with the core composed of a helix bundle, is not found in Ber e 1. Instead the C-terminal region is participating in the formation of the core between helix 3, 4 and 5. The dynamic properties of Ber e 1 were investigated using 15N relaxation experiments and data was analyzed using the model-free approach. The analysis showed that a few residues in the loop between helix 2 and 3 experience decreased mobility, compared to the rest of the loop. This is consistent with NOE data as long range NOEs were found from the loop to the core region of the protein. The anchoring of this loop is a unique feature of Ber e 1, as it is not found in any other structures of 2S albumins. Chemical shift mapping of Ber e 1 upon the addition of lipid extract from Brazil nut identified 4 regions in the protein where chemical shift perturbations were detected. Interestingly, all four structural clusters align along a cleft in the structure formed by helix 1-3 on one side and helix 4-5 on the other. This cleft is big enough to encompass a lipid molecule. It is therefore tempting to speculate whether this cleft is the lipid binding epitope in Ber e 1.

This thesis presents the results of a comprehensive multi-wavelength study of the high mass X-ray binary 2S 0114+650. This enigmatic source has previously been proposed to be the first in a new class of super-slow X-ray pulsars, containing a neutron star revolving once every 2.7 h. The 11.6 d orbital period of this system has been well established in both X-ray and optical wavelengths. During the initial stages of the research presented in this thesis we discovered an additional 30.7 d ???super-orbital??? modulation in the X-ray emission from this source. While similar periodicities seen in other X-ray binaries are commonly attributed to the precession of a warped accretion disc, the observational evidence suggests the absence of such a disc in 2S 0114+650. The purpose of this project is thus to determine the nature of the super-orbital modulation and to better constrain the astrophysical parameters of this system. To investigate the long-term variability we analysed ~8.5 yr of archived data from the Rossi X-ray Timing Explorer space telescope. The problem of the spurious ~24 h periods in this data was solved as a by-product of these studies. Follow-up pointed observations were obtained with this satellite in order to examine the spectral and temporal behaviour over the spin, orbital and super-orbital timescales. Independent confirmation of the super-orbital modulation was performed using ~2 yr of data from the INTEGRAL satellite obtained during a long-term monitoring campaign of the Cassiopeia region. The evolution of the spin, orbital and super-orbital periods over ~10 yr was examined using archived data from the Rossi X-ray Timing Explorer satellite. Radio observations were performed with the Giant Meterwave Radio Telescope to search for any radio emission associated with this source and to determine whether it is variable over the known periodicities. Near infrared observations were performed with the Mt Abu telescope to determine whether a Be star nature can be ruled out for the optical component. Finally, a statistical analysis of the properties of the confirmed super-orbital X-ray binaries was performed in order to search for commonalities between these systems.

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Yes / The single enantiomer (1S,2S)-2-benzyl-2,3-dihydro-2-(1H-inden-2-yl)-1H-inden-1-ol (2), has recently been synthesized and isolated from its corresponding diastereoisomer (1). The molecular and crystal structures of this novel compound have been fully analyzed. The relative and absolute configurations have been determined by using a combination of analytical tools including X-ray crystallography, X-ray Powder Diffraction (XRPD) analysis and Nuclear Magnetic Resonance (NMR) spectroscopy. / Wellcome Trust