The first three, pure, single-isomer, 6-O-sulfo a-cyclodextrins, the sodium salts of hexakis(6-O-sulfo)-a-CD (HxS), hexakis(2,3-di-O-methyl-6-O-sulfo)-a-cyclodextrin (HxDMS) and hexakis(2,3-di-O-acetyl-6-O-sulfo)-a-cyclodextrin (HxDAS) have been synthesized, analytically characterized and utilized as chiral resolving agents in capillary electrophoresis. The purity of each synthetic intermediate and of the final product was determined by HPLC-ELSD and indirect UV-detection capillary electrophoresis. The structural identity of each intermediate and final product was verified by 1D and 2D NMR, and mass spectrometry.HxS, HxDMS and HxDAS have been used to separate a series of neutral, basic, ampholytic and acidic enantiomers in pH 2.5 and pH 9.5 aqueous and acidic methanol background electrolytes using capillary electrophoresis. Rapid separations with satisfactory peak resolution values were obtained for most of the analytes, indicating that HxS, HxDAS and HxDMS can serve as chiral resolving agent for a wide range of analytes. The observed separation patterns follow the predictions of the CHArged Resolving agent Migration (CHARM) model. The separation patterns observed with HxS, HxDAS and HxDMS as chiral resolving agent were compared with those of (1) b-cyclodextrin analogues, such as, heptakis(6-O-sulfo)-b-cyclodextrin (HS), heptakis(2,3-di-O-acetyl-6-O-sulfo)-b-cyclodextrin (HDAS) and heptakis(2,3-di-O-methyl-6-O-sulfo)-b-cyclodextrin (HDMS); (2) g-cyclodextrin analogues, such as, octakis(6-O-sulfo)-g-cyclodextrin (OS), octakis(2,3-di-O-acetyl-6-Osulfo)- g-cyclodextrin (ODAS) and octakis(2,3-di-O-methyl-6-O-sulfo)-g-cyclodextrin (ODMS). The effects of the structure of the analytes, and those of the pH and the solvent of the background electrolyte were also studied.

Purpose/Aims: The American Thoracic Society (ATS) published recommendations for alpha-1 antitrypsin deficiency (AATD) testing in 2003. This descriptive project evaluates the outcomes of ATS AATD guideline use in the setting of the pulmonary function testing (PFT) lab.The specific aims met by this descriptive project describe the prevalence of AATD cases and carriers in the sample, examine to what degree the established clinical guideline promoted accurate patient selection for the alpha-1 test in the sample, and aimed to determine whether alpha-1 antitrypsin blood levels are reduced in current smokers compared to former or never smokers.Background: Alpha-1 antitrypsin prevents lung tissue breakdown by attenuating excess elastase released from neutrophils during the inflammatory response. Smoking impairs alpha-1 antitrypsin protection at the site of lung inflammation promoting emphysema development. In the case of genetic mutation, protective alpha-1 antitrypsin levels are reduced, causing emphysema even in non-smokers. Significantly reduced protective levels of alpha-1 antitrypsin increase the odds for morbidity and early mortality from emphysema. The literature provides support for targeted testing in the population most affected.Sample/Methods: The sample population included adults 21 through 79 years completing pulmonary function testing over 18 months in a metropolitan pulmonary medicine practice and was retrospectively reviewed.Of the 521 in the sample, 190 were tested for AATD, and 24 were found to carry an abnormal genotype. However, using Table 11 from the ATS CPG failed to provide structured, consistent guidance in selecting patients for AATD testing. Still, the prevalence of the abnormal genotypes MS, MZ, SZ, and ZZ was increased in this pulmonary population compared to the published estimated prevalence for the general population.A structured decision-tree, developed from the original guideline for diagnostic testing, may provide superior guidance for AATD test patient selection in this setting. Increased case finding by targeted testing of patients in the setting of the pulmonary function lab can serve to integrate this clinical practice guideline in a consistent streamlined fashion.In this sample, no difference between AAT blood levels among ever, never, and current tobacco smokers was detected. A more powerful sample is needed.

The Alpha Particle X-ray Spectrometer (APXS) on the Mars Exploration Rovers possesses the ability to detect carbon and oxygen within martian samples via Rutherford backscattering principles. Several consecutive measurements of the martian atmosphere by Spirit, paralleled by Monte Carlo simulations, provided an energy calibration to mitigate the absence of an alpha-mode calibration pre-flight. Data from a pre-flight thermal acceptance test agreed with this energy calibration, confirming the presence of an unexpected offset. Correcting a bug in the APXS firmware resulted in a temperature-independent energy scale. A model was developed and applied to all atmospheric data illustrating a dip in atmospheric peak areas, potentially arising from a week-long weather event on Mars. An early expansion of this model to solid samples has not yet been able to detect any hydrated minerals or carbonates. Preliminary investigations into determining martian atmospheric pressure and potential elemental layering within samples shows promise.

Thèse diffusée initialement dans le cadre d'un projet pilote des Presses de l'Université de Montréal/Centre d'édition numérique UdeM (1997-2008) avec l'autorisation de l'auteur.

Plusieurs études animales et cellulaires suggèrent un rôle protecteur pour l’oestrogène dans les maladies inflammatoires intestinales. L'oestrogène agit principalement via l’interaction avec ses récepteurs ER? et ER?. Puisqu'ER? est l’isoforme prédominant dans l'épithélium colique, celui-ci devrait y être le médiateur des effets de l'oestrogène. Dans cette étude, nous avons tout d'abord démontré que la perte d’ER? chez la souris augmente les signes d'inflammation dans un modèle murin de colite. Également, l’activation sélective d’ER? avec le diarylpropionitrile (DPN) réduit les signes de colite dans le même modèle. L'expression de la cytokine inflammatoire TNF? était réduite alors que celle des cytokines anti-inflammatoires TGF?1, TGF?2 et TGF?3 était augmentée dans le côlon des souris traitées au DPN. Des études réalisées dans les cellules cancéreuses du côlon LS1034 ont démontré que le DPN augmente la signalisation du TGF? et que cette régulation contribue en partie à l’effet anti-inflammatoire du DPN dans ces mêmes cellules. Bref, ces résultats suggèrent que l'activation pharmacologique d’ER? réduit l’inflammation intestinale en partie via la régulation de la voie du TGF? dans les cellules épithéliales intestinales. Le récepteur nucléaire ERR?, un régulateur clé du métabolisme énergétique, possède une forte homologie avec les ERs bien que celui-ci ne peut lier l’oestrogène. Son activité est plutôt régulée par la disponibilité des coactivateurs ainsi que par des modifications post-traductionnelles. La découverte d'un variant d'épissage pour ERR? (ERR? ?5) a soulevé un nouveau mécanisme de régulation d'ERR?. En effet, puisqu'ERR? ?5 ne pourrait interagir avec les coactivateurs, il pourrait donc agir comme dominant négatif sur l’activité d'ERR?. Dans cette étude, nous avons démontré qu'ERR? et ERR? ?5 colocalisent au noyau en plus d’interagir physiquement De plus, ERR? ?5 réduit l'activité transcriptionnelle d'ERR? ainsi que l'expression de ses gènes cibles. Tout comme ERR?, ERR? ?5 est exprimé dans une multitude de lignées normales et cancéreuses du côlon ainsi que dans des cellules ou tissus provenant de d'autres organes et de d’autres espèces animales. Par contre, la protéine ERR? ?5 est exprimée plus faiblement qu'ERR? puisqu’elle est instable et rapidement dégradée par le protéasome. Néanmoins, la surexpression d'ERR? ?5 réduit la croissance cellulaire et possiblement l'adhésion cellulaire. Bref, nos résultats démontrent qu’ERR? ?5 agit comme dominant négatif sur ERR? et que son expression transitoire pourrait donc affecter l’activité d'ERR? ainsi que ses fonctions biologiques. [symboles non conformes]

Systemic fungal infection is a life-threatening problem. Anti-fungal drugs are the most effective clinical strategy to cure such infections. However, most current anti-fungal drugs either have high toxicity or have a narrow spectrum of effect. Meanwhile, anti-fungal drugs are losing their clinical efficacy due to emerging drug resistance. To protect us from these deadly pathogenic fungi, scientists need to study new drug targets and to solve problems related to drug resistance. The cell wall is essential for fungal cell survival and is absent from animal cells, so it is a promising reservoir for screening safe and effective drug targets. Alpha-1,3-glucan is one of the major cell wall carbohydrates and is important for the virulence of several pathogenic fungi. In this thesis, molecular biology and microscopy techniques were used to investigate the function and the synthesis process of α-1,3-glucan in the model fungus A. nidulans. My results showed that α-1,3-glucan comprises about 15% of A. nidulans cell wall dry weight, but also that α-1,3-glucan does not have an important role in cell wall formation and cell morphology. Deletion of α-1,3-glucan only affects conidial adhesion and cell sensitivity to calcofluor white. In contast, elevated α-1,3-glucan content can cause severe phenotypic defects. To study the α-1,3-glucan synthesis process, I systematically characterized four proteins, including two α-1,3-glucan synthases (AgsA and AgsB) and two amylase-like proteins (AmyD and AmyG). Results showed AgsA and AgsB are both functional synthases. AgsB is the major synthase due to its constant expression. AgsA mainly functions in conidiation stages. AmyG is a cytoplasmic protein that is critical for α-1,3-glucan synthesis, likely being required for an earlier step in the synthesis process. In contrast to the other three proteins, AmyD has a repressive effect on α-1,3-glucan accumulation. These results shed light on therapeutic strategies that might be developed against α-1,3-glucan. I also developed a strategy to investigate drug resistance mutations. The tractability of A. nidulans and the power of next generation sequencing enabled an easy approach to isolate single mutation strains and to identify the causal mutations from a genome scale efficiently. I suggest this strategy has applications to study the drug resistance mechanisms of current anti-fungal drugs and even possibly future ones.

Neonatal ventral hippocampus (nVH) lesioned animals show molecular and behavioral abnormalities analogous to those described in schizophrenia. As an extension to previous studies that showed an increase in ligand binding of cortical alpha-1 adrenergic receptors (AR) and a dysfunction in alpha-1 AR regulation of mesolimbic dopamine functions in post-pubertal nVH lesioned rats, we investigated the subcellular expression and activity of protein kinase C (PKC)---a second messenger in alpha-1 AR signaling---in the prefrontal cortex (PFC) and nucleus accumbens (NAcc) of post-pubertal nVH lesioned rats. Western blot analysis of membrane and cytosolic fractions showed complex changes in lesioned animals in the expression of different PKC subtypes following saline or alpha-1 AR agonist (cirazoline i.p.) injection. Among these changes, nVH lesioned animals showed a significant increase in membrane bound PKC alpha and phospho-PKC, and a decrease in cytosolic PKC gamma and PKC betaII in the PFC in comparison to sham-lesioned controls following saline. Cirazoline increased membrane bound PKC alpha in controls but decreased it in lesioned animals. In the NAcc, lesioned animals showed an increase in membrane bound and cytosolic PKC epsilon and PKC lambda levels following saline. Following cirazoline, lesioned animals showed a decrease in membrane bound PKC epsilon and PKC lambda, while controls showed an increase in cytosolic and membrane fractions of PKC epsilon with no change in PKC lambda. In vitro PKC activity assays showed increased basal activity in PFC slices of lesioned animals compared to controls, with no difference in NAcc slices. alpha-1 AR stimulation by the agonist phenylephrine (PE) increased PKC activity in PFC of controls while decreasing activity substantially in lesioned animals. In the NAcc, high concentrations of PE increased activity in controls, but decreased activity in lesioned animals. This abnormal expression and activity of PKC in the PFC and NAcc of nVH lesioned animals may be related to abnormal alpha-1 AR functions and may modulate some of the abnormal neuronal functions in these animals, such as working memory deficits and hyper neuronal excitability of the PFC and the NAcc.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 4 uppsatser.

Thesis (doctoral)--Lund University, 1998. / Added t.p. with thesis statement inserted.

Thesis (doctoral)--Lund University, 1998. / Added t.p. with thesis statement inserted. Includes bibliographical references.