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Investigation of micro-mechanical applications of amorphous carbon filmsTsai, Kun-Chao January 2003 (has links)
No description available.
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Biochemical studies on the dopamine receptor in the central nervous systemSenior, K. A. January 1984 (has links)
No description available.
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Expression et fonctionnalité des Toll-Like récepteurs par les cellules de leucémie lymphoïde chronique B / Expression and functionality of Toll-Like receptors by B-chronic lymphocytic leukemia cellsGrandjenette, Cindy 06 March 2008 (has links)
La leucémie lymphoïde chronique reste une pathologie incurable même si les nouveaux moyens thérapeutiques permettent d’obtenir des réponses de bonne qualité. Les déterminants idiotypiques des lymphocytes B représentent une cible idéale pour la génération de cellules T spécifiques dirigées contre le clone malin. Dans ce cadre, l’induction d’une réponse immunitaire efficace semble donc intéressante. L’objectif principal de cette étude était d’augmenter l’immunogénicité des cellules tumorales de LLC-B. Pour cela, la possibilité d'une activation de ces cellules par des agonistes de Toll-like récepteurs (TLRs) a été explorée. Tout d’abord, le profil d’expression protéique des TLRs a été recherché au niveau des lymphocytes B sains et pathologiques par cytométrie de flux. La mise en évidence d'une expression significative de TLR-7 et TLR-9 a ensuite permis d'analyser l’effet d'une stimulation par des ligands de ces TLRs par immunophénotyage, cytomorphologie, analyse de l’apoptose par la méthode TUNEL et étude de la prolifération par marquage CFSE. Le profil cytokinique des suspensions cellulaires stimulées ou non a également été examiné par cytométrie de flux. Enfin, une analyse fonctionnelle a été réalisée dans un modèle de stimulation allogénique afin de mesurer la capacité de ces cellules stimulées à activer des lymphocytes T naïfs. Les résultats obtenus montrent un profil d’expression des TLR-4, -7, -9 et -10 similaire pour les cellules B de LLC et les lymphocytes B sains avec toutefois une expression plus importante de TLR-9 dans les cellules tumorales. L’engagement de TLR-4 a pour conséquence une faible activation de HLA-DR sur les cellules B saines et tumorales. L’activation de TLR-7 via les plus fortes concentrations d'imiquimod R837 induit surtout une apoptose rapide et importante des cellules de LLC-B ainsi qu'une augmentation modérée de l'expression des marqueurs de costimulation et de présentation des cellules B saines et tumorales. La stimulation de TLR-9 via les ODN CpG entraîne une activation des cellules B saines et tumorales et protège les cellules tumorales de l'apoptose. Les profils cytokiniques des suspensions cellulaires ont montré que ces ligands de TLRs orientent les réponses cellulaires vers un profil pro-inflammatoire, éventuellement propice à la génération de réponses cellulaires, potentiellement anti-tumorales via la production d'IL-8 et de TNF. Enfin, il est apparu que les cellules amygdaliennes et tumorales stimulées avec les ODN CpG n'étaient pas capables d'activer des lymphocytes T naïfs allogéniques, ce qui pourrait être lié à la faible production d'IL-2 ou à la production d'IL-10 par ces cellules. En conclusion, TLR-7 et TLR-9 représentent des cibles thérapeutiques intéressantes dans le traitement de la LLC-B. L’imiquimod R837 pourrait aider à la clairance tumorale dans les LLC-B. L’activation cellulaire induite par via les ODN CpG M362 pourrait par contre être utilisée pour augmenter le potentiel de cellules présentatrices d’antigènes des cellules B de LLC à condition de parvenir à contrecarrer l'environnement tolérogène et à différencier les cellules B en cellules activatrices de réponses Th1 et cytotoxiques. / thesis
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Impact of asthma, environmental exposures and ethnicity on functional responsiveness to Toll-like receptor (TLR) stimulation in childrenLissitsyn, Yuriy V 31 August 2007 (has links)
TLRs play a key role in initiating innate immunity and in regulating the nature of the adaptive immune response. We hypothesized that functional responsiveness to TLR stimulation differs in clinically; environmentally; ethnically distinct pediatric populations.
PBMC obtained from 272 children were stimulated with a panel of TLR ligands. Levels of pro- and anti-inflammatory, Th1-, Th2-associated cytokines were quantified by ELISA.
We demonstrate that use of threshold concentrations of TLR4 and TLR2 ligands reveal striking differences in cytokine responses between asthmatic and non-atopic children. Specifically, non-atopic controls produce higher levels of pro-inflammatory cytokines, whereas asthmatics exhibit increased anti-inflammatory IL-10 responses.
Asthmatic children exposed to environmental tobacco smoke (ETS) demonstrated elevated levels of chemokines relative to non-ETS exposed asthmatics and controls.
First Nation children favor anti-inflammatory IL-10 responses, whereas Caucasian population respond to TLR activation by production of more robust pro-inflammatory and Th1 biased cytokine and chemokine responses. / October 2007
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Structural study of polyglutamine and molecular mechanism of toll-like receptor signalingLiu, Zhuyun 15 May 2009 (has links)
Huntington’s disease (HD) is caused by the expansion of a CAG repeats
encoding polyglutamine (polyQ) in the first exon of Huntingtin (Htt) gene. In HD
patients, polyQ contains 36-183 glutamine residues, whereas normal individuals have a
polyQ of only 8-35 residues. To elucidate this threshold phenomenon of polyQ
aggregation, fluorescence proteins CFP and YFP were attached to both ends of polyQ of
different lengths. FRET (fluorescence resonance energy transfer) was conducted to
characterize the conformation of polyQ in the pre-aggregation state. Our FRET data
show that both the normal and expanded polyQ tracts reveal the same extended structure
in low concentration. Longer polyQ has multiple cooperative binding sites with higher
avidity. PolyQ tracts form aggregates when proteins exceed a critical concentration. The
antibody MW1 Fv fragment binds to polyQ, breaks apart polyQ oligomer and stabilizes
it in a more extended conformation.
The addition of polyproline to the C-terminus inhibits polyQ aggregation by
inducing PPII-like Helix structure. To understand how the flanking sequence affects the
polyQ structure, the structure of Q10P10 peptide in complex with MW1 Fv was determined by protein crystallography and compared with Q10/Fv crystal structure.
Q10P10 peptide bound to Fv has a similar extended structure as Q10 peptide when a
polyproline tract adopts PPII helical structure sticking out of the complex.
Toll-like receptors are transmembrane receptors on different kinds of leukocytes.
They can recognize the structural conserved molecular motifs derived from microbes.
On the upstream of the TLR signal pathway, TLRs recruit the adaptor protein-MyD88
through TIR/TIR domain interaction, and MyD88 recruits the downstream kinases
IRAK4 and IRAK1 through death domain/death domain interaction. Pellino1, a newly
identified E3 ubiquitin ligase, is also involved in TLR signaling by adding polyubiquitin
chain to IRAK1 in conjugation with Ubc13/Uev1a E2 complex. TIR/TIR and DD/DD
binding motifs were studied with techniques including mutagenesis, analytical gel
filtration, NMR spectroscopy and crystallography. We identified a MyD88DD
(E52QR62S) double-mutant that attenuates protein aggregation without interrupting the
binding with IRAK4. This double mutant is a good candidate for structure determination
by NMR spectroscopy. Our ubiquitination assay showed Pellino1 catalyzes
polyubiquitination in the presence of Ubc13/Uev1a in vitro. Needle cluster-shaped
crystals of Pellino1/Ubc13/ Uev1a protein complex were obtained by “hanging drop”
method of vapor diffusion. Once the crystallization conditions are optimized, we will be
able to collect X-ray diffraction data for this E2/E3 complex.
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Impact of asthma, environmental exposures and ethnicity on functional responsiveness to Toll-like receptor (TLR) stimulation in childrenLissitsyn, Yuriy V 31 August 2007 (has links)
TLRs play a key role in initiating innate immunity and in regulating the nature of the adaptive immune response. We hypothesized that functional responsiveness to TLR stimulation differs in clinically; environmentally; ethnically distinct pediatric populations.
PBMC obtained from 272 children were stimulated with a panel of TLR ligands. Levels of pro- and anti-inflammatory, Th1-, Th2-associated cytokines were quantified by ELISA.
We demonstrate that use of threshold concentrations of TLR4 and TLR2 ligands reveal striking differences in cytokine responses between asthmatic and non-atopic children. Specifically, non-atopic controls produce higher levels of pro-inflammatory cytokines, whereas asthmatics exhibit increased anti-inflammatory IL-10 responses.
Asthmatic children exposed to environmental tobacco smoke (ETS) demonstrated elevated levels of chemokines relative to non-ETS exposed asthmatics and controls.
First Nation children favor anti-inflammatory IL-10 responses, whereas Caucasian population respond to TLR activation by production of more robust pro-inflammatory and Th1 biased cytokine and chemokine responses.
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GLP-1 effects on pancreatic b-cell linesSinclair, Elaine M. January 2002 (has links)
The aims of this thesis were to establish a suitable model system for the study of glucose and nutrient regulation of insulin secretion and biosynthesis. This would serve as a basis for investigating the GLP-1 effects on pancreatic b-cell biology. This thesis shows that two b-cell model systems, namely MIN6 and INS-1 cells, respond to increasing concentrations of glucose, by increasing insulin secretion and cell proliferation in a physiological manner. The MIN6 cell line also responds to other cellular nutrients, L-arginine and L-leucine in a manner similar to primary islet cells. The MIN6 cells however, fail to consistently increase insulin secretion in response to GLP-1, a known potentiator of insulin secretion in b-cells, despite the presence of the GLP-1 receptor. Incubation of GLP-1 with INS-1 cells, increases insulin secretion in a glucose-dependent manner, and causes a small increase in cell proliferation. GLP-1 is also known to increase cAMP levels within the cell and interact with cAMP response elements (CRE) via activation protein kinase A (PKA). Using a luciferase reporter gene construct containing 4 copies of the CRE, glucose, GLP-1 and forskolin failed to increase luciferase activity in MIN6 cells, suggesting that a defect in cAMP signalling may explain the inconsistent effect of GLP-1 in MIN6 cells. A stimulatory effect of GLP-1 and forskolin was observed in the INS-1 cells. Using both a rat insulin I and human insulin gene promoter construct, a stimulatory effect of GLP-1 on insulin gene transcription was observed in INS-1 cells. An insight into the signalling pathways involved in GLP-1 stimulation of the rat insulin I gene was gained through the use of protein kinase inhibitors, which inhibit signalling of known signal transduction cascades. It was found that an inhibitor of protein kinase A (H-89) was effective in blocking the increase in insulin promoter activity induced by GLP-1 using the rat insulin I promoter construct. Interestingly, the p38/SAPK2 inhibitor, SB203580, further increased the GLP-1 stimulation of rat insulin I promoter activity, indicating that this pathway usually invokes an inhibitory effect on insulin promoter activity.
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Molecular cloning and nucleotide sequencing of sacbrood virus of the honey beeGhosh, Ratan Chandra January 1999 (has links)
No description available.
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Impact of asthma, environmental exposures and ethnicity on functional responsiveness to Toll-like receptor (TLR) stimulation in childrenLissitsyn, Yuriy V 31 August 2007 (has links)
TLRs play a key role in initiating innate immunity and in regulating the nature of the adaptive immune response. We hypothesized that functional responsiveness to TLR stimulation differs in clinically; environmentally; ethnically distinct pediatric populations.
PBMC obtained from 272 children were stimulated with a panel of TLR ligands. Levels of pro- and anti-inflammatory, Th1-, Th2-associated cytokines were quantified by ELISA.
We demonstrate that use of threshold concentrations of TLR4 and TLR2 ligands reveal striking differences in cytokine responses between asthmatic and non-atopic children. Specifically, non-atopic controls produce higher levels of pro-inflammatory cytokines, whereas asthmatics exhibit increased anti-inflammatory IL-10 responses.
Asthmatic children exposed to environmental tobacco smoke (ETS) demonstrated elevated levels of chemokines relative to non-ETS exposed asthmatics and controls.
First Nation children favor anti-inflammatory IL-10 responses, whereas Caucasian population respond to TLR activation by production of more robust pro-inflammatory and Th1 biased cytokine and chemokine responses.
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Galanin-like peptide : a molecular link between energy homeostasis and reproduction /Krasnow, Stephanie Maxwell. January 2004 (has links)
Thesis (Ph. D.)--University of Washington, 2004. / Vita. Includes bibliographical references (leaves 129-156).
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