Plusieurs paramètres participent à l'homéostasie du système immunitaire. On peut notamment citer : la production lymphocytaire (les lymphocytes B dans la moelle osseuse ou les lymphocytes T (LyT) dans le thymus), la prolifération et la survie des lymphocytes périphériques et enfin, la compétition entre les différents clones de lymphocytes ou les différentes populations de lymphocytes pour des ressources limitées en particulier les cytokines. Les mécanismes de régulation homéostatique sont différents selon la sous-population de lymphocyte considérée. Dans le cadre de cette thèse, nous avons étudié le rôle de l'interleukine-7 (IL-7) dans l'homéostasie des lymphocytes et plus particulièrement les lymphocytes T ΑΒ. Disposant de souris surexprimant l'IL-7 dans un état lymphopénique plus ou moins sévère, nous avons pu décortiquer les effets de l'IL-7 sur le développement précoce des LyT et leur survie périphérique. L'analyse du développement précoce des lymphocytes n'a pas révélé d'effet notable de la sur-expression de l'IL-7 sur le développement thymique des LyT. En revanche, une expansion considérable des LyT périphériques a été observée. A partir d'expérience de transferts adoptifs, nous avons montré que l'IL-7 favorisait la prolifération et la survie des LyT périphériques. La quantité d'IL-7 disponible module donc le devenir des LyT présents dans les organes lymphoïdes lors d'un épisode lymphopénique.

Any lipoproteins and cells present in the inflammatory environment of atherosclerotic plaques are likely to be exposed to high levels of oxidative stress. As 7,8-dihydroneopterin (7,8-NP) is synthesized by interferon-γ (IFN-γ)-activated macrophages, this pteridine is also thought to exist at sites of inflammation. 7,8-NP s in vivo role remains controversial, but numerous in vitro studies have identified a radical scavenging activity. The possibility of 7,8-NP protecting against oxidative damage in inflammatory environments like plaque was investigated in this thesis. Both human monocyte-derived macrophages (HMDMs) and low density lipoprotein (LDL) were used as substrates. The extent of protein hydroperoxide formation in each model, and 7,8-NP s effect on this process, were specifically studied since most previous research has focussed on lipid rather than protein peroxidation. For the first time, neopterin (including oxidized 7,8-NP) was also directly detected by high performance liquid chromatography in the inflammatory environments of 19 pus and two atherosclerotic plaque samples. Peak concentrations even reached the low micromolar range. The positive correlation identified in the pus between neopterin and a well known antioxidant, vitamin E, further hinted at a potential antioxidant function. However, no significant association was noted between neopterin and markers of protein or lipid oxidation. Exposure of HMDMs to the AAPH peroxyl radical generator resulted in significant quantities of lipid hydroperoxides but not protein hydroperoxides, as detected by the FOX assays. This is likely due to the large accumulation of polyunsaturated fatty acidrich lipid in the primary HMDMs during differentiation in 10% human serum and is of relevance to atherosclerotic plaque, where macrophages also become lipid-loaded. The addition of up to 200μM 7,8-NP failed to prevent AAPH-induced lipid peroxidation and was also unable to inhibit a loss of cellular thiols or viability. This lack of effect suggests the damaging peroxyl radicals are not being scavenged by 7,8-NP. The high lipid content of HMDM cells appears to cause the AAPH and/or 7,8-NP to localize to a cellular site, where they are unable to interact. Macrophage-mediated oxidation of LDL in iron(II)-supplemented Hams F10 was associated with the formation of 30-40 moles of protein hydroperoxides per mole of LDL. The close parallel between protein and lipid peroxidation supports the theory that lipid-derived radicals are involved in protein hydroperoxide formation on LDL and indicates that protein hydroperoxides are an early product of LDL oxidation. Their detection during exposure of LDL to both the THP-1 macrophage cell line and primary HMDM cells confirms that protein hydroperoxides are also a normal consequence of macrophage-mediated LDL oxidation. Incubation of LDL with micromolar 7,8-NP prevented macrophage-mediated protein hydroperoxide formation in a concentration-dependent manner. Lipid oxidation and vitamin E loss were similarly inhibited by 7,8-NP during the cell-mediated attack of LDL. Kinetic analysis revealed protection due to extension of the lag phase, with 7,8-NP depletion and initiation of the propagation phase coinciding. This supports a radical scavenging activity for 7,8-NP, resulting in protection of the entire LDL particle. By contrast, the release of nanomolar quantities of 7,8-NP by IFN-γ-stimulated THP-1 macrophages failed to prevent LDL oxidation. HMDMs activated by IFN-γ did significantly inhibit LDL oxidation, including protein hydroperoxide formation, for up to 48 hours but this antioxidant effect was not due to the de novo synthesis of 7,8-NP. These results indicate that both the prevalence of protein hydroperoxides, and the ability of 7,8-NP to act as an antioxidant, depend on the system under investigation. Neopterin exists in inflammatory environments but, considering the lack of protection against AAPH-mediated HMDM oxidation and the 7,8-NP concentration required to inhibit macrophage-mediated LDL oxidation, strong evidence for an antioxidant activity of 7,8-NP in atherosclerotic plaque is currently lacking.

No description available.

No description available.

The host-guest chemistry between cucurbit[7]uril (CB[7]) and cationic organic guests of medicinal and biological interest are described in this thesis. In the first part, three cationic steroidal neuromuscular blockers (SNMBs) were studied, along with guests that model their monocationic N-alkyl-N-methylheterocyclic (morpholinium, pyrrolidinium and piperidinium) terminal groups of the SNMBs, and dicationic guests in which the two N-methylheterocyclic rings are linked by a decamethylene chain, modelling a variety of NMBs. Other cationic drugs related to acetylcholine processes in neuromuscular blockage were also studied. In the second part, the amino acids lysine, and its mono-, di- and trimethylated and acetylated Nε derivatives, and arginine, and mono- and (symmetric and asymmetric) dimethylarginine, were investigated as guests, along with analogs of arginine. The nature and strength of the complexation between CB[7] and these guests in aqueous solution were determined by 1H NMR spectroscopy and ESI mass spectrometry. The CB[7] showed high binding affinity (KCB[7] = 106-109 M-1) towards the N-alkyl-N-methylheterocyclic cations with a trend of piperidinium > pyrrolidinium > morpholinium, which reflects the relative hydrophobicities of the guests. The CB[7] forms 1:1 and 2:1 host-guest complexes with dicationic model guests, with the CB[7] initially encapsulating the decamethylene chain. The second CB[7] binds to a terminal site, resulting in electrostatic repulsions with the first CB[7], which are resolved by the translocation of the first CB[7] to the opposite terminal site. This 2:1 binding mode is also observed with CB[7] and the SNMBs, and the trend in KCB[7] with these SNMB terminal sites is comparable to that observed for the monocationic model guests. The other cationic drugs also form stable host-guest complexes with CB[7], and the binding constants displayed dependences on the size, charge, and hydrophobicity of the guests. The CB[7] exhibits significant selectivity towards different lysine and arginine derivatives, which can be related to the relative hydrophobicity afforded by the methyl substituents and the positioning of the guest within the CB[7] cavity. The 3500-fold selectivity for Nε,Nε,Nε-trimethyllysine over lysine by CB[7] is the highest observed for a synthetic macrocyclic receptor, while a modest selectivity of symmetrical over asymmetrical dimethylarginine by CB[7] is observed. / Thesis (Ph.D, Chemistry) -- Queen's University, 2013-09-26 14:33:40.063

My essay has three foci. One is the presentation of a survey I gave to two groups of pupils, one grade 7 and the other grade 8 about their attitude to reading books in school, and their attitude to reading books in general. I was interested in how the result would correspond to what many investigations on this subject had concluded. One such result was that girls were usually not only more willing to read, but they were also better readers than boys. Therefore, the answerers must tell if they were boys or girls. The result of my investigation rather corroborated what many others had already said: Swedish young people are not enthusiastic book readers, but when reading, the girls are the ones that take the lead. Another focus is in a way historical. I used a novel by a Swedish author telling about poor people on the Swedish countryside in the nineteenth century dreaming of being able to emigrate from their home country and come to America. What interests me is the story telling of the attitude to reading, both in the characters and the authorities. There is much about reading incapacities, but also of lust for, and skill in reading. The attitudes from the authorities is dominated by churchly paternalism: reading skill in ordinary people has one primary function, to make them good Christians. This common value-system is fixed by the didactic curricula of the time. My third focus is the Swedish curricula especially in what they say about reading and literature. I thought I found that they have at least something in common with the curricula of mid-nineteenth century than might be expected. The older ones have what could be seen as an instrumental attitude to reading and such an attitude I thought could be perceived also in the modern ones: they seem to be eager to point out the usefulness of reading in general and even of reading literature. Another point would be the great importance attached to the strengthening of the common value-system, here, not Christian belief, but a democratic ground of values. In my study of what is said about reading in school by people discussing the subject, and when looking at the result of my own investigation by the questionnaire I very often find a similar instrumental attitude: all reading, be it fact or fiction, is expected to lead to something useful not just pleasure.

The Arizona Child Acoustic Database consists of longitudinal audio recordings from a group of children over a critical period of growth and development (ages 2-7 years). The goal of this database is to 1) document acoustic changes in speech production that may be related to physical growth 2) inform development of a model of speech production for child talkers. This work was funded by NSF BSC-1145011 awarded to Kate Bunton, Ph.D. and Brad Story, Ph.D, Principal Investigators. This database contains longitudinal audio recordings of 55 American English speaking children between the ages of 2-7 at 3-month intervals. Since children began the study at different ages, some children have fewer recording sessions than others. The database can also be used to provide cross-sectional data for children of a specific age. Please refer to the subject data table for information on specific sessions available here http://arizona.openrepository.com/arizona/handle/10150/316065. All children were recorded using the same protocol; therefore, task numbers are consistent across children and sessions. A calibration tone is included as Record 1 for all sessions. The speech protocol focused on production of English monopthong and diphthong vowels in isolation, sVd, hVd, and monosyllabic real words. In addition, the protocol includes several nonsense vowel-to-vowel transitions. Speakers were prompted either verbally by investigators or by graphical prompts. Details of the protocol with reference to task numbers can be found in the protocol spreadsheet available here http://arizona.openrepository.com/arizona/handle/10150/316065. Details on data recording: All samples were recorded digitally using an AKG SE 300B microphone with a mouth to mic distance of approximately 10 inches. Signals were recorded digitally using a Marantz PMD671, 16 bit PCM (uncompressed) at 44.1KHz. Recordings are made available in .wav format. Individual zip files contain all recordings from a single session.

The Arizona Child Acoustic Database consists of longitudinal audio recordings from a group of children over a critical period of growth and development (ages 2-7 years). The goal of this database is to 1) document acoustic changes in speech production that may be related to physical growth 2) inform development of a model of speech production for child talkers. This work was funded by NSF BSC-1145011 awarded to Kate Bunton, Ph.D. and Brad Story, Ph.D, Principal Investigators. This database contains longitudinal audio recordings of 55 American English speaking children between the ages of 2-7 at 3-month intervals. Since children began the study at different ages, some children have fewer recording sessions than others. The database can also be used to provide cross-sectional data for children of a specific age. Please refer to the subject data table for information on specific sessions available here http://arizona.openrepository.com/arizona/handle/10150/316065. All children were recorded using the same protocol; therefore, task numbers are consistent across children and sessions. A calibration tone is included as Record 1 for all sessions. The speech protocol focused on production of English monopthong and diphthong vowels in isolation, sVd, hVd, and monosyllabic real words. In addition, the protocol includes several nonsense vowel-to-vowel transitions. Speakers were prompted either verbally by investigators or by graphical prompts. Details of the protocol with reference to task numbers can be found in the protocol spreadsheet available here http://arizona.openrepository.com/arizona/handle/10150/316065. Details on data recording: All samples were recorded digitally using an AKG SE 300B microphone with a mouth to mic distance of approximately 10 inches. Signals were recorded digitally using a Marantz PMD671, 16 bit PCM (uncompressed) at 44.1KHz. Recordings are made available in .wav format. Individual zip files contain all recordings from a single session.

Many cancerous solid tumors metastasize to the bone and induce pain (cancer-induced bone pain [CIBP]). Cancer-induced bone pain is often severe because of enhanced inflammation, rapid bone degradation, and disease progression. Opioids are prescribed to manage this pain, but they may enhance bone loss and increase tumor proliferation, further compromising patient quality of life. Angiotensin-(1-7) (Ang-(1-7)) binds and activates the Mas receptor (MasR). Angiotensin-(1-7)/MasR activation modulates inflammatory signaling after acute tissue insult, yet no studies have investigated whether Ang-(1-7)/MasR play a role in CIBP. We hypothesized that Ang-(1-7) inhibits CIBP by targeting MasR in a murine model of breast CIBP. 66.1 breast cancer cells were implanted into the femur of BALB/cAnNHsd mice as a model of CIBP. Spontaneous and evoked pain behaviors were assessed before and after acute and chronic administration of Ang-(1-7). Tissues were collected from animals for ex vivo analyses of MasR expression, tumor burden, and bone integrity. Cancer inoculation increased spontaneous pain behaviors by day 7 that were significantly reduced after a single injection of Ang-(1-7) and after sustained administration. Preadministration of A-779 a selective MasR antagonist prevented this reduction, whereas pretreatment with the AT(2) antagonist had no effect; an AT(1) antagonist enhanced the antinociceptive activity of Ang-(1-7) in CIBP. Repeated Ang-(1-7) administration did not significantly change tumor burden or bone remodeling. Data here suggest that Ang-(1-7)/MasR activation significantly attenuates CIBP, while lacking many side effects seen with opioids. Thus, Ang-(1-7) may be an alternative therapeutic strategy for the nearly 90% of patients with advanced-stage cancer who experience excruciating pain.

Renal cell carcinoma accounted for an estimated 57,760 new cases and estimated 12,980 deaths in the United States in 2009. Current treatment options for systemic renal cell carcinoma yield markedly low response percentages; however, recent cytokine therapy experiments have produced promising results. A novel adenovirus, Ad.5/3-mda-7, has been synthesized to efficiently infect renal cancer cells with the mda-7 gene. This gene encodes for the cytokine MDA-7/IL-24 that has the ability to specifically target transformed cells. Assays performed with this adenovirus resulted in an increased percentage of cell death in renal cancer cells when compared to infection with Ad.5/3-cmv empty vector. Further assays that combined Ad.5/3-mda-7 infection with treatments of small molecule inhibitors increased the percentages of cell death by upregulating JNK and p38 MAPK pathways, downregulating the ERK1/2 MAPK pathway, and downregulating the PI3K pathway. Western blots confirmed upregulation and downregulation of these pathways by probing for key proteins. Renal cancer cells responded best to infection with Ad.5/3-mda-7 and treatment with PD184352, PX866, and Rapamycin. This combinatorial treatment caused a greater percentage of cell death than the sum of the two individual treatments, suggesting a synergistic inhibition of cell growth pathways. These findings suggest that the combination of Ad.5/3-mda-7 and specific small molecule inhibitors has developmental potential as a novel and more efficient treatment option for systemic renal cell carcinoma.