Evidence for, and characterization of, a lipopolysaccharide-inducible adenosine A2 receptor in human tracheal gland serous cells

AbstractHuman tracheal glands are considered as the principle secretory structures in the bronchotracheal tree. In earlier studies, we successfully performed primary cultures of human tracheal gland (HTG) serous cells and noted that these cells were responsive to many secretagogues including purinergic agonists but not to the inflammatory mediator adenosine. In this study, we demonstrate that adenosine was capable of inducing stimulation of protein secretion by HTG serous cells which had previously been cultured in pro-inflammatory conditions (induced by lipopolysaccharide (LPS)). This stimulation was inhibited by 8-phenyltheophylline but not by dipyridamole, which is indicative of a P1 purinoceptor. This inducible receptor is of the adenosine A2 subtype [rank potency order: 5′-(N-ethyl)-carboxamidoadenosine (NECA) > adenosine > N6-(phenylisopropyl)-adenosine (PIA); and stimulation of adenylyl cyclase]. The adenosine-induced protein secretion was concentration-dependent, however, increased intracellular cyclic adenosine monophosphate (cAMP) was not dependent on the concentration of adenosine. The adenosine-induced secretion and the ATP-induced secretion were shown to be additive. This study concludes that there is evidence of a LPS-inducible adenosine A2 receptor in human tracheal gland serous cells

Single and biphase magnetic microwires: Microwave behavior and temperature dependence

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Física de la Materia Condensada. Fecha de lectura: 08-04-2015This work is has been devoted to the study of single and biphase magnetic microwires, from fabrication to their magnetic characterization at low but particularly at microwave frequencies. As single phase microwires we consider glass-coated amorphous microwires with very soft magnetic behavior (both, positive magnetostriction Fe-based and vanishing magnetostriction CoFe-based samples). In biphase microwires, we grow an external layer of polycrystalline character with either soft (FeNi) or medium-hard (CoNi) magnetic character so that, we consider soft/soft and soft/hard magnetic biphase systems. Microwires have been fabricated by combined glass-coating rapid-solidification technique (the amorphous core) and sputtering/electrodeposition technique (the external shell). After their fabrication, the microwires were characterized from a geometry, composition and structure points of view. Quite a number of different samples were produced as a function of core and external shell magnetic phases as well as for different thickness of intermediate insulating Pyrex thickness and external layer. In addition, selected samples were selected for studies on the influence of thermal annealing at temperatures up to reaching first steps of crystallization at 600ºC. A first magnetic characterization was performed at low-frequency magnetic fields in a Vibrating Sample Magnetometer, VSM, to determine the magnetization processes and main magnetic magnitudes as coercivity and remanence of each magnetic phase. This has been performed for all the series of samples mentioned above. Studies have been extended to different measuring temperature particularly in the range up to 900ºC; where after partial crystallization significant deterioration of properties is confirmed. The study about the coercive field of individual and bimagnetic phase systems is analyzed in view of the relevance for technological applications of bimagnetic microwires. Also, the magnetic phase transitions (ferro to paramagnetic) of some individual phases are first identified. The main part of the study has been devoted to the magnetic properties at high frequency, in the microwave regime. The ferromagnetic resonance (FMR) spectra have been determined for single and biphase samples with different geometry and compositional characteristics as indicated above. FMR experiments in wires can be basically divided into two categories. The former is done by means of network analyzer up to around 15 GHz and as a function of applied magnetic field. The latter has been performed in a coaxial or microstrip microwave circuit making use of classical FMR spectrometers and waveguide microwave techniques. A first study was focused on the influence of glass Pyrex on single and biphase microwires. For single-phase microwires, the increase of the Pyrex thickness results in a continuous strengthening of the circular magnetoelastic anisotropy of the CoFe-based core as deduced from FMR and confirmed by low-frequency measurements. For biphase microwires three absorption peaks are observed: two of them can be ascribed to each magnetic phase since FMR frequencies obey the Kittel condition for a thin film. A third absorption peak is observed at lower frequencies that does not follow such an equation and can be ascribed to a pure geometrical effect of these biphase microwires. Then, the effect of annealing treatment at temperatures up to 700ºC on high frequency behavior was investigated. FMR behavior was analyzed by fitting to Kittel equation for annealed samples. The observed different absorption peaks are correlated with the presence of the soft core and the peaks presented at low frequency do not obey such Kittel condition. In this regards, it has been confirmed that the peaks observed at lower frequency corresponded to a capacitance effect. Additional studies have been presented investigating the microwave absorption phenomena of single and biphase magnetic microwires with soft magnetic behavior using two alternative techniques: (i) absorption measurements in the temperature range of -271_25ºC using a spectrometer operating at X-band frequency, at 9.5GHz, and (ii) room temperature, RT, ferromagnetic resonance measurements in a network analyzer up to 15 GHz. For single phase CoFeSiB microwire, a single absorption is observed, whose DC field dependence of resonance frequency at RT fits to a Kittel law behavior for in-plane magnetized thin film. The temperature dependence behavior shows a monotonic increase in the resonance field, Hr, with temperature. For biphase,CoFeSiB/FeNi, microwires, the absorption phenomena at RT also follow the Kittel condition. The observed opposite evolution with temperature of resonance field, Hr, in two different thick FeNi samples is interpreted considering the opposite sign of magnetostriction of the respective FeNi layers

High lysosomal activities in cystic fibrosis tracheal gland cells corrected by adenovirus-mediated CFTR gene transfer

AbstractHuman tracheal gland serous (HTGS) cells are now believed to be a major target of cystic fibrosis (CF) gene therapy. To evaluate the efficiency of adenovirus-mediated gene transfer in these cells we tested the adenovirus construction containing β-galactosidase cDNA. We observed that the endogenous β-galactosidase activity in cultured CF-HTGS cells was too strong to allow us to detect any exogenous β-galactosidase activity. Immunohistological study on sections of human tracheal tissue confirmed the presence of β-galactosidase in the serous component of the submucosal glands. We then looked for other lysosomal activities in normal and CF-HTGS cells. We showed that normal cells already have elevated enzyme values and that CF-HTGS cells contained 2–4-fold more β-galactosidase, α-fucosidase, α-mannosidase and β-glucuronidase activities than normal cells. An analysis of their kinetic constants has shown that this difference could be attributed to a lower Km of CF lysosomal enzymes. More importantly, these differences are eliminated after adenovirus-mediated CFTR gene transfer and not after β-galactosidase gene transfer

Abnormal spatial diffusion of Ca2+ in F508del-CFTR airway epithelial cells

<p>Abstract</p> <p>Background</p> <p>In airway epithelial cells, calcium mobilization can be elicited by selective autocrine and/or paracrine activation of apical or basolateral membrane heterotrimeric G protein-coupled receptors linked to phospholipase C (PLC) stimulation, which generates inositol 1,4,5-trisphosphate (IP₃) and 1,2-diacylglycerol (DAG) and induces Ca²⁺release from endoplasmic reticulum (ER) stores.</p> <p>Methods</p> <p>In the present study, we monitored the cytosolic Ca²⁺transients using the UV light photolysis technique to uncage caged Ca²⁺or caged IP₃into the cytosol of loaded airway epithelial cells of cystic fibrosis (CF) and non-CF origin. We compared in these cells the types of Ca²⁺receptors present in the ER, and measured their Ca²⁺dependent activity before and after correction of F508del-CFTR abnormal trafficking either by low temperature or by the pharmacological corrector miglustat (N-butyldeoxynojirimycin).</p> <p>Results</p> <p>We showed reduction of the inositol 1,4,5-trisphosphate receptors (IP₃R) dependent-Ca²⁺response following both correcting treatments compared to uncorrected cells in such a way that Ca²⁺responses (CF+treatment <it>vs </it>wild-type cells) were normalized. This normalization of the Ca²⁺rate does not affect the activity of Ca²⁺-dependent chloride channel in miglustat-treated CF cells. Using two inhibitors of IP₃R1, we observed a decrease of the implication of IP₃R1 in the Ca²⁺response in CF corrected cells. We observed a similar Ca²⁺mobilization between CF-KM4 cells and CFTR-cDNA transfected CF cells (CF-KM4-reverted). When we restored the F508del-CFTR trafficking in CFTR-reverted cells, the specific IP₃R activity was also reduced to a similar level as in non CF cells. At the structural level, the ER morphology of CF cells was highly condensed around the nucleus while in non CF cells or corrected CF cells the ER was extended at the totality of cell.</p> <p>Conclusion</p> <p>These results suggest reversal of the IP₃R dysfunction in F508del-CFTR epithelial cells by correction of the abnormal trafficking of F508del-CFTR in cystic fibrosis cells. Moreover, using CFTR cDNA-transfected CF cells, we demonstrated that abnormal increase of IP₃R Ca²⁺release in CF human epithelial cells could be the consequence of F508del-CFTR retention in ER compartment.</p

Azithromycin reduces spontaneous and induced inflammation in ΔF508 cystic fibrosis mice

BACKGROUND: Inflammation plays a critical role in lung disease development and progression in cystic fibrosis. Azithromycin is used for the treatment of cystic fibrosis lung disease, although its mechanisms of action are poorly understood. We tested the hypothesis that azithromycin modulates lung inflammation in cystic fibrosis mice. METHODS: We monitored cellular and molecular inflammatory markers in lungs of cystic fibrosis mutant mice homozygous for the ΔF508 mutation and their littermate controls, either in baseline conditions or after induction of acute inflammation by intratracheal instillation of lipopolysaccharide from Pseudomonas aeruginosa, which would be independent of interactions of bacteria with epithelial cells. The effect of azithromycin pretreatment (10 mg/kg/day) given by oral administration for 4 weeks was evaluated. RESULTS: In naive cystic fibrosis mice, a spontaneous lung inflammation was observed, characterized by macrophage and neutrophil infiltration, and increased intra-luminal content of the pro-inflammatory cytokine macrophage inflammatory protein-2. After induced inflammation, cystic fibrosis mice combined exaggerated cellular infiltration and lower anti-inflammatory interleukin-10 production. In cystic fibrosis mice, azithromycin attenuated cellular infiltration in both baseline and induced inflammatory condition, and inhibited cytokine (tumor necrosis factor-α and macrophage inflammatory protein-2) release in lipopolysaccharide-induced inflammation. CONCLUSION: Our findings further support the concept that inflammatory responses are upregulated in cystic fibrosis. Azithromycin reduces some lung inflammation outcome measures in cystic fibrosis mice. We postulate that some of the benefits of azithromycin treatment in cystic fibrosis patients are due to modulation of lung inflammation

La interculturalitat dins de les aules d'Educació Primària

[cat] En la societat actual, s’observen molts de moviments migratoris al voltant de tot el món i això provoca societats cada vegada més multiculturals. Concretament a Espanya, durant els darrers anys s’ha viscut l’arribada de persones procedents de diferents països del món. Aquest fet fa que a les escoles hi trobem alumnat de diferents nacionalitats i cultures minoritàries, els quals poden tenir problemes en el seu desenvolupament de la socialització i integració amb els seus companys a causa de les diferències culturals. A les escoles és important treballar aquesta diversitat cultural i fomentar una educació intercultural dins de les aules. L’objectiu d’aquesta educació intercultural és la socialització de la diversitat cultural de l’alumnat i fer que la cultura majoritària conegui de manera més propera les altres cultures afavorint un ambient de compassió, convivència i comprensió de les diverses cultures. Considerant la importància de la educació intercultural, concretament a les aules de primària, he realitzat aquest treball sobre aquests conceptes en el qual es troba una part teòrica sobre aquests diversos conceptes i posteriorment una proposta d’intervenció a les aules d’Educació Primària.[eng] In today's society, many migratory movements are observed around the world and this causes increasingly multicultural societies. Specifically in Spain, the arrival of people from different countries of the world has been experienced in recent years. This makes schools available to students of different nationalities and minority cultures, which may have problems in their development of socialization and integration with their classmates due to cultural differences. In schools it is important to work on this cultural diversity and promote intercultural education within classrooms. The objective of this intercultural education is the socialization of the cultural diversity of the students and make the majority culture know more closely the other cultures, favoring an atmosphere of compassion, coexistence and understanding of the different cultures. Considering the importance of intercultural education, specifically in elementary classrooms, I have done this work on these concepts in which there is a theoretical part about these various concepts and subsequently a proposal for intervention in the Primary Education classrooms

Etude de la précipitation, de la microstructure et de leurs influences sur le taux et la stabilité de l'austénite résiduelle dans des aciers TRIP

Les aciers TRIP (TRansformation Induced Plasticity) of&ent des propriétés très intéressantes qui les rendent idéals pour l'emboutissage des tôles minces destinées à l'industrie automobile. Sous déformation, l'austenite résiduelle (fraction volumique - 5 à 15 % ) se transforme en martensite, donnant ainsi une ductilité remarquable et, en final, un acier plus dur. Néanmoins, pour certaines applications, ii est souhaitable d'augmenter encore la dureté de ces aciers en rajoutant un ou plusieurs éléments dispersoides, comme le titane, qui durcit îa matrice molle (ferrite) par une précipitation fine. L'objectif de ce travail de thèse est de mieux comprendre l'effet de la précipitation (consommation du carbone) et de la microstructure (répartition du carbone) sur le taux et la stabilité de l'austenite résiduelle dans cinq aciers TRIP différents. Les outils principaux sont la microscopie électronique en transmission et à balayage et leurs techniques associées. Pour la préparation des échantillons MET, le Focused Ion Beam (FIB) s'est avéré être la technique la plus adaptée pour une localisation précise de l'austenite résiduelle. Nous avons montré que même si la précipitation à base de titane a participé d'une manière considérable à l'augmentation des résistances mécaniques, elle n'a pas d'effet direct sur le taux et la stabilité de l'austenite résiduelle. En effet, la stabilité de cette dernière est étroitement liée à la nature des phases formées lors de l'entrée dans le domaine bainitique. Enfin, une corrélation microstructure/propriétés a montré que la réponse mécanique de l'austenite résiduelle est le facteur déterminant de la limite d'élasticité et donc de l'aptitude à l'emboutissage. Effectivement, pour une précipitation identique et une morphologie bainitique et microstructurale équivalente, mais une austénite résiduelle à deux taux de carbone différents, la limite d'élasticité peut varier de plusieurs centaines de MPa.AIX-MARSEILLE3-BU Sc.St Jérô (130552102) / SudocSudocFranceF