Massive OB stars are very influential objects in the ecology of galaxies like our own. Current catalogues of Galactic OB stars are heavily biased towards bright (g < 13) objects, only typically including fainter objects when found in prominent star clusters (Garmany et al., 1982; Reed, 2003; Maíz-Apellániz et al., 2004). Exploitation of the VST Photometric Hα Survey (VPHAS+) allows us to build a robust catalogue of photometrically-selected OB stars across the entire Southern Galactic plane, both within clusters and in the field, down to ~20th magnitude in g. For the first time, a complete accounting of the OB star runaway phenomenon becomes possible. Along with making the primary selection using VPHAS+ colours, I have performed Markov-Chain Monte Carlo fitting of the spectral energy distributions of the selected stars by combining VPHAS+ u, g, r, i with published J, H, K photometry. This gives rough constraints on effective temperature and distance, whilst delivering much more precise reddening parameters A0 and RV - allowing us to build a much richer picture of how extinction and extinction laws vary across the Galactic Plane. My thesis begins with a description of the method of photometric selection of OB star candidates and its validation across a 2 square degree field including the well- known young massive star cluster Westerlund 2 (Mohr-Smith et al., 2015)1. Following on from this I present spectroscopy with AAOmega of 283 candidates identified by our method, which confirms that ~ 94% of the sample are the expected O and early B stars. I then develop this method further and apply it to a Galactic Plane strip of 42 square-degrees that runs from the Carina Arm tangent region to the much studied massive cluster in NGC 3603. A new aspect I attend to in this expansion of method is tightening up the uniform photometric calibration of the data, paying particular attention to the always-challenging u band. This leads to a new and reliable catalogue of 5915 OB stars. As well as increasing the numbers of identified massive stars in this large region of the sky by nearly an order of magnitude, a more complete picture of massive star formation in the Carina Arm has emerged. I have found a broad over-density of O stars around the highly luminous cluster NGC 3603 and have uncovered two new candidate OB clusters/associations. I have also paired up the ionization sources of a number of HII regions catalogued by the RMS survey. It is also shown that the OB star scale- height can serve as a roughly standard ruler, leading to the result that the OB star layer shows the onset of warping at RG ~ 10kpc. My results confirm that this entire region requires a non-standard (3.5 < RV < 4.0) reddening law for distances greater than ~2kpc. The methods developed in this study are ready to roll out across the rest of the VPHAS+ footprint that has been observed to date. This extension will take in a strip ~ ±2 degrees across the entire Southern Galactic mid-plane (a sky area of over 700 square degrees), within which we expect to find the majority of massive OB stars. This will result in the largest catalogue of Galactic OB stars to date.

Abstract The CAS-OB (composition adjustment by sealed argon bubbling - oxygen blowing) process is used in secondary steelmaking to adjust the composition and temperature of the steel melt. The steel melt can be heated by oxidizing aluminium in process which feeds aluminium particles and oxygen to the melt surface. Oxygen is in fed by a top lance, which is an important part of many metallurgical processes and is typically used to deliver oxygen to steel melt surface by supersonic blowing. Because observing and measuring the metallurgical processes is challenging due to the high temperature, numerical models predicting the processes are especially important. In this thesis, both top lances and the heat-up stage of the CAS-OB process were studied, and numerical models were constructed. CFD (computational fluid dynamics) were used to study top lances. A turbulence model was adjusted for supersonic flows with experimental data from literature. The CAS-OB process model involves chemical reactions and fluid flows. In order to keep the computation times reasonable, a full fluid flow calculation is not included in the model but is calculated in advance. Heat and mass transfer correlations are calculated with CFD, and the results are then used in the process simulation model. Chemical reactions are calculated based on the law of mass action and thermodynamics. The results were validated with industrial measurements. The CAS-OB heat-up stage model can be used in its current state in process development, and in the future for online control of the process. The CFD model for the top lance can be applied to a lance in any other process. / Tiivistelmä CAS-OB-prosessia (composition adjustment by sealed argon bubbling - oxygen blowing) käytetään teräksen valmistuksessa sulan teräksen koostumuksen ja lämpötilan säätämiseen. Terässulaa voidaan tarvittaessa lämmittää syöttämällä alumiinikappaleita ja happea sulan pinnalle. Hapen syöttö tapahtuu yliäänilanssilla, jota käytetään monissa metallurgisissa prosesseissa, yleensä toimittamaan happea sulan pinnalle yliäänisellä puhalluksella. Metallurgisten prosessien havainnointi ja mittaaminen ovat haastavia korkeiden lämpötilojen vuoksi, joten numeeriset mallit ovat erityisen tärkeitä prosessien ennustamisessa. Tässä työssä on tutkittu yliäänilansseja ja CAS-OB-prosessin lämmitysvaihetta ja luotu niille numeeriset mallit. Yliäänilanssien tutkimiseen käytettiin numeerista virtauslaskentaa (CFD, computational fluid dynamics). Lanssien mallinnusta varten olemassa olevaa turbulenssimallia muokattiin paremmin yliäänivirtausta kuvaavaksi kirjallisuudesta löytyvän mittaustiedon perusteella. CAS-OB-prosessimallissa huomioidaan virtaus ja kemialliset reaktiot. Koska laskenta-ajat haluttiin pitää käytännöllisinä, virtauslaskentaa ei suoriteta mallissa, vaan se tehdään etukäteen. Aineen- ja lämmönsiirtokertoimet lasketaan CFD-laskennalla, ja tuloksia käytetään prosessimallissa. Kemialliset reaktiot lasketaan perustuen massavaikutuksen lakiin ja termodynamiikkaan. CAS-OB-mallin tulokset on validoitu terästehtaalla tehtyjen kokeiden perusteella. Mallia voidaan käyttää nykyisessä muodossaan prosessin kehityksessä ja tulevaisuudessa myös prosessin ohjauksessa. Yliäänilanssin CFD-mallia voidaan soveltaa myös muihin metallurgisiin prosesseihin.

Abstract Composition Adjustment by Sealed argon bubbling – Oxygen Blowing (CAS-OB) process is a ladle treatment process that was developed for chemical heating and alloying of steel. The main stages of the process are heating, (possible) alloying and reduction of slag. The CAS-OB process aims for homogenization and control of the composition and temperature of steel. In this dissertation, a mathematical reaction model was developed for the slag reduction stage of the CAS-OB process. Slag reduction is carried out by applying vigorous bottom stirring from porous plugs at the bottom of the ladle. Due to the bottom stirring a circular steel flow is induced which disrupts top slag layer and due to shear stress at the steel-slag interface small slag droplets are detached. This induces an immense increase in the interfacial area between steel and slag which provides favourable conditions for the reduction reactions. In order to model reduction reactions, a sub-model for describing the interfacial area was needed. For this the slag droplet formation was studied using computational fluid dynamics (CFD) and based on these studies, a sub-model for droplet formation was developed. The model for the reduction stage of the CAS-OB process takes into account not only the reaction during the reduction but also the heat transfer between the phases, ladle and surroundings. The reduction stage model was validated using validation data obtained from the CAS-OB station of the SSAB Raahe steel plant in Finland. It was concluded that the model was able to predict steel and slag compositions as well as the steel temperature very well. / Tiivistelmä CAS-OB -prosessi on sulametallurgiassa käytettävä senkkakäsittelyprosessi, joka on kehitetty teräksen kemialliseen lämmittäseen ja seostukseen. CAS-OB-prosessin pääprosessivaiheet ovat lämmitysvaihe, mahdollinen seostusvaihe ja kuonan pelkistysvaihe. CAS-OB -prosessilla tavoitellaan teräksen koostumuksen homogenisointiin ja lämpötilan kontrollointiin. Tässä tutkimuksessa kehitettiin matemaattinen reaktiomalli CAS-OB -prosessin kuonan pelkistysvaiheen kuvaamiseen. Kuonan pelkistys tapahtuu senkan pohjassa olevien huuhtelutiilien avulla suoritettavan voimakkaan kaasuhuuhtelun avulla. Pohjahuuhtelu aiheuttaa kiertävän teräsvirtauksen senkassa. Teräsvirtaus irrottaa teräksen päällä olevasta kuonakerroksesta pisaroita ja kuonan ja teräksen välinen reaktiopinta-ala kasvaa voimakkaasti. Tämä tarjoaa suotuisat olosuhteet pelkistysreaktiolle senkassa. Pelkistysreaktioiden mallintamiseksi tässä työssä kehitettiin CFD-simulaatioiden avulla alimalli, jonka avulla voidaan kuvata teräksen ja kuonan välisen pinta-alan suuruutta. Pelkistysvaiheen mallissa huomioidaan reaktioiden lisäksi myös systeemissä tapahtuva lämmösiirto. Pelkistysmalli validoitiin mittausdatalla, joka hankittiin SSAB Raahen terässulaton CAS-OB -asemalla järjestetyssä validointikampanjassa. Tutkimuksessa havaittiin, että malli kykenee hyvin ennustamaan teräksen ja kuonan koostumuksen sekä teräksen lämpötilan.

No description available.

Die meisten Cyanobakterien und Chloroplasten von Rotalgen verfügen über Phycobilisomen, große lichtsammelnde Multiproteinkomplexe, die an der cytoplasmatischen Seite der Thylakoidmembran gebunden vorliegen. Unter stickstofflimitierten Bedingungen werden die Phycobilisomen proteolytisch abgebaut. Dieser Prozess schützt vor Fotoschäden unter den gegebenen Stressbedingungen und liefert gleichzeitig einen großen Vorrat an Stickstoff enthaltenden Substanzen. Das Gen nblA, welches in allen Phycobilisomen enthaltenden Organismen vorkommt, kodiert für ein Polypeptid von ungefähr 7 kDa, das eine Schlüsselrolle im Abbau der Phycobilisomen einnimmt. Die Wirkungsweise von NblA dabei wird jedoch bisher kaum verstanden. Ein Selenomethioninderivat von NblA des filamentösen Cyanobakteriums Anabaena sp. PCC 7120 wurde in Escherichia coli rekombinant hergestellt, gereinigt und kristallisiert. Die Röntgenkristallstruktur von NblA wurde mithilfe der single-wavelength anomalous dispersion-Methode bis zu einer Auflösung von 1,8 Angstrom bestimmt. Das finale Modell verfügt über einen kristallographischen R-Wert von 18,2% und einen freien R-Wert von 21,7%. Das kleine NblA-Protein von 65 Aminosäuren besteht aus zwei alpha-Helices, die in einem ca. 37°-Winkel in einer antiparallelen, V-förmigen Anordnung zueinander stehen. Zwei dieser Monomere bilden die grundlegende strukturelle Einheit von NblA, ein vier-Helix-Bündel, bei dem sich die Spitzen der "Vs" auf der gleichen Seite des Dimers überlagern und über eine nicht-kristallographische zweizählige Achse miteinander verknüpft sind. Auf der Grundlage von Bindungsstudien und der Kenntnis der NblA-Struktur konnte ein Modell für die Bindung von NblA an die Phycobilisomenstruktur postuliert werden. / Cyanobacterial light harvesting complexes, the phycobilisomes, are proteolytically degraded when the organisms are starved for combined nitrogen, a process referred to as chlorosis or bleaching. Gene nblA, present in all phycobilisome-containing organisms, encodes a protein of about 7 kDa that plays a key role in phycobilisome degradation. To gain deeper insights into the mode of action of NblA in this degradation process the crystal structure of NblA was determined and a model of its binding to phycobilisomes was proposed. For this purpose, NblA from Anabaena sp. PCC 7120 was produced as selenomethionine NblA derivative in Escherichia coli B834 (DE3), purified and crystalized. NblA crystals grew as long, but thin rods and belong to the monoclinic space group P2(1) with cell parameters of a = 43.2 A, b = 95.9 A, c = 104.8 A and Beta = 97.0°. They contain twelve NblA monomers in the asymmetric unit. The crystal structure with a resolution of 1.8 A was determined using the single-wavelength anomalous dispersion (SAD) technique and refined to final values for Rwork and Rfree of 0.182 and 0.217, respectively. The small NblA polypeptide of 65 amino acids consists of two alpha-helices which are assembled at a ~37° angle in an antiparallel, V-shaped arrangement. Two NblA monomers form the basic structural unit of NblA, a four-helix bundle with the tips of the V superimposing on the same side of a dimer. The dimer is formed by two molecules related by a non-crystallographic dyad axis. Based on the crystal structure presented here, pull-down experiments, and peptide scan data a model of binding of NblA to phycobilisomes was proposed. Considering the entire phycobilisome structure, NblA is predicted to bind via its amino acids Leu51 and Lys53 to the trimer-trimer interface of the phycobiliprotein hexamers.

<p>Oscillations in cytoplasmic Ca²⁺ concentration ([Ca²⁺]_i) is the key signal in glucose-stimulated β-cells governing pulsatile insulin release. The glucose response of mouse β-cells is often manifested as slow oscillations and rapid transients of [Ca²⁺]_i. In the present study, microfluorometric technique was used to evaluate the role of amino acids, glucagon, ryanodine and caffeine on the generation and maintenance of [Ca²⁺]_i oscillations and transients in individual murine β-cells and isolated mouse pancreatic islets. The amino acids glycine, alanine and arginine, at around their physiological concentrations, transformed the glucose-induced slow oscillations of [Ca²⁺]_i in isolated mouse β-cells into sustained elevation. Increased Ca²⁺ entry promoted the reappearance of the slow [Ca²⁺]_i oscillations. The [Ca²⁺]_i oscillations were more resistant to amino acid transformation in intact islets, supporting the idea that cellular interactions are important for maintaining the oscillatory activity. Individual rat β-cells responded to glucose stimulation with slow [Ca²⁺]_i oscillations due to periodic entry of Ca²⁺ as well as with transients evoked by mobilization of intracellular stores. The [Ca²⁺]_i oscillations in rat β-cells had a slightly lower frequency than those in mouse β-cells and were more easily transformed into sustained elevation in the presence of glucagon or caffeine. The transients of [Ca²⁺]_i were more common in rat than in mouse β-cells and often appeared in synchrony also in cells lacking physical contact. Depolarization enhanced the generation of [Ca²⁺]_i transients. In accordance with the idea that β-cells have functionally active ryanodine receptors, it was found that ryanodine sometimes restored oscillatory activity abolished by caffeine. However, the IP3 receptors are the major Ca²⁺ release channels both in β-cells from rats and mice. Single β-cells from ob/ob mice did not differ from those of lean controls with regard to frequency, amplitudes and half-widths of the slow [Ca²⁺]_i oscillations. Nevertheless, there was an excessive firing of [Ca²⁺]_i transients in the β-cells from the ob/ob mice, which was suppressed by leptin at close to physiological concentrations. The enhanced firing of [Ca²⁺]_i transients in ob/ob mouse β-cells may be due to the absence of leptin and mediated by activation of the phospholipase C signaling pathway.</p>

Oscillations in cytoplasmic Ca2+ concentration ([Ca2+]i) is the key signal in glucose-stimulated β-cells governing pulsatile insulin release. The glucose response of mouse β-cells is often manifested as slow oscillations and rapid transients of [Ca2+] i. In the present study, microfluorometric technique was used to evaluate the role of amino acids, glucagon, ryanodine and caffeine on the generation and maintenance of [Ca2+] i oscillations and transients in individual murine β-cells and isolated mouse pancreatic islets. The amino acids glycine, alanine and arginine, at around their physiological concentrations, transformed the glucose-induced slow oscillations of [Ca2+] i in isolated mouse β-cells into sustained elevation. Increased Ca2+ entry promoted the reappearance of the slow [Ca2+] i oscillations. The [Ca2+] i oscillations were more resistant to amino acid transformation in intact islets, supporting the idea that cellular interactions are important for maintaining the oscillatory activity. Individual rat β-cells responded to glucose stimulation with slow [Ca2+] i oscillations due to periodic entry of Ca2+ as well as with transients evoked by mobilization of intracellular stores. The [Ca2+] i oscillations in rat β-cells had a slightly lower frequency than those in mouse β-cells and were more easily transformed into sustained elevation in the presence of glucagon or caffeine. The transients of [Ca2+] i were more common in rat than in mouse β-cells and often appeared in synchrony also in cells lacking physical contact. Depolarization enhanced the generation of [Ca2+] i transients. In accordance with the idea that β-cells have functionally active ryanodine receptors, it was found that ryanodine sometimes restored oscillatory activity abolished by caffeine. However, the IP3 receptors are the major Ca2+ release channels both in β-cells from rats and mice. Single β-cells from ob/ob mice did not differ from those of lean controls with regard to frequency, amplitudes and half-widths of the slow [Ca2+] i oscillations. Nevertheless, there was an excessive firing of [Ca2+] i transients in the β-cells from the ob/ob mice, which was suppressed by leptin at close to physiological concentrations. The enhanced firing of [Ca2+] i transients in ob/ob mouse β-cells may be due to the absence of leptin and mediated by activation of the phospholipase C signaling pathway.

Diese Arbeit beinhaltet Untersuchungen zu den unterschiedlichen Isoformen des Leptinrezeptors und dessen Regulation in primären humanen Hepatozyten. Leptin und der Leptinrezeptor nehmen in der Physiologie des menschlichen Energiehaushaltes eine wesentliche Funktion ein und sind an der Pathogenese der Adipositas mit Folgeerkrankungen wie der Entwicklung einer Fettleber beteiligt. Es wird erstmalig geprüft inwieweit das Modellsystem primärer humaner Hepatozyten für Analysen der Leptinrezeptor-Expression und der Abspaltung von löslichem Leptinrezeptor geeignet ist. Weiterhin werden untersucht, welchen Einfluss endokrine Regulatoren wie Dexamethason, Leptin und Glucagon auf die isoformspezifischen Rezeptormengen in primären Hepatozyten haben und wie der Rezeptor unter Apoptose reguliert ist, welche durch die lipotoxischen Effekte der freien Fettsäure Palmitat und den Apoptoseinduktor Staurosporin induziert wird. Hierdurch können Rückschlüsse auf eine möglicherweise veränderte Wirksamkeit des Leptins in der Leber gezogen werden.

No description available.

Les études cliniques et in vitro suggèrent que la sclérose de l’os sous-chondral due aux ostéoblastes (Ob) anormaux est impliquée dans la progression de l’ostéoarthrose (OA). Les Ob OA humains isolés à partir d’os sous-chondral sclérosé montrent un phénotype altéré, un niveau réduit de signalisation Wnt/β-caténine canonique et une minéralisation in vitro réduite. Il existe également deux voies non-canoniques, Wnt/PKC et Wnt/PCP qui ont étés décrites dans la littérature. Cependant, il n’existe aucune étude qui traite de ces deux voies dans les Ob OA. Ces voies sont activées après qu’un ligand Wnt non-canonique tel que Wnt-5a se lie à un récepteur Wnt couplé à des corécepteurs de la voie non-canonique. Ceci enclenche, respectivement pour la voie Wnt/PKC-Ca2+ et Wnt/PCP, la phosphorylation de PKC (p-PKC) et la phosphorylation de JNK (p-JNK) et agit sur les cibles en aval. Nous avons voulu déterminer s’il était possible de constater des altérations dans les voies Wnt non-canoniques dans les Ob OA. Nous avons préparé des cultures primaires d’ostéoblastes sous-chondral humains à partir de plateaux tibiaux de patients OA subissant une arthroplastie totale du genou, ainsi qu’à partir de plateaux tibiaux recueillis à l’autopsie de patients « normaux ». L’expression des gènes impliqués dans les voies Wnt/PKC et Wnt/PCP a été évaluée par RT-qPCR et la production par Western Blot des protéines, ainsi que celle de p-PKC et p-JNK et que l’activité des facteurs NFAT et AP-1 utilisés par ces deux voies. L’activité phosphatase alcaline (ALPase) et la quantité d’ostéocalcine (OC) ont étés évaluées respectivement à l’aide d’hydrolyse de substrat et d’ELISA. Le niveau de minéralisation a été évalué par la coloration au rouge Alizarine. Nos résultats montrent que l’expression et la production de Wnt-5a étaient augmentées dans les Ob OA comparées aux Ob N et LGR5 était significativement plus élevée. De plus, l’expression de LGR5 est directement régulée via la stimulation ou la diminution de Wnt-5a, à la fois au niveau de l’ARNm et des protéines. Par ailleurs, Wnt-5a a stimulé la phosphorylation de JNK et de PKC ainsi que l’activité NFAT et AP-1. Les niveaux de minéralisation ainsi que d’activité ALPase et de sécrétion d’OC ont aussi été affectés par les changements du niveau de Wnt-5a. Ces résultats suggèrent que Wnt-5a, qui est augmentée dans les OA Ob, peut stimuler les voies Wnt non-canoniques et affecter le phénotype et la minéralisation des OA Ob humains. / Clinical and in vitro studies suggest that subchondral bone sclerosis due to abnormal osteoblasts (Ob) is involved in the progression and/or onset of osteoarthritis (OA). Human Ob isolated from sclerotic subchondral OA bone tissue show an altered phenotype, a decreased canonical Wnt/ß-catenin signaling pathway (cWnt), and a reduced mineralization in vitro. Besides the cWnt pathway, at least two non-canonical signaling pathways, the Wnt/PKC and Wnt/PCP pathway have been described. These pathways are activated when a non-canonical Wnt ligand like Wnt-5a binds to a Wnt receptor coupled with non-canonical co-receptors. This activates, respectively for Wnt/PKC-Ca2+ and Wnt/PCP, the phosphorylation of PKC (pPKC) and the phosphorylation of JNK (pJNK) and their effect on downstream targets. However, there are no reports of either pathway in OA Ob. Here, we studied if alterations of the non-canonical pathways could be observed in OA Ob. We prepared primary human subchondral Ob using the tibial plateaus of OA patients undergoing total knee arthroplasty, or from tibial plateaus of normal individuals at autopsy. The expression of genes involved in Wnt/PKC and Wnt/PCP was evaluated by RT-qPCR and their protein production by Western blot analysis, in addition to p-PKC and p-PCP and NFAT and AP-1 activity with luciferase. Alkaline phosphatase activity and osteocalcin levels were evaluated respectively by substrate hydrolysis and ELISA respectively, and mineralization levels were evaluated with Alizarin red staining. OA Ob showed an increased alkaline phosphatase activity and osteocalcin release. The expression of Wnt5a was increased in OA Ob compared to normal. The expression of LGR5 was significantly increased in these cells. Moreover, the expression and production of LGR5 is directly modulated via the stimulation or inhibition of Wnt5a. However, Wnt5a did not stimulate the expression of LGR4. Wnt5a increased the phosphorylation of PKC and JNK as well as NFAT and AP-1 activity. Mineralization levels as well as alkaline phosphatase activity and osteocalcin secretion levels were also linked with changes in Wnt-5a levels. These data indicate that Wnt5a, which is increased in OA Ob, can directly stimulate the Wnt/PKC and Wnt/PCP pathways and this can affect the phenotype and mineralization observed in human OA Ob.