Die Arbeit beschäftigt sich mit dem Problem der Fettlebererkrankung. In der Einleitung wird auf die aktuelle Relevanz der Gesundheitsstörung und Therapiemöglichkeiten eingegangen, insbesondere durch das, in der Therapie des Diabetes mellitus Typ 2 gebräuchliche Biguanid Metformin. Der Bezug zu molekularbiologischen Signalwegen wird hergestellt und verschiedene in vitro Modellsysteme werden vorgestellt. Anschließend wird auf die Herkunft und genetische Besonderheiten der verwendeten primären Maushepatozyten und Hepatomzellen eingegangen, bevor die angewandten Methoden vorgestellt werden. Zum Einsatz kam in dieser Arbeit vor allem die Lipidmessung mittels Fettrot, um das Ausmaß an Steatosis quantifizierbar zu machen. Im Ergebnisteil folgen zuerst Versuche zur Zytotoxizität der einzelnen Chemikalien und deren Einfluss auf intrazelluläre Energieniveaus, bevor der Einfluss auf die hepatozellulären Fetteinlagerungen im Detail untersucht wird. Unterstützt werden die Ergebnisse durch mikroskopische Bilder der Hepatozyten, welche die beschriebenen Effekte verdeutlichen. Insgesamt konnten folgende Thesen aufgestellt werden: • Zwischen primären Hepatozyten von Wildtyp- und Knockout-Mäusen, bestehen nach 24 stündiger Kultivierung Unterschiede bezüglich des intrazellulären Lipidgehaltes, welche sich nach 72 stündiger Kultivierungszeit nivellieren. • Metformin- und Fructoseinkubation senken den intrazellulären ATP-Gehalt, gleichzeitige Anwesenheit von Metformin und Glucose vermindern den Effekt. • Durch 72-stündige Inkubation der primären Hepatozyten und Behandlung mit Metformin konnte der intrazelluläre Lipidgehalt um circa 40% gesenkt werden. • Durch 72-stündige Inkubation der primären Hepatozyten mit Glucose konnte der intrazelluläre Lipidgehalt um circa 100% gesteigert werden. • Bei humanen Hepatomzellen (HuH7) konnte kein Metformin- und kein Glucoseeffekt beobachtet werden. • Der LXR-Agonist TO901317 wirkt auf den intrazellulären Lipidgehalt Metformin entgegen.

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Bakgrund: Postoperativa sårinfektioner orsakar lidande för den drabbade patienten och kostnader för patienten, vården och samhället. Intraoperativ kontamination av operationsinstrumenten är en av källorna till dessa infektioner. Operationssjuksköterskan är ansvarig för aseptiken och operationsinstrumenten.   Syfte: Syftet med denna pilotstudie var att mäta graden av bakteriell kontamination över tid på operationsinstrument som används vid öppen neurokirurgi samt observera utvalda källor till kontamination under samma tid. Metod: Denna studie designades som en kvantitativ icke-experimentell observationsstudie. Datainsamlingen skedde under hösten 2015 och bestod av omhändertagandet av specifika instrument som använts av operatören under operationen efter förutbestämd åtgången knivtid. Bakterieodlingar på dessa instrument skedde sedan på laboratorium enligt en förutbestämd metod och graden av kontamination mättes genom observation av mängden colony forming units (CFU). Resultaten presenteras i form av förändringar av mängden CFU. Resultat: Fem operationer observerades och tio odlingar utfördes på tio instrument. Totalt tio CFU registrerades under samtliga observationer. Ingen signifikant ökning av antal CFU kunde observeras från en till två timmars knivtid (P=0,156). Antal dörröppningar ökade från 3,4 till 9,0 och antal personer på operationssalen ökade från 5,4 till 5,8. Slutsats: Trots det ringa antal observationer som ingick i denna studie tyder resultaten på god aseptik och minimal bakteriell kontamination på operationsinstrumenten under operationens två första timmar. Ytterligare forskning med längre observationer, fler observationer och observationer kring annan typ av kirurgi behövs. / Background: Post-operative surgical site infections (SSI) cause suffering for the afflicted patient and is a great cost for the patient, the health care system and society. Intra-operative contamination of surgical instruments is one of the sources of these infections. The operating room nurse is responsible for preventing infections. Purpose: The purpose of this pilot-study was to measure the degree of bacterial contamination over time on surgical instruments used in open neurosurgery as well as to observe specific sources of contamination during this time. Method: The design was a quantitative nonexperimental observational study. The data collection took place during autumn 2015 and consisted of the sampling of specific instruments used by the surgeon during surgery after preset elapsed operating times. Bacterial culturing on these instruments was then carried out at a laboratory according to a preset routine and the degree of contamination was measured by observing the amount of colony forming units (CFU). The results are presented as changes in CFU. Results: Five operations were observed and ten bacterial culturings were conducted on ten instruments. A total of ten CFU was registered during all observations. No significant increase in the number of CFU could be observed from one to two hours of elapsed surgery (P=0,156). The amount of door openings increased from 3,4 to 9,0 and the number of personnel in the operating room increased from 5,4 to 5,8. Conclusion: Despite the small number of observations included in this study the results indicate good aseptics and minimal bacterial contamination on the surgical instruments during the first two hours of surgery. Further research with longer observations, more observations and observations on other forms of surgery is needed.

Concern regarding the health effects of indoor air quality has grown in recent years, due to the increased prevalence of many diseases, as well as the fact that many people now spend most of their time indoors. While numerous studies have reported on the dynamics of aerosols indoors, the dynamics of bioaerosols in indoor environments are still poorly understood and very few studies have focused on fungal spore dynamics in indoor environments. Consequently, this work investigated the dynamics of fungal spores in indoor air, including fungal spore release and deposition, as well as investigating the mechanisms involved in the fungal spore fragmentation process. In relation to the investigation of fungal spore dynamics, it was found that the deposition rates of the bioaerosols (fungal propagules) were in the same range as the deposition rates of nonbiological particles and that they were a function of their aerodynamic diameters. It was also found that fungal particle deposition rates increased with increasing ventilation rates. These results (which are reported for the first time) are important for developing an understanding of the dynamics of fungal spores in the air. In relation to the process of fungal spore fragmentation, important information was generated concerning the airborne dynamics of the spores, as well as the part/s of the fungi which undergo fragmentation. The results obtained from these investigations into the dynamics of fungal propagules in indoor air significantly advance knowledge about the fate of fungal propagules in indoor air, as well as their deposition in the respiratory tract. The need to develop an advanced, real-time method for monitoring bioaerosols has become increasingly important in recent years, particularly as a result of the increased threat from biological weapons and bioterrorism. However, to date, the Ultraviolet Aerodynamic Particle Sizer (UVAPS, Model 3312, TSI, St Paul, MN) is the only commercially available instrument capable of monitoring and measuring viable airborne micro-organisms in real-time. Therefore (for the first time), this work also investigated the ability of the UVAPS to measure and characterise fungal spores in indoor air. The UVAPS was found to be sufficiently sensitive for detecting and measuring fungal propagules. Based on fungal spore size distributions, together with fluorescent percentages and intensities, it was also found to be capable of discriminating between two fungal spore species, under controlled laboratory conditions. In the field, however, it would not be possible to use the UVAPS to differentiate between different fungal spore species because the different micro-organisms present in the air may not only vary in age, but may have also been subjected to different environmental conditions. In addition, while the real-time UVAPS was found to be a good tool for the investigation of fungal particles under controlled conditions, it was not found to be selective for bioaerosols only (as per design specifications). In conclusion, the UVAPS is not recommended for use in the direct measurement of airborne viable bioaerosols in the field, including fungal particles, and further investigations into the nature of the micro-organisms, the UVAPS itself and/or its use in conjunction with other conventional biosamplers, are necessary in order to obtain more realistic results. Overall, the results obtained from this work on airborne fungal particle dynamics will contribute towards improving the detection capabilities of the UVAPS, so that it is capable of selectively monitoring and measuring bioaerosols, for which it was originally designed. This work will assist in finding and/or improving other technologies capable of the real-time monitoring of bioaerosols. The knowledge obtained from this work will also be of benefit in various other bioaerosol applications, such as understanding the transport of bioaerosols indoors.

A sister group of the Arthropoda, the Tardigrada are micrometazoans that occupy a diversity of niches in freshwater, marine, and terrestrial habitats. Commonly called water bears because of their slow, lumbering gait, these molting lobopods have four pairs of legs, usually terminating in claws. Most are less than 1 mm in length, with a complete digestive tract, a dorsal gonad with one or two gonoducts, and a dorsal lobed brain with a ventral nerve cord and four bilobed ganglia, one per leg-bearing metamere. The body cavity (hemocoel) functions in respiration and circulation. Over 1200 species have been described based primarily on the morphology of the claws and buccal-pharyngeal apparatus. Individuals may be either gonochoric, unisexual, or hermaphroditic, with fertilized or unfertilized eggs deposited either freely or within the shed exuvium. Parthenogenesis occurs frequently in limnic and terrestrial tardigrades, allowing them to colonize new territories by passive dispersal of a single individual. Cryptobiosis (anhydrobiosis, anoxybiosis, cryobiosis, and osmobiosis) and diapause (encystment and resting eggs) occur during the life history. Active adults (surrounded by water) and cryptobiotic adults and eggs are primarily dispersed passively, but some active dispersal can also occur. Due to the characteristic patchy distributions of tardigrade populations, little is known about their population dynamics and trophic relationships. Improved methods for collection, microscopy, culturing, and molecular analyses have been have contributed much to our knowledge of tardigrades.

Otic neuronal precursors are specified in the otic placode but interestingly do so only in the anterior domain of the otic placode, the proneural domain. In the present study, we have explored why only this territory has the competence to undergo neurogenesis, this means the early events of otic proneural regionalization and neural commitment. The proneural and non-neural domains presented complementary gene expression patterns of transcription factors and members of the Notch pathway. Overall, we propose that proneural character is acquired in the anterior territory by the action of localized ectodermal FGF8-FGF10 signaling that enhances Sox3 function. FGF signals through Sox3 activity would be essential for the specification of the proneural domain versus a non-neural territory, while Notch would be involved in refining this early regionalization. / Els precursors neurals de la oïda són especificats dins un subdomini de la placoda acústica, el domini proneural. En aquesta tesis doctoral, s'ha explorat la raó per la qual la competència neural només és adquirida en aquest subdomini. Els dominis neural i no-neural del territori presenten dominis d'expressió genètica complementaris. Aquest treball proposa que el caràcter proneural és adquirit en el subdomini anterior del territori per l'acció de FGF8 i FGF10 localitzats en l�ectoderm �tic que promouen l�acci� de Sox3. De manera que, les senyals FGF i l�activitat Sox3 s�n essencials per l�especificaci� del domini proneural, mentre que la via Notch �s necess�ria per refinar la regionalitzaci� final entre un domini neural complementari a un domini no-neural.

We now report an in depth analysis of the successful in vitro enzymatic synthesis of PHB utilizing the three-enzyme system from the bacteria Cupriavidus necator. Using HPLC methodology developed in this laboratory, and by adding each enzyme in a step-wise manner, we follow each individual stage in the three-enzyme route for PHB synthesis and delineate all stoichiometric relationships. We report the construction of the first metabolic model developed specifically for analyzing in vitro enzymatic PHB synthesis. We developed a hands-on student laboratory for culturing, producing, isolating, and purifying the bacterial biopolyesters PHB. We now report the first structural characterizations of iso-CoA, acetyl-iso-CoA, acetoacetyl-iso-CoA, and beta-hydroxybutyryl-iso-CoA using MS, MS/MS, and homo- and hetero-nuclear NMR analyses.We describe HPLC methodology to separate the isomers of several iso-CoA-containing compounds and report the first examples of iso-CoA-containing compounds acting as substrates in enzymatic acyl-transfer reactions. We describe a simple regioselective synthesis of iso-CoA from CoA. We also demonstrate a plausible mechanism, which accounts for the existence of iso-CoA isomers in commercial preparations of CoA-containing compounds. Herein we report that phenylaminoethyl selenide compounds protect DNA from peroxynitrite-mediated single-strand breaks. The mechanism of protection against peroxynitrite mediated DNA damage was investigated by HPLC. The chemistry of the reaction between peroxynitrite and HOMePAES was investigated using HPLC and HPLC/MS. The unique chemistry of the reaction between peroxynitrite and HOMePAES was investigated using HPLC and HPLC/MS. We report the development of novel CDB derivatives, which are selective COX-II inhibitors. A series of compounds were assayed with an in vitro colorimetric inhibitor screening and with a whole blood ELISA screening and the results indicate that MST is a selective inhibitor of COX-II.