Algae are photosynthetic microorganisms that convert carbon dioxide and sunlight into biomass that can be used for biofuel production. Although they are usually cultivated in suspension, these microorganisms are capable of forming productive biofilms over substrata given the right conditions. This dissertation focuses on algal biofilms and their application in biofuel feedstock production. In particular it reports the construction and performance of an algae biofilm photobioreactor, the physico-chemical surface properties of different algal species and adhesion substrata, and cell-surface interactions based on experimental results and theoretical models. A novel algae biofilm photobioreactor was constructed and operated (i) to demonstrate the proof of concept, (ii) to analyze the performance of the system, and (iii) to determine the key advantages and short comings for further research. The results indicated that significant reductions in water and energy requirements were possible with the biofilm photobioreactor. Although the system achieved net energy ratio of about 6, the overall productivity was low as Botryococcus branunii is notoriously slow growing algae. Thus, further studies were focused on identification of algal species capable of biofilm growth with larger biomass and lipid productivities. Adhesion of cells to substrata precedes the formation of all biofilms. A comprehensive study has been conducted to determine the interactions of a planktonic and a benthic algal species with hydrophilic and hydrophobic substrata. The physico-chemical surface properties of the algal cells and substrata were determined and using these data, cell-substrata interactions were modeled with the thermodynamic, Derjaguin, Landau Verwey, Overbeek (DLVO) and Extended Derjaguin, Landau, Verwey, Overbeek (XDLVO) approaches and critical parameters for algal adhesion were identified. Finally, the adhesion rate and strength of algal species were quantified with parallel plate flow chamber experiments. The results indicated that both cell and substrata surface hydrophobicity played a critical role for the adhesion rate and strength of the cells and XDLVO approach was the most accurate model. Finally, based on these findings the physico-chemical surface properties of ten algal species and six substrata were quantified and a screening was done to determine algae species substratum couples favoring adhesion and biofilm formation. / text

Die Plasmamembranspannung von eukaryotischen Zellen soll maßgeblich zur Regulation von zellulären Prozessen wie der Zellmigration, Mitose, Endo- und Exozytose, Membranreparatur, Osmoregulierung und Zellspreiten beitragen, welche zu einer Veränderung der Membranfläche und ihrer Deformation führt. In dieser Arbeit wurde die epitheliale Zelllinie MDCK II (Madin-Darby Canine Kidney) benutzt, um spannungsgesteuerte Oberflächenregulierung zu untersuchen. Indentationsexperimente kombiniert mit dem Herausziehen von Membrannanoröhren wurden mit Hilfe des Rasterkraftmikroskops (Atomic Force Microskope, AFM) durchgeführt, um lokale Variationen in der Membranspannung und überschüssiger Membranfläche als Funktion von äußeren Reizen abzuschätzen. Die verwendeten externen Stimuli beinhalten eine Veränderung der Funktionalität des Actomyosin-Cortexes durch die Wirkung von Blebbistatin und Cytochalasin D, sowie die Manipulation der Zytoskelett-Membran Adhäsionspunkte durch Einzel-Mikroinjektion. Die Injektion von Neomycin verhindert die Anbindung von ERM-Proteinen an das Lipid Phosphatidylinositol-(4,5)-bisphosphat (PIP2) und bewirkt somit die Abkopplung des Zytoskeletts von der Plasmamembran. Als Gegenexperiment diente die Injektion des Lipids PIP2 selbst, welches zur Erhöhung der Anzahl der Zytoskelett-Membran Adhäsionspunkte führte. Weiterhin wurden die als Membranreservoire dienenden Mikrovilli durch den Entzug von Cholesterol entfernt. Auswirkung auf das Vorhandensein von Membranreservoiren hat ebenfalls die Veränderung des osmotischen Drucks innerhalb der Zellen. Zusätzlich wurden die elastischen Eigenschaften von apikalen Zellmembran-Fragmenten von konfluenten MDCK II Zellen untersucht, welche Aufschluss über die intrinsischen Membraneigenschaften ohne den Einfluss des Zytosols und Zytoskeletts geben konnten. Abschließend wurde die Mechanik von adhärierenden und spreitenden Zellen untersucht. Zusammenfassend kann gesagt werden, dass die Plasmamembran, bestehend aus einer Phospholipiddoppelschicht, lateral schwer ausdehnbar ist aufgrund ihrer flüssig-kristallinen Natur. Durch das Vorhandensein von dynamischen Membranreservoiren wie Mikrovilli, die schnell auf Veränderungen der Membranspannung durch Membranhomöostase reagieren, werden zellulare Prozesse wie die Zellmotilität oder die Anpassung an osmotischen Stress ermöglicht. In der vorliegenden Arbeit gelang es gleichzeitig, die Membranspannung und die Verfügbarkeit von Membranfläche von adhärenten konfluenten als auch von adhärierenden und spreiten Zellen zu messen. Die durchgeführten Experimente ergaben ein detailliertes Bild wie sich die zelluläre Oberflächenregulierung in der Membranmechanik widerspiegelt.

Repeated exposure to amphetamine (AMPH) induces sensitization to its behavioral-activating effects. The development of sensitization depends on (1) the direct actions of AMPH in the ventral tegmental area (VTA), the cell body region of the mesocorticolimbic DA system, and (2) AMPH-induced glutamatergic neurotransmission in this region. Moreover, sensitization is accompanied by morphological changes in mesocorticolimbic DA circuitry. During development, the DA system is organized, at least in part, by the netrin-1 family of guidance cues. Both netrin-1 and its DCC and UNC-5 receptors continue to be expressed in the mesocorticolimbic DA system of the adult brain. Importantly, netrin-1 receptor deficient mice do not develop sensitization to AMPH, implicating an involvement of netrin-1 signaling in AMPH-induced plasticity of the DA system. To explore this possibility, adult rats were pretreated with repeated AMPH or saline, and DCC and UNC-5 receptor expression was examined in DA cell body and terminal regions using western blot. Striking AMPH-induced increases in the expression of DCC and UNC-5 were observed in the VTA only. Remarkably, these changes depended on NMDA-mediated glutamatergic neurotransmission. This is the first demonstration that repeated AMPH pretreatment regulates netrin-1 receptor expression in the adult brain and suggests that netrin-1 receptor regulation may be involved in the development of AMPH-induced sensitization.

Flocculation, the process whereby yeast cells attach in groups and sediment to the top or bottom of a fermenter, is industrially important in many fermentation batch operations. These batch operations include wine, distilled spirits, cider, bio-ethanol and production of commercial yeast metabolites. In the case of brewing yeast, it has been determined that flocculation occurs due to three forces called hydrophobic interactions, zymolectin binding and to a lesser extent, surface charge neutralization. This project sought to more closely study hydrophobic interactions and zymolectin binding. Earlier studies had shown that certain hydrophobic carboxylic acids, 3-OH oxylipins, formed in brewing yeast at flocculation onset. Therefore, these compounds showed potential as an indicator of overall cell surface hydrophobicity, and it was believed that flocculation level, cell surface hydrophobicity and oxylipin level would increase in unison in the yeast cell during brewing fermentations. During lab scale fermentations in shaker flasks and in a miniature fermentation assay setup, both flocculation level and cell surface hydrophobicity increased coincidently. However, 3-OH oxylipins could not be isolated from whole cells or cell wall isolates grown in the shaker flasks or whole cells grown in the miniature fermentation assay at detection limits approximated as 50 ng/0.5 g wet yeast. Due to their minute levels in brewing yeast cells, it was proposed that 3-OH oxylipins may mediate flocculation and aggregation via a quorum sensing mechanism instead of by increasing cell surface hydrophobicity. A disagreement exists in the literature where certain researchers believe zymolectin activity is induced, while others believe it is constitutive. The second part of this study attempted to address this by measuring zymolectin density during lab scale fermentations with a flow cytometer. Because of flow cytometry’s capacity for multiparametric analysis, large amounts of data were produced which gave information on not only zymolectin density, but also cell size and cellular complexity. Upon statistical analysis of the data, it was not possible to either refute or confirm the claim that zymolectin activity is induced or constitutive. However, the results suggested there could have been a population of cells with fewer zymolectins, and this certainly warrants further investigation. During the lab scale fermentations, cell size measured by a flow cytometer appeared to be correlated with manual measures of cell size. Furthermore, cell size tended towards uniformity during the fermentation which has also been observed in similar studies employing flow cytometry. Conversely, the cellular complexity of the yeast in this study did not change as in other studies by this may have been due either to strain differences or the methods used herein.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2002. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2002. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 4 uppsatser.

Thesis (Ph. D.)--West Virginia University, 2009. / Title from document title page. Document formatted into pages; contains x, 192 p. : ill. (some col.). Vita. Includes abstract. Includes bibliographical references.