Mainz, Universiẗat, Diss., 2001.

The ocean, especially the deep ocean, dominates this world; it is the largest single habitat on the planet, a habitat whose inhabitants constitute the most common forms of life on this planet. By its immense influence on the global climate systems, this vast realm continually shapes life on the land. It is the least understood realm on the planet, home to a system of life that we did not know existed ? nor was it one we could even have imagined - only found by accident in the late twentieth century. We are tampering blindly with this vast realm, destroying segments of the intricate and complex systems of life within it. We plunder its riches and only return our waste. We need to know the ocean; it just may control our fate. <br /><br /> Presently, there is a gap in our ability to study this realm: we can no longer only sit on the surface, peering in from time to time; we need to look beneath the ocean's obscuring surface, at any point, for extended periods. Small research submersibles and self-contained diving gear only become available in the later half of the twentieth century, allowing us to venture beneath the ocean's surface. However, these have severe limitations, in their endurance (usually measured in hours) and operational conditions. The heyday for underwater research was the late nineteen-sixties; at that time there were, around the world, over fifty fixed undersea habitats operated by half a dozen countries. Their complexity, and their large on- and off- shore support requirements, eventually lead their sponsors to abandon of most of these habitats. There are only two left operating today, both of which are just off the coast of Florida, with one converted to a dive-access hotel in a coastal lagoon and the other anchored well offshore, the last remaining active undersea research habitat in the world. <br /><br /> We need a new type of ocean-going research vessel that will operate as an observation post on the deep ocean. Scientists need to collect a variety of data, over scales ranging from millimetres to kilometres and time spans ranging from seconds to days, years, and even decades; do this through a continuous, comprehensive, long-term, manned presence on and in the ocean, down to the seafloor, instead of trying to piece together processes by taking intermittent snapshots of a relatively few places and events; and keep this whole endeavour open and accessible to the entire world. A vessel that bridges the surface that isolates the two separate but intricately linked worlds, above and below, would enable researchers to be in both places at once. What such a vessel would be like, how it would function, and what challenges it would deal with; such a vessel is the focus of this thesis.

The ocean, especially the deep ocean, dominates this world; it is the largest single habitat on the planet, a habitat whose inhabitants constitute the most common forms of life on this planet. By its immense influence on the global climate systems, this vast realm continually shapes life on the land. It is the least understood realm on the planet, home to a system of life that we did not know existed ? nor was it one we could even have imagined - only found by accident in the late twentieth century. We are tampering blindly with this vast realm, destroying segments of the intricate and complex systems of life within it. We plunder its riches and only return our waste. We need to know the ocean; it just may control our fate. <br /><br /> Presently, there is a gap in our ability to study this realm: we can no longer only sit on the surface, peering in from time to time; we need to look beneath the ocean's obscuring surface, at any point, for extended periods. Small research submersibles and self-contained diving gear only become available in the later half of the twentieth century, allowing us to venture beneath the ocean's surface. However, these have severe limitations, in their endurance (usually measured in hours) and operational conditions. The heyday for underwater research was the late nineteen-sixties; at that time there were, around the world, over fifty fixed undersea habitats operated by half a dozen countries. Their complexity, and their large on- and off- shore support requirements, eventually lead their sponsors to abandon of most of these habitats. There are only two left operating today, both of which are just off the coast of Florida, with one converted to a dive-access hotel in a coastal lagoon and the other anchored well offshore, the last remaining active undersea research habitat in the world. <br /><br /> We need a new type of ocean-going research vessel that will operate as an observation post on the deep ocean. Scientists need to collect a variety of data, over scales ranging from millimetres to kilometres and time spans ranging from seconds to days, years, and even decades; do this through a continuous, comprehensive, long-term, manned presence on and in the ocean, down to the seafloor, instead of trying to piece together processes by taking intermittent snapshots of a relatively few places and events; and keep this whole endeavour open and accessible to the entire world. A vessel that bridges the surface that isolates the two separate but intricately linked worlds, above and below, would enable researchers to be in both places at once. What such a vessel would be like, how it would function, and what challenges it would deal with; such a vessel is the focus of this thesis.

Syftet med mitt examensarbete har varit att skriva, arrangera och spela in musik till Tritons kommande debutskiva, Beneath Our Feet. Arbetet består av två delar, en skriftlig del där jag redogör för dels en sammanfattning av den förberedande kursen 2012, som legat till grund och möjliggjort mitt arbete. Dels också huvudarbetet under 2013 där jag i egenskap av projektledare, producent och musiker genomfört den studioinspelning som utgjort vårt mål.Den andra, konstnärliga och klingande delen av mitt arbete gestaltas av de 8 låtinspelningarna jag tillsammans med Triton åstadkommit i Studio 2 på KMH. / <p>Bilaga: 2 CD</p>

The Grey-bellied Pygmy Mouse (Mus triton) has been for a long time considered as a single species, although validity of the single species status was questioned. In order to revise current taxonomy of M. triton, I analyzed sequences of one mitochondrial (cytochrome b) and two nuclear genes (IRBP and Intron 7 of the fibrinogen) from specimens collected across the most of its known distributional range. Four well-supported phylogroups at species level, differentiated during the Plio-Pleistocene, were evidenced. Divergence dating suggests that the diversification of "triton" species complex was likely caused by Plio-Pleistocene climatic oscillations together with highly diverse topography of Eastern Africa

Mushroom (Agaricus bisporus) polyphenoloxidase (PPO) (EC 1.14.18.1) was isolated and purified using an aqueous two phase system composed of octyl phenol (ethyleneglycol) 7-8 ether (Triton X-114/TX-114). TX-114 is a non-ionic surfactant which thermoseparates in water and forms an aqueous two phase system with a surfactant-depleted top phase and a surfactant-enriched bottom phase. Critical micelle concentration (CMC) and cloud point of the surfactant are 0.17 mM and 22&ordm / C respectively. The partitioning behavior of mushroom PPO in water/TX-114 aqueous two-phase systems was studied and the effects of TX-114 concentration, ionic strength, pH, temperature and crude extract preparation on PPO partitioning were studied. PPO generally partitioned to the surfactant-depleted top phase / whereas many other hydrophobic proteins and molecules partitioned to the surfactantenriched bottom phase. When two-step ultrafiltration was used as a pretreatment, complete enzyme recovery was achieved with all studied TX-114 concentrations. Moreover, about 5 fold purification was achieved by using 8% TX-114. The purification increased to 10 fold by using polyvinylpolypyrolidone (PVPP) at pH 7.0 with a recovery of 72%. However changing pH from 7.0 to 6.0 increased the purification factor and enzyme recovery to 15 fold and 100%, respectively. Addition of potassium or sodium salts caused PPO molecules to partition in the surfactantenriched bottom phase. Finally, crude enzyme can be concentrated besides being purified and recovered by doing aqueous two-phase separation at room temperature.

Universiẗat, Diss., 2001--Mainz.

Coenzyme Q10 is a lipid-soluble coenzyme, synthesized in mammalian tissue to support energy production, and also act as an antioxidant. Certain medication, stress and age may deplete the body’s endogenous Coenzyme Q10 store. Numerous disease conditions have been shown to benefit from Coenzyme Q10 supplementation. It is a lipid-soluble component of virtually all cell membranes, and is located in the hydrophobic domain of the phospholipid bilayer of cellular membranes. It is also the only known lipid-soluble antioxidant that animal cells can synthesize de novo, and for which there exist enzymatic mechanisms which can regenerate it from its oxidized product formed in the course of its antioxidant function. The aim of this study was to investigate the cellular effects of Coenzyme Q10 and Triton X-100 on primary chicken embryo heart and muscle cell cultures. Triton X-100, a well known membrane disrupter, extensively used by cell biologists for that purpose, was used to investigate whether Coenzyme Q10 might offer protection to cell membranes exposed to disruption. Due to the correlation found between the chemical structures of nonylphenol and Triton X-100, it was decided to determine whether Triton X-100 possess estrogenic properties. Using the Recombinant Yeast Screen Assay for estrogenic activity, it was found that Triton X-100 induced weak estrogenic activity. The primary heart and skeletal muscle cell cultures were established by harvesting skeletal muscle tissue and hearts from 13 day old chicken embryos. After establishment of the cell cultures, the concentrations of Coenzyme Q10 and Triton X-100 were tested for cytotoxicity using the MTT, NR, and CV assays, in the form of a combined colorimetric cytotoxicity assay. The MTT assay revealed an increase in cell viability in both cell cultures upon exposure to Triton X-100 and Coenzyme Q10, alone, and in combination. Triton X-100 and Coenzyme Q10, alone, and in combination, caused a decrease in lysosomal membrane integrity, as measured by the NR assay, and both substances, alone, and in combination, had no effect on cellular proteins, as measured by the CV assay. Scanning electron microscopy (SEM) was done to determine the cellular effect of heart and skeletal muscle cell cultures on the external surface, more specifically the membranes, of cells in culture. Triton X-100 in the concentrations used in the study, caused membrane disruption, ranging from complete membrane lyses at the highest concentrations to membrane ruptures and apoptotic blebbing in lower concentrations. SEM revealed that no adverse effects were caused by Coenzyme Q10 on the membrane structure, in dissimilarity, cell differentiation and proliferation, including myoblast formation were seen in the presence of all the concentrations of Coenzyme Q10. Numerous ion channels were observed on cellular surfaces exposed to Coenzyme Q10. Upon exposure to 0.005% Triton X-100, after pre-treatment with Coenzyme Q10, SEM revealed a “membrane patch” formation on membranes disrupted by Triton X-100. Damage to cell membranes in the presence of Triton X-100, were less severe when cells were pre-treated with Coenzyme Q10. Confocal microscopy was utilized to investigate intracellular occurrences in the presence of Triton X-100 and Coenzyme Q10. Using Mito Tracker Red to stain active respiring mitochondria and DAPI to stain nuclei, confocal microscopy confirmed the observations made by SEM, that Coenzyme Q10 enhance cell proliferation and differentiation, and that the adverse effects to cells exposed to Triton X-100 are less severe after pre-treatment with Coenzyme Q10. ROS generation was detected, using dichlorodihydrofluorescein diacetate, in cultures exposed to Triton X-100, and none in the presence of Coenzyme Q10. In the presence of Triton X-100, after pre-treatment with Coenzyme Q10, ROS generation was remarkably lower. The study provided apparent evidence that Coenzyme Q10 offer protection to cardiac and skeletal muscle cells in culture after exposure to relatively low concentrations of the membrane disrupter Triton X-100. Coenzyme Q10 also promotes the process of proliferation and differentiation in primary chicken embryonic cultures of heart and skeletal muscle cells. / Dissertation (MSc)--University of Pretoria, 2008. / Anatomy / unrestricted

Two-neutron pick up reactions have been performed on targets of 149Sm and radioactive 151Sm using 18 MeV protons. The outgoing tritons from the 151Sm target were analyzed with a magnetic spectrograph at 16 angles between 6 degrees and 8 degrees. Unlike the two-neutron transfer data on neighbouring even-even targets, the angular distributions indicated l=o transitions to many levels in the final nucleus. Partial angular distributions for the 149Sm (p, t) 147Sm reaction were obtained, but showed only one strong l=o transition populating the 147Sm ground state. In addition, spectra from the 152Sm (p, t) reaction were measured at several angles in order to provide normalization to previous results. / Thesis / Master of Science (MSc)

Foi investigada a utilização de Sistema Micelar de Duas Fases Aquosas (SMDFA) para remoção de lipolissacarídeos (LPS) de preparações contendo proteínas recombinantes de interesse farmacêutico, como a proteína verde fluorescente (GFPuv). Os SMDFA são constituídos por soluções de tensoativos contendo micelas e oferecem ambientes hidrofóbico e hidrofílico, que possibilitam seletividade na partição de biomoléculas de acordo com sua hidrofobicidade, permitindo a remoção de LPS contaminante. Neste trabalho, foi realizada a implementação do método para a quantificação de LPS em amostras contaminadas e a obtenção de LPS e GFPuv puros a partir de cultivo de E. coli recombinante. Além disso, foi estudada a influência do Triton X-114 na metodologia de quantificação de LPS, e a adição de MgSO4, CaCl2, KI e (NH4)2SO4 na partição de GFPuv e LPS puros em SMDFA. E ainda, realizou-se um planejamento experimental (22) para avaliar os maiores KGFPuv e %RECGFPuv. O homogeneizado celular de E. coli foi testado nas melhores condições obtidas com o planejamento experimental. E finalmente, o processo por cromatografia de afinidade por íons metálicos (IMAC) foi empregado para investigar a adsorção de LPS em matriz IDA-Ca2+. Conforme os resultados obtidos, o TX-114 causou elevada interferência no método cinético cromogênico, em função da similaridade desta molécula com os LPS. Os LPS apresentaram partição preferencial para a fase concentrada em micelas, com altos valores de remoção, %REMLPS>98,0%. Ao contrário, a GFPuv foi recuperada preferencialmente na fase diluída, na qual existe maior volume disponível, resultando em valores de KGFPuv>1. A adição de sais ocasionou diminuição nos valores KGFPuv, provavelmente por causa da carga negativa que GFPuv adquiriu nas condições avaliadas. Os resultados do planejamento experimental mostraram que a melhor condição de partição obtida foi na região do ponto central, 4,0% (p/p) a 60,0°C, com KGFPuv>10. O processo por IMAC apresentou as maiores capacidades de adsorção de LPS-IDA-Ca+2 nas condições de menor pH e maior força iônica 4,0 e 1,0 mol/L, respectivamente. O processo de purificação por SMDFA empregado para a remoção de LPS contaminante presente em meio fermentado contendo GFPuv, demonstrou ser eficiente em recuperar a biomolécula-alvo na fase diluída e separar o principal contaminante na fase rica em micelas. Portanto, pode ser empregado como primeira etapa para a remoção de altas concentrações de LPS na purificação de proteínas hidrofílicas como a GFPuv. / The Aqueous Two-Phase Micellar System (ATPMS) was investigated for endotoxin (LPS) removal from preparations containing recombinant proteins of pharmaceutical interest, such as the green fluorescent protein (GFPuv). These systems usually consist of micellar surfactants solutions and offer both hydrophobic and hydrophilic environments, providing selectivity to the biomolecules partitioning according to its hydrophobicity. In this work, the implementation of the method for LPS quantification in contaminated samples was accomplished, as well as the obtaining of pure LPS and GFPuv from recombinant E. coli. Furthermore, the influence of Triton X-114 in the methodology for LPS quantification was studied, as the addition of MgSO4, CaCl2, KI, and (NH4)2SO4 into the partition of pure GFPuv and LPS in ATPMS. In addition, a statistical design (22) was carried out to evaluate the highest KGFPuv and %RECGFPuv. The E. coli cell lysate was tested under optimum conditions obtained with the statistical design. And, finally, the process by ionmetal affinity chromatography (IMAC) was used to investigate the adsorption of LPS in IDA-Ca2+ matrix. The results showed that the TX-114 caused high interference in the kinetic chromogenic method, according to the similarity of this molecule to LPS. The LPS showed preferential partitioning to the micellerich phase, with high values of removal, %REMLPS>98.0%. In the other hand, the GFPuv was preferentially recovered in the micelle-poor phase, in which there is greater volume available resulting in values of KGFPuv>1. The addition of salts caused a reduction in the values KGFPuv, probably because of the negative charge that the GFPuv acquired at the conditions evaluated. The results of the statistical design showed that the best partitioning condition obtained was in the central point region, 4.0% (wt/wt) at 60.0°C, with KGFPuv>10. The process by IMAC showed the highest adsorption of LPS-IDA-Ca+2 capacities at the conditions of lower pH and higher ionic strength 4.0 and 1.0 mol/L, respectively. The purification process for the LPS contaminant removal from E. coli fermentation containing GFPuv by ATPMS proved to be efficient in the recovering of target biomolecule in the micelle-poor phase, and separating the main contaminant in the micelle-rich phase. Furthermore, this system can be exploited as the first step for removal of higher LPS concentrations from hydrophilics protein purification.