The role that each of the Notch receptors play in controlling alveolar development and cell fate determination in the mouse mammary gland has remained unclear. By utilizing a cre-conditional constitutively active intracellular Notch1 knock-in I define, in vivo, that ectopic Notch1 activation is sufficient to inhibit ductal outgrowth, cause the formation of alveolar-like cell accumulations, and promote Elf5+/ER- cell fate, at the expense of ER+ cell fate, in the mammary gland of pubescent mice. Furthermore, ectopic Notch1 in the pregnant mammary gland is sufficient to promote the formation of pregnancy/lactation-dependent lactating adenomas. These lactating adenomas consist of differentiated secretory cells and normally regress during involution but progress into non-regressing tumours after multiple pregnancies. These lactating adenomas exhibit decapitation secretions characteristic of apocrine differentiation. Together these results suggest that Notch1 may function to promote Elf5+/ER- cell fate and may be misregulated in pregnancy-associated masses and apocrine-carcinoma of the breast in humans.

The role that each of the Notch receptors play in controlling alveolar development and cell fate determination in the mouse mammary gland has remained unclear. By utilizing a cre-conditional constitutively active intracellular Notch1 knock-in I define, in vivo, that ectopic Notch1 activation is sufficient to inhibit ductal outgrowth, cause the formation of alveolar-like cell accumulations, and promote Elf5+/ER- cell fate, at the expense of ER+ cell fate, in the mammary gland of pubescent mice. Furthermore, ectopic Notch1 in the pregnant mammary gland is sufficient to promote the formation of pregnancy/lactation-dependent lactating adenomas. These lactating adenomas consist of differentiated secretory cells and normally regress during involution but progress into non-regressing tumours after multiple pregnancies. These lactating adenomas exhibit decapitation secretions characteristic of apocrine differentiation. Together these results suggest that Notch1 may function to promote Elf5+/ER- cell fate and may be misregulated in pregnancy-associated masses and apocrine-carcinoma of the breast in humans.

<i>Pseudomonas fluorescens</i> BRG100 produces secondary metabolites with herbicidal activity to the grass weeds wild oat, Avena fatua, and green foxtail, Setaria viridis. The green fluorescence protein (gfp) gene was introduced into P. fluorescens BRG100 from Escherichia coli S17-1¥ë via a Tn5 mini transposon suicide vector system. Colony morphology, growth rate in liquid media, weed biocontrol efficacy (plant growth pouch), carbon utilization (Biolog GN) and root colonization of green foxtail by several P. fluorescens BRG100gfp transformants were determined to be the same as the wild type. <i>Pseudomonas fluorescens</i> BRGgfp-15 was found to be most similar to the wild-type in all of the above characteristics and was thus used in subsequent experiments. Note: all strains of Pseudomonas fluorescens will be referred to by only their strain throughout (ie. BRGgfp-15 and BRG100). <p>It was determined by population dynamics per section of root with spiral plating on culture medium, epi-fluorescence and confocal microscopy that BRGgfp-15 colonized all areas of the root, but showed a preference for the proximal 1/3 section and the seed. In the proximal section the mean number of viable cells per gram dry weight was log109.06 and log109.31, when applied as liquid inoculum and as the pesta granular formulation, respectively. With liquid inoculum there was only log107.53 viable cells/g in the middle 1/3 section and log107.01 viable cells/g in the distal 1/3 section. The number of viable cells/g with pesta granules was log107.61 and log107.34, for the middle and distal sections, respectively. The root hairs, root tip, and ventral portion of the seed were all areas of heavy colonization relative to the other areas of the root. <p>Survival of BRGgfp-15 in the pesta formulation was examined in 2 soil types, clay and clay loam, in a thermogradient plate apparatus by a factorial randomized design complete block experiment. The experiment included: 3-12 hour diurnal temperature regimes: 5-15¨¬C, 15-25¨¬C, and 25-35¨¬C and 3 moisture levels: 25, 50 and 75% of soil moisture holding capacity. Sampling was carried out after 0, 14, 28 and 42 days. The highest numbers of viable BRGgfp-15 cells/g were found in the pesta granules in soil subjected to the lowest diurnal temperature regime and moisture content. The lowest numbers of viable cells/g were found in the pesta granules incubated in the highest diurnal temperature and moisture. This suggests lower soil temperature and moisture enhances survival of BRGgfp-15 in pesta and/ or higher soil temperature and moisture enhances the release and dissemination of BRGgfp-15 from pesta granules. When subjected to a 5-15¨¬C-temperature regime the number of viable cells/g was log109.80. When subjected to 15-25¨¬C the viable cells/g was log108.96 and with 25-35¨¬C it was log107.33. The mean number of viable cells/g was log109.36, log108.86, and log107.87, for 25, 50, and 75% soil moisture holding capacity, respectively. There was also a significantly higher number of viable cells/g in the clay soil collected from Saskatoon, log109.00, as compared to the clay loam soil collected from Scott, which was log108.40. <p>These results suggest that Pseudomonas fluorescens BRG100 has considerable potential as a bioherbicide because of its successful root colonization of green foxtail and wheat. <i>Pseudomonas fluorescens</i> BRGgfp-15 survived well under various environmental conditions when formulated into pesta granules, proving the pesta formulation was an excellent formulation. In addition, gfp was shown to be an excellent conservative marker for monitoring the root colonization and survival of <i>P. fluorescens</i> BRG100.

Passive air samplers (PAS) using XAD-resin were deployed at a wide variety of sites around the world for four years to asses the spatial and temporal trends of legacy organochlorine pesticides (OCPs) and current use pesticides (CUPs) in the global atmosphere. Legacy OCPs are prevalent in developing countries, whereas certain CUPs dominate in North America and Europe. OCP levels are declining world wide. Concentrations from the XAD-based PAS agreed with those from polyurethane foam (PUF) disk PAS within a factor of 4 for most sites. The comparison revealed that the sampling rate of the PUF-based PAS is more dependent on wind speed, whereas that of the XAD-PAS has a higher dependence on temperature. Analysis of PAS deployed across arid, subtropical Botswana showed that recent use has more impact on present day air concentrations than historical use. Year-long measurements by high volume air sampling in Botswana yielded higher HCB levels in winter, and higher endosulfan levels in summer. Those variations are neither related to temperature fluctuation nor seasonal hydrological events, and are therefore more likely caused by pesticide usage pattern. Very low levels of OCPs were found in the warm, dry, low organic matter soils of Botswana, including in soils from historical use areas. Such soils appear to have a low long-term storage capacity for pesticides allowing for rapid volatilization. They are thus not long-term sources of pesticides to the atmosphere. Endosulfan sulfate levels were observed to increase in tropical soils with increasing elevation. Water samples from high altitude cloud forests in Costa Rica contained very low concentrations of CUPs and it is unlikely that that those levels pose a threat to amphibians in cloud forests. Laboratory experiments and literature analysis showed that the inert gas stripping method for the determination of air-water partitioning equilibria is susceptible to surface sorption artifacts for chemicals with a interface-air partition coefficient log (KIA/m) > -3. Using larger bubbles reduces the surface-to-volume ratio and produces accurate air-water partition coefficients for chemicals with log (KIA/m) < -1.2.

In my first chapter, I will discuss the mark of Cain at greater length and introduce the idea of daemon. In the second chapter, I will discuss how it functions as part of Nietzschean active nihilism. Finally, in the third chapter, I will explore the problem of somberness and the problem of fanaticism, and see how Hesse tries to resolve these two problems.

Passive air samplers (PAS) using XAD-resin were deployed at a wide variety of sites around the world for four years to asses the spatial and temporal trends of legacy organochlorine pesticides (OCPs) and current use pesticides (CUPs) in the global atmosphere. Legacy OCPs are prevalent in developing countries, whereas certain CUPs dominate in North America and Europe. OCP levels are declining world wide. Concentrations from the XAD-based PAS agreed with those from polyurethane foam (PUF) disk PAS within a factor of 4 for most sites. The comparison revealed that the sampling rate of the PUF-based PAS is more dependent on wind speed, whereas that of the XAD-PAS has a higher dependence on temperature. Analysis of PAS deployed across arid, subtropical Botswana showed that recent use has more impact on present day air concentrations than historical use. Year-long measurements by high volume air sampling in Botswana yielded higher HCB levels in winter, and higher endosulfan levels in summer. Those variations are neither related to temperature fluctuation nor seasonal hydrological events, and are therefore more likely caused by pesticide usage pattern. Very low levels of OCPs were found in the warm, dry, low organic matter soils of Botswana, including in soils from historical use areas. Such soils appear to have a low long-term storage capacity for pesticides allowing for rapid volatilization. They are thus not long-term sources of pesticides to the atmosphere. Endosulfan sulfate levels were observed to increase in tropical soils with increasing elevation. Water samples from high altitude cloud forests in Costa Rica contained very low concentrations of CUPs and it is unlikely that that those levels pose a threat to amphibians in cloud forests. Laboratory experiments and literature analysis showed that the inert gas stripping method for the determination of air-water partitioning equilibria is susceptible to surface sorption artifacts for chemicals with a interface-air partition coefficient log (KIA/m) > -3. Using larger bubbles reduces the surface-to-volume ratio and produces accurate air-water partition coefficients for chemicals with log (KIA/m) < -1.2.

<i>Pseudomonas fluorescens</i> BRG100 produces secondary metabolites with herbicidal activity to the grass weeds wild oat, Avena fatua, and green foxtail, Setaria viridis. The green fluorescence protein (gfp) gene was introduced into P. fluorescens BRG100 from Escherichia coli S17-1¥ë via a Tn5 mini transposon suicide vector system. Colony morphology, growth rate in liquid media, weed biocontrol efficacy (plant growth pouch), carbon utilization (Biolog GN) and root colonization of green foxtail by several P. fluorescens BRG100gfp transformants were determined to be the same as the wild type. <i>Pseudomonas fluorescens</i> BRGgfp-15 was found to be most similar to the wild-type in all of the above characteristics and was thus used in subsequent experiments. Note: all strains of Pseudomonas fluorescens will be referred to by only their strain throughout (ie. BRGgfp-15 and BRG100). <p>It was determined by population dynamics per section of root with spiral plating on culture medium, epi-fluorescence and confocal microscopy that BRGgfp-15 colonized all areas of the root, but showed a preference for the proximal 1/3 section and the seed. In the proximal section the mean number of viable cells per gram dry weight was log109.06 and log109.31, when applied as liquid inoculum and as the pesta granular formulation, respectively. With liquid inoculum there was only log107.53 viable cells/g in the middle 1/3 section and log107.01 viable cells/g in the distal 1/3 section. The number of viable cells/g with pesta granules was log107.61 and log107.34, for the middle and distal sections, respectively. The root hairs, root tip, and ventral portion of the seed were all areas of heavy colonization relative to the other areas of the root. <p>Survival of BRGgfp-15 in the pesta formulation was examined in 2 soil types, clay and clay loam, in a thermogradient plate apparatus by a factorial randomized design complete block experiment. The experiment included: 3-12 hour diurnal temperature regimes: 5-15¨¬C, 15-25¨¬C, and 25-35¨¬C and 3 moisture levels: 25, 50 and 75% of soil moisture holding capacity. Sampling was carried out after 0, 14, 28 and 42 days. The highest numbers of viable BRGgfp-15 cells/g were found in the pesta granules in soil subjected to the lowest diurnal temperature regime and moisture content. The lowest numbers of viable cells/g were found in the pesta granules incubated in the highest diurnal temperature and moisture. This suggests lower soil temperature and moisture enhances survival of BRGgfp-15 in pesta and/ or higher soil temperature and moisture enhances the release and dissemination of BRGgfp-15 from pesta granules. When subjected to a 5-15¨¬C-temperature regime the number of viable cells/g was log109.80. When subjected to 15-25¨¬C the viable cells/g was log108.96 and with 25-35¨¬C it was log107.33. The mean number of viable cells/g was log109.36, log108.86, and log107.87, for 25, 50, and 75% soil moisture holding capacity, respectively. There was also a significantly higher number of viable cells/g in the clay soil collected from Saskatoon, log109.00, as compared to the clay loam soil collected from Scott, which was log108.40. <p>These results suggest that Pseudomonas fluorescens BRG100 has considerable potential as a bioherbicide because of its successful root colonization of green foxtail and wheat. <i>Pseudomonas fluorescens</i> BRGgfp-15 survived well under various environmental conditions when formulated into pesta granules, proving the pesta formulation was an excellent formulation. In addition, gfp was shown to be an excellent conservative marker for monitoring the root colonization and survival of <i>P. fluorescens</i> BRG100.

<p>Nanomaterials often exhibit enhanced reactivity relative to their larger colloidal counterparts because of the high specific surface area and number of imperfections on the crystal lattice at the nanoscale. Management of ecosystems, remediation of contaminated waters, and assessment of the potential risks from the industrial use on nanomaterials requires an understanding of the environmental factors that control the reactivity and bioavailability of natural and manufactured nanomaterials. Dissolved organic matter (DOM) acts as a moderator of reactivity and bioavailability for dissolved and particulate moieties in natural waters. DOM consists of a range of low and high molecular weight species that are complex and heterogeneous. It has been historically categorized based on operational definitions, rather than physical properties. In order to understand the effect of DOM on nanomaterials, there is an urgent need for information regarding specific properties of DOM, such as ligand groups. </p><p>The goal of this research was to study how cysteine, a low molecular weight metal-binding ligand, affects the composition and reactivity of nanoparticulate zinc sulfide and metallic silver. Zinc sulfide was used as a representative of nanoparticulate metal sulfide which occurs naturally in sulfidic environments. Metallic silver nanoparticles were also studied because of its wide use in consumer products. Both types of nanomaterials contain metal constituents (zinc and silver) that are expected to strongly bind to sulfhydryl-containing ligands (such as cysteine) in the environment. Serine is structurally similar to cysteine, with the only difference of a hydroxyl group in the place of the sulfhydryl group of cysteine. Therefore, serine was used for comparison as a hydroxyl-containing analogue to cysteine. </p><p>The aggregation kinetics of zinc and other metal sulfide nanoparticles in the presence of cysteine and serine were investigated using dynamic light scattering. Cysteine decreased aggregation rates of the particles, while serine had no effect on their aggregation behavior. Further experiments revealed that the mechanism of stabilization occurred through the adsorption of cysteine on zinc sulfide, which induced electrostatic charge on the particles surface. A direct link was established between the amount of cysteine sorbed and attachment efficiency, an indicator of the tendency of particles to aggregate. These results shed light on discrepancies in the literature between metal sulfide precipitation experiments conducted in our lab and work on the formation and aggregation of zinc sulfide nanoparticles on biofilms of sulfate reducing bacteria. </p><p>The early-stage growth and aggregation kinetics of zinc sulfide nanoclusters in the presence of cysteine was studied in detail using a suite of complementary techniques. Growth and aggregation experiments have been traditionally difficult to conduct due to instrumental precision issues, but newly developed analytical tools and software products have made it possible to study the early-stage formation of nanoclusters. Experiments with small angle X-ray scattering, X-ray diffraction, dynamic light scattering, and X-ray absorption spectroscopy at the extended fine structure range showed that cysteine controlled the growth and aggregation of zinc sulfide nanoclusters. The molar ratio between zinc, sulfide, and cysteine was a determining factor in the precipitation process. When zinc and sulfide were in equimolar concentrations with cysteine, very small nanoclusters of about 2.5 nm formed within 12 hours and aggregated to structures with hydrodynamic diameter larger than 100 nm. When cysteine was in excess of zinc and sulfide, aggregation was held to a minimum, but monomer nanoclusters were able to grow to about 5 nm in 12 hours. Overall, these results indicate the importance of thiol ligands on the monomer size, extent of aggregation, and aggregate structure of zinc sulfides. </p><p>The effect of metal ligands on metal bearing particle surfaces is of particular interest for manufactured nanoparticles, because they are typically coated with an organic coating during the production process. These coatings are sorbed on the particles surface and are likely to interfere between the metallic surface and the ligand. Dissolution experiments using citrate and polyvinylpyrrolidone (PVP) coated zero valent silver nanoparticles in the presence of cysteine and serine showed that cysteine dissolved both types of particles, while serine did not. Dissolution rates depended on the aggregation state of the particles exposed to cysteine. As indicated by zeta potential and adsorption measurements, cysteine replaced the coating on the particles surface and altered their aggregation pattern. X-ray absorption spectroscopy near the absorption edge showed partial oxidation of silver and formation of Ag(+I)-sulfur bonds, indicating that the thiol group in cysteine formed chemical bonds with oxidized surface silver atoms. A comparison between the two coatings showed that citrate coated particles dissolved approximately three times faster than PVP coated particles. Overall, these results show that metal binding ligands can drastically change the fate of manufactured silver nanoparticles in the environment and that this effect is moderated by surface coatings. </p><p>The results of this study suggest that cysteine, a metal binding ligand was able to induce and control transformations, such as growth, aggregation, dissolution, and surface reactivity of zinc sulfide and metallic silver nanoparticles. Cysteine adsorbed on metal sites on both ZnS and Ag particles, inducing changes on their surface charge. Aggregation of ZnS particles was slowed because of a net decrease in zeta potential compared to the bare particles. On the contrary, cysteine enhanced the aggregation of Ag particles, by replacing the citrate and PVP coatings on the particles surface. Finally, the cysteine-Ag(+I) bonds caused strong polarization on the particles surface and lead to the oxidative dissolution of the particles. </p><p>Overall, this research provides a better understanding of the fate of natural and manufactured nanoparticles in anaerobic waters, where thiols are present in significant amounts. It may also be used for risk assessment of manufactured nanomaterials and the production of safer and environmentally responsible materials.</p> / Dissertation

published_or_final_version / Psychology / Master / Master of Philosophy

Diplominiame darbe nagrinėjamas žmogaus likimo suvokimas filosofiniame kontekste bei ženklo, simbolio ištakos ir vartojimas Vakarų kultūroje. Darbo tikslas – analizuoti lemties nuotaikų, nuojautų raišką fotografijoje. Šio tikslo siekiama remiantis tokių fotografų kaip A. Ostašenkovas kūryba, kuriai būdingas fatališkumas, egzistencinis laikinumas. Remiantis C. G. Jung‘o kolektyvinės pasąmonės ir archetipų teorija, taip pat Vakarų kultūroje vyraujančiais prietarais, fotografijose ieškoma lemties ženklų, simbolių. Kūrybiniame darbe įprasminamos teorinės dalies žinios. Pasiriktoje erdvėje – ligoninėje, ieškoma lemties ženklų, jie fiksuojami, analizuojami. / This study analyzes the perception of humans destiny in philosophical context and Western cultures understanding and usage of signs, symbols. The aim of this work is to analyze fateful moods and foreboding feelings in photography. The purpose is being attained by referring to creations of photographers like A. Ostašenkovas, for their characteristic fatality, existential temporality. Referring to C. G. Jungs theory of collective consciousness and archetypes, also to dominant superstitions of Western culture, it is being searched for signs, symbols of fate in photography.