Дисертації з теми "Granite genesis"

Abstract The gold mineralization event within Archaean granite-greenstone terranes occurred during the late Archaean, and followed the intrusion of syn- to late-tectonic granitic plutons into previously deformed greenstone belts. An Archaean granite/gneiss-hosted gold deposit, in terms of this project, is classified as having a gold-assay cutoff of 1g/metric ton over widths of at least several metres, or higher grades over narrower widths and/or verbal descriptions that indicate such values. Fluid inclusion studies and isotopic data identify two possible origins for the auriferous fluids; namely magmatic and metamorphic. The exploration target according to the magmatic model, is a late-Archaean, hydrothermally altered, mineralized and fractured granitic intrusion preferably with a granodioritic or quartz-dioritic composition. The exploration target according to the metamorphic replacement model is a granitic stock that has intruded a zone of crustal weakness such as a shear zone, active during the late Archaean. Alternatively, the granitic intrusion should be affected by regionally extensive late-Archaean shearing. It should be hydrothermally altered, deformed and mineralized. Five areas within the Pietersburg granite-greenstone terrane were selected for the 'Regional Area Selection' phase of exploration for Archaean granite/gneiss-hosted gold deposits; namely Roodepoort, Waterval, Ramagoep, Moletsie and Matlala. Roodepoort contains a known granodiorite-hosted gold deposit; the Knight's Pluton, and served as an orientation survey for this project. The use and interpretation of LANDSAT images formed an integral part of exploration techniques; to assess their usefulness in the exploration of Archaean granite/gneiss-hosted gold deposits. Area selection criteria for granite/gneiss-hosted gold mineralization at Roodepoort are the major ENE-trending shear zone, the NNW-trending lineament and hydrothermal alteration, shearing, quartz-stockworks and sulphide mineralization within the Knight's Pluton. The origin of the gold within the Knight's Pluton is uncertain; both magmatic and metamorphic models are possibilities. Ongoing exploration is in progress at Roodepoort. The only area selection criterion for granite/gneiss-hosted gold mineralization at Waterval is the sericitized, subcropping granites located within trenches. Gold mineralization is insignificant. No area selection criteria for Archaean granite/gneiss-hosted gold mineralization were located at Ramagoep, Matlala and Moletsie. No further exploration is recommended for all these areas. The MES image interpretations were successful in identifying lineaments, granitic outcrops, greenstones, vegetation and soil cover. The Clay-iron images adequately differentiated betweeen iron-rich and clay-bearing areas. However, not all clay-bearing areas were associated with hydrothermal alteratian; field checks were necessary to discriminate between weathered granites and hydrothermally altered granites. The Wallis images served to locally enhance the contrasts of the MES and Clay-iron images.

Thesis advisor: J. Christopher Hepburn
The Nashoba terrane is a highly metamorphosed and sheared Paleozoic tectonic block in eastern Massachusetts. The metamorphic rocks that compose the terrane are intruded by a series of diorites, tonalites, and granites. The Andover Granite is a complex multiphase granitic suite found in the northern part of the Nashoba terrane and is composed of both foliated and unfoliated granites as well as a granodiorite phase. The Sgr Group of granites is a series of unfoliated granites exposed along the Nashoba-Avalon terrane boundary. New crystallization ages for the foliated Andover Granite and the Sudbury Granite, southernmost body of the Sgr Group of granites, are presented. CA-TIMS U-Pb geochronology on zircons collected from these granites yielded 419.43 ± 0.52 Ma and 419.65 ± 0.51 Ma crystallization ages for the foliated Andover Granite and a 420.49 ± 0.52 Ma crystallization age for the Sudbury Granite. Geochemical and petrographic analysis of these granites indicate that the foliated Andover Granite is a high-K calc-alkaline, peralmuminous, S-type, biotite + muscovite granite and the Sudbury granite is high-K calc-alkaline, metaluminous to slightly peraluminous, I-type, biotite granite. These two granites are interpreted to have formed from the anatexis of either Nashoba terrane metasedimentary rocks and/or its underlying basement just prior to the Acadian orogeny. It is proposed that when Silurian diorite/tonalite magmas intruded into the Nashoba terrane, the influx of magmatic heat was sufficient to trigger crustal melting and promote granite genesis. This petrogenetic scenario fits well with regional tectonic models showing the Silurio-Devonian convergence of Avalonia towards Ganderia (which formed the eastern side of composite Laurentia at the time) in the northern Appalachians. Prior to the collision of Avalonia to composite Laurentia, mafic and intermediate composition arc magmas intruded the eastern Ganderian margin. The large amount of heat that accompanied these intrusions is believed to have contributed to Acadian metamorphism and influenced the formation of granitic plutons along the margin. It is therefore proposed that the plutonic record of the Nashoba terrane shows that by the Late Silurian - Early Devonian, Avalonia was still outboard of Laurentia in the vicinity of southern New England
Thesis (MS) — Boston College, 2014
Submitted to: Boston College. Graduate School of Arts and Sciences
Discipline: Earth and Environmental Sciences

The Trans-Suture Suite (TSS) of granitic plutons located in Northern Britain span the Iapetus Suture and represent a particularly enigmatic stage of post-Caledonian Devonian magmatism. Despite calc-alkaline affinities, proximity to the Iapetus Suture precludes a direct relationship to active subduction. Furthermore, the absence of inherited zircons distinguishes the TSS from plutons of a similar age throughout the Scottish Highlands, and is not easily reconciled with the abundance of peraluminous and S-type granites. Micro-analytical techniques are employed to analyse accessory zircon and apatite from three TSS plutons (Criffell, Fleet and Shap) in order to investigate connections between magmatic and tectonic processes within a continental suture zone. Accessory minerals contain a robust and accessible record of magma evolution. However, their trace element compositions are shown to document the final stages of pluton emplacement only, and are determined primarily by competitive crystallisation of other accessory phases at shallow crustal levels. By contrast, whole-rock compositions record an earlier stage of magma evolution that occurred in deeper and open-system crustal hot zones. The absence of inherited zircon in the final crystal assemblage reflects resorption during rapid and potentially adiabatic ascent of super-liquidus and water-rich magmas from the crustal hot zone. The concentrations of REE in apatite inclusions decrease with increasing crystallisation of other accessory minerals and in some samples have been distinguished on the basis of their host phase. In metaluminous granodiorites, no preferred crystallisation sequence is observed between host phases. In peraluminous samples, zircon-hosted apatite compositions appear more primitive compared to those hosted by other phases and reflects earlier saturation of zircon in these magmas. The isotopic record of zircon is shown to be biased towards earlier stages of magma evolution in peraluminous magmas and provides a means of assessing zircon isotope compositions in the context of often protracted histories during silicic magma evolution. The Mn content of apatite varies independently to whole-rock composition, correlating positively with decreasing oxygen fugacity and indices of increasing reduction. Apatite is proposed as a robust and effective redox proxy with application to magmatic, detrital and lunar studies of redox conditions. Zircon O-HfU- Th-Pb isotope compositions have identified increasing degrees of compositional heterogeneity in more silicic components of the TSS, including the involvement of more mafic magmas in the generation of the large S-type pluton of Fleet. Compositional trends between zircons from plutons emplaced on both sides of the Iapetus Suture are used to identify a common Avalonian component beneath the Southern Uplands and English Lake District related to Avalonian underthrusting beneath the Laurentian margin during the late Caledonian. New geochronological dating indicates that pluton emplacement occurred during periods of both pre- and post-Acadian transtension. The distinguishing characteristics of these plutons relative to other Caledonian plutons reflect their unique emplacement into the hydrated lithosphere of the Iapetus Suture Zone. Oxygen isotope disequilibrium between the magmatic compositions recorded by zircon and those of the whole-rock and some quartz crystals reflect hydrothermal alteration of the latter two archives. Hydrothermal alteration and exchange caused by 18O-rich magmatic fluids has elevated the 18O compositions of the whole-rock and some quartz crystals, with most disequilibrium observed around the outer margins of the plutons due to further fluid interaction with local 18O-rich sediments. The hydrothermal history of the TSS is markedly different from that of the British Tertiary Igneous Province where hydrothermal alteration resulted from circulation of meteoric water, reflecting significantly different magmatic and emplacement histories.

The previously undifferentiated, "Melmoth Granite-Greenstone Remnant" (MGGR¹) crops out over an area of about 360 km² in northern Natal, South Africa. The greenstone sequence is comprised mainly of mafic metalavas with lesser serpentinite, talc schist, dacitic tuff, quartz-muscovite schist, quartzite and calc-silicate rocks. The greenstones are intruded by syntectonic trondhjemitic gneisses, late-tectonic granodioritic gneisses and post-tectonic granite dykes. Four phases of deformation and metamorphism are recognized. Epigenetic, disseminated and quartz vein-hosted gold mineralization is associated with D₂ shearing - a positive correlation existing between the intensity of the shearing, the thickness of the shear zone and the grade of ore it contains. Auriferous quartz veins are distinguished from an earlier generation of barren vein quartz on the basis of mineralogy, texture and relationship to the s-fabric. The mineralization occurs in zones of dilation associated with shear zone refraction. Associated wall rock alteration includes sericitization, argillization and chloritization. An ore genesis model based on the aforementioned parameters, is proposed. Finally, an exploration programme has been devised in order to locate undiscovered gold deposits in the MGGR. The programme could probably be applied, with minor modifications, to shear zone-hosted gold deposits in other granite-greenstone remnants in northern Natal. ¹- Name suggested by writer.

Wheat (Triticum aestivum) is the world's most widely grown and economically important crop. It is both a staple food for humans and a raw material for many industrial processes. World trade in wheat is important for economic stability and an ability to grow wheat is a valuable national resource. Wheat is Australia's major crop with an annual production of about 23 million tonnes. One-quarter of this is used domestically and meets all of Australia's requirements; the remaining three-quarters is exported. Therefore, Australia's wheat industry provides both the national staple food source and the basis of an export industry worth almost 2 billion dollars. There is great potential for further genetic improvement of wheat, not only by increasing grain yields by improved resistance to pathogens and tolerance to adverse environmental conditions, but also by improving functional quality. For example, one can change the physical properties of the storage components, starch and protein, to increase their usefulness in conventional applications and for novel uses. Some examples of the physical properties of starch affecting its uses are, the large starch granules from wheat that are suitable to make carbonless copy paper (Bligh, 1999), the small starch granules from rice that are used as a fat substitute because the a comparable mouthfeel, the high-amylose starches that have film-forming properties desirable for fried food coating batters and some forms of plastics, and the low-amylose starch that swells more in water and can be used for soft foods such as Asian noodles. Continued improvement of wheat is vital to meet the quantity and quality demands of the local and international wheat markets. One specialty market for Western Australian wheat, is export to Japan and South Korea for the production of Japanese white salted Udon noodles, an export market worth more than $200M pa (Garlinge, 1996). Udon noodles have specific eating qualities including a light, creamy, uniform colour, a 'bright' appearance to the noodle, a soft but elastic texture to the noodle, and a smooth 'mouthfeel', all of which result from the quality of the wheat flour starch they are made from(Crosbie, 1991; Batey et al., 1997; Zeng et al., 1997). The Australian Standard White Noodle (ASWN) wheat that Australia exports to produce Udon noodles is soft-grained, white coloured, contains between 9.5% and 11.5% protein, and produces flour of fine particle size with little starch damage (V. Reck, DAWA, pers. comm.). The flour also has good starch-swelling characteristics, moderate dough strength and good dough extensibility. The good starch-swelling characteristics of the flour result, for the most-part, from containing relatively less of the starch amylose than other varieties (22-23% compared to 25%), a property controlled by the GBSS genes (Nelson and Rines, 1962; Garlinge, 1996). When less amylose is present in the starch granule as it is heated in water, the amylopectin matrix inside the granule can swell, causing the finished Udon noodle to be soft. When more amylose is present in the starch granule, the amylopectin matrix cannot swell as much, and the finished noodle is too hard to have the desired 'mouthfeel' of an Udon noodle. The amylose fraction of starch is produced by the granule-bound starch synthase (GBSS) enzymes, encoded by the GBSS genes. The overall aim of the research described in this thesis was to investigate the genomic organization of the GBSS genes of wheat. Since the GBSS genes influence wheat starch quality, an understanding of the action of these genes is needed for future improvement of starch quality in noodle-wheats. There are three loci for GBSS genes in wheat, and these are located on chromosomes 4A, 7A and 7D. Both wild-type alleles and non-functional 'null' alleles exist at each locus. At the start of the project, these alleles had not been sequenced and the molecular differences between the alleles were not known. Other GBSS alleles were also thought to exist in Australian varieties that had yet to be identified and characterised. GBSS genes from a selection of wheat varieties, and from all three GBSS loci, were sequenced searching for DNA polymorphisms that were different between the different alleles. If any DNA polymorphisms were found to result in GBSS protein sequence differences, or differences in GBSS enzyme expression, they could influence the functional characteristics of the starch. Identifying GBSS allelic variants would enable molecular markers to be developed to detect the alleles and investigate their potential effects upon starch quality. Different PCR-based methods and one non-PCR-based method were used to investigate the genomic organization of the GBSS genes in a selection of genetically diverse wheat varieties. The 31 wheat varieties studied included noodle-wheat varieties from the ASWN classification, varieties with similar genetic background to ASWN wheat varieties but of unsuitable quality for noodle production, unrelated varieties of Australian Standard White wheat, and were compared with those 'Chinese Spring' varieties described in the literature. Most of the varieties are grown in the Western Australian wheatbelt and southern regions, either for export and the production of Asian noodles, or for the production of domestic baked-goods. A 500bp section from the middle of the GBSS genes was amplified, from a selection of wheat varieties, and sequenced to search for polymorphisms. Twenty-one single nucleotide differences were found between genes at the three loci and two PCR-based tests were designed to validate these differences as Single Nucleotide Polymorphisms (SNPs). A novel microsatellite was also discovered in intron 4 of the GBSS 7A genes. This (TGCCG)n microsatellite was variable between wheat varieties and so defines a novel allele in the Australian germplasm present at a frequency of 40%. A PCR-based test was developed to identify this variable locus. However, the new GBSS allele was not linked to Flour Swelling Volume (FSV) quality properties. The variable microsatellite locus Xsun1 (Shariflou and Sharp, 1999) in the 3' untranslated region of the GBSS genes and linked to GBSS allelic variation was used to genotype a wheat breeding population for its GBSS status. The population (n=69) contained combinations of wild-type and null alleles at the 7A and 7D loci. Once genotyped using this marker, the GBSS alleles were assessed for possible likage to starch variation. Although the trend suggested that the presence ofnull alleles increased the FSV, the size of the population tested was too small for the differences in FSV between wild-type and partially-waxy wheats to be statistically significant. The linkage between the Xsun1 microsatellite variation and the (TGCCG)n microsatellite variation from intron 4 of the GBSS 7A genes was studied. By combining these two microsatellite loci, which are closely linked to the GBSS coding regions, GBSS genes at the 7A locus could be separated into 12 allelic groups. Although none of these groups could be linked to specific changes in starch qualities, they can be analysed further for functional differences. In order to access a larger section of the GBSS genes using PCR, new PCR primers were designed and optimized to amplify segments of the GBSS genes. Primers for GBSS genes tend to generate many PCR products, but many of these were shown to be non-specific. These artifacts could be reduced by increasing the annealing temperatures, and non-specific priming was repressed by the presence of the second primer in the PCR reaction. Using one primer set, a nearly 2000bp segment of the GBSS 7A genes from wheat varieties 'Kulin' and 'Eradu' was amplified and sequenced. These sequences indicated the presence of single nucleotide differences that resulted in changed amino acids in the protein when compared to published GBSS sequences. The sequencing should be repeated to validate this result, which indicates that these are novel alleles, but it does suggest that allelic variation for GBSS exists in Australian wheat varieties and that these alleles are different from those described internationally. The EcoR1, HindIII and BamH1 restriction enzyme sites surrounding the GBSS genes were identified using Southern hybridisation. This provided the potential to access the entire GBSS gene, including the promoter and untranscribed regions, by restriction enzyme mediated cloning of genomic DNA. However, attempts to clone the genomic GBSS genes into both plasmid and viral vectors were not successful.The potential existence of pseudogene copies of the GBSS genes in the wheat genome was investigated using both PCR and Southern hybridisation techniques. No evidence of GBSS pseudogenes was found, and this suggests that the wheat genome does not contain them. This result was unexpected since organisms with large genomes, such as wheat, normally contain repeated sequences and pseudogenes. However, the absence of repeated sequences and pseudogenes should be beneficial in molecular wheat breeding because it suggests that there will not be interference from non-coding GBSS sequences in identifying molecular markers to GBSS genes. The GBSS genes present in Australian wheat varieties were similar enough to those described internationally that Australian breeders can make full use of research and molecular tests for GBSS genes developed elsewhere. However, enough variation exists between overseas and domestic varieties to warrant further investigation of novel GBSS alleles in domestic wheat, which may relate to differences in functionality.

Wheat (Triticum aestivum) is the world’s most widely grown and economically important crop. It is both a staple food for humans and a raw material for many industrial processes. World trade in wheat is important for economic stability and an ability to grow wheat is a valuable national resource. Wheat is Australia’s major crop with an annual production of about 23 million tonnes. One-quarter of this is used domestically and meets all of Australia’s requirements; the remaining three-quarters is exported. Therefore, Australia’s wheat industry provides both the national staple food source and the basis of an export industry worth almost 2 billion dollars. There is great potential for further genetic improvement of wheat, not only by increasing grain yields by improved resistance to pathogens and tolerance to adverse environmental conditions, but also by improving functional quality. For example, one can change the physical properties of the storage components, starch and protein, to increase their usefulness in conventional applications and for novel uses. Some examples of the physical properties of starch affecting its uses are, the large starch granules from wheat that are suitable to make carbonless copy paper (Bligh, 1999), the small starch granules from rice that are used as a fat substitute because the a comparable mouthfeel, the high-amylose starches that have film-forming properties desirable for fried food coating batters and some forms of plastics, and the low-amylose starch that swells more in water and can be used for soft foods such as Asian noodles. Continued improvement of wheat is vital to meet the quantity and quality demands of the local and international wheat markets. One specialty market for Western Australian wheat, is export to Japan and South Korea for the production of Japanese white salted Udon noodles, an export market worth more than $200M pa (Garlinge, 1996). Udon noodles have specific eating qualities including a light, creamy, uniform colour, a ‘bright’ appearance to the noodle, a soft but elastic texture to the noodle, and a smooth ‘mouthfeel’, all of which result from the quality of the wheat flour starch they are made from(Crosbie, 1991; Batey et al., 1997; Zeng et al., 1997). The Australian Standard White Noodle (ASWN) wheat that Australia exports to produce Udon noodles is soft-grained, white coloured, contains between 9.5% and 11.5% protein, and produces flour of fine particle size with little starch damage (V. Reck, DAWA, pers. comm.). The flour also has good starch-swelling characteristics, moderate dough strength and good dough extensibility. The good starch-swelling characteristics of the flour result, for the most-part, from containing relatively less of the starch amylose than other varieties (22-23% compared to 25%), a property controlled by the GBSS genes (Nelson and Rines, 1962; Garlinge, 1996). When less amylose is present in the starch granule as it is heated in water, the amylopectin matrix inside the granule can swell, causing the finished Udon noodle to be soft. When more amylose is present in the starch granule, the amylopectin matrix cannot swell as much, and the finished noodle is too hard to have the desired ‘mouthfeel’ of an Udon noodle. The amylose fraction of starch is produced by the granule-bound starch synthase (GBSS) enzymes, encoded by the GBSS genes. The overall aim of the research described in this thesis was to investigate the genomic organization of the GBSS genes of wheat. Since the GBSS genes influence wheat starch quality, an understanding of the action of these genes is needed for future improvement of starch quality in noodle-wheats. There are three loci for GBSS genes in wheat, and these are located on chromosomes 4A, 7A and 7D. Both wild-type alleles and non-functional ‘null’ alleles exist at each locus. At the start of the project, these alleles had not been sequenced and the molecular differences between the alleles were not known. Other GBSS alleles were also thought to exist in Australian varieties that had yet to be identified and characterised. GBSS genes from a selection of wheat varieties, and from all three GBSS loci, were sequenced searching for DNA polymorphisms that were different between the different alleles. If any DNA polymorphisms were found to result in GBSS protein sequence differences, or differences in GBSS enzyme expression, they could influence the functional characteristics of the starch. Identifying GBSS allelic variants would enable molecular markers to be developed to detect the alleles and investigate their potential effects upon starch quality. Different PCR-based methods and one non-PCR-based method were used to investigate the genomic organization of the GBSS genes in a selection of genetically diverse wheat varieties. The 31 wheat varieties studied included noodle-wheat varieties from the ASWN classification, varieties with similar genetic background to ASWN wheat varieties but of unsuitable quality for noodle production, unrelated varieties of Australian Standard White wheat, and were compared with those ‘Chinese Spring’ varieties described in the literature. Most of the varieties are grown in the Western Australian wheatbelt and southern regions, either for export and the production of Asian noodles, or for the production of domestic baked-goods. A 500bp section from the middle of the GBSS genes was amplified, from a selection of wheat varieties, and sequenced to search for polymorphisms. Twenty-one single nucleotide differences were found between genes at the three loci and two PCR-based tests were designed to validate these differences as Single Nucleotide Polymorphisms (SNPs). A novel microsatellite was also discovered in intron 4 of the GBSS 7A genes. This (TGCCG)n microsatellite was variable between wheat varieties and so defines a novel allele in the Australian germplasm present at a frequency of 40%. A PCR-based test was developed to identify this variable locus. However, the new GBSS allele was not linked to Flour Swelling Volume (FSV) quality properties. The variable microsatellite locus Xsun1 (Shariflou and Sharp, 1999) in the 3’ untranslated region of the GBSS genes and linked to GBSS allelic variation was used to genotype a wheat breeding population for its GBSS status. The population (n=69) contained combinations of wild-type and null alleles at the 7A and 7D loci. Once genotyped using this marker, the GBSS alleles were assessed for possible likage to starch variation. Although the trend suggested that the presence ofnull alleles increased the FSV, the size of the population tested was too small for the differences in FSV between wild-type and partially-waxy wheats to be statistically significant. The linkage between the Xsun1 microsatellite variation and the (TGCCG)n microsatellite variation from intron 4 of the GBSS 7A genes was studied. By combining these two microsatellite loci, which are closely linked to the GBSS coding regions, GBSS genes at the 7A locus could be separated into 12 allelic groups. Although none of these groups could be linked to specific changes in starch qualities, they can be analysed further for functional differences. In order to access a larger section of the GBSS genes using PCR, new PCR primers were designed and optimized to amplify segments of the GBSS genes. Primers for GBSS genes tend to generate many PCR products, but many of these were shown to be non-specific. These artifacts could be reduced by increasing the annealing temperatures, and non-specific priming was repressed by the presence of the second primer in the PCR reaction. Using one primer set, a nearly 2000bp segment of the GBSS 7A genes from wheat varieties ‘Kulin’ and ‘Eradu’ was amplified and sequenced. These sequences indicated the presence of single nucleotide differences that resulted in changed amino acids in the protein when compared to published GBSS sequences. The sequencing should be repeated to validate this result, which indicates that these are novel alleles, but it does suggest that allelic variation for GBSS exists in Australian wheat varieties and that these alleles are different from those described internationally. The EcoR1, HindIII and BamH1 restriction enzyme sites surrounding the GBSS genes were identified using Southern hybridisation. This provided the potential to access the entire GBSS gene, including the promoter and untranscribed regions, by restriction enzyme mediated cloning of genomic DNA. However, attempts to clone the genomic GBSS genes into both plasmid and viral vectors were not successful.The potential existence of pseudogene copies of the GBSS genes in the wheat genome was investigated using both PCR and Southern hybridisation techniques. No evidence of GBSS pseudogenes was found, and this suggests that the wheat genome does not contain them. This result was unexpected since organisms with large genomes, such as wheat, normally contain repeated sequences and pseudogenes. However, the absence of repeated sequences and pseudogenes should be beneficial in molecular wheat breeding because it suggests that there will not be interference from non-coding GBSS sequences in identifying molecular markers to GBSS genes. The GBSS genes present in Australian wheat varieties were similar enough to those described internationally that Australian breeders can make full use of research and molecular tests for GBSS genes developed elsewhere. However, enough variation exists between overseas and domestic varieties to warrant further investigation of novel GBSS alleles in domestic wheat, which may relate to differences in functionality.

Germ cells are a totipotent stem cell line where meiosis occurs; without germ cell formation sexual and types of asexual reproduction cannot occur. Drosophila germ cell formation depends on a specialized cytoplasm which is at the posterior of early embryos. The germ plasm in Drosophila contains electron dense structures called polar granules. Similar electron dense structures are found in the germ line of many metazoan species. This thesis examines the polar granule component, TUDOR and its molecular function in Drosophila germ cell formation and polar granule assembly. I created a null allele of tud and found that without TUDOR, embryos are incapable of forming germ cells, while some embryos had normally patterned abdomens. As TUDOR was dispensable for somatic functions but central to germ cell formation, I isolated TUDOR containing complexes from embryos. I then compared the proteins found in my TUDOR complexes to those in VASA complexes to determine which TUDOR complex components are polar granule constituents. I examined five proteins in the TUDOR and VASA complexes, TER94, AUBERGINE, Me31B, eIF4A and Pyruvate Kinase. Reduction of VASA or TUDOR from the early embryo along with TER94, Me31B, eIF4A or AUBERGINE results in a decrease in germ cell formation. As well, TER94, Me31B, eIF4A and AUBERGINE all localize to polar granules. Three of the polar granule components we isolated, Me31B, eIF4A and AUBERGINE, are proteins typical of P bodies, sites of mRNA metabolism, suggesting that polar granules might be related to P bodies specific. Interestingly, immuno-electron micrographs of TER94, an endoplasmic reticulum (ER) component, showed that polar granules can associate with ER. We also show Pyruvate Kinase, a glycolytic protein
Les cellules germinales sont des cellules souches, totipotentes, dans lesquelles se produit la méiose et qui sont nécessaires à la reproduction sexuelle et certains cas de reproduction asexuelle. Chez la drosophile, la formation des cellules germinales est liée à l'assemblage du cytoplasme germinal. Ce cytoplasme distinct contient des granules électron-denses appelées granules polaires. Les études présentées dans cette thèse examinent le rôle de TUDOR, un composant des granules polaires, au cours de l'assemblage des granules polaires et lors de la formation des cellules germinales. J'ai créé un allèle nul de tud qui m'a permis de déterminer que TUD est nécessaire à l'établissement des cellules germinales alors qu'il est partiellement dispensable à la formation de l'abdomen. Due à l'importance de TUDOR durant l'établissement des cellules germinales, j'ai d'isolé le complexe de protéines associées à TUDOR. J'ai comparé les protéines ainsi isolées à celles du complexe de VASA afin d'identifier des composants des granules polaires. J'ai ainsi étudié cinq protéines appartenant aux complexes de TUDOR et VASA soit TER94, AUBERGINE, Me31B, eIF4A et Pyruvate Kinase. La réduction de TUDOR ou de VASA en combinaison avec la réduction de TER94, Me31B, eIF4A ou AUBERGINE cause une baisse dans le nombre de cellules germinales formées. De plus, TER94, Me31B, eIF4A et AUBERGINE sont observés dans les granules polaires. Trois de ces composants soit Me31B, eIF4A et AUBERGINE sont des protéines également trouvées dans les « P-bodies », sites de métabolisme d'ARNm, ce qui suggère que les granules polaires sont un type de « Pbodies ». Des images de microscopie immuno-électronique de TER94, une protéine

Many epithelial cell types, particularly those in glands, employ distinct regulated and constitutive secretory pathways to perform their functions. Understanding how membrane trafficking and secretion are coupled to other cellular functions like growth, which is also thought to be linked to secretory and endocytic functions, is fundamental to cell biology and to certain pathologies such as diabetes and cancer. During my DPhil, I have been developing the secondary cells (SCs), which reside in the male Drosophila accessory gland, as a new in vivo system to dissect out the genetic regulation of membrane trafficking and its link to secretion and cell growth. SCs are epithelial cells located at the distal tip of each accessory gland. They contain a number of large intracellular compartments (>5 μm diameter) allowing membrane trafficking to be easily visualised. This in turn makes scoring specific trafficking phenotypes in normal and genetically manipulated cells very reliable. Using imaging of both live and fixed cells, I show that SC compartments have specific Rab signatures characteristic of both secretory and endolysosomal compartments. The late endosomal and lysosomal marker, Rab7, is predominantly found on acidic compartments whilst Rab11, a slow recycling endosomal marker, is localised on the membranes of about ten non-acidic compartments containing dense-core granules (DCGs) and the protease angiotensin-converting enzyme (ANCE). I show that Rab GTPases regulate the maturation and number of compartments, the secretion of intraluminal vesicles, called exosomes, and the growth of SCs. Rab11 and Rab7 appear to control the balance between large non-acidic and acidic compartments in SCs, while Rab35, which coats smaller fast-recycling endosomal compartments (~1 μm), also controls the number of these large compartments. BMP signalling regulates SC growth and membrane trafficking. Using knockdown experiments, I show that all components associated with BMP signalling are involved in this process. Interestingly, I demonstrate that as well as ANCE, Dpp, the fly homologue of mammalian BMP 2 and 4, is localised inside DCGs of SCs. Dpp is the predominant ligand controlling SC growth and exosome secretion via autocrine and paracrine mechanisms. Interestingly, mating elicits the secretion of DCG compartments (DCGs), a process dependent on the Synaptosomal-associated protein 24kDa, SNAP24. BMP pathway activation is also increased in a Dpp-dependent fashion, presumably as a result of Dpp secretion, and it accelerates the rate of DCG biogenesis. I propose a model in which Dpp regulates the replenishment of DCGs in response to release via an autocrine feedback loop.

Tese (doutorado)-Universidade de Brasília, Instituto de Ciências Humanas, 2011.
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A história política do mundo helenístico não era formada unicamente por comandantes sêniores do exército de Alexandre, o Grande; sob os Diádocos contava-se uma série de ex-oficiais de poder menor ou aventureiros que desempenhavam funções importantes em regiões controladas pelos ex-generais do rei macedônico ou mesmo em territórios que não haviam sido previamente subjugados. Juntamente com os Diádocos, tais comandantes de poder menor moldaram a política do mundo helenístico e transformaram de maneira relevante a sociedade na qual estavam inseridos, contribuindo para a formação da imitatio Alexandri e de seu impacto na arte da guerra do período helenístico. Este era o caso de Agátocles de Siracusa e, em seguida, Pirro do Epiro (com poder obviamente maior que o de Agátocles), que, a despeito das diferenças na esfera de poder e do vínculo com Alexandre, tiveram papel fundamental na concretização das inovações políticas e militares no ocidente helenístico. Esta tese de doutorado apresenta duas hipóteses: em primeiro lugar, que a monarquia de Agátocles era de natureza “helenística”, e que essa “inovação” política se dirigiu às suas tropas mercenárias e não à cidade de Siracusa, onde sua magistratura compulsória era uma simples formalidade; em segundo lugar, que a expedição africana de Agátocles e a experiência militar de Pirro na Magna Grécia e na Sicília provocaram inovações militares em Cartago, primeiramente em nível estratégico e, em seguida, já na invasão africana liderada pelos romanos, em nível tático. Tais inovações, por fim, teriam transformado o exército cartaginês numa autêntica arma helenística. _________________________________________________________________________________ ABSTRACT
The political history of the Hellenistic world was not composed only of the senior commanders of Alexander the Great; under the Diadochi both a number of his minor officers and adventurers played important roles. They were active both in territories ruled by former generals of Alexander and even in territories which had not been subdued by the Macedonian King. With the Diadochi, such commanders with minor power molded the politics of the Hellenistic world and shaped their society, thus contributing to the imitatio Alexandri as well as to its impact on the art of war in Hellenistic period. This was the case of Agathocles of Syracuse, and after him Pyrrhos of Epirus, both fundamental to the concretization of political and military innovations in the western Hellenistic world, despite their differences in power and their connection with Alexander. This DPhil thesis presents two hypotheses. First of all, Agathocles‟ monarchy was of “Hellenistic” nature, and such political “innovation” was proposed to his mercenary troops instead of the city of Syracuse, where his power was a mere formality. Secondly, the African expedition led by Agathocles and the military experience of Pyrrhos in Magna Graecia and Sicily fomented military innovations in Carthage. These innovations were, in the first place, at the logistical level, and after that, during the African invasion led by the Romans, at the tactical level. Such innovations, in the end, would have changed the Carthaginian army into an authentic Hellenistic weapon.

Secretory processes underpin the emergence of cellular diversity in complex multicellular organisms. However, our understanding of the basic mechanisms controlling the different secretory and endosomal compartments involved remains surprisingly incomplete. During my DPhil I have studied a specialised epithelial cell type in the male Drosophila accessory glands, the secondary cell, which contains unusually large intracellular compartments that are accessible to detailed morphological study. I characterise the organisation, ultrastructure and molecular composition of this cell's secretory and endosomal compartments, and I employ specific Rab GTPases, conserved coordinators of membrane trafficking and identity, to define multiple compartmental subtypes. By developing super-resolution and time-lapse microscopy approaches in these cells, I show that numerous intraluminal vesicles (ILVs) are formed within Rab11-labelled secretory compartments and released into the accessory gland lumen as exosomes, the first clear demonstration in eukaryotic cells of exosome biogenesis within a non-late endosomal compartment. Biogenesis of these ILVs is dependent on evolutionarily conserved Endosomal Sorting Complexes Required for Transport (ESCRT) 0-III genes and involves loading of compartment-specific cargoes. Work by others, some in collaboration with me, has shown that these novel mechanisms are conserved in human cells. I show that dense-core granules, the structures employed to package proteins and other molecules destined for regulated secretion, form within large non-cored Rab6- positive compartments, in a process that seems to involve inputs from both the Golgi and recycling endosomal pathways. Further analysis has revealed roles for specific Rabs, for ILVs, and for the conserved fibrillar protein Mfas/TGFBI in different aspects of DCG formation. I also show that DCGs are not only secreted, but can also be degraded by fusion to acidic endosomal compartments. Remarkably, there is evidence that mammalian cells may employ all of these mechanisms and defects in these processes may be linked to diseases like cancer, diabetes and neurodegenerative disorders. Hence my work has established a new system to study complex secretory mechanisms, which can now be developed to model specific disease processes in the future. In summary, I have discovered several novel cell biological mechanisms controlling exosome biology, dense-core granule biogenesis, regulated secretion, and endolysosomal trafficking. Some of these already appear relevant to human health and disease, suggesting that the secondary cell system has considerable further potential for unravelling the fundamental processes underlying eukaryotic secretion in the future.

Dentro da proposta de estudo do desenvolvimento, a presente pesquisa propõe a análise da gênese do empresário no Rio Grande do Sul sob a perspectiva institucional, mais propriamente segundo os modelos de matriz institucional e de construção mental de Douglass North. Para tanto, recorre-se a teoria de Schumpeter para caracterizar esse empresário, bem como o seu papel no sistema capitalista. Os primeiros empresários gaúchos são imigrantes ou descendentes de germânicos e, ao contrário do que acontece em São Paulo, na sua maioria são descapitalizados. Boa parte exerce também a função de capitalista, e assume-se a tese de Pesavento de que o capital industrial é acumulado na atividade comercial, prioritariamente vinculada à economia colonial. Deste modo, apesar de ser a economia pecuária-charqueadora a mais rentável durante a formação econômica do estado, não é dela que surgem o capital e a mão-de-obra da indústria, nem o empresário. Visando compreender o ambiente tradicional não propício ao surgimento do empresário vis-à-vis o capitalismo moderno, são utilizadas as teses de Weber e Veblen sobre o desenvolvimento das sociedades, relacionando as instituições com as trajetórias seguidas. Ainda, para o estudo dessa dicotomia entre instituições tradicionais e modernas, são apresentadas as análises de Florestan Fernandes e Sérgio Buarque de Holanda para a sociedade brasileira. Assume-se que o legado cultural dos imigrantes alemães, tanto a ética protestante, quanto a superação das adversidades, foi um dos vetores de estímulo da matriz institucional que permitiu o surgimento do empresário no Rio Grande. Ainda, a dominação do capital e a disciplina do trabalho, bem como a aliança com o Estado e a constituição da classe empresarial, são os três outros vetores da matriz institucional que se forma na transição capitalista no estado, durante a República Velha. Na aplicação do modelo de construção mental são estudadas as biografias de alguns empresários do escalão regional onde se verifica um comportamento diferenciado que procurava aproveitar as oportunidades econômicas, na tentativa de mostrar como a leitura do ambiente institucional desses atores é influenciada pela herança cultural.
Within development study purpose, this present research proposes to analyze the genesis of entrepreneur in Rio Grande do Sul under institutional perspective, more specifically following institutional matrix and mental construction models created by Douglass North. It is based in the theory of Schumpeter to characterize this entrepreneur as well as its role in the capitalist system. The first gaúchos entrepreneurs are Germanic immigrants or descendants and, unlike what happens in São Paulo, most of them are undercapitalized. Much of them also had capitalist functions, and it is assumed Pesavento's thesis that industrial capital is accumulated in commercial activity, primarily linked to the colonial economy. Thus, although the livestock-dry beef economy was the most profitable activity during the economic formation of the state, is not from it that that appears the capital and labor of industry neither the entrepreneur. To understand the traditional environment not auspicious to the emergence of the entrepreneur vis-à-vis modern capitalism, are used Weber and Veblen thesis on the development of societies, relating institutions with the paths followed. To the study of this dichotomy between traditional and modern institutions, are presented the analysis of Florestan Fernandes and Sergio Buarque de Holanda to Brazilian society. It is assumed that the cultural legacy of German immigrants (both Protestant ethic, and overcoming adversity) was one of the vectors to stimulate institutional matrix that allowed the emergence of the entrepreneur in the Rio Grande. Still, the domination of capital and labor discipline, as the alliance with the State and the constitution of a business class are the three other vectors of institutional matrix that forms in capitalist transition during the República Velha in the state. In the application of mental construction model are studied the biographies of some entrepreneurs from regional level where there is a differentiated behavior that seek to take advantage of economic opportunities in an attempt to show how the perception of the institutional environment of these actors is influenced by cultural heritage.

Las patologías más importantes del páncreas exocrino, como la pancreatitis crónica (PC) o el cáncer de páncreas, representan un gran problema de salud pública en Europa. En la PC, el tejido acinar es substituido por complejos ductales. Además, es difícil mantener el fenotipo diferenciado de las células acinares en cultivo ya que sufren una transdiferenciación acinar-ductal.

Las células madre embrionarias (ES) de ratón han sido utilizadas en la última década para generar in vitro células completamente diferenciadas de varios linajes celulares. No obstante, la capacidad de las células ES a diferenciarse a tipos celulares de origen endodérmico es muy limitada. El objetivo principal de este proyecto ha consistido en desarrollar estrategias para diferenciar células ES de ratón a células pancreáticas acinares con una elevada eficiencia mediante 1) la optimización de las condiciones de cultivo con tal de activar vías de señalización implicadas en el desarrollo/diferenciación pancreáticas; 2) la sobreexpresión de factores transcripcionales maestros utilizando vectores virales con el fin de recapitular específicamente un programa de diferenciación acinar; 3) la selección genética de las células comprometidas al linaje acinar con el objetivo de purificar las células acinares diferenciadas.

Mediante la integración de estos abordajes, hemos conseguido aislar células que comparten características fenotípicas con células acinares inmaduras según la expresión de marcadores de diferenciación y la respuesta funcional a secretagogos.
Exocrine pancreatic diseases such as chronic pancreatitis (PC) or pancreatic cancer are major health issues in Europe. In CP, the acinar tissue is substituted by ductal complexes. In addition, it is difficult to maintain the differentiated phenotype of the acinar cells in culture as within few days an acinar-ductal transdifferentiation takes place.

In the last decade, mouse embryonic stem cells (mES) have been used to generate differentiated cells of a variety of cellular lineages in vitro. However, the ability of ES cells to differentiate into endodermal lineages is limited. The main objective of this project has focused on the development of strategies to differentiate mES to pancreatic acinar cells with high efficiency by means of: 1) Optimization of cell culture conditions to activate signalling pathways involved in pancreatic differentiation/development; 2) the overexpression of master transcription factors involved in pancreas development using viral vectors in order to recapitulate specific acinar differentiation program; 3) the genetic selection of cells committed to the acinar linage in order to purify the differentiated cells.

The integration of these different strategies allowed us to isolate cells that share phenotypic features with immature acinar cells according to the expression of differentiation markers and the functional response to acinar secretegogues.

La mondialisation et les changements globaux actuels ont un impact considérable sur la distribution des espèces et la composition des communautés. Lorsque ces espèces sont impliquées dans une interaction hôte-parasite les changements dans leur répartition peuvent entraîner la (ré)émergence de maladies infectieuses. La fasciolose, maladie causée par les grandes douves du foie (Fasciola hepatica et Fasciola gigantica) est réémergente dans de nombreux points du globe. Il est difficile de mettre en place des programmes de contrôle parce que (1) les hôtes intermédiaires, des mollusques d’eau douce de la famille des Lymnaeidae, sont composés d’un groupe d’espèces cryptiques difficilement identifiables et (2) plusieurs espèces impliquées dans cette maladie sont invasives. L’objectif de cette thèse était d’étudier les facteurs écologiques et évolutifs à large échelle de l’interaction entre la grande douve du foie et ses hôtes intermédiaires susceptibles de favoriser une (ré)émergence de la fasciolose. J’ai, dans un premier temps, développé des approches moléculaires pour reconnaître les espèces cryptiques tant du côté de la grande douve du foie que du côté des Lymnaeidae. L’utilisation d’une des techniques développées sur Galba schirazensis, Galba cubensis et Galba truncatula, trois espèces de limnées, m’a permis d’identifier leur distribution respective et de modéliser leur niches bioclimatiques grâce à l’utilisation de modèles de niches écologiques. Cette approche de modélisation de niches permet d’inférer la distribution potentielle des trois espèces et nous amène à discuter des avantages potentiels de ces modèles dans la gestion de la fasciolose. Dans un second temps, je me suis intéressée à la structuration de la diversité génétique chez des espèces invasives du système grande douve du foie-limnée, par des approches de génétique des populations et de phylogénie. J’ai pu retracer l’histoire de colonisation, les dynamiques démographiques et le système de reproduction chez P. columella, G. schirazensis et G. cubensis. J’ai montré que ces trois espèces font préférentiellement de l’autofécondation entrainant des pertes drastiques de diversité génétiques sur le front d’invasion. G. cubensis présente cependant une coexistence de plusieurs génotypes dans les aires anciennement colonisées. Les différences génétiques entre ces trois espèces sont discutées à la lumière de ce qu’on sait de leur écologie. Enfin ces résultats m’ont permis de discuter de l’avantage d'être autofécondant en cas d’invasion biologique. Pour conclure l’invasion par ces populations d’hôtes intermédiaires dépourvus de diversité génétique pourrait représenter un risque épidémiologique. En effet un parasite a plus de probabilité de circuler dans une population hôte clonale que dans une population polymorphe. Cette thèse fait le lien entre écologie, interactions hôtes-parasites et génétique de l’invasion afin de mieux comprendre les facteurs d’expansion de la fasciolose à échelle globale
Globalization and the current global change have significant impacts on species distribution and community composition. When these species are involved in a host-parasite interaction, changes in species range distribution can result in the (re)emergence of infectious diseases. Fasciolosis, a disease caused by the liver flukes (Fasciola hepatica and Fasciola gigantica) is reemerging in many parts of the world. It is difficult to implement control programs because (1) the intermediate hosts, freshwater molluscs of the Lymnaeidae family, are composed by a group of cryptic species (2) several species involved in this system are invasive. The objective of this thesis was to study the ecological and evolutionary factors at a large scale in the interaction between the liver fluke and its intermediate hosts that might drive to fasciolosis (re)emergence. First, I developed molecular approaches to quickly identify cryptic species on the two liver flukes and on three Lymnaeidae species. Using one of these techniques, I identify the respective distribution of Galba schirazensis, Galba cubensis and Galba truncatula and infer their respective climatic envelope by ecological niche modelling. We then modelled and projected the potential species distribution ranges. We discussed the contribution of models to predict the species distribution in space and time giving a considerable advantage to control fasciolosis. I then study the genetic diversity structuration in invasive snails involved in the transmission of F. hepatica, using population genetics and phylogeny. I could infer colonization history, population dynamics and reproductive system of Pseudosuccinea columella, G. schirazensis and G. cubensis. I showed that these three species preferentially make inbreeding causing drastic losses of genetic diversity in the invasion front. However G. cubensis presents a coexistence of several genotypes in formerly colonized areas. Genetic differences between these three species are discussed in the light of what we know about their ecology. Overall, these results illustrate how dramatic the reduction in genetic diversity can be for hermaphrodite animals. Finally, we discuss the epidemiological risk related to the invasion by intermediate hosts depleted in genetic diversity. Indeed, a parasite might circulate easily in a clonal host population than in a polymorphic population. In my thesis I linked ecology, host-parasite interactions with genetics of the invasion to better understand the expansion of fasciolosis at a global scale

The Pyrenean frog (Rana pyrenaica) is a Pyrenees endemic species, mostly distributed between Navarra and Aragón, and recently was described in few streams of France. The typical habitat of the species is mountain streams. It is cataloged as an Endangered, by IUCN, however we not know some aspects about it biology, natural history, as well as the current populations stats and precise distribution range. In this thesis we were done an intensive fieldwork that has allowed us collect data of distribution and density population, genetics, special connectivity and the real populations status of rana pirenaica (Rana pyrenaica) populations. About conservation status, we have increased the known distribution range of the species with new localities, even if the species was disappeared in many historical localities, for not known reasons. The estimated population size is low in many localities, so local extinctions probabilities is higher. The rana pirenaica is genetically homogeneous, both using mitochondrial genomes and nuclear marker (SNPs), and has suffered a bottleneck with subsequent geographical expansion since the Last Glacial Maximum. The differences between western and east populations cores are minimal for Mitochondrial DNA, and there has been a recent gene flow between populations through the pre-Pyrenees. In Nuclear genes is observed a population division during the Holocene in five or six genetically differentiation cores. The landscape genetics analyses suggest that big rivers and pronounced unevenness have acted as barriers that allowed this differentiation, and exist an effectiveness separation between the species eastern and western cores. The lethal amphibian fungi Batrachoquitrium dendrobatidis was massively detected throughout the occurrence area of R. pyrenaica, even though the fungi impact on the specie is not known. The forecast climate models predict that growth chytrid conditions will be better in high elevation under Global Change scenarios, that high elevation populations of many amphibians’ species may be affected. The obtained data confirmed rana pirenaica as an Endangered species. A series of management measures of in situ conservation is proposed, as well as conservation unities in the whole distribution area. This work has made possible identify management measures for these species as well as assessing their current conservation status, which we hope will help the conservation of the species in Navarra and Aragón and especially in the Ordesa and Monte Perdido National Park, which is the only one that harbors Pyrenean frog populations.
La rana pirenaica (Rana pyrenaica) es una especie endémica de los Pirineos, que se distribuye principalmente entre Navarra y Aragón, y habita en ríos de montaña. Ha sido catalogada por la IUCN como una especie en Peligro de Extinción, si bien se desconocen muchos aspectos de su biología, historia natural, así como el estado actual de sus poblaciones y distribución precisa. En esta tesis se ha realizado un trabajo de campo intenso que nos ha permitido recabar datos de distribución y densidad poblacional, genética, conectividad espacial y estado de las poblaciones de la rana pirenaica (Rana pyrenaica). En cuanto al estado de conservación, hemos incrementado el área de distribución conocida de esta especie con nuevas localidades, si bien varias poblaciones históricas han desaparecido por causas desconocidas. El tamaño poblacional estimado es bajo en muchas localidades, por lo que la probabilidad de extinción local es elevada. La rana pirenaica es muy homogénea genéticamente, tanto empleando genomas mitocondriales como con marcadores nucleares (SNPs), y ha sufrido un cuello de botella con posterior expansión geográfica desde la última glaciación. Las diferencias entre el núcleo oriental y occidental son mínimas en ADN mitocondrial, y ha existido un flujo genético reciente entre poblaciones a través del pre-Pirineo. En genes nucleares se observa una división poblacional durante el Holoceno en 5 o 6 núcleos genéticamente diferenciados. Los análisis de genética del paisaje sugieren que los principales ríos y desniveles pronunciados han actuado como barreras que permitieron esta diferenciación, y existe una separación efectiva entre el núcleo oriental y occidental de la especie. Se ha detectado la presencia masiva del hongo Batrachoquitrium dendrobatidis en todo el área de distribución de rana pirenaica, si bien su impacto en la especie es desconocido. Los modelos de clima futuro predicen que las condiciones de crecimiento del quítrido serán mejores en altitudes mayores bajo escenarios de cambio global, de forma que las poblaciones de altitud de muchas especies de anfibios se podrán ver afectadas. Los datos obtenidos confirman a rana pirenaica como en Peligro de Extinción. Se proponen una serie de medidas de gestión para la conservación in situ de la especie, así como unidades de gestión en toda el área de distribución. Este trabajo ha permitido identificar medidas de gestión de estas especies así como valorar su situación actual de conservación, las cuales esperamos que sirvan de ayuda a la gestión de la especie en Navarra y Aragón y especialmente en el Parque Nacional de Ordesa y Monte Perdido que es el único que alberga poblaciones de rana pirenaica.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
O continente sul-americano é o mais biodiverso da Terra, sendo palco da interação de complexos processos climáticos e geológicos que moldaram sua biota de forma muito heterogênea. Um crescente numero de estudos estudos de filogeografia de especies Sul Americanos tem auxiliado no entendimento das respostas evolutivas envolvidas em tal diversificação. A família Bromeliaceae é caracterizada por extensa radiação adaptativa, apresenta heterogeneidade de estratégias reprodutivas e padrões distintos de fluxo gênico e estrutura genética. Tillandsia aeranthos (Lois.) L.B. Smith é uma bromeliácea epífita que habita matas ciliares por toda região dos Pampas. Sua ocorrência em densas populações ao longo de ambientes geograficamente distintos a torna um bom modelo para a estudos sobre a influência de fatores geoclimáticos e ecológicos no padrão de distribuição da variabilidade genética e decorrentes processos de especiação ou manutenção da integridade da espécie. Esta dissertação foi dividida em dois manuscritos a fim de fornecer dados e análises úteis para a compreensão da evolução desta espécie neotropical. No Capítulo 1 foi realizada a amplificação heteróloga em Tillandsia aeranthos e Tillandsia recurvata de marcadores microssatélites nucleares previamente desenvolvidos para outras espécies de Bromeliaceae. Conjuntos de sete e seis marcadores apresentaram índices satisfatórios de polimorfismos em T. aeranthos e T. recurvata, respectivamente. A análise dos dados em duas populações de 20 indivíduos de cada espécie apresentou resultados compatíveis com sistemas reprodutivos distintos de cada espécie: fecundação cruzada predominante em T. aeranthos e auto-fecundação predominante em T. recurvata. No Capítulo 2 investigamos os padrões de variabilidade e estrutura genética e sistema reprodutivo de Tillandsia aeranthos ao longo da distribuição geográfica da espécie. Um total de 203 indivíduos de 13 localidades foi analisado a partir de sete marcadores microssatélites nucleares; 12 indivíduos tiveram 13 regiões universais plastidiais sequenciadas; e 74 indivíduos com 543 flores foram submetidos a experimentos de polinização manual. Os dados de microssatélites nucleares apontam altos níveis de diversidade genética em T. aeranthos (HE=0,806; HO=0,745) apesar de todas as regiões plastidiais sequenciadas terem sido monomórficas, sem diferenciação haplotípica. Foi observada também baixa diferenciação populacional (FST=0,031) sem correlação significativa entre as distâncias genéticas e geográficas das populações (isolamento-por-distância). Sinais moderados de eventos recentes de gargalo genético foram detectados em somente quatro das 13 populações, indicando que a maior parte das populações apresentou estabilidade demográfica durante o último máximo glacial. Os experimentos de manipulação polínica evidenciaram auto-incompatibilidade total em T. aeranthos. Em conclusão, os resultados demonstram altos níveis de diversidade genética e estabilidade demográfica na espécie, com fluxo gênico ocorrendo sem barreiras geográficas evidentes dentro da área de ocorrência de Tillandsia aeranthos.
South America is the most biodiverse subcontinent of the planet, bearing interactions between complex geoclimatic processes that heterogeneously molded its biota. An increasing number of Phylographic studies in South American species have helped us to understand the evolutionary responses that gradually formed such great biodiversity. Bromeliaceae is a family of herbaceous plants characterized by extreme adaptive radiation, its species present a wide range of reproductive strategies and distinct patterns of gene flow and genetic structure. Tillandsia aeranhos (Lois.) L.B. Smith is an epiphyte that inhabits mainly riparian forests of the Pampas biome. It occurs in dense populations across distinct habitats and topographic profiles, which makes it a good model species in studies about the influence of geo-climatic and ecologic factors over patterns of genetic variability and structure, as well as subsequent evolutionary processes of speciation or species cohesion maintenance. This dissertation presents two manuscripts aiming to provide data and analysis that will allow a better comprehension of T. aeranthos evolutionary history. In Chapter 1, we performed cross-amplifications of several nuclear microsatellite loci developed for other bromeliad species in Tillandsia aeranthos and T. recurvata. Sets of seven and six markers amplified satisfactorily and were polymorphic in T. aeanthos an T. recurvata respectively. The following analysis were carried in two populations of 20 individuals for each species and results were in accordance to opposite breeding sytems of each species: predominant cross-pollination in T. aeranthos and predominant self-pollination in T. recurvata. In Chapter 2, we investigated patterns of genetic diversity, phylogeographic structure and breeding system in T. aeranthos across most of its geographic distribution. Altogether, 203 individuals were analyzed from seven microsatellite markers; 12 individuals were analyzed from 13 chloroplast regions; and controlled pollinatin experiments were carried in 74 individuals bearing 543 flowers. Nuclear microsatellite data suggests very high levels of genetic diversity (HE=0,806; HO=0,74). Contrastingly, all chloroplast regions were monomorphic, with no haplotype differentiation. Genetic structure was very low (FST=0,031)) and isolation-by-distance hypothesis was refuted. Moderated signs of recent bottleneck events were detected in four out of 13 populations, suggesting that most populations were demographically stable since the last glacial maximum. Controlled pollination experiments showed complete self-incompatibility in T. aeranthos. In conclusion, our results sow high levels of genetic diversity and demographic stability in the species, with gene flow occurring freely without evidence of geographic barriers across the species geographic distribution.
FAPESP: 2016/03777-4
FAPESP: 2014/15586-6

Esta pesquisa, cujo objetivo principal é a análise da construção da função do tutor no âmbito do Curso de Pedagogia Licenciatura na modalidade a distância da Universidade Federal do Rio Grande do Sul (PEAD), é um estudo de caso e segue um modelo qualitativo de investigação. A análise dos dados foi feita com base na Epistemologia Genética de Piaget, buscando-se o modo pelo qual a Tomada de Consciência se processa. Para a análise dos dados, operou-se com os níveis de Tomada de Consciência em três categorias: apropriação tecnológica, estratégias de intervenção nos espaços de tutoria e compreensão das mudanças proposta pelo Curso. A análise foi realizada a partir do levantamento dos registros dos tutores nos ambientes do curso. Com este estudo foi possível concluir que a partir das interações com as professoras-alunas1 e com os estudos no curso de especialização, os tutores foram constituindo a tomada de consciência da própria função da tutoria, o que implicou em transformações nas suas maneiras de entender a proposta do curso, as professoras-alunas e as tecnologias, qualificando, por conseguinte o trabalho pedagógico.
This research, whose main objective is to examine the construction of the function of tutor within the Graduate Course of Pedagogy in the distance mode of the Federal University of Rio Grande do Sul (PEAD, in Portuguese) is a case study and follow the model of qualitative research. The data analysis was based on the genetic epistemology of Piaget, seeking for the way how Become Awareness (Consciousness) takes place. For the analysis of the data, it was operated with the levels of Consciousness in three categories: technology ownership, strategies of intervention in areas of tutorial and understanding of the changes proposed by the course. The analysis was conducted from a survey of the registers of the tutors in environments of the course. With this study it was possible to conclude that from interactions with the teachers-students 2and with studies in the specialization course, tutors were forming the awareness of the very same function of tutorial, which resulted in changes in their ways to understand the proposal of the course, the teachers-students and the technologies, qualifying, therefore the pedagogical work.

Esta dissertação é o resultado de um intenso processo de imersão em um dos espaços mais íntimos do fazer científico: o laboratório. O objetivo interposto foi o de realizar uma pesquisa de caráter etnográfico em um laboratório de pesquisa em genética de populações humanas, vinculado ao Instituto de Biociências, ao Departamento de Genética e ao Programa de Pós-Graduação em Genética e Biologia Molecular (PPGBM) da Universidade Federal do Rio Grande do Sul (UFRGS). O objeto deste estudo não foi o laboratório de pesquisa em si, mas as práticas científicas levadas a cabo por este coletivo ao ser fazer do Consórcio para Análise da Diversidade e Evolução na América Latina (CANDELA), um consórcio de pesquisa multi-cêntrico que, como o próprio nome indica, procura dar conta da diversidade étnico-racial em diversos países na América Latina. Buscou-se etnografar o que acontecia, sobretudo, entre instrumentos e práticas laboratoriais, durante a realização deste consórcio de pesquisa, abordando as escolhas práticas e conceituais que foram adotadas no cotidiano científico durante pouco mais de seis meses. Tentou-se não perder de vista as associações mais amplas que foram estabelecidas nesse contexto, de forma que o laboratório foi o ponto de partida e não o ponto de chegada. Nesse sentido, o que se realizou é o que se denomina aqui de “etnografia a partir do laboratório”. No plano teórico-epistemológico, a proposta é a de colocar em questão dicotomias clássicas da ciência moderna, como cultura-natureza, a partir do estudo etnográfico de um projeto de pesquisa que estaria na fronteira entre as ditas ciências naturais e ciências sociais, contribuindo para a ampliação da discussão em torno da agência dos não humanos e de quanto isso se faz central em uma pesquisa de cunho etnográfico que leve a sério não só o que dizem nossos interlocutores humanos, mas também aqueles que emergem a partir da fala destes e da observação da prática científica. Além disso, ao mesmo tempo que esta dissertação procura mostrar a centralidade dos não humanos na prática científica principalmente através de um evento ocorrido ao longo do trabalho de campo, ela aponta para a possibilidade de interlocução entre as ciências biológicas e as ciências sociais.
This dissertation is the result of an intense immersion in one of the most intimate spaces of the scientific practice: the laboratory. The goal brought was to conduct an ethnographic research in a laboratory in population genetics, linked to the Instituto de Biociências, of Departamento de Genética and to Programa de Pós-Graduação em Genética e Biologia Molecular (PPGBM) at the Universidade Federal do Rio Grande do Sul (UFRGS). The object of this study was not to research the laboratory itself, but the scientific practices undertaken by this collective as being part of the Consortium for Analysis of Diversity and Evolution in Latin America (CANDELA), a multi-center research consortium, as its name implies, sought to account for the racial-ethnic diversity in several countries in Latin America. It tries to give an account of what happened, especially among instruments and laboratory practice, within this research consortium, tackling the everyday practical and conceptual scientific choices ocurred during the over six months of fieldwork research. It intends to not lose sight of the broader associations that were established in this context, so that the laboratory was the starting point and not the ending point. In this sense, what took place is what is called here an “ethnography from the laboratory”. In a theoretical-epistemological scheme, the proposal is to discuss traditional dichotomies of modern science, such as culture and nature, from ethnographic study of a research project that was on the border between natural sciences and social sciences, contributing to expanding the discussion on the agency of nonhumans and how this is done in an ethnographic research that takes seriously not only what is said by our human counterparts, but also those that emerge from these talks and from the observation of scientific practice. Moreover, while this dissertation seeks to show the centrality of non-human in scientific practice mainly through an event that occurred over the fieldwork, it points to the possibility of dialogue between the natural sciences and the social sciences.

Essa pesquisa investiga os impactos da proposta teórico-metodológica da interdisciplina Desenvolvimento e Aprendizagem sob o Enfoque da Psicologia II sobre o processo de reconstrução e tomada de consciência das concepções de aprendizagem das alunas do curso de Licenciatura em Pedagogia - modalidade a distância, da UFRGS. Trata-se de um estudo de caso e segue o modelo qualitativo de investigação através de estudo de caso. A análise foi feita com base na Epistemologia Genética de Piaget, buscando-se o modo pelo qual a Tomada de Consciência se processa em relação à construção da concepção de aprendizagem. A pesquisa foi realizada com um grupo de oito professoras-alunas, tomando por base os registros das atividades realizadas por elas na interdisciplina Desenvolvimento e Aprendizagem sob o Enfoque da Psicologia II, cujas postagens foram feitas nos ambientes virtuais de aprendizagens utilizados no próprio curso. Os resultados da pesquisa apontam que os desenvolvimentos de atividades que tiveram propostas mais abertas e flexíveis, permitindo aos indivíduos se posicionarem a partir das suas vivências pessoais tiveram maiores impactos no processo de construção conceitual na medida em que possibilitam a efetiva articulação entre teoria e prática. Observou-se também que as intervenções problematizadoras propostas por professores e tutores foram significativas para o processo de reflexão e reconstrução conceitual na medida em que auxilia o aluno a refletir sobre suas ideias, contextualizando-as nos referenciais teóricos piagetianos abordados na interdisciplina.
This search investigates the impacts of the proposal theoretical and methodological aspects of interdisciplinary Development and Learning under de Focus of Psychology II on the reconstruction process and decision of conscience and awareness of concepts of students’s learning in Degree in Pedagogy – distance mode, from UFRGS. It is a case study and follows a model of qualitative research through case study. The analysis was based on Piaget’s Genetic Epistemology, searching for the way in which the Consciousness proceeds regarding the construction of the conception of learning. The search was performed with a group of eight teachers, students, buildind on the records of activicties undertaken by them in interdisciplinary Development and Learning under the Focus of Psychology II , whose posts have been made in virtual learning environments used in the actual course. The survey results indicate that the Development of the activicties had proposed more open and flexible, allowing individuals to position themselves from their personal livings to get greater impacts in process for building conceptual insofar as possible the effective articulation linkage between theory and practice. There was also observed that the problematizing interventions proposed by teachers and tutors were significant to the process of reflection and conceptual reconstruction in that it helps the student to reflect on his/her ideas, contextualizing him/her in the piagetion theoretical framework discussed in interdisciplinary.

Les systèmes de transport sont caractérisés de manière dynamique non seulement par des interactions non linéaires entre les différents composants, mais également par des boucles de rétroaction entre l'état du réseau et les décisions des utilisateurs. En particulier, la congestion du réseau impacte à la fois la répartition de la demande locale en modifiant les choix d’itinéraire et la demande multimodale globale. Selon les conditions du réseau, ils peuvent décider de changer, par exemple, leur mode de transport. Plusieurs équilibres peuvent être définis pour les systèmes de transport. L'équilibre de l'utilisateur correspond à la situation dans laquelle chaque utilisateur est autorisé à se comporter de manière égoïste et à minimiser ses propres frais de déplacement. L'optimum du système correspond à une situation où le coût total du transport de tous les utilisateurs est minimal. Dans ce contexte, l’étude vise à calculer les modèles de flux d'itinéraires dans un réseau prenant en compte différentes conditions d’équilibre et à étudier l’équilibre du réseau dans un contexte dynamique. L'étude se concentre sur des modèles de trafic capables de représenter une dynamique du trafic urbain à grande échelle. Trois sujets principaux sont abordés. Premièrement, des méthodes heuristiques et méta-heuristiques rapides sont développées pour déterminer les équilibres avec différents types de trafic. Deuxièmement, l'existence et l'unicité des équilibres d'utilisateurs sont étudiées. Lorsqu'il n'y a pas d'unicité, la relation entre des équilibres multiples est examinée. De plus, l'impact de l'historique du réseau est analysé. Troisièmement, une nouvelle approche est développée pour analyser l’équilibre du réseau en fonction du niveau de la demande. Cette approche compare les optima des utilisateurs et du système et vise à concevoir des stratégies de contrôle afin de déplacer la situation d'équilibre de l'utilisateur vers l'optimum du système
Transport systems are dynamically characterized not only by nonlinear interactions between the different components but also by feedback loops between the state of the network and the decisions of users. In particular, network congestion affects both the distribution of local demand by modifying route choices and overall multimodal demand. Depending on the conditions of the network, they may decide to change for example their transportation mode. Several equilibria can be defined for transportation systems. The user equilibrium corresponds to the situation where each user is allowed to behave selfishly and to minimize his own travel costs. The system optimum corresponds to a situation where the total transport cost of all the users is minimum. In this context, the study aims to calculate route flow patterns in a network considering different equilibrium conditions and study the network equilibrium in a dynamic setting. The study focuses on traffic models capable of representing large-scale urban traffic dynamics. Three main issues are addressed. First, fast heuristic and meta-heuristic methods are developed to determine equilibria with different types of traffic patterns. Secondly, the existence and uniqueness of user equilibria is studied. When there is no uniqueness, the relationship between multiple equilibria is examined. Moreover, the impact of network history is analyzed. Thirdly, a new approach is developed to analyze the network equilibrium as a function of the level of demand. This approach compares user and system optimums and aims to design control strategies in order to move the user equilibrium situation towards the system optimum

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Various igneous bodies have intruded the Palmer area throughout the Delamerian Orogeny. The earliest, the Rathjen Gneiss, intruded either before or during D1 which gave it the prominent foliation. D1 was also responsible for crenulations in migmatite veins throughout the area. These crenelated migmatite veins are in areas folded by D2 mesoscale folds. Some pegmatite veins are also folded by D2 folds. The Palmer Granite intruded during D2 as is seen by shearing in a semi-crystalline state and a tectonic foliation that has been folded. The ballooning of the granite during emplacement deforms the surrounding sediments and the pre-granite folds hence their axes lie parallel to the contact of the granite. The effect of the granite intruding during the deformation has lead to the axis of the D2 folds forming after the granite to have a degree of randomness about their axis. Migmatite grade was reached again after the intrusion of the granite causing melt veins to develop to disrupt the foliation. D3 formed a regional syncline of the area combined with some small scale folding within the granite, however a foliation did not form. The emplacement of the granite and some other igneous bodies throughout the area has been controlled by using the bedding plane of the Kanmantoo. The geochemical trends throughout the Palmer Granite is formed by two different groups fractionally crystallising zircon, amphibole and biotite. This results in a decrease of normally incompatible elements. The two groups form by one group from a homogeneous source and the other a heterogeneous source. The xenoliths crystallised from a mafic magma. The amphibolites form two groups according to their differentiation and genetic relationship. They both form by fractional crystallisation however U and Pb are decreasing cannot be explained by this. Another possible mechanism is liquid un-mixing. To tie all of the groups together a model of a mafic pluton that crystallises the xenoliths as a chilled margin. The mafic magma evolves some of the Palmer Granite whilst turbulently convecting hence homogenising the magma. A magma recharge forms the more evolved mafic and this forms more Palmer Granite which convects in a laminar fashion forming heterogeneities. Part of the mafics evolve enough to be caught up in the Palmer Granite and as it does not crystallise zircons all the fractional crystallisation of the Palmer Granite must have occurred in the mafic plution.
Thesis (B.Sc.(Hons)) -- University of Adelaide, School of Earth and Environmental Sciences, 1993

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Two ideologies of thought exist when models of granite genesis are considered. Do they represent the products of direct fractionation of a basaltic mantle melt, or, do they form in accordance with the restite model of White and Chappell (1977)? Assimilation and fractional crystallization (AFC) modelling of Nd - and Sr - isotopic data from the Anabama Granite, of this study, and data from the granites of the southern Adelaide Fold Belt, Antarctica and the Lachlan Fold Belt of New South Wales, all of approximately the same age, appears to reflect mixed sources with components derived both from an average Delamerian basalt composition and an average Archean crust composition. Results indicate that the Anabama Granite mostly represents primitive Delamerian basalt, contaminated by 12- 14 % Archean crustal material. Field relationships of the Anabama Granite indicate that it was the site of multiple magmatic intrusions, between approximately 490- 425 Ma. These intrusions are represented by several episodes of hydrothermal alteration and crosscutting dykes. A long-lived thermal source, not represented in the southern Adelaide Fold Belt, may be responsible for this ongoing magmatic activity. Examples of these dykes are the lamprophyre dyke, dated at 457 ± 18 Ma, which is similar in composition and appearance to the lamprophyres near Truro (South Australia) and the dacite porphyry dyke which crosscuts all other lithologies and was dated at 425 ± 13Ma. This age corresponds to the onset of thermal activity in the Lachlan Fold Belt, and therefore, leads to the suggestion that the region where the Anabama Granite outcrops may represent the western margin of the thermal perturbation responsible for the production of granitic melts in the Lachlan Fold Belt at around 400 Ma. Differences in source regions for the Anabama Granite, the granites of Antarctica and those of the Lachlan Fold Belt are recognized by the different Nd- and Sr - isotopic ratios, although all granites may represent the same process of formation, that being AFC. The dacite porphyry's isotopic signature indicates a more primitive source than that suggested for the Anabama Granite, and therefore its genesis does not represent a remelting of the Anabama Granite or of its source region. Geochemically, the Anabama Granite is similar to the Reedy Creek Granodiorite of the southern Adelaide Fold Belt and the Wanda Granodiorite of western Victoria. It can also be classified as an I-type granite using the criteria established by Chappell and White (1974). Geophysical gravity modelling of the Anabama Granite was carried out and it was found that the granite extends to a depth of approximately 15 km and dips uniformly to the north west. Thus giving an indication that fracture propagation, rather than plutonism, is the mechanism of granitic melt transport through the upper crust for the Anabama Granite and granites of the southern Adelaide Fold Belt.
Thesis (B.Sc.(Hons)) -- University of Adelaide, School of Physical Sciences, 1992

Thesis (Ph. D.)--University of Wisconsin--Madison, 1987.
Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Closely related populations or species often occupy ecologically disparate habitats. Adaptation to new habitats can maintain genetic variation within a species or eventually lead to speciation. Local adaptation to different environments has been repeatedly demonstrated in plants and animals, however the traits and genes that underlie this adaptation are poorly understood. This is because many traits differ between divergent populations and species. One way to solve this problem is to separate a trait from its genetic background through genetic manipulation and look for differences in fitness between genetically manipulated individuals.

My dissertation focuses on investigating the traits and genes that allow two species of Monkey flower, Mimulus laciniatus and Mimulus filicifolius, to survive in a unique habitat. Most closely related Mimulus species, such as M. guttatus, occur in streams and seeps, but M. laciniatus and M. filicifolius have each colonized a harsh granite outcrop environment. Another unique characteristic that both these species share is a lobed leaf shape. Because of the physiological properties of lobed leaves they should be adaptive in a dry, exposed granite outcrop. M. laciniatus also flowers earlier than nearby M. guttatus and is a small flowered self-fertilizing species while M. guttatus has large flowers and is highly outcrossing. Early flowering allows plants to escape the onset of seasonal drought while a self-fertilizing mating system and small flower size is often correlated with the occupation of harsh habitats.

In chapter one I describe a new granite outcrop endemic species of Mimulus, M. filicifolius based on morphological divergence from M. laciniatus. M. filicifolius was previously categorized as M. laciniatus but it is geographically disjunct and its leaves are more finely dissected (Sexton, Ferris, and Schoenig 2013). In the second chapter I explore whether M. filicifolius is genetically divergent and reproductively isolated from M. laciniatus using genetic sequence, microsattelite, and hybrid fertility data from four members of the M. guttatus species complex with highly overlapping geographic ranges: M. guttatus, M. nasutus, M. lacinaitus, and M. filicifolius. In the third chapter I investigate the genetic basis of leaf shape differences in three members of the M. guttatus species complex, M. laciniatus, M. nudatus, and M. guttatus using bulk segregant analysis to map quantitative trait loci. In the fourth and final chapter I examine the genetic basis of flowering time, floral size, and leaf shape divergence between sympatric M. guttatus and M. laciniatus populations in a common garden using quantitative trait locus (QTL) mapping, phenotypic selection on flowering time, flower size, and leaf shape in M. laciniatus x M. guttatus hybrids in a reciprocal transplant experiment in the field, and whether QTL's from my common garden experiment overlap fitness QTL's in the field by genotyping hybrid individuals that survived to flower in the field.

Dissertation

Bibliography: leaves 186-210.
v, 242 leaves, [19] leaves of plates : ill. (chiefly col.), map ; 30 cm.
Title page, contents and abstract only. The complete thesis in print form is available from the University Library.
The Granites gold deposits of The Granites-Tanami Inlier are the principal interest of the thesis.
Thesis (Ph.D.)--University of Adelaide, Dept. of Geology and Geophysics, 1998

Bibliography: leaves 186-210.
v, 242 leaves, [19] leaves of plates : ill. (chiefly col.), map ; 30 cm.
The Granites gold deposits of The Granites-Tanami Inlier are the principal interest of the thesis.
Thesis (Ph.D.)--University of Adelaide, Dept. of Geology and Geophysics, 1998

Mutations in HNF1A cause Maturity Onset Diabetes of the Young type 3, the second most frequent form of diabetes caused by single gene mutation. We generated human stem cell-derived pancreatic endocrine cells with clinically pathogenic mutations in HNF1A and show that HNF1A deficiency impairs endocrine cell fate, insulin granule maturation and the secretion of insulin in response to glucose. Single-cell RNA sequencing reveals that HNF1A orchestrates a network of genes involved in β-cell fate, granule maturation, glucose metabolism, calcium ion binding and hormone exocytosis. In both patients and stem cell-derived β-cells, HNF1A deficiency altered the stoichiometry of secreted insulin to c-peptide. Sulfonylurea, used in the treatment of these patients, restored both insulin secretion and stoichiometry. The uncoupling of insulin and c-peptide secretion as described here questions the common practice of using c-peptide as a proxy to evaluate β-cell function. We also demonstrate that correction of the HNF1A mutations restores function, providing a path to cell-based replacement therapy.

Indiana University-Purdue University Indianapolis (IUPUI)
Toxoplasma gondii is an obligate intracellular eukaryotic pathogen that causes severe neurologic disease in immunocompromised adults and congenitally infected neonates. Events critical to the propagation of T. gondii, such as invasion and egress, are regulated by calcium-dependent signaling. In order to identify unique components of the parasite’s calcium signaling networks, members of the Arrizabalaga laboratory have used a forward genetics approach to isolate mutants with altered sensitivity to the calcium ionophore A23187. Exposing extracellular parasites to A23187 induces protein secretion, motility and cytoskeletal rearrangements and prolonged treatment causes exhaustion of factors required for invasion, which results in what is referred to as ionophore induced death (iiDeath). Mutants capable of surviving this treatment were isolated from a chemically mutagenized population. Whole genome sequencing of one such mutant, MBD2.1, identified a nonsense mutation in a protein of unknown function (TGGT1_069070, ToxoDBv7.2) Complementation of MBD 2.1 with a wild-type copy of TGGT1_069070 restored sensitivity to iiDeath treatment. Endogenous tagging of this locus revealed that the encoded protein is secreted from a unique parasite secretory organelle known as the dense granule into the parasitophorous vacuole, leading to its designation as TgGRA41. Complete knockout of TgGRA41 recapitulates the resistance to iiDeath observed in MBD2.1 but also exhibits a dramatic decrease in propagation in tissue culture not seen in the original mutant. The knockout shows defects in multiple steps of the lytic including compromised invasion efficiency and premature egress of parasites from host cells. Cytosolic calcium measurements of extracellular parasites show enhanced uptake of calcium in the knockout strain as compared to parental and complemented, suggesting that the loss of TgGra41 results in calcium dysregulation. Together, these results provide a novel insight into the role that the parasitophorous vacuole of T. gondii plays in calcium homeostasis and calcium-dependent signaling processes.

The abstract of the dissertation Forcing-theoretic framework for the Fraïssé theory Ziemowit Kostana 15.03.2021 Subject of the dissertation The subject of the dissertation is the study of possible generalizations of the Fraïssé theory, using the method of forcing. In 1954 Roland Fraïssé discovered that many classes of finite models, like graphs or linear orders, can be canonically assigned certain infinite models. These infinite models are universal – they contain isomorphic copies of all finite models from the class – and homogeneous – each isomorphism between finite substructures can be extended to an automorphism of the whole structure. An infinite structure with these two properties is called a Fraïssé limit. This correspondence is reversible – given a countable, homogeneous model, one can recover the class of finite models from which it was built – it is exactly the class of its finite substructures. The Fraïssé theory studies this correspondence. The Fraïssé limit of a class K has a natural connection with the forcing Fn (!;K; !) = fA 2 Kj F(A) 2 [!] !, the generic structure is typically rigid, i.e. has no automorphisms other than the identity. By slightly modifying the forcing, we can add structures together with automorphisms. Recall, that a linear order is !1-dense if each nonempty open subset has size !1, and separable if has a countable dense subset. Theorem (Kostana). The following is consistent with ZFC: There exists an !1-dense, separable linear order (A; ) together with an automorphism : A ! A, such that • 8a 2 A (a) > a; • Aut (A; ) = f kj k 2 Zg: In particular Aut (A; ) ' (Z; +). Chapter 5 We continue the study of models from Chapter 4, focusing on the models of size !1. Applying the ideas of Avraham, Rubin, and Shelah from the 80s, we show that in many cases the generically added structures become homogeneous after suitably extending the model of set theory, so that it satisfies Martin’s Axiom. In this extended models, they also have certain Ramsey-like properties. Theorem (Avraham-Shelah). The following is consistent with ZFC +MA!1 : There exists an !1-dense separable linear order L with the property that each uncountable partial function f L L is monotone on an uncountable set. Theorem (Kostana). The following is consistent with ZFC +MA!1 : There exists a separable metric space X of size !1 with the property that each uncountable partial 1-1 function f X X is an isometry on an uncountable set. Moreover, distances between points of the space X are rational, and X has a dense copy of the rational Urysohn space. 2