Genetic Analysis of Mitotic Recombination in Saccharomyces cerevisiae

Mitotic genome instability can occur during the repair of double-strand breaks (DSBs) in DNA, which arise from endogenous and exogenous sources. Studying the mechanisms of DNA repair in the budding yeast, Saccharomyces cerevisiae has shown that Homologous Recombination (HR) is a vital repair mechanism for DSBs. HR can result in a crossover event, in which the broken molecule reciprocally exchanges information with a homologous repair template. The current model of double-strand break repair (DSBR) also allows for a tract of information to non-reciprocally transfer from the template molecule to the broken molecule. These “gene conversion” events can vary in size and can occur in conjunction with a crossover event or in isolation. The frequency and size of gene conversions in isolation and gene conversions associated with crossing over has been a source of debate due to the variation in systems used to detect gene conversions and the context in which the gene conversions are measured.

In Chapter 2, I use an unbiased system that measures the frequency and size of gene conversion events, as well as the association of gene conversion events with crossing over between homologs in diploid yeast. We show mitotic gene conversions occur at a rate of 1.3x10-6 per cell division, are either large (median 54.0kb) or small (median 6.4kb), and are associated with crossing over 43% of the time.

DSBs can arise from endogenous cellular processes such as replication and transcription. Two important RNA/DNA hybrids are involved in replication and transcription: R-loops, which form when an RNA transcript base pairs with the DNA template and displaces the non-template DNA strand, and ribonucleotides embedded into DNA (rNMPs), which arise when replicative polymerase errors insert ribonucleotide instead of deoxyribonucleotide triphosphates. RNaseH1 (encoded by RNH1) and RNaseH2 (whose catalytic subunit is encoded by RNH201) both recognize and degrade the RNA in within R-loops while RNaseH2 alone recognizes, nicks, and initiates removal of rNMPs embedded into DNA. Due to their redundant abilities to act on RNA:DNA hybrids, aberrant removal of rNMPs from DNA has been thought to lead to genome instability in an rnh201Δ background.

In Chapter 3, I characterize (1) non-selective genome-wide homologous recombination events and (2) crossing over on chromosome IV in mutants defective in RNaseH1, RNaseH2, or RNaseH1 and RNaseH2. Using a mutant DNA polymerase that incorporates 4-fold fewer rNMPs than wild type, I demonstrate that the primary recombinogenic lesion in the RNaseH2-defective genome is not rNMPs, but rather R-loops. This work suggests different in-vivo roles for RNaseH1 and RNaseH2 in resolving R-loops in yeast and is consistent with R-loops, not rNMPs, being the the likely source of pathology in Aicardi-Goutières Syndrome patients defective in RNaseH2.

(Table T4) Final depth vs. velocity and two-way traveltime data, ODP Site 178-1103

Site 1103 was one of a transect of three sites drilled across the Antarctic Peninsula continental shelf during Leg 178. The aim of drilling on the shelf was to determine the age of the sedimentary sequences and to ground truth previous interpretations of the depositional environment (i.e., topsets and foresets) of progradational seismostratigraphic sequences S1, S2, S3, and S4. The ultimate objective was to obtain a better understanding of the history of glacial advances and retreats in this west Antarctic margin. Drilling the topsets of the progradational wedge (0-247 m below seafloor [mbsf]), which consist of unsorted and unconsolidated materials of seismic Unit S1, was very unfavorable, resulting in very low (2.3%) core recovery. Recovery improved (34%) below 247 mbsf, corresponding to sediments of seismic Unit S3, which have a consolidated matrix. Logs were only obtained from the interval between 75 and 244 mbsf, and inconsistencies on the automatic analog picking of the signals received from the sonic log at the array and at the two other receivers prevented accurate shipboard time-depth conversions. This, in turn, limited the capacity for making seismic stratigraphic interpretations at this site and regionally. This study is an attempt to compile all available data sources, perform quality checks, and introduce nonstandard processing techniques for the logging data obtained to arrive at a reliable and continuous depth vs. velocity profile. We defined 13 data categories using differential traveltime information. Polynomial exclusion techniques with various orders and low-pass filtering reduced the noise of the initial data pool and produced a definite velocity depth profile that is synchronous with the resistivity logging data. A comparison of the velocity profile produced with various other logs of Site 1103 further validates the presented data. All major logging units are expressed within the new velocity data. A depth-migrated section with the new velocity data is presented together with the original time section and initial depth estimates published within the Leg 178 Initial Reports volume. The presented data confirms the location of the shelf unconformity at 222 ms two-way traveltime (TWT), or 243 mbsf, and allows its seismic identification as a strong negative and subsequent positive reflection.

Commodification of health care systems: The Catalan case

Commodification of the public healthcare system has been a growing process in recent decades, especially in universal healthcare systems and in high-income countries like Spain.  There are substantial differences in the healthcare systems of each autonomous region of Spain, among which Catalonia is characterized by having a mixed healthcare system with complex partnerships and interactions between the public and private healthcare sectors.  Using a narrative review approach, this article addresses various aspects of the Catalan healthcare system, characterizing the privatization and commodification of health processes in Catalonia from a historical perspective with particular attention to recent legislative changes and austerity measures.  The article approximates, the eventual effects that commodification and austerity measures will have on the health of the population and on the structure, accessibility, effectiveness, equity and quality of healthcare services.

"Ad nostram scientiam referatur": la supervisión administrativa imperial de la aplicación de las leyes durante la dinastía constantiniana

Constantino y sus sucesores, Constancio II, Constante y Juliano, expresaron su particular interés por supervisar la aplicación de determinadas leyes imperiales mediante el uso de la fórmula ad nostram scientiam referatur y de otras similares. Constantino fue el príncipe que la empleó con más frecuencia, especialmente durante el período de 313-324, exceptuando el intervalo de 315-316, cuando la confrontación con Licinio le obligó a renunciar a atender directamente las cuestiones administrativas. La progresiva disminución de la inclusión de estas cláusulas en los textos legales imperiales a partir de la unificación del Imperio en 324 se debe poner en relación con la aparición de los curiosi y luego de los agentes in rebus, como oficiales encargados de vigilar la observancia de las normas vigentes.

Self‐cleaning perovskite type catalysts for the dry reforming of methane

Gas-to-liquid processes are generally used to convert natural gas or other gaseous hydrocarbons into liquid fuels via an intermediate syngas stream. This includes the production of liquid fuels from biomass-derived sources such as biogas. For example, the dry reforming of methane is done by reacting CH4 and CO2, the two main components of natural biogas, into more valuable products, i.e., CO and H2. Nickel containing perovskite type catalysts can promote this reaction, yielding good conversions and selectivities; however, they are prone to coke laydown under certain operating conditions. We investigated the addition of high oxygen mobility dopants such as CeO2, ZrO2, or YSZ to reduce carbon laydown, particularly using reaction conditions that normally result in rapid coking. While doping with YSZ, YDC, GDC, and SDC did not result in any improvement, we show that a Ni perovskite catalyst (Na0.5La0.5Ni0.3Al0.7O2.5) doped with 80.9 ZrO2 15.2 CeO2 gave the lowest amount of carbon formation at 800 °C and activity was maintained over the operating time.

Standardisation of the Selling Process in Franchising : A Take on Sales Funnel Management

This paper addresses the two opposing extremes of standardisation in franchising and the dynamics of sales in search of a juncture point in order to reduce franchisees’ uncertainties in sales and improve sales performance. A conceptual framework is developed based on both theory and practice in order to investigate the sales process of a specific franchise network. The research is conducted over a period of six weeks in form of a customised sales report considering the sales funnel concept and performance indicators along the sales process. The received quantitative data is analysed through descriptive statistics and logistic regressions in respect to what variations in the sales process can be discovered and what practices yield higher performance. The results indicate an advantage of a prioritisation guideline regarding the activities and choices to make as a salesperson over strict standardisation. Defining the sales funnel plus engaging in the process of monitoring sales in itself has proven to be a way of reducing uncertainty as the franchisor and franchisees alike inherently gain a greater understanding of the process. The extended knowledge gained from this research allowed for both practical as well as theoretical implications and expands the knowledge on standardisation of sales and the appropriateness of the sales funnel and its management for dealing with the dilemma between standardisation and flexibility of sales in franchising contexts.

Greener methodology for the synthesis of alpha-diazocarbonyl compounds and a novel approach to dioxinone derivatives

This thesis outlines a more environmentally benign approach to diazo transfer, and the investigation of the reactivity of -diazocarbonyl compounds when subjected to transition metal and lanthanide catalysis. Extensive studies were carried out to find the optimum conditions for a greener diazo transfer methodology, and this was also applied to a continuous process for the synthesis of -diazo--ketoesters. The first chapter includes a literature review of the synthesis and subsequent reactivity of -diazocarbonyl compounds. An overview of the applications of flow chemistry for the synthesis of hazardous intermediates is also included. The applications of lanthanide catalysts in organic synthesis is also discussed. The second chapter outlines the extensive studies undertaken to determine the optimum conditions for a greener diazo transfer methodology, including base and solvent studies. Use of water as a viable solvent for diazo transfer was successfully investigated. Diazo transfer to a range of -diazo--ketoesters was achieved using 5 mol% triethylamine or DMAP in water with high conversions. Polystyrene-supported benzenesulfonyl azide as an alternative diazo transfer reagent was also explored, as well as investigations into cheaper generation of this safer reagent. This polymer-supported benzenesulfonyl azide was used with 25 mol% of base in water to achieve successful diazo transfer to a range of -diazo--ketoesters. The third chapter describes the application of the new methodology developed in Chapter 2 to a continuous processing approach. Various excellent conditions were identified for both batch and flow reactions. A series of -diazo--ketoesters were synthesised with excellent conversions using 25 mol% triethylamine in 90:10 acetone water using flow chemistry. Successful diazo transfer was also achieved using a polymer-supported benzenesulfonyl azide in water under flow conditions. The fourth chapter discusses the reactivity of -diazo--ketoesters under transition metal and lanthanide catalysis. This chapter describes the synthesis of a range of -ketoesters via transesterification, which were used to synthesise a range of novel -diazo--ketoesters that were used in subsequent decomposition reactions. A novel route to dioxinones via rhodium(II) catalysis is reported. Attempted OH and SH insertion reactions in the presence of various lanthanide(II) catalysts are outlined, leading to some unexpected and interesting rearrangement products. The experimental details, including spectroscopic and analytical data for all compounds prepared, are reported at the end of each chapter.

Analyse comparée de l'efficacité des programmes de conversion de titres de dette en prise de participation en Argentine, au Chili et au Mexique comme solution partielle à leur niveau d'endettement critique

Cet ouvrage comporte d'abords une mise en situation décrivant le contexte économique qui mena à la crise du début des années 1980 et à la mise sur pieds de nouveaux mécanismes financiers visant le ré-équilibre des économies en difficulté. Parmi l'ensemble des nouveaux types de transactions, les conversions de titres de dette en prise de participation (Debt for Equity Swaps: DES) sont étudiées en particulier à cause des nombreux avantages qu'elles comportent. Dans un contexte de déséquilibres interne et externe graves tel ceux vécus par certains pays d'Amérique latine, ces conversions semblent en effet très appropriées grâce à leur contribution à réduire l'encours de la dette extérieure et à augmenter le niveau des investissements, tout en limitant les pertes des banques prêteuses. L'analyse des programmes DES de l'Argentine, du Chili et du Mexique est faite à trois niveaux: le comportement des principaux indicateurs économiques est observé avant et après la mise en application des programmes, les avantages que les investisseurs et les pays débiteurs ont pu tirer des transactions sont ensuite mesurés en termes monétaires, et finalement, une étude des clauses particulières à chaque programme permet de terminer l'étude de l'efficacité des démarches de chaque pays pour retrouver un certain équilibre économique. Chacun des trois pays étudiés a démontré une certaine force en particulier. L'Argentine est le pays qui a le mieux profité de la décote de sa dette sur le marché secondaire et ainsi réussit à réduire son encours avec des escomptes considérables. Le volume des transactions fut cependant minime. C'est le Chili qui a pu radier la plus forte proportion de sa dette totale grâce aux transactions DES. Finalement, le Mexique s'est démarqué par l'importance des avantages qu'il a offerts aux investisseurs lors de la conversion des titres. L'étude permet de conclure que les transactions DES ont en effet un potentiel très intéressant comme outil de redressement pour un pays ne pouvant plus assumer le niveau de sa dette, mais ce, à condition que la mise sur pieds des programmes coïncide avec une prise en charge globale des économies. Le succès d'un programme dépend en grande partie de la confiance des investisseurs internationaux et des banques en la volonté de chaque état à adopter les mesures d'ajustement qu'impose leur situation critique.

Estimation of the Crustal Bulk Properties Beneath Mainland Portugal from P-Wave Teleseismic Receiver Functions

In this work, we present results from teleseismic P-wave receiver functions (PRFs) obtained in Portugal, Western Iberia. A dense seismic station deployment conducted between 2010 and 2012, in the scope of the WILAS project and covering the entire country, allowed the most spatially extensive probing on the bulk crustal seismic properties of Portugal up to date. The application of the H-kappa stacking algorithm to the PRFs enabled us to estimate the crustal thickness (H) and the average crustal ratio of the P- and S-waves velocities V (p)/V (s) (kappa) for the region. Observations of Moho conversions indicate that this interface is relatively smooth with the crustal thickness ranging between 24 and 34 km, with an average of 30 km. The highest V (p)/V (s) values are found on the Mesozoic-Cenozoic crust beneath the western and southern coastal domain of Portugal, whereas the lowest values correspond to Palaeozoic crust underlying the remaining part of the subject area. An average V (p)/V (s) is found to be 1.72, ranging 1.63-1.86 across the study area, indicating a predominantly felsic composition. Overall, we systematically observe a decrease of V (p)/V (s) with increasing crustal thickness. Taken as a whole, our results indicate a clear distinction between the geological zones of the Variscan Iberian Massif in Portugal, the overall shape of the anomalies conditioned by the shape of the Ibero-Armorican Arc, and associated Late Paleozoic suture zones, and the Meso-Cenozoic basin associated with Atlantic rifting stages. Thickened crust (30-34 km) across the studied region may be inherited from continental collision during the Paleozoic Variscan orogeny. An anomalous crustal thinning to around 28 km is observed beneath the central part of the Central Iberian Zone and the eastern part of South Portuguese Zone.

Culturing the African lungfish in Uganda: effects of exogenous fish feed on growth performance in tanks

The availability of African lungfish (Protopterus aethiopicus) in many communities in Uganda is declining. Indigenous efforts to culture this fish usually produce poor yields and depend on feeding fish fry, minced meat, and leftover food. This study evaluates three formulated diets (diet-1, diet-2, diet-3) fed to wild caught lungfish fingerlings reared in indoor tanks for 77 days. Experimental fish gradually accepted sinking pellets, and marginal increases in average body weight were observed. Mean (± SE) final weight (15.86 ± 0.80 g) for fish fed on diet-3 was significantly higher (p < 0.05) than fish fed diet-1 and diet-2. Specific growth rates (SGR) for diet- 3 were significantly higher (p < 0.05) than diet-1, and marginally more than diet-2 (0.37 ± 0.04 %/ d). Feed conversions were similar (p >0.05), ranging from 1.61 ± 0.26 to 2.07 ± 0.11. Survivals after an 11-week culture were relatively low (< 60%), but generally increased (R2 = 0.667, P = 0.007) with increasing dietary proteins. Diet-3 had a significant higher survival rate (p< 0.05) than diet-1 and diet-2. Significant growth performance was attained with diet-3. This study demonstrated that sinking fish feed pellets can be used to culture wild-caught African fingerlings in captivity.

Congenital adrenal hyperplasia: focus on the molecular basis of 21-hydroxylase deficiency

Congenital adrenal hyperplasia (CAH) is an autosomal recessive disorder caused by defects in one of several steroidogenic enzymes involved in the synthesis of cortisol from cholesterol in the adrenal glands. More than 90% of cases are caused by 21-hydroxylase deficiency, and the severity of the resulting clinical symptoms varies according to the level of 21-hydroxylase activity. 21-Hydroxylase deficiency is usually caused by mutations in the CYP21A2 gene, which is located on the RCCX module, a chromosomal region highly prone to genetic recombination events that can result in a wide variety of complex rearrangements, such as gene duplications, gross deletions and gene conversions of variable extensions. Molecular genotyping of CYP21A2 and the RCCX module has proved useful for a more accurate diagnosis of the disease, and prenatal diagnosis. This article summarises the clinical features of 21-hydroxylase deficiency, explains current understanding of the disease at the molecular level, and highlights recent developments, particularly in diagnosis.

Processing Argo oxygen data at the DAC level cookbook

This document does NOT address the issue of oxygen data quality control (either real-time or delayed mode). As a preliminary step towards that goal, this document seeks to ensure that all countries deploying floats equipped with oxygen sensors document the data and metadata related to these floats properly. We produced this document in response to action item 14 from the AST-10 meeting in Hangzhou (March 22-23, 2009). Action item 14: Denis Gilbert to work with Taiyo Kobayashi and Virginie Thierry to ensure DACs are processing oxygen data according to recommendations. If the recommendations contained herein are followed, we will end up with a more uniform set of oxygen data within the Argo data system, allowing users to begin analysing not only their own oxygen data, but also those of others, in the true spirit of Argo data sharing. Indications provided in this document are valid as of the date of writing this document. It is very likely that changes in sensors, calibrations and conversions equations will occur in the future. Please contact V. Thierry (vthierry@ifremer.fr) for any inconsistencies or missing information. A dedicated webpage on the Argo Data Management website (www) contains all information regarding Argo oxygen data management : current and previous version of this cookbook, oxygen sensor manuals, calibration sheet examples, examples of matlab code to process oxygen data, test data, etc..

Síntese e caracterização de biodiesel metílico e etílico a partir de blendas dos óleos de tungue e de soja

No presente trabalho foi investigada a transesterificação de blendas dos óleos de soja e de tungue com metanol ou etanol empregando catalisador alcalino (NaOH ou KOH). Foi investigado o tempo reacional, a proporção da blenda, a concentração e o tipo de catalisador, tipo de álcool e razão molar, temperatura e metodologia empregada no tratamento da reação. Nas reações com metanol obtiveram-se melhores conversões com tempo reacional de 1,5h; temperatura de 60°C; proporção blenda dos óleos de soja e de tungue de 90:10 (m/m); concentração de NaOH de 0,5% em relação a massa da blenda e razão molar metanol:blenda de 6:1. O tratamento dos ésteres metílicos produzidos na reação foi realizado por lavagem com água a 60°C após o processo de decantação das fases, metodologia C. O rendimento de ésteres metílicos foi superior a 96% e, o teor de mono-, di- e triacilglicerídeos, glicerol livre e total ficou abaixo dos limites estabelecidos pela ANP, indicando boa conversão (> 96,5%). Nas reações com etanol verificou-se que as melhores condições reacionais foram com uma concentração de catalisador de 0,8% de NaOH em relação a massa da blenda, razão molar etanol:blenda de 9:1, tempo de 1,5h e temperatura de 60°C. O tratamento dos produtos da reação foi realizado por lavagem com água a 60°C após o processo de remoção do etanol e decantação das fases, metodologia D. A concentração do catalisador foi um fator determinante na separação das fases. Uma maior concentração de catalisador favorece a saponificação, dificultando a separação das fases e afetando o rendimento do biodiesel sintetizado, tanto para o metílico quanto o etílico. O índice de acidez, tanto para o biodiesel metílico como o etílico, para qualquer proporção da blenda dos óleos de soja e tungue, ficaram dentro das normas da ANP, com valores abaixo de 0,5 mg.g-1 de KOH.

Active site engineering for heterogeneous catalysis using emerging nano-structured materials

The growing concern about the depletion of oil has spurred worldwide interest in finding alternative feedstocks for important petrochemical commodities and fuels. On the one hand, the enormous re-serves found (208 trillion cubic feet proven1), environmental sustainability and lower overall costs point to natural gas as the primary source for energy and chemicals in the near future.2 Nowadays the transformation of methane into useful chemicals and liquid fuels is only feasible via synthesis gas, a mixture of molecular hydrogen and carbon monoxide, that is further transformed to methanol or to hydrocarbons under moderate reaction conditions (150-350 °C and 10-100 bar).3 For a major cost reduction and in order to valorize small natural gas sources, either more efficient "syngas to products" catalysts should be produced or the manner in which methane is initially activated should be changed, ideally by developing catalysts able to directly oxidize methane to interesting products such as methanol. On the other hand, from the point of view of CO2 emissions, the use of the re-maining fossil resources will further contribute to global warming. In this scenario, the development of efficient routes for the transformation of CO2 into useful chemicals and fuels would represent a considerable step forward towards sustainability. Indeed, the environmental and economic incen-tives to develop processes for the conversion of CO2 into fuels and chemicals are great. However, for such conversions to become economically feasible, considerable research is necessary. In this lecture we will summarize our recent efforts into the design of new catalytic systems, based on MOFs and COFs, to address these challenges. Examples include the development of new Fe based FTS catalysts, electrocatalysts for the selective conversion of CO2 into syngas, the development of efficient catalysts for the utilization of formic acid as hydrogen storage vector and the development of new enzyme inspired systems for the direct transformation of methane to methanol under mild reaction conditions. References (1) http://www.clearonmoney.com/dw/doku.php?id=public:natural_gas_reserves. (2) Derouane, E. G.; Parmon, V.; Lemos, F.; Ribeiro, F. R. Sustainable Strategies for the Up-grading of Natural Gas: Fundamentals, Challenges, and Opportunities; Springer, 2005. (3) Rofer-DePoorter, C. K. Chemical Reviews. ACS Publications 1981, pp 447–474.

Síntese e caracterização de NiO-CGO para anodo e eletrólitos sólidos e base de Céria para SOFC

The direct use of natural gas makes the Solid Oxide Fuel Cell (SOFC) potentially more competitive with the current energy conversions technologies. The Intermediate Temperature SOFC (IT-SOFC) offer several advantages over the High Temperature SOFC (HT-SOFC), which includes better thermal compatibility among components, fast start with lower energy consumption, manufacture and operation cost reduction. The CeO2 based materials are alternatives to the Yttria Stabilized Zirconia (YSZ) to application in SOFC, as they have higher ionic conductivity and less ohmic losses comparing to YSZ, and they can operate at lower temperatures (500-800°C). Ceria has been doped with a variety of cations, although, the Gd3+ has the ionic radius closest to the ideal one to form solid solution. These electrolytes based in ceria require special electrodes with a higher performance and chemical and termomechanical compatibility. In this work compounds of gadolinia-doped ceria, Ce1-xGdxO2-δ (x = 0,1; 0,2 and 0,3), used as electrolytes, were synthesized by polymeric precursors method, Pechini, as well as the composite material NiO - Ce0,9Gd0,1O1,95, used as anode, also attained by oxide mixture method, mixturing the powders of the both phases calcinated already. The materials were characterized by X ray diffraction, dilatometry and scanning electronic microscopy. The refinement of the diffraction data indicated that all the Ce1-xGdxO2-δ powders were crystallized in a unique cubic phase with fluorite structure, and the composite synthesized by Pechini method produced smaller crystallite size in comparison with the same material attained by oxide mixture method. All the produced powders had nanometric characteristics. The composite produced by Pechini method has microstructural characteristics that can increase the triple phase boundaries (TPB) in the anode, improving the cell efficiency, as well as reducing the mass transport mechanism effect that provokes anode degradation