Made available in DSpace on 2016-07-18T17:51:08Z (GMT). No. of bitstreams: 1 dissertacao.pdf: 391866 bytes, checksum: 86924f659a38a06a2128bcb727df7899 (MD5) Previous issue date: 2012-08-23 / The aim of this study was to evaluate the development during the cycle and yield of cane sugar, RB 86 7515, led the field with different irrigation water by drip irrigation. The experiment was conducted at the experimental area of experimental UNOESTE, from October 2008 to September 2009 in Presidente Prudente - SP, latitude 22o07´04 "S and longitude 51o22´05" W, altitude 435.5m. The climate is the classification of Köppen Aw mesothermal, with prevalence of hot summers and dry winters. We used a variety RB 86 7515, the most cultivated in the West Paulista by high productivity, rapid growth and tolerance to soils with low / medium yield. The planting was done in furrows 30 cm deep with cuttings of 40 cm in length on October 10, 2008. The experiment was arranged in bands consisting of four irrigation levels, with six replicates, resulting in 24 plots of 31.5 m2. The plots consisted of three rows of seven meters floor area being considered as five meters from the centerline. Irrigation was performed in an irrigation interval of seven days from planting to harvest to 360 days after planting. After the planting was done to ensure a full irrigation starting the treatments with soil at field capacity. Reference evapotranspiration (ETo) was obtained by water balance using daily data of a pan evaporation Class A installed in front of the planting area. The water depth was defined in percentage terms in relation to crop evapotranspiration (ETc) obtained by equation 2: T1 - no irrigation (0% ETc - control), T2 - 50% ETc, T3 - 100% ETc -150% ETc and T4. The height of stem and tillering were affected by irrigation. The leaves had the highest negative natural increase with higher irrigation. ATR per hectare was higher with 50% ETc. / O objetivo do presente trabalho foi avaliar a brotação durante o ciclo e a produtividade de cana-de-açúcar, cultivar RB 86 7515, conduzidas com diferentes lâminas de irrigação por gotejamento superficial. O experimento foi realizado na área experimental da UNOESTE, no período de outubro de 2008 a setembro de 2009, em Presidente, SP, 22o07 04 S e 51o22 05 W, de 435,5 m. O clima é, pela classificação de Köppen, Aw mesotérmico, com verões quentes e invernos secos. Utilizou-se a cultivar RB 86 7515, a mais utilizada no Oeste Paulista por apresentar média produtividade, crescimento rápido e tolerante a solos de baixa/média produtividade. Foi plantado em sulco de 30 cm de profundidade, quarenta e cinco metros de comprimento e com toletes de 40 cm de comprimento, no dia 10 de outubro de 2008. O delineamento experimental utilizado foi em faixas constando de quatro lâminas de irrigação, com seis repetições, resultando em 24 parcelas de 31,5 m2. As parcelas foram constituídas de três linhas de sete metros sendo considerados como área útil os cinco metros da linha central. As irrigações foram realizadas em um espaço de tempo de sete dias desde o plantio até a colheita aos 360 dias após a emergência. Logo após o plantio foi realizada uma irrigação plena para garantir o inicio dos tratamentos com o solo em capacidade de campo, que consiste na quantidade de água retida no solo. A evapotranspiração de referência (ETo) foi obtida utilizando dados diários do Tanque Classe A da estação meteorológica da Unoeste, que fica localizada em frente a área do experimento. As lâminas de irrigação foram definidas em termos percentuais, em relação à evapotranspiração da cultura (ETc) obtidas pela equação 2, sendo: T1 testemunha sem irrigação (0%ETc - testemunha), T2 50% ETc, T3 - 100% ETc e T4 150% ETc. A altura do colmo e o perfilhamento foram afetados pela irrigação. As folhas novas demonstraram maior crescimento vegetativo com maiores lâminas de irrigação. Açúcares Teoricamente Recuperáveis - ATR por hectare foi maior com 50% ETc.

From gametogenesis until death an organism’s genome is under constant bombardment from endogenous and exogenous sources of DNA damage. To maintain genomic integrity amid this damage, cells have evolved responses which allow them to either preserve viability for recovery or initiate self-destructive pathways depending on the severity of DNA damage. One protein involved in initiating and carrying out these responses is the protein kinase ataxia telangiectasia and Rad3-related (ATR). ATR is known primarily for its regulatory role in initiating the checkpoint-signaling cascade following DNA damage and replicative stress. These signaling events lead to cell cycle arrest, DNA repair, or apoptosis when damage is too extreme. In addition to these kinase-dependent roles, ATR also is capable of directly blocking the intrinsic apoptotic pathway through structural sequestration of the proapoptotic protein tBid. The sum of these regulatory events is a delicate balancing act resulting in either cell death or cell survival depending on the severity of the damage and the differentiation state of the cell in question. In the following studies, we sought to investigate the complex interplay of ATR’s kinase and structural roles in determining cellular fate. First, we investigated the structural role of prolyl isomerization of ATR across development by using mouse models of two isomerically locked forms of ATR which were previously shown to lock cytoplasmic ATR into a single isomer. Studies showed that ATR which is locked in ATR-L (trans-ATR, hATR-P429A/mATR-P432A) is embryonically lethal and that heterozygotes tend to have neurological and other developmental abnormalities. This contrasts with ATR-H (cis-ATR, hATR-S428A/mATR-S431A), which is viable, but naturally prone to cancer development. Next, we used various in vitro stroke-like conditions to test if ATR inhibition could serve as a therapeutic target for stroke. We found that ATR inhibition is protective in non-dividing neuron-like cells; whereas, it potentiates death in cycling glial and immune-like cycling cells. Thus, ATR inhibition could likely be a target for both neuron sparing and immunosuppressive anti-stroke therapeutic strategies. Taken together, these studies provide insightful information into the structural and pathological roles of ATR in development and disease.

This master thesis deals with characterization of two black and white products based on ethylene-propylene-diene rubber (EPDM) used for skating rink. Products marked with A and are different in their diameter of circular tubes trough which cooling medium passes and also in the distance of these tubes. Both of materials were characterized in order to determine whether it is the EPDM and how are they different. The characterization methods were used: differential scanning calorimetry, thermogravimetric analysis, attenuated total reflectance Fourier transform infrared spectroscopy, tensile test, swelling test, thermooxidative test. The composition of both materials, including fillers and others additives is almost the same but difference is in structure of EPDM and probably also in interaction with fillers, what exhibits different mechanical properties, thermooxidative stability and thermal capacity. These differences which have been found are essential for long-term use properties of both materials.

The history of African Independent Churches (AICs) in Southern Africa goes back for more than a hundred years. They have proliferated geographically and demographically in Africa more than the mainline churches could ever have imagined. They have grown to be as widespread and as influential as the African mainline churches. The reason for this growth is that the AICs are the churches of African indigenous people. They are launched by Africans from a background of an African traditional and cultural frame of reference. The most significant reason is that the founders of these churches are not Westerners, but Africans. Western missionaries find it difficult to understand the AICs from their perspective. Thus the Western churches describe the AICs as sectarian, separatist, syncretist, nativitist, and so on. Nevertheless, some scholars are attempting to view the AICs in positive ways. The fact that these two different churches have never acknowledged each other as true churches is a big challenge for Christian missions in Namibia. Each group has been viewing and judging the other party through suspicious eyes from their own perspective, each driving the other to block the channel of reconciliation before the presence of God. With the aim of solving this problem, this thesis attempts to answer the following questions about the AICs in Namibia: • What are the reasons that the AICs in Namibia have been seceded from mission churches? • What are the activities in civil society in which the AICs in Namibia are currently involved? • Do the AICs engage in any activities which go against the Word of God? • What causes other churches to be suspicious of the AICs? • What level of enculturation is inherent to the AICs in Namibia? In other words, what is the relationship between the liturgies of the AIC and African traditional religion and African culture? • What makes the AICs in Namibia regard themselves as a church? Would it be possible for the AICs and the mainline churches in Namibia to cooperate in Christian missionary work? • What is a possible Korean missionary perspective on this particular situation? This will be dealt throughout this thesis from a Korean missionary missional perspective. / Thesis (PhD)--University of Pretoria, 2014. / tm2015 / Science of Religion and Missiology / PhD / Unrestricted

En esta tesis se describe un estudio espectroelectroquímico in-situ de la adsorción y reactividad, sobre electrodos de oro, de pequeñas moléculas modelo derivadas de la urea (tiourea (TU) e hidroxiurea (HU)) y compuestos relacionados con las mismas (disulfuro de formamidina, (FDS)), cianato y ácido cianúrico (CYA)). La caracterización voltamperométrica del comportamiento de estas moléculas se complementó con el estudio mediante espectroscopía infrarroja in-situ (IRRAS y ATR-SEIRAS) y Raman (SERS). Además se realizaron cálculos teóricos (DFT) para interpretar los espectros de especies adsorbidas a partir de las geometrías optimizadas y las frecuencias armónicas correspondientes. El trabajo experimental se realizó principalmente con electrodos nanoestructurados de oro, comparando su comportamiento con el obtenido con electrodos monocristalinos Au(111) y Au(100). La caracterización microscópica, voltamperometrica y espectroscópica de los electrodos nanoestructurados de oro, preparados por evaporación térmica sobre sustratos de Si, ha mostrado para los mismos una orientación preferente (111). En el estudio del comportamiento de la tiourea, se ha demostrado la existencia de un proceso de adsorción irreversible de la misma a través del átomo de azufre, con el enlace C-S inclinado con respecto a la normal a la superficie. A potenciales suficientemente altos, la TU adsorbida se desprotona oxidativamente dando lugar a capas mixtas de TU y aniones tioureato, los cuales se enlazan de forma bidentada a través de átomo de azufre y el nitrógeno desprotonado de uno de los grupos amino. Este tipo de capas también puede formarse por disociación homolítica del FDS, producto de la oxidación de la TU. Los cálculos DFT para el FDS indican que esta especie se fisisorbe con una orientación paralela a la superficie de oro. La hidroxiurea presenta un perfil voltamperométrico de oxidación irreversible complejo sobre los electrodos de oro. Esto sugiere la existencia de procesos que involucran especies adsorbidas, con una rápida acumulación de las mismas y un bloqueo de sitios superficiales que produce la inhibición de la reacción de oxidación de HU. En los experimentos espectroscópicos se han detectado bandas asociadas a la formación de CO2, óxido nitroso y aniones cianato adsorbidos, los cuales podrían dar lugar a la formación de ácido isociánico al pH al que se realizan los experimentos. Se han detectado además otras especies adsorbidas que dan lugar a una estructura de bandas de absorción compleja entre 1300 y 1800 cm-1 en los espectros ATR-SEIRAS. Cálculos DFT han demostrado que no hay contribución de la propia HU a estas bandas, planteándose la presencia de otros adsorbatos que conservan el esqueleto NCN y que se forman en etapas sucesivas de desprotonación oxidativa. En particular, una banda alrededor de 1800 cm-1 podría estar relacionada con el grupo carbonilo de la nitrosoformamida, un intermedio propuesto para la oxidación química de la HU. Se ha realizado un estudio espectroelectroquímico paralelo del comportamiento en medio neutro de la adsorción, sobre electrodos de oro, de los aniones cianato. Se han obtenido perfiles voltamperométricos sobre superficies monocristalinas de oro que muestran procesos de adsorción/desorción reversibles con respecto al potencial. Los espectros IRRAS presentan bandas de adsorción en un amplio rango de frecuencias entre 2150 y 2250 cm-1 que, de acuerdo con los cálculos DFT, corresponden a aniones cianato adsorbidos a través de átomo de nitrógeno en una variedad de sitios de adsorción y con frecuencias que dependen de los acoplamientos entre especies adsorbidas que aumentan al hacerlo el grado de recubrimiento. La contribución de posibles formas protonadas (ácidos ciánico e isociánico) puede considerarse como despreciable ya que estas especies interaccionan muy débilmente con la superficie electródica. Por otra parte, experimentos con capas finas de oro en contacto con disoluciones neutras de cianato, muestran bandas adicionales entre 1300 y 1800 cm-1 en los espectros ATR-SEIRA que plantean la existencia de otras especies adsorbidas formadas a partir de cianato adsorbido. La observación de estas bandas en la región de los grupos carbonilo, sugiere la posibilidad de una proceso de trimerización del cianato adsorbido, vía su transformación en ácido isociánico, para formar el anión monocianurato adsorbido. Esta hipótesis se ve refrendada por los espectros obtenidos con los electrodos de capa fina de oro en disoluciones de ácido cianúrico, similares, salvo la banda del cianato adsorbido, a los que se obtienen en disoluciones de cianato, y por los cálculos DFT, que ratifican que el anión monocianurato se adsorbe con su plano molecular perpendicular a la superficie mientras que el ácido cianúrico lo haría de forma paralela a la superficie de Au(111).

Hyperactivation of PARP1 is known to be a major cause of necrotic cell death by depleting NAD+/ATP pools during Ca2+ overload which is associated with many ischemic diseases. However, little is known about how PARP1 hyperactivity is regulated during calcium overload. In this study we show that ATR kinase, well known for its role in DNA damage responses, suppresses ionomycin, glutamate, or quinolinic acid-induced necrotic death of cells including SH-SY5Y neuronal cells. We found that the inhibition of necrosis requires the kinase activity of ATR. Specifically, ATR binds to and phosphorylates PARP1 at Ser179 after the ionophore treatments. This site-specific phosphorylation inactivates PARP1, inhibiting ionophore-induced necrosis. Strikingly, all of this occurs in the absence of detectable DNA damage and signaling up to 8 hours after ionophore treatment. Furthermore, little AIF was released from mitochondria/cytoplasm for nuclear import, supporting the necrotic type of cell death in the early period of the treatments. Our results reveal a novel ATR-mediated anti-necrotic mechanism in the cellular stress response to calcium influx without DNA damage signaling.

Homologous recombination (HR) and nonhomologous end joining (NHEJ) are two distinct DNA double-stranded break (DSB) repair pathways. Here, we report that DNA-dependent protein kinase (DNA-PK), the core component of NHEJ, partnering with DNA-damage checkpoint kinases ataxia telangiectasia mutated (ATM) and ATM- and Rad3-related (ATR), regulates HR repair of DSBs. The regulation was accomplished through modulation of the p53 and replication protein A (RPA) interaction. We show that upon DNA damage, p53 and RPA were freed from a p53-RPA complex by simultaneous phosphorylations of RPA at the N-terminus of RPA32 subunit by DNA-PK and of p53 at Ser37 and Ser46 in a Chk1/Chk2-independent manner by ATR and ATM, respectively. Neither the phosphorylation of RPA nor of p53 alone could dissociate p53 and RPA. Furthermore, disruption of the release significantly compromised HR repair of DSBs. Our results reveal a mechanism for the crosstalk between HR repair and NHEJ through the co-regulation of p53-RPA interaction by DNA-PK, ATM and ATR.

Homologous recombination (HR) and nonhomologous end joining (NHEJ) are two distinct DNA double-stranded break (DSB) repair pathways. Here, we report that DNA-dependent protein kinase (DNA-PK), the core component of NHEJ, partnering with DNA-damage checkpoint kinases ataxia telangiectasia mutated (ATM) and ATM- and Rad3-related (ATR), regulates HR repair of DSBs. The regulation was accomplished through modulation of the p53 and replication protein A (RPA) interaction. We show that upon DNA damage, p53 and RPA were freed from a p53-RPA complex by simultaneous phosphorylations of RPA at the N-terminus of RPA32 subunit by DNA-PK and of p53 at Ser37 and Ser46 in a Chk1/Chk2-independent manner by ATR and ATM, respectively. Neither the phosphorylation of RPA nor of p53 alone could dissociate p53 and RPA. Furthermore, disruption of the release significantly compromised HR repair of DSBs. Our results reveal a mechanism for the crosstalk between HR repair and NHEJ through the co-regulation of p53-RPA interaction by DNA-PK, ATM and ATR.

The genetic diseases Hutchinson-Gilford progeria syndrome (HGPS) and restrictive dermopathy (RD) arise from accumulation of farnesylated prelamin A because of defects in the lamin A maturation pathway. Both of these diseases exhibit symptoms that can be viewed as accelerated aging. The mechanism by which accumulation of farnesylated prelamin A leads to these accelerated aging phenotypes is not understood. Here we present evidence that in HGPS and RD fibroblasts, DNA damage checkpoints are persistently activated because of the compromise in genomic integrity. Inactivation of checkpoint kinases Ataxia-telangiectasia-mutated (ATM) and ATR (ATM- and Rad3-related) in these patient cells can partially overcome their early replication arrest. Treatment of patient cells with a protein farnesyltransferase inhibitor (FTI) did not result in reduction of DNA double-strand breaks and damage checkpoint signaling, although the treatment significantly reversed the aberrant shape of their nuclei. This suggests that DNA damage accumulation and aberrant nuclear morphology are independent phenotypes arising from prelamin A accumulation in these progeroid syndromes. Since DNA damage accumulation is an important contributor to the symptoms of HGPS, our results call into question the possibility of treatment of HGPS with FTIs alone.

This work reports the elucidation of the structure of a copolymer generated by the cross- catalyzed reactions of PF and pMDI prepolymers.  The electronic behavior of phenolic monomers as perturbed by alkali metal hydroxides in an aqueous environment was studied with 1H and 13C NMR.  Changes in electronic structure and thus reactivity were related to solvated ionic radius, solvent dielectric constant, and their effect on ion generated electric field strength. NMR chemical shifts were used to predict order of reactivity for phenolic model compounds with phenyl isocyanate with good success.  As predicted, 2-HMP hydroxymethyl groups were more reactive than 4-HMP in forming urethane bonds under neutral conditions and 2-HMP hydroxymethyl groups were more reactive than 4-HMP in forming urethane bonds under alkaline conditions. The structure of the reaction products of phenol, benzyl alcohol, 2-HMP, and 4-HMP with phenyl isocyanate were studied using 1H and 13C NMR under neutral organic and aqueous alkaline conditions.  Reactions in THF-d8 under neutral conditions, without catalyst, were relatively slow, resulting in residual monomer and the precipitation of 1,3-diphenyl urea from the carbamic acid reaction.  The reactions of phenol, 2-HMP, and 4-HMP in the presence of TEA catalyst favored the formation of phenyl urethanes (PU). Reactions with benzyl alcohol, 2-HMP, and 4-HMP in the presence of DBTL catalyst favored the formation of benzyl urethanes (BU).  Reactions of 2-HMP and 4-HMP led to formation of benzylphenyldiurethane (BPDU).  DBTL catalysts favored formation of BDPU strictly by a benzyl urethane pathway, while TEA favored its formation mostly via phenyl urethane, although some BU was also present.  Under aqueous alkaline conditions, 2-HMP was more reactive than 4-HMP, exhibiting an enhanced reactivity that was attributed to intramolecular hydrogen bonding and a resulting resonance stabilization of the phenolic aromatic ring. ATR-FTIR spectroscopic studies generated real time structural information for model compound reactions of the cross-catalyzed system, differentiating among reaction peaks generated by the carbamic acid reaction, PU and BU formation.  ATR-FTIR also permitted monitoring of propylene carbonate hydrolysis and accelerated alkaline PF resole condensation.  ATR-FTIR data also showed that the overall reaction stoichiometry between the PF and pMDI components drove copolymer formation.  Benzyl urethane formation predominated under balanced stoichiometric conditions in the presence of ammonium hydroxide, while phenyl urethane formation was favored in its absence.  Accelerated phenolic methylene bridge formation became more important when the PF component was in excess in the presence of sufficient accelerator.  A high percentage of free isocyanate was present in solid copolymer formed at ambient temperature. The combination of ammonium hydroxide and tin (II) chloride synergistically enhanced the reactivity of the materials, reducing the residual isocyanate. From 13C CP/MAS NMR of the copolymer, the presence of ammonium hydroxide and tin (II) chloride and the higher PF concentration resulted in substantial urethane formation.  Ammonium hydroxide favored formation of benzyl urethane from the 2-hydroxymethyl groups, while phenyl urethane formed in its absence.  The low alkalinity PF resole with ammonium hydroxide favored benzyl urethane formation.  Comparison of these results with the 13C NMR model compound reactions with phenyl isocyanate under alkaline conditions confirmed high and low alkalinity should favor phenyl and benzyl urethane formation respectively.  These cross catalyzed systems are tunable by formulation for type of co-polymer linkages, reactivity, and cost. / Ph. D.