Submitted by Alice Araujo (alice.caraujo@ufpe.br) on 2018-06-20T17:52:58Z No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) TESE Kennedy Nascimento de Jesus.pdf: 2821633 bytes, checksum: ad7413c76f1a1fd792f8d226e55211d0 (MD5) / Made available in DSpace on 2018-06-20T17:52:58Z (GMT). No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) TESE Kennedy Nascimento de Jesus.pdf: 2821633 bytes, checksum: ad7413c76f1a1fd792f8d226e55211d0 (MD5) Previous issue date: 2017-02-02 / CAPES / A conversão de áreas de vegetação nativa em áreas destinadas à exploração agropecuária é apontada como um dos principais fatores responsáveis pelas emissões de Gases de Efeito Estufa para a atmosfera. Essas mudanças no uso da terra causam na paisagem um mosaico de áreas com diferentes usos e coberturas do solo. Porém, ainda é limitado o conhecimento da dinâmica do carbono (C) e do Nitrogênio (N) do solo nesses sistemas, principalmente na região Nordeste do Brasil. Nesse sentido, o objetivo desse trabalho foi determinar as alterações dos estoques e da dinâmica do C e N no solo devido às mudanças de uso da terra nas diferentes regiões fisiográficas e nas principais classes de solo de Pernambuco. Foram realizados três estudos, que auxiliaram no alcance do objetivo proposto: 1) no primeiro, foi realizado um levantamento e compatibilização da literatura sobre os estoques de C na camada superficial (0-30 cm) dos solos de Pernambuco. 2) no segundo estudo, foram quantificados os estoques de carbono (in situ) e sua distribuição nas camadas ao longo do perfil do solo (0-100 cm) em áreas de vegetação nativa, agrícolas e pasto, nas principais regiões fisiográficas e classes de solo do Estado; e 3) no terceiro, foram avaliados a dinâmica do C e do N, através da abundância natural de δ13C e δ15N e das concentrações de C e N em perfis (0-100 cm) de Argissolos ao longo de um gradiente climático em Pernambuco em áreas representativas de floresta nativa, de pasto e agrícolas. Em todos os estudos, as áreas de vegetação nativa foram tomadas como condição original do solo de acordo com cada região estudada. Nos estudos 2 e 3 quantificou-se as concentrações de C e N por meio de analisador elementar CHN e no estudo 3 foram quantificadas a abundância natural de δ 13C e δ 15N nos Argissolos do Estado, por meio de espectrômetro de massa. Aproximadamente 368 Tg de C estão estocados nos solos de Pernambuco de 0-30 cm. De uma forma geral, os maiores estoques de carbono total ocorreram nas vegetações nativas densas e decresceram da região úmida à semiárida Oeste, com grandes variações entre as classes de solos. A substituição da vegetação nativa para implantação de áreas de pastagens ou agrícolas acarretou mudanças no sinal δ13C do solo, sendo maiores nas áreas úmidas e subúmidas do estado em relação às áreas semiáridas. O enriquecimento de δ15N encontrado na superfície do solo das regiões semiáridas em relação às áreas úmidas e subúmidas estão associados as menores concentrações de C e N presentes nos Argissolos dessa região, em decorrência das maiores temperaturas e menores precipitações pluviais. Fica evidente que a intervenção humana nessas áreas através de práticas agropecuárias, reduz os estoques e modificam a dinâmica do C e do N no solo, e para se contrapor a esta situação no Estado, são necessárias medidas mitigatórias, como a adoção de sistemas conservacionistas do solo e da água, através de políticas públicas de incentivo às práticas conservacionistas em substituição à agricultura de baixo C. / The conversion of native vegetation areas into agricultural areas is pointed out as one of the main factors responsible for the emissions of greenhouse gases to the atmosphere. Changes in land use create a mosaic of areas in the landscape with different soil coverages and knowledge of the dynamics of C and N in these systems is limited. The objective of this work was to determine the changes in soil carbon (C) and nitrogen (N) stocks and dynamics due to changes in land use in the different physiographic regions and in the main soil classes of Pernambuco state. Three studies were carried out: 1) the first was a survey and compatibilization of the literature on C stocks in the surface layer (0-30 cm) of the soils of Pernambuco; 2) the second study was a quantification of the C stocks along the different layers of the soil profile (0-100 cm) in areas under native vegetation, agriculture and pasture, in the main physiographic regions and soil classes of Pernambuco; and 3) the third study evaluated soil C and N dynamics through the natural abundance of δ13C and δ15N and the concentrations of C and N in profiles of Ultisols (0-100 cm) along a climatic gradient in Pernambuco. In all studies, dense native vegetation areas were taken as the original, reference soil condition. In studies 2 and 3 the concentrations of C and N were quantified using CHN elemental analyzer. In study 3 the natural abundance of δ13C and δ15N were quantified by mass spectrometer. Approximately 368 Tg of carbon are stored in the soils of Pernambuco in the 0-30 cm layer. The largest total carbon stocks occurred under dense native vegetation and decreased from the humid region towards the semi-arid western state region, with large variations among soil classes. The replacement of the native vegetation by agriculture or pasture caused changes in the soil δ13C signal, being greater in the humid areas and subhumid regions than in the semi-arid region. The enrichment of δ15N in the soil surface layer of the semi-arid regions in relation to the humid and subhumid regions is associated with the lower concentrations of C and N in these regions, due to their higher temperatures and lower rainfall. It is evident that Human intervention in these areas through conventional farming practices reduces soil carbon stocks at levels lower than those found in native areas, and modifies the C and N dynamics in the soil in these regions. To counteract this situation in the State, mitigating measures are necessary, such as the adoption of soil and water conservation systems, through public policies to encourage conservation practices in place of low carbon agriculture.

Magister Scientiae - MSc / As a means of further localising the interaction, truncated fragments derived from the C-terminal region of YB-1, were tested for their interaction with the RING finger domain of RBBP6 using three different assays: a directed yeast 2-hybrid assay, co-immunoprecipitation and NMR chemical shift perturbation analysis. Our results suggest that the entire 62 amino acid region at the C-terminal domain of YB-1 may be involved in the interaction with RBBP6. Using chemical shift perturbation analysis, this study provides an indication of where YB-1 binds to the RING finger. This represents the first step towards the design of therapeutics aimed at modulating the interaction between RBBP6 and YB-1 as a means of targeting the oncogenic effects of YB-1. In order to identify E2 enzymes involved in the ubiquitination of YB-1, we examined the efficiencies of selected E2s in an in vitro ubiquitination assay. UbcH5c and UbcH7 were both found to catalyse the ubiquitination of YB-1 in conjuction with RBBP6, whereas Ubc13 was not. Finally, we show using NMR that two single-point mutations of the RING finger-like domain are sufficient to abolish homodimerisation of the domain. These will be used in future studies to investigate the requirement for homodimerisation on the ubiquitination activity of RBBP6. / South Africa

Magister Scientiae - MSc / Retinoblastoma binding prot ein 6 (RBBP6) is a 250 kDa human splicing-associated protein that is also known to interact with tumour suppresso r proteins p53 and pRb and to mediate ubiquitination of p53 via its intera ction with Hdm2. RBBP6 is highly up regulated in oesophageal cancer, and has been shown to be a promising target for immunotherapy against the disease. RBBP6 is also known to play a role in mRNA splicing, cell cycle control and apoptosis. / South Africa

Una fertilización nitrogenada racional debe contemplar no sólo el aporte de una dosis ajustada a las necesidades del cultivo, sino que a su vez debe considerar la correcta distribución de ésta durante el período de abonado. Sin embargo, no se dispone de suficiente información del efecto que la distribución estacional del fertilizante tiene sobre la absorción y la movilización del N acumulado en los órganos de reserva de los cítricos. El objetivo del presente trabajo es evaluar en plantas jóvenes de cítricos el efecto de la distribución estacional diferencial del abonado nitrogenado sobre la absorción del N y su reparto en los distintos órganos, la movilización del N acumulado en los órganos viejos de reserva hacia los óganos en desarrollo, así como su repercusión en la fructificación. De este modo, se profundizará tanto en el conocimiento de la dinámica del N en el sistema planta-suelo en los cítricos, como en los posiles factores implicados en este proceso, con el fin de ampliar las bases sobre las que descansan los criterios del abonado nitrogenado y optimizar así la aplicación estacional de los fertilizantes. Para la consecución de estos objetivos se recurrió al empleo de la técnica de dilución isotópica mediante la incorporación al sistema planta-suelo de un fertilizante marcado con el isótopo estable 15N. Éste se aplicó desde el inicio de la actividad vegetativa (principios de marzo) hasta el completo desarrollo del fruto (final de octubre) siguiento tres distribuciones estacionales. Se comparó una distribución simétrica en la que se aplicó igual fracción de la dosis desde el inicio del abonado hasta principio de julio (final de la caída fisiológica) y desde ese momento en adelante, con otras dos en las que el máximo aporte del abono (75% de la dosis) se adelantó con respecto a ésta en los meses comprendidos entre marzo y julio, ó se retrasó el periodo de julio a octubre. / Martínez Alcántara, B. (2010). Estudio de la absorción y translocación del nitrógeno en cítricos en función del aporte estacional del abono nitrogenado mediante el uso de la técnica de dilución isotópica [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/8380 / Palancia

The gut microbiome and its metabolic processes are dynamic systems. Surprisingly, our understanding of gut microbiome dynamics is limited. Here we report a metaproteomic workflow that involves protein stable isotope probing (protein-SIP) and identification/quantification of partially labeled peptides. We also developed a package, which we call MetaProfiler, that corrects for false identifications and performs phylogenetic and time series analysis for the study of microbiome dynamics. From the stool sample of five mice that were fed with 15-N hydrolysate from Ralstonia eutropha, we identified 15,297 non-redundant unlabeled peptides of which 10,839 of their heavy counterparts were quantified. These peptides revealed incorporation profiles over time that were different between and within taxa, as well as between and within clusters of orthologous groups (COGs). Our study helps unravel the complex dynamics of protein synthesis and bacterial dynamics in the mouse gut microbiome.

>Magister Scientiae - MSc / Retinoblastoma binding protein 6 (RBBP6) is a 250 kDa RING finger-containing protein whose function is known to be mediated through interaction with other proteins. RBBP6 plays a role in the regulation of the tumour suppressor protein p53 and is also thought to be involved in mRNA splicing although its role has yet to be characterised. A recent study utilising a yeast 2-hybrid screen identified the cancer-associated protein known as YB-l as an interacting partner of RBBP6, and showed that RBBP6 ubiquitinates YB-I, leading to its degradation in the proteasome.Human Y-box binding protein 1 (YB-I) is member of the cold-shock domain family of proteins, which regulates a number of growth related genes through both transcriptional and translational mechanisms. YB-l is a cell-survival factor whose expression is increased in proliferating normal and cancer cells. It also protects cells against p53-mediated apoptosis by repressing the p53- promoter and down-regulating endogenous p53. The interaction between RBBP6 and YB-l involves the RING finger-like domain ofRBBP6 and the C-terminal62 amino acids ofYB-l. As a means of further localising the interaction, truncated fragments derived from the C-terminal region of YB-I, were tested for their interaction with the RING finger domain of RBBP6 using three different assays: a directed yeast 2-hybrid assay, co-immunoprecipitation and NMR chemical shift perturbation analysis. Our results suggest that the entire 62 amino acid region at the C-terminal domain ofYB-l may be involved in the interaction with RBBP6. Using chemical shift perturbation analysis, this study provides an indication of where YB-l binds to the RING fmger. This represents the first step towards the design of therapeutics aimed at modulating the interaction between RBBP6 and YB-l as a means of targeting the oncogenic effects ofYB-l. In order to identify E2 enzymes involved in the ubiquitination of YB-I, we examined the efficiencies of selected E2s in an in vitro ubiquitination assay. UbcH5c and UbcH7 were both found to catalyse the ubiquitination of YB-l in conjuction with RBBP6, whereas Ubc 13 was not. Finally, we show using NMR that two single-point mutations of the RING finger-like domain are sufficient to abolish homodimerisation of the domain. These will be used in future studies to investigate the requirement for homodimerisation on the ubiquitination activity of RBBP6. www.etd.

Motiviert durch die Zunahme der weltweiten Nachfrage nach hochwertigen Humusstoffen wurde am Institut für Holz- und Pflanzenchemie ein neuartiger Humusersatzstoff entwickelt. Dieses N-modifizierte Produkt wird auf Basis eines patentierten Normaldruckverfahrens der oxidativen Ammonolyse (OA) aus ligninhaltigen Ausgangssubstanzen, im vorliegenden Fall Lausitzer Braunkohle, hergestellt. Ziel war, die bisher vorliegenden Erkenntnisse der strukturellen Eigenschaften solcher Humusstoffe zu erweitern. Im Fokus standen die Gehalte und Charakteristik der Huminstofffraktionen (Humine, Humin- und Fulvosäuren). Der Stickstoff und die Art seines Einbaus in die organische Substanz spielt hierbei eine besondere Rolle. Für die Huminstoffisolierung kam die hinsichtlich der Ausbeuten und des Zeitaufwandes optimierte IHSS-Methode zum Einsatz. Wesentliche Merkmale N-modifizierter Substanzen sind erhöhte Huminsäurengehalte, zeitlich differenziert wirksame N-Bindungsformen und damit eine Diversifizierung der Funktionalität. Zusätzlich mit dem geringen Aschegehalt unterscheidet dies N-modifizierte Lausitzer Braunkohle von anderen auf Braunkohlebasis erzeugten Bodenverbesserungsmitteln des Marktes. Es konnte gezeigt werden, dass Huminsäuren hinsichtlich der Bereitstellung von Stickstoff und Funktionalität (z.B. Austauscherplätze) eine Schlüsselrolle einnehmen. Die Veränderungen durch die OA basieren stofflich auf der Bildung regenerierter Huminsäuren und chemisch u.a. auf der oxidativen Ringspaltung von Aromaten an nicht veretherten phenolischer OH-Gruppen von Methoxyphenolstrukturen. Dabei ist diese Reaktion entgegen früherer Annahmen nicht an eine Demethoxylierung gebunden und erfordert auch keine verschärften Reaktionsbedingungen (z.B. erhöhter Druck). Diese und weitere Reaktionsmechanismen führen zu ammoniumartig, amidartig und fest organisch gebundenen N-Bindungsformen. 15N-NMR-Spektroskopische Untersuchungen an einem 15N-angereicherten Produkt, Py-GC/MS-Untersuchungen und nasschemische Experimente zeigen, dass in der Vergangenheit die Bedeutung heterocyclischer, insbesondere heteroaromatischer N-Bindungsformen überschätzt wurde. Amidartiger Stickstoff präsentiert sich dabei als außerordentlich heterogen hinsichtlich seiner Hydrolysestabilität bzw. Pflanzenverfügbarkeit. Für die effektive Beurteilung möglicher Ausgangssubstanzen hinsichtlich ihrer Eignung für die Herstellung von Humusdüngestoffen nach dem Prinzip der OA kann der Huminsäurengehalt herangezogen werden. Er wird mit Hilfe eines degradativen Verfahrens ermittelt. Die mit dem Huminsäurengehalt in Verbindung stehenden Stoffeigenschaften von Kohlen korrelieren gut mit dem N-Einbau. Aus Sicht der Verfahrensführung hängt die Intensität des N-Einbaus eng mit dem Oxidationsregime zusammen. Durch Anwendung von reinem Sauerstoff anstelle von Luft als sehr einfach und günstig zu realisierende Maßnahme kann die Reaktionszeit halbiert werden. / The development of a novel artificial humus material at the Institute of Wood- and Plant Chemistry was accounted by the increasing demand for high grade humic matter. This N-modified product base on a patented ambient pressure technology of the oxidative ammonolysis. of lignin containing substances, in particular Lusitian lignite. Objective of the work was to broadening the current knowledge of structural properties with focus on the contents and characteristics of humic substances fractions (humins, humic acids, fulvic acids). Especially attention was given to nitrogen and its way of incorporation into organic matter. For isolating humic substances the IHSS method was used. The procedure was adapted in order to increase the yield of humic acids and decrease time needed. Main characteristics of N-modified substances are higher contents of humic acids and subtly differenciateted effective N-binding forms with leads to higher diversity of the chemical functionality. This and the low ash content makes the differences to other brown coal based soil improving agents available on the marked. It has been shown that humic acids give the main part providing nitrogen and functionality (e.g. cation exchange capacity). The oxidative ammonolysis leads to regenerated humic acids as well as e.g. the clevage of aromatic structures by reactions of free phenolic groups. In contrast to former assumptions the cleavage is not strongly related to demethoxylation or strong reaction conditions like high pressure. This and other reactions lead to short, middle, and long lasting N-binding forms. 15N-NMR-spectroscopic investigations on a 15N-enriched product, Py-GC/MS-investigations and conventional investigations show an overestimation of heterocyclic, in particular heteroaromatic N-binding forms up to now. Consequently amide like nitrogen reveals as extraordinary according to its persistent behaviour to hydrolysis and plant availability respectively. Corresponding the correlation of the N-incorporation and humic acid content of raw materials the latter can be used for evaluating possible raw material for its usage for N-modification. It can be easy obtained in a degratadive way. From the process engineering point of view the success of N-incorporating is strongly correlated to the oxidation conditions during processing. Using pure oxygen instead of air shorts the needed reaction at 50%.

Alfalfa overseeded into warm-season perennial grasses such as bahiagrass or bermudagrass has the potential to increase forage quality and dry matter yield as well as extend the growing season. Many of the challenges associated with alfalfa in mixtures have been observed in warm-season grasses, but few studies have attempted to explain the dynamics between these grasses and alfalfa when grown together. Two studies were designed to note these relationships: The objective of the first study was to observe the effect of seeding rate and sod preparation on alfalfa and bermudagrass persistence over three years by using minimum tillage and alfalfa seeding rates of 17, 22, 28, and 39 kg ha-1. Results indicated that sod preparation had no effect on establishment compared to no-tillage. Seeding rates resulted in similar biomass yields and species composition by the second year suggesting that increased seeding rates were unnecessary. Forage quality and yield were improved throughout the year, but alfalfa composition generally decreased towards the end of summer. The objective of the second study was to quantify the amount of nitrogen recovered when alfalfa was overseeded into bahiagrass and bermudagrass and fertilized with ammonium nitrate during late summer. This was accomplished by using two nitrogen rates of 15N labeled fertilizer (28 and 56 kg N ha-1) and two application times (after 2nd and 3rd harvest) and in two separate planting environments (planting I and II). It was found that bahiagrass was more competitive than bermudagrass when overseeded with alfalfa. Alfalfa/grass mixtures did not increase forage dry matter yield, nutritive value or grass composition with N application. Nitrogen fertilizer recovery was variable between plantings due to a large variation of alfalfa persistence within different soil types. Where alfalfa was limited, grass fertilizer recovery was over 60%, but when grass composed less than 10% in the mix, then alfalfa had the potential to recover over 40% of the applied fertilizer. The greatest N recovery by grass were favored during late N application. On the other hand, alfalfa N recovery was favored by early fertilizer application.

Stable isotope analysis is often used to evaluate elements of the lives of past peoples, such as diet and health status, at a societal level. Analysis at an individual level is exceptionally rare, and has not been conducted using a variety of tissues representing both early life and life approximate to death. In this study, δ 13C and δ15N isotope signatures are used to create life histories for single individuals from Romano-Christian period Kellis 2 cemetery in the Dakhleh Oasis, Egypt. Samples are obtained from several different tissues, including tooth dentin, bone collagen, hair, nail, skin, and gut content, all of which have been previously researched, but have not been studied at such an individualistic level. By using data and previous research conducted by Drs. Tosha Dupras and Lana Williams, this research uses isotopic values from the aforementioned tissues, and the differing turnover rates of these tissues, to develop lifetime timetables for 15 individuals (female, male, and juvenile). Results show that individual analysis is possible, informative, and can enlighten researchers not just concerning the individual, but about the population as a whole. The methods presented can serve as a model for reconstructing individual life histories using isotope data from multiple tissues.

No description available.