Immobilization of Aspergillus oryzae alpha-galactosidase in gelatin and its application in removal of flatulence-inducing sugars in soymilk

Raffinose oligosaccharides (RO) are the major factors responsible for flatulence following ingestion of soybean-derived products. Removal of RO from seeds or soymilk would then have a positive impact on the acceptance of soy-based foods. In this study, alpha-galactosidase from Aspergillus oryzae was entrapped in gelatin using formaldehyde as the hardener. The immobilization yield was 64.3% under the optimum conditions of immobilization. The immobilized alpha-galactosidase showed a shift in optimum pH from 4.8 to 5.4 in acetate buffer. The optimum temperature also shifted from 50 degrees C to 57 degrees C compared with soluble enzyme. Immobilized alpha-galactosidase was used in batch, repeated batch and continuous mode to degrade RO present in soymilk. In the repeated batch, 45% reduction of RO was obtained in the fourth cycle. The performance of immobilized alpha-galactosidase was tested in a fluidized bed reactor at different flow rates and 86% reduction of RO in soymilk was obtained at 25 ml h(-1) flow rate. The study revealed that immobilized alpha-galactosidase in continuous mode is efficient in reduction of RO present in soymilk.

Synthesis of new Aβ-ligands useful in diagnosis of Alzheimer's disease

Alzheimer’s disease is a chronic progressive neurodegenerative disease and is the most common form of dementia (estimated 50−60% of all cases), associated with loss of memory (in particular episodic memory), cognitive decline, and behavioural and physical disability, ultimately leading to death. Alzheimer’s disease is a complex disease, mostly occurring sporadically with no apparent inheritance and being the age the main risk factor. The production and accumulation of amyloid-beta peptide in the central nervous system is a key event in the development of Alzheimer’s disease. This project is devoted to the synthesis of amyloid-beta ligands, fluorophores and blood brain barrier-transporters for diagnosis and therapy of Alzheimer’s disease. Different amyloid-beta ligands will be synthesized and their ability to interact with amyloid-beta plaques will be studied with nuclear magnetic resonance techniques and a process of lead optimization will be performed. Many natural and synthetic compounds able to interact as amyloid-beta ligands have been identified. Among them, a set of small molecules in which aromatic moieties seem to play a key role to inhibit amyloid-beta aggregation, in particular heteroaromatic polycyclic compounds such as tetracyclines. Nevertheless tetracyclines suffer from chemical instability, low water solubility and possess, in this contest, undesired anti-bacterial activity. In order to overcome these limitations, one of our goals is to synthesize tetracyclines analogues bearing a polycyclic structure with improved chemical stability and water solubility, possibly lacking antibacterial activity but conserving the ability to interact with amyloid-beta peptides. Known tetracyclines have in common a fourth cycle without an aromatic character and with different functionalisations. We aim to synthesize derivatives in which this cycle is represented by a sugar moiety, thus bearing different derivatisable positions or create derivatives in which we will increase or decrease the number of fused rings. In order to generate a potential drug-tool candidate, these molecules should also possess the correct chemical-physical characteristics. The glycidic moiety, not being directly involved in the binding, it assures further possible derivatizations, such as conjugation to others molecular entities (nanoparticles, polymeric supports, etc.), and functionalization with chemical groups able to modulate the hydro/lipophilicity. In order to be useful such compounds should perform their action within the brain, therefore they have to be able to cross the blood brain barrier, and to be somehow detected for diagnostic purposes.

Critique praxéologique d’une exposition sur le « vrai » visage de Jésus : un essai de théologie trash

Ce mémoire s'inscrit dans la méthode de praxéologie pastorale. Il analyse en qua-tre temps la mise en œuvre d'une exposition picturale qui oppose une figure alié-nante (conventionnelle et doucereuse) de Jésus à une vision trash (c'est à dire sub-versive et provocante) qui se veut plus fidèle à l'évangile. Le premier temps (ob-servation), qui présente le premier projet d'exposition ainsi que ses sources d'inspi-ration (personnelle, culturelle et artistique), amène à cerner deux problématique : la question de la vérité et l'orientation subversive (trash) du Jésus des évangiles. Le deuxième temps (interprétation) creuse tout d'abord la question de la vérité avec Hans Georg Gadamer, ce qui pousse à dépasser l'affirmation violente de la vérité du premier projet d'exposition pour le réorienter dans une perspective de questionnement. Par ailleurs, un modèle de dynamique trash permet de fonder la figure subversive et provocante de Jésus dans une relecture des évangiles, d’où un troisième temps (intervention) : l’amendement du premier projet d'exposition. En guise de conclusion, le quatrième temps (prospective) exporte des pointes de ré-flexion au-delà de l'interprétation concrète, à savoir au plan des questions de l'art et du public, du deuil de la vérité, de la dynamique trash et de la foi.

Abrupt climate change: ice sheets and oceans in action

Large temperature variations on land, in the air, and at the ocean surface, and highly variable flux of ice-rafted debris (IRD) delivered to the North Atlantic Ocean show that rapid climate fluctuations took place during the last glacial period. These quasi-periodic, high-amplitude climate variations followed a sequence of events recognized as a rapid warming, followed by a phase of gradual cooling, and terminating with more rapid cooling and increased flux of IRD to the north Atlantic Ocean. Each cycle lasted ˜1500 years, and was followed by an almost identical sequence. These cycles are referred to as Dansgaard/Oechger cycles (D/O cycles), and approximately every fourth cycle culminated in a more pronounced cooling with a massive discharge of IRD into the north Atlantic Ocean over an interval of ˜500 years. These massive discharges of IRD are known as Heinrich layers. “Heinrich events” are thus characterized as a rapid transfer of IRD from a “source,” the bed of the Laurentide Ice Sheet (LIS), to a “sink,” the North Atlantic.

O ensino de óptica no nível fundamental: uma proposta de ensino aprendizagem contextualizada para a oitava série

This dissertation presents the investigation and possible interference in the current teaching of physics, specially of optics, in the eighth grade/ ninth year / fourth cycle of junior high school. The context of the current science teaching is also presented. Besides, the following aspects are discussed: the preparation of the eighth grade teachers, their professional needs, the pedagogical practices and the theoreticmethodological difficulties faced by them. Another important issue in Science Teaching, which is also discussed in this dissertation, is the need for an efficient scientific literacy so that the citizen may express value judgment about Science and Technology issues. The data about the context were gathered from questionaires answered by the teachers. In value of this information, lesson plans were elaborated and implemented. The goal was to point out alternatives for the development of a teaching-learning evaluation which would be both pleasant for the teachers in relation to the implemented methodology, end meaningful for the students by adding elements such as: interdisciplinary approach, contextualization and preparation by competences. Furthermore it was carried out a deeper analysis of the Brazilian curriculum references (LDB, DCN, PCN, RCB) and content aspects (concepts, procedures e attitudes) to be executed by that branch of human knowledge, aiming at a really Contextualized Science Teaching, qualified and pleasant for the eighth grade level. It is also presented in this paper the profile or the Professional which teach this level at state public schools in Russas, state of Ceará, Vale do Jaguaribe Region. The efficacy of the lesson plans were also analysed by using evaluation procedures with the eighth grade students of the following schools: CAIC Senador Carlos Jereissati and EEF Manuel Matoso Filho, from which the concepts acquired during the implementation of the classes became evident in the students

Atlas escolar interativo: desenvolvimento de um protótipo para o quarto ciclo do ensino fundamental

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

PCN de história na escola: caminho e descaminhos para a construção da cidadania

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Representação social de ciência de estudantes do ensino fundamental da rede municipal de Belém

Este trabalho relata uma pesquisa descritiva quanti-qualitativa realizada entre estudantes do ensino fundamental da rede municipal de Belém, com os objetivos de identificar a representação social de Ciência desses alunos, além de compreender os conteúdos e identificar as tendências de núcleo central reveladas nessas representações. A pesquisa teve como sujeitos 124 estudantes de uma escola da rede municipal de Belém, dos quais 65 encontravam-se cursando 3o ciclo e 59 o 4o ciclo do ensino fundamental. Dos 124, 52 são do sexo masculino e 72 são do sexo feminino, distribuídos entre as faixas etárias de 10 a 17 anos. Tais representações são discutidas tendo como referências teórico-metodológicas a teoria das representações sociais de Serge Moscovici, e suas teorias complementares, como a Teoria do Núcleo Central de Jean-Claude Abric. Os dados coletados através de questionários, compostos por uma questão aberta e uma de evocação livre foram analisados utilizando-se a técnica de análise de conteúdo e de Vergès, respectivamente. A análise dos dados nos revelou a vinculação restrita da Ciência à disciplina escolar Ciências e a alguns de seus conteúdos, como corpo humano e meio ambiente, entre outros. Esses resultados nos fazem concluir que tais representações estão fortemente vinculadas à abordagem adotada pelo ensino de Ciências, ainda predominantemente prescritivo e propedêutico. Com essa conclusão evidencia-se a necessidade de incentivo à formação dos professores de Ciências, na expectativa de se viabilizar a implementação de propostas de ensino que pretendam romper, com esse modelo tradicional, como Alfabetização Científica e Ciência, Tecnologia e Sociedade (CTS), dessa forma contribuindo para formação de cidadãos atuantes de maneira crítica e criativa em nossa sociedade.

Adubação nitrogenada e potássica em goiabeiras ‘paluma’: II. efeito no estado nutricional das plantas

The proper mineral fertilization of guava orchards under intensive management, is a major factor in productivity and monitoring the nutritional status of the plants contributes to the efficiency of this agronomic practice. The aim of this study was to investigate different doses of nitrogen and potassium in the nutritional status of guava 'Paluma'. The experiment was conducted at Vista Alegre do Alto, Sao Paulo, in a 7 year old irrigated orchard, managed with fruiting pruning during four consecutive growing seasons. The soil is a dystrophic Ultisol. The experimental design was randomized blocks, in factorial, with four nitrogen doses (0, 0.5, 1 and 2 kg of N plant(-1)) and four of potassium (0, 0.55, 1.1 and 2.2 kg of K2O plant(-1)), with three replications. Nitrogen fertilization promoted increased levels of leaf N and Mn and the decrease in the levels of P and B; effects observed from the second to the fourth production cycle. Nitrogen fertilization increased the Ca and Mg levels, respectively, in the second and third cycle. Except for the first production cycle, K and Mn foliar concentrations increased with increasing potassium fertilization, whereas the levels of Mg, in the second and the fourth cycle, decreased as a function of fertilization.

Growth and flowering of five mango cultivar under subtropics conditions of Brazil

The evaluation of the new cultivars adaptation and yield potential of Mangifera indica L. provides tools to assist and improve the mango production in different climates conditions. This study aimed to evaluate the phenological and reproductive development of five mango cultivars (Bourbon, Haden, Palmer, Parwin and Tommy Atkins), on climate conditions in dry land and four production cycles (from 08/2007 to 01/2011), at São Manuel, São Paulo state, Brazil. The variables were total plant height, trunk height, stem diameter and the first insertion diameter, fruits physical characteristics, yield and harvest period, flowering seasonality and morphological characterization of the inflorescences. It was found that the four-year-old plants of Haden cv. reach 4.0 m high and 0.20 m of trunk diameter. Bourbon cultivar had the highest flowering period, from April to October. Tommy Atkins cv. had productivities of 14779.07 kg∙ha−1 in the fourth cycle and the productivity of Bourbon, Haden and Palmer cultivars was alternated.

Enzyme engineering of aminotransferases for improved activity and thermostability

The production by biosynthesis of optically active amino acids and amines satisfies the pharmaceutical industry in its demand for chiral building blocks for the synthesis of various pharmaceuticals. Among several enzymatic methods that allow the synthesis of optically active aminoacids and amines, the use of minotransferase is a promising one due to its broad substrate specificity and no requirement for external cofactor regeneration. The synthesis of chiral compounds by aminotransferases can be done either by asymmetric synthesis starting from keto acids or ketones, and by kinetic resolution starting from racemic aminoacids or amines. The asymmetric synthesis of substituted (S)-aminotetralin, an active pharmaceutical ingredient (API), has shown to have two major factors that contribute to increasing the cost of production. These factors are the raw material cost of biocatalyst used to produce it and product loss during biocatalyst separation. To minimize the cost contribution of biocatalyst and to minimize the loss of product, two routes have been chosen in this research: 1. To engineer the aminotransferase biocatalyst to have greater specific activity, and 2. Improve the engineering of the process by immobilization of biocatalyst in calcium alginate and addition of cosolvents. An (S)-aminotransferase (Mutant CNB03-03) was immobilized, not as purified enzyme but as enzyme within spray dried cells, in calcium alginate beads and used to produce substituted (S)-aminotetralin at 50 °C and pH 7 in experiments where the immobilized biocatalyst was recycled. Initial rate of reaction for cycle 1 (6 hr duration) was determined to be 0.258 mM/min, for cycle 2 (20 hr duration) it decreased by ~50% compared to cycle 1, and for cycle 3 (20 hr duration) it decreased by ~90% compared to cycle 1 (immobilized preparation consisted of 50 mg of spray dried cells per gram of calcium alginate). Conversion to product for each cycle decreased as well, from 100% in cycle 1 (About 50 mM), 80% in cycle 2, and 30% after cycle 3. This mutant was determined to be deactivated at elevated temperatures during the reaction cycle and was not stable enough to allow multiple cycles in its immobilized form. A new mutant aminotransferase was isolated by applying error-prone polymerase chain reaction (PCR) on the gene coding for this enzyme and screening/selection: CNB04-01. This mutant showed a significant improvement in thermostability in comparison to CNB03-03. The new mutant was immobilized and tested under similar reaction conditions. Initial rate remained fairly constant (0.2 mM/min) over four cycles (each cycle with a duration of about 20 hours) with the mutant retaining almost 80% of initial rate in the fourth cycle. The final product concentrations after each cycle did not decrease during recycle experiments. Thermostability of CNB04-01 was much improved compared to CNB03-03. Under the same reaction conditions as stated above, the addition of co-solvents was studied in order to increase substituted tetralone solubility. Toluene and sodium dodecylsulfate (SDS) were used. SDS at 0.01% (w/v) allowed four recycles of the immobilized spray dried cells of CNB04-01, always reaching higher product concentration (80-85 mM) than the system with toluene at 3% (v/v) -70 mM-. The long term activity of immobilized CNB04-01 in a system with SDS 0.01% (w/v) at 50 °C, pH 7 was retained for three cycles (20 to 24 hours each one), reaching always final product concentration between 80-85 mM, but dropping precipitously in the fourth cycle to a final product concentration of 50 mM. Although significant improvement of immobilization on productivity and stability were observed using CNB04-01, another observation demonstrated the limitations of an immobilization strategy on reducing process costs. After analyzing the results of this experiment it was seen that a sudden drop occurred on final product concentration after the third recycle. This was due to product accumulation inside the immobilized preparation. In order to improve the economics of the process, research was focused on developing a free enzyme with an even higher activity, thus reducing raw material cost as well as improving biomass separation. A new enzyme was obtained (CNB05-01) using error-prone PCR and screening using as a template the gene derived from the previous improved enzyme. This mutant was determined to have 1.6 times the initial rate of CNB04-01 and had a higher temperature optimum (55°). This new enzyme would allow reducing enzyme loading in the reaction by five-fold compared to CNB03-03, when using it at concentration of one gram of spray dried cells per liter (completing the reaction after 20-24 hours). Also this mutant would allow reducing process time to 7-8 hours when used at a concentration of 5 grams of spray dried cells per liter compared to 24 hours for CNB03-03, assuming that the observations shown before are scalable. It could be possible to improve the economics of the process by either reducing enzyme concentration or reducing process time, since the production cost of the desired product is primarily a function of both enzyme concentration and process time.

Redefining life cycle for a building sustainability assessment framework

Understanding the differences between the temporal and physical aspects of the building life cycle is an essential ingredient in the development of Building Environmental Assessment (BEA) tools. This paper illustrates a theoretical Life Cycle Assessment (LCA) framework aligning temporal decision-making with that of material flows over building development phases. It was derived during development of a prototype commercial building design tool that was based on a 3-D CAD information and communications technology (ICT) platform and LCA software. The framework aligns stakeholder BEA needs and the decision-making process against characteristics of leading green building tools. The paper explores related integration of BEA tool development applications on such ICT platforms. Key framework modules are depicted and practical examples for BEA are provided for: • Definition of investment and service goals at project initiation; • Design integrated to avoid overlaps/confusion over the project life cycle; • Detailing the supply chain considering building life cycle impacts; • Delivery of quality metrics for occupancy post-construction/handover; • Deconstruction profiling at end of life to facilitate recovery.

The Neurofibromatosis 2 tumor suppressor merlin in cytoskeleton organization and cell cycle regulation

Neurofibromatosis 2 (NF2) is a dominantly inherited disorder, which predisposes to multiple tumours of the nervous system, typically schwannomas and meningiomas. Biallelic inactivation of the NF2 gene occurs both in sporadic and NF2-related schwannomas and in most meningiomas. The NF2 gene product merlin (or schwannomin) is structurally related to the ERM proteins, ezrin, radixin and moesin, which act as molecular linkers between the actin cytoskeleton and the plasma membrane. Merlin is a tumor suppressor that participates in cell cycle regulation. Merlin s phosphorylation status appears to be associated with its tumour suppressor activity, i.e. non-phosphorylated merlin functions as a tumour suppressor, whereas protein phosphorylation results in loss of functional activity. This thesis study was initiated to investigate merlin s role as a tumor suppressor and growth inhibitor. These studies show, that like many other tumor suppressors, also merlin is targeted to the nucleus at some stages of the cell cycle. Merlin s nuclear localization is regulated by cell cycle phase, contact inhibition and adhesion. In addition, a potential nuclear binding partner for merlin was identified, Human Enhancer of Invasion 10 (HEI10), a cyclin B interacting protein. Many tumor suppressors interact with microtubules and this thesis work shows that also merlin colocalizes with microtubules in mitotic structures. Merlin binds microtubules directly, and increases their polymerization in vitro and in vivo. In addition, primary mouse Schwann cells lacking merlin displays disturbed microtubule cytoskeleton. Fourth part of this thesis work began from the notion that PKA phosphorylates an unidentified site from the merlin N-terminus. Our studies show that serine 10 is a target for PKA and modulation of this residue regulates cytoskeletal organization, lamellipodia formation and cell migration. In summary, this thesis work shows that merlin s role is much more versatile than previously thought. It has a yet unidentified role in the nucleus and it participates in the regulation of both microtubules and the actin cytoskeleton. These studies have led to a better understanding of this enigmatic tumor suppressor, which eventually will aid in the design of specific drugs for the NF2 disease.

A novel DNA intercalator, butylamino-pyrimido[4',5':4,5]selenolo(2,3-b)quinoline, induces cell cycle arrest and apoptosis in leukemic cells

DNA intercalators are one of the most commonly used chemotherapeutic agents. Novel intercalating compounds of pyrimido[4',5':4,5]selenolo(2,3-b)quinoline series having a butylamino or piperazino group at fourth position (BPSQ and PPSQ, respectively) are studied. Our results showed that BPSQ induced cytotoxicity whereas PPSQ was cytostatic. The cytotoxicity induced by BPSQ was concentration- and time-dependent. Cell cycle analysis and tritiated thymidine assay revealed that BPSQ affects the cell cycle progression by arresting at S phase. The absence of p-histone H3 and reduction in the levels of PCNA in the cells treated with BPSQ further confirmed the cell cycle arrest. Further, annexin V staining, DNA fragmentation, nuclear condensation and changes in the expression levels of BCL2/BAD confirmed the activation of apoptosis. Activation of caspase 8 and lack of cleavage of caspase 9, caspase 3 and PARP suggest the possibility of BPSQ triggering extrinsic pathway for induction of apoptosis, which is discussed. Hence, we have identified a novel compound which would have clinical relevance in cancer chemotherapeutics.

Climate warming–related strengthening of the tropical hydrological cycle

The authors estimate climate warming–related twenty-first-century changes of moisture transports from the descending into the ascending regions in the tropics. Unlike previous studies that employ time and space averaging, here homogeneous high horizontal and vertical resolution data from an Intergovernmental Panel on Climate Change Fourth Assessment Report (IPCC AR4) climate model are used. This allows for estimating changes in much greater detail (e.g., the estimation of the distribution of ascending and descending regions, changes in the vertical profile, and separating changes of the inward and outward transports). Low-level inward and midlevel outward moisture transports of the convective regions in the tropics are found to increase in a simulated anthropogenically warmed climate as compared to a simulated twentieth-century atmosphere, indicating an intensification of the hydrological cycle. Since an increase of absolute inward transport exceeds the absolute increase of outward transport, the resulting budget is positive, meaning that more water is projected to converge in the moist tropics. The intensification is found mainly to be due to the higher amount of water in the atmosphere, while the contribution of weakening wind counteracts this response marginally. In addition the changing statistical properties of the vertical profile of the moisture transport are investigated and the importance of the substantial outflow of moisture from the moist tropics at midlevels is demonstrated.