Functional expression of Yarrowia lipolytica acetyl- CoA carboxylase in Saccharomyces cerevisiae

Malonyl coenzyme A (Malonyl-CoA) is an important precursor for a range of economically valuable compounds such as biodiesel, 3-hydroxypropionic acid, fatty alcohols, flavonoids, stilbenoids and polyketides. Even though Saccharomyces cerevisiae is one of the most used cell factories for the production of a variety of compounds, it is limited in its ability to generate malonyl-CoA and derived products. In this yeast, the only source of malonyl-CoA is the ATP-dependent carboxylation of acetyl coenzyme A (acetyl-CoA) catalyzed by acetyl-CoA carboxylase (ACC), the rate-limiting step in fatty acid biosynthesis. In this work, expression of Yarrowia lipolytica ACC in S. cerevisiae was attempted with the objective of establishing an alternative malonyl-CoA production system in this yeast. A S. cerevisiae strain with a tetracycline repressible ACC1 was used for functional testing. Since ACC1 is an essential gene, this strain is not capable of growing in medium supplemented with tetracycline. Expression of S. cerevisiae or Yarrowia lipolytica ACC1 from a plasmid complemented the conditional phenotype and enabled growth in tetracycline containing medium. Expression of ACCs from plasmids caused prolonged lag phase, and specific growth rates were considerably lower than the ones obtained for the wild type strain. This result is consistent with previous studies where ACC activity was increased and likely results from the metabolic imbalance this increase might cause. Results of in-vivo measurements of malony—CoA levels in the recombinant strain will be discussed.FatVal POCI-01-0145-FEDER-032506. Ph.D. scholarship SFRH/BD/140039/201

A fundamentação substancial das decisões judiciais como garantia do Estado democrático de direito

Divulgação dos SUMÁRIOS das obras recentemente incorporadas ao acervo da Biblioteca Ministro Oscar Saraiva do STJ. Em respeito à Lei de Direitos Autorais, não disponibilizamos a obra na íntegra.Localização na estante: 347.95(81) L864

The role of MAPK signalling pathways in acetic acid-induced cell death of Saccharomyces cerevisiae

Dissertação de mestrado em Genética MolecularMitogenic Activated Protein Kinase (MAPK) cascades are important signalling pathways that allow yeast cells to swiftly adapt to changing environmental conditions. Previous studies suggested that the High Osmolarity Glycerol (HOG) MAPK pathway and ceramide production are involved in acetic-acid induced apoptosis in yeast. Evidence that changes in the levels of endogenous ceramides can affect yeast cell fate has also been put forth. However, knowledge on the molecular basis of acetic acid induced cell death and ceramide-induced cell changes, as well as the signalling pathways involved in these processes, is still lacking. In this work, we tried to elucidate the role of MAPK signalling pathways in the cell response to ceramide and in acetic acid-induced cell death. Cells treated with the soluble ceramide analog N-acetyl-D-sphingosine (C2-ceramide) maintained plasma membrane integrity and did not produce reactive oxygen species detectable by DHE staining. A subpopulation of cells died when exposed to C2-ceramide but the surviving part eventually grew, so no reduction in plate counts was observed. Preliminary results indicated that deletion mutants in MAPK components showed smaller sensitive subpopulations than the reference strain and were therefore deemed to be more resistant to C2-ceramide, namely wsc3 . The MAPK mutants wsc2 , wsc3 , ste20 , slt2 and mkk1 /mkk2 were significantly more resistant to acetic acid than the reference strain. These mutants also displayed a lower percentage of cells with increased accumulation of ROS and a higher percentage of cells with preserved plasma membrane integrity than the wild type strain. The WSC2 and WSC3 genes encode sensors in the cell wall integrity MAPK pathway, suggesting a role for this signalling pathway in both ceramide and acetic acid induced cell death. The link between acetic acid-induced cell death and cell wall integrity is not known, but lipid peroxidation by acetic acid may be a plausible candidate.As cascatas “Mitogenic Activated Protein Kinase” (MAPK) são vias de sinalização importantes que permitem que as células de levedura se adaptem rapidamente às mudanças das condições ambientais. Estudos anteriores sugeriram que a via “High Osmolarity Glycerol” (HOG) MAPK e a produção de ceramida estão envolvidas na morte celular apoptótica de leveduras induzida por ácido acético. Há também evidências que alterações nos níveis de ceramidas endógenas podem afectar o destino de células de levedura. No entanto, ainda não há um conhecimento exacto sobre a base molecular da morte celular induzida por ácido acético, sobre as mudanças induzidas ao nível celular pela ceramida, bem como as vias de sinalização envolvidas nestes processos. Neste trabalho, procurou-se elucidar o papel das vias de sinalização MAPK na resposta celular à ceramida e na morte celular induzida por ácido acético. Células tratadas com o análogo solúvel da ceramida N-acetil-D-esfingosina (C2-ceramida) mantiveram a integridade da membrana plasmática e não produziram espécies reactivas de oxigénio detectadas através da incubação com DHE. Numa cultura exposta a C2-ceramida, uma subpopulação das células morreu, enquanto outra parte da cultura cresceu, não se observando assim na placa uma redução de Cfus em relação ao tempo zero. Alguns mutantes deletados em componentes das vias MAPK mostraram subpopulações menos sensíveis do que a estirpe selvagem e assim consideraram-se como sendo estirpes mais resistentes à C2-ceramida, nomeadamente o mutante wsc3 . Os mutantes wsc2 , wsc3 , ste20 , slt2 e mkk1 /mkk2 foram significativamente mais resistentes ao ácido acético do que a estirpe selvagem. Esses mutantes mostraram também uma menor percentagem de células exibindo uma acumulação de ROS e uma maior percentual de células com integridade da membrana plasmática preservada do que a estirpe selvagem. Os genes WSC2 e WSC3 codificam sensores membranares na via MAPK da integridade da parede, sugerindo um papel para esta via de sinalização tanto na morte celular induzida por ácido acético como nas alterações induzidas por ceramida. Não é conhecida a ligação entre morte celular induzida por ácido acético, mas a peroxidação lipídica pode ser uma hipótese provável.Fundação para a Ciência e a Tecnologia (FCT) - PTDC/BIA-BCM/69448/200

Warfarin-induced gastric intramural hematoma

Federal University of São Paulo Department of Internal MedicineFederal University of São Paulo Department of RadiologyFederal University of São PauloUNIFESP, Department of Internal MedicineUNIFESP, Department of RadiologyUNIFESPSciEL

Política nacional de resíduos sólidos (Lei no 12.305/2010) e educação ambiental

DOI: 10.24859/fdv.2017.1007 O presente trabalho discute a necessária efetivação da Lei no 12.305/2010, destacando aspectos como a gestão integrada dos resíduos sólidos e a responsabilidade compartilhada entre governos e sociedade. Argumenta que uma política pública que se propõe a modificar comportamentos e práticas enraizadas, no tocante ao manejo e gerenciamento do antes considerado lixo, não se constrói apenas com a homologação de uma lei. Conclui-se que, para se alcançar a etapa do engajamento popular, é necessária a efetivação de políticas públicas de educação ambiental, uma vez que tudo começa na separação e descarte adequado dos resíduos na fonte geradora, o que evita a perda de qualidade dos materiais recicláveis. A população precisa ser educada adequadamente de modo que possa mudar seus hábitos no momento do descarte do lixo

A local meshless analysis of dynamics problems / Uma análise local desordenada dos problemas dinâmicos

This paper is concerned with new formulations of local meshfree numerical method, for the solution of dynamic problems in linear elasticity, Integrated Local Mesh Free (ILMF) method. The key attribute of local numerical methods is the use of a modeling paradigm based on a node-by-node calculation, to generate the rows of the global system of equations of the body discretization. In the local domain, assigned to each node of a discretization, the work theorem is kinematically formulated, leading thus to an equation of mechanical equilibrium of the local node, that is used by local meshfree method as the starting point of the formulation. The main feature of this paper is the use of a linearly integrated local form of the work theorem. The linear reduced integration plays a key role in the behavior of local numerical methods, since it implies a reduction of the nodal stiffness which, in turn, leads to an increase of the solution accuracy. As a consequence, the derived meshfree and finite element numerical methods become fast and accurate, which is a feature of paramount importance, as far as computational efficiency of numerical methods is concerned. The cantilever beam was analyzed with this technique, in order to assess the accuracy and efficiency of the new local numerical method for dynamic problems with regular and irregular nodal configuration. The results obtained in this work are in perfect agreement with Mesh-Free Local Petrov-Galerkin (MLPG) and the Finite Element Method (FEM) solutions

Eco-friendly recycled polypropylene matrix composites incorporated with geopolymer concrete waste particles

Civil construction wastes have been incorporated into polymers for recycling as novel engineering composites. In the present work eco-friendly composites with recycled polypropylene (rPP) matrix incorporated with geopolymer concrete waste particles, wither plain (GCW) or surface-modified with oleic acid (AGC) were investigated. The geopolymer concrete waste particles were mixed with polymer powder to provide an effective dispersion between the different materials. Composites were produced by an initial reactive extrusion processing followed by injection molding. These novel composites with amount of 20, 40 and 50 wt% of GCW particles, both plain as-received and surface-modified, were technically evaluated by tensile tests, statistically analyzed by ANOVA, as well as by water absorption as per ASTM standards. Surface dispersion of nanoparticles was revealed by atomic force microscopy. Microstructural analysis was performed by scanning electron microscopy. The results indicated that these sustainable GCW particles incorporated into rPP matrix exhibit superior processability and water absorption less than 0.01%. The rPP/AGC composites present relatively higher elastic modulus, 629 MPa, as compared to the neat rPP, with 529 MPa. These properties suggest potential sustainable applications in building construction using waste materials.Peer reviewe

Atelectasias durante anestesia: fisiopatología y tratamiento

JUSTIFICATIVA E OBJETIVOS: O colapso pulmonar intra-operatório é uma complicação de elevada incidência em pacientes submetidos à intervenção cirúrgica sob anestesia geral com relaxamento/paralisia da musculatura. Essa complicação está associada à piora das trocas gasosas no intra-operatório e, em alguns casos, necessidade de suporte respiratório prolongado no período pós-operatório. Os objetivos deste estudo foram revisar os aspectos fisiopatológicos da formação de atelectasias durante anestesia geral e as possíveis manobras terapêuticas para prevenir e tratar essa complicação. CONTEÚDO: Nesta revisão, os conceitos sobre a incidência de atelectasias intra-operatórias, os fatores relacionados com o seu desenvolvimento, tanto mecânicos quanto associados ao ajuste do respirador durante procedimento cirúrgico, os aspectos do diagnóstico e as estratégias de prevenção e tratamento foram abordados de maneira sistemática. CONCLUSÕES: A compreensão dos mecanismos relacionados com o desenvolvimento do colapso pulmonar durante o período intra-operatório, assim como o seu tratamento, pode contribuir para a redução da incidência de complicações pulmonares pós-operatórias, o tempo de recuperação e os custos hospitalares.BACKGROUND AND METHODS: The incidence of intraoperative pulmonary collapse is elevated in patients undergoing surgery under general anesthesia with muscle relaxation/paralysis. This complication is associated with worsening intraoperative gas exchange and, in some cases, the need for prolonged postoperative respiratory support. The objective of this report was to review the pathophysiological aspects of atelectasis during general anesthesia and possible therapeutic maneuvers that could prevent and treat this complication. CONTENTS: This review discusses the concepts about the incidence of intraoperative atelectasis, factors that influence their development, both mechanical and those related to mechanical ventilator settings during the surgery, diagnostic criteria, and strategies to prevent and treat this complication. CONCLUSIONS: Understanding of the mechanisms related with the development of intraoperative pulmonary collapse, as well as its treatment, can contribute to reduce the incidence of postoperative pulmonary complications, the length of recovery and hospital costs.JUSTIFICATIVA Y OBJETIVOS: El colapso pulmonar intraoperatorio es una complicación de elevada incidencia en pacientes sometidos a la intervención quirúrgica bajo anestesia general con relajamiento/parálisis de la musculatura. Esta complicación está asociada al empeoramiento de los cambios de gas en el intraoperatorio y en algunos casos, necesidad de soporte respiratorio prolongado en el período postoperatorio. Los objetivos de este estudio fueron los de revisar los aspectos fisiopatológicos de la formación de atelectasias durante anestesia general y las posibles maniobras terapéuticas para prevenir y tratar esa complicación. CONTENIDO: En esta revisión, los conceptos sobre la incidencia de atelectasias intraoperatorias, los factores relacionados a su desarrollo, tanto mecánicos como los relacionados al ajuste del respirador durante el procedimiento quirúrgico, los aspectos del diagnóstico y las estrategias de prevención y tratamiento fueron abordados de manera sistemática. CONCLUSIONES: La comprensión de los mecanismos relacionados al desarrollo del colapso pulmonar durante el período intraoperatorio, como también su tratamiento, pueden contribuir para la reducción de la incidencia de complicaciones pulmonares postoperatorias, el tiempo de recuperación y los costes de las internaciones en los hospitales

Low-Level Laser Therapy Attenuates the Myeloperoxidase Activity and Inflammatory Mediator Generation in Lung Inflammation Induced By Gut Ischemia and Reperfusion: A Dose-Response Study

Introduction: Intestinal ischemia and reperfusion (i-I/R) is an insult associated with acute respiratory distress syndrome (ARDS). Herein we evaluate the dose-response effect of low-level laser therapy (LLLT) on lung inflammation induced by i-I/R.Methods: Mice were subjected to mesenteric artery occlusion (45 min) and killed after clamp release and intestinal reperfusion (2h). Increasing doses (1, 3, 5 and 7,5 J/cm2) of laser irradiation (660 nm) was carried out on the mice skin over the upper bronchus for 5 min after initiating reperfusion. Neutrophils activation was determined by myeloperoxidase (MPO) activity. The mRNA expression and protein concentration of inflammatory mediators IL-1β, IL-6, TNF and IL-10 in lung were measured by RT-PCR and ELISA, respectively.Results: With exception of 1J/cm2, LLLT reduced MPO activity as well as IL-1β levels in the lungs from inflamed mice. LLLT was also markedly effective in reducing both IL-6 and TNF expression and levels in the lungs from mice submitted to i-I/R in all laser doses studied. Otherwise, LLLT significantly increased the protein levels of IL-10 in inflamed mice by i-I/R; however only in the dose of 1J/cm2.Conclusion: We conclude that the LLLT is able to control the neutrophils activation and pro-inflammatory cytokines release into the lungs in a model of i-I/R in mice