The two-component system of a novel copper resistant operon of Marinobacter hydrocarbonoclasticus

Dissertation for the Master’s Degree in Structural and Functional BiochemistryThe majority of bacterial heavy metal resistance systems are regulated by twocomponent signal transduction systems. Stimuli from the environment interact with the histidine kinase, which in turn activates the response regulator by phosphorylation. The effector domain of the response regulator then binds to DNA, eliciting the specific response. Analysis of the Marinobacter hydrocarbonoclasticus genome revealed the presence of genes, copXAB, that code for proteins associated with copper response. The biochemical characterization of the two-component signal transduction system, copSR, is of interest due to the vital role it plays in the regulation of expression of the copXAB operon. The genes that encode for the CopR and CopS_C (cytosolic sensor domain of CopS) proteins were heterologously expressed in E.coli and expression was optimized for the production of soluble protein using LB medium. Due to solubility problems, the genes that code for these proteins were cloned as hexahistidine or glutathione S-transferase fusion proteins. CopR and its domains were optimally expressed at 16°C for 16 and 3 h after induction, respectively, whilst CopS_C was expressed at 37°C during 3 h after induction. Proteins were purified using different chromatographic strategies, most of them using affinity chromatography. The yields of pure protein per liter of growth culture obtained after complete purification from the soluble cellular extract were: 0.14 to 0.23 mg/L for CopR; 0.42 mg/L, CopR_NHis6; for the CopR_CHis6 it was 0.16 mg/L and 4.2 mg/L of CopS_C. The molecular mass of each protein was determined by gel filtration, 31 kDa for CopR, 17.5 kDa for CopR_NHis6, 15.1 kDa for CopR_CHis6 and 38.2 kDa for CopS_C. In the case of CopS_C there is the possibility that a dimer is formed, which should be evaluated. From the evaluation of disulfide bonds, using SDS PAGE and PAGE gels, all proteins or protein domains appeared to be monomers when in the presence of β-MEtOH. Circular dichroism evaluated the state of folding of the CopS_C and CopR proteins, which were shown to be folded in which the α-helix structures predominate. A model structure for CopR was also determined which agrees with this analysis. However, in the case of the CopR domains, the data obtained merely indicate folding, due to the low concentrations of the proteins. Phosphorylation and electrophoresis mobility shift assays of the CopR protein were, for the most part, inconclusive. However, in the absence of BSA, formation of the CopR:DNA complex in a gel filtration column is observed, though requires additional evaluation

Effects of reading fluency stimulation with emphasis on prosody

PURPOSE: To characterize the reading ability of children with five years of education according to temporal standards, as well as to compare the reading fluency performance of these children before and after a reading program based on prosody standards. METHODS: Thirty two fourth-grade children participated in this study. They performed reading aloud and picture description tasks in order to verify reading rate, speech rate, text comprehension and the adequacy of prosody variation. Afterwards, it was carried out a reading stimulation program with emphasis on prosody, composed of five 15-minute sessions of reading tasks. At the end of the program, children were re-assessed, in order verify their performance after stimulation. RESULTS: Changes were observed in reading rate, number of words misread, and quality of prosody during the reading task. CONCLUSIONS: The reading program promoted positive changes in reading fluency measuresOBJETIVO: Caracterizar a leitura de crianças com cinco anos de escolaridade segundo os parâmetros temporais, e comparar o desempenho da fluência de leitura pré e pós-estimulação de leitura por meio de programa baseado em padrões de prosódia. MÉTODOS: Participaram 32 crianças do 5º ano do Ensino Fundamental I. As crianças realizaram leituras de textos e descrição de uma figura. Foram observados: taxa de leitura, velocidade de fala, compreensão de textos e adequação da variação da prosódia durante a leitura. Em seguida, foi aplicado o programa de estimulação de leitura com ênfase na prosódia, composto por cinco sessões de 15 minutos cada, com tarefas de leitura. Ao final do programa, foi realizado o mesmo procedimento de avaliação, a fim de se verificar o desempenho pós-estimulação. RESULTADOS: Houve modificações quanto às médias da taxa de leitura, ao número de palavras lidas de forma incorreta e à qualidade da prosódia realizada durante a atividade de leitura. CONCLUSÃO: O programa de estimulação promoveu modificações positivas nas medidas de fluência de leituraUniversidade Federal de São Paulo (UNIFESP) Programa de Pós-graduação (Mestrado) em Distúrbios da Comunicação HumanaSanta Casa de São Paulo Faculdade de Ciências MédicasUNIFESP, Programa de Pós-graduação (Mestrado) em Distúrbios da Comunicação HumanaSciEL

IGF-1 Regulates the extracellular level of active MMP-2 and promotes Müller glial cell motility

In ischemic proliferative retinopathies, Müller glial cells (MGCs) acquire migratory abilities. However, the mechanisms that regulate this migration remain poorly understood. In addition, proliferative disorders associated with enhanced activities of matrix metalloproteinases (MMPs) also involve insulin-like growth factor (IGF)-1 participation. Therefore, the main interest of this work was to investigate the IGF-1 effect on the extracellular proteolytic activity in MGCs.Methods: Cell culture supernatants and cell lysates of the human MGC line MIO-M1 stimulated with IGF-1 were analyzed for MMP-2 by zymographic and Western blot analysis. The MGCs´ motility was evaluated by scratch wound assay. The MMP-2, β1-integrin, and focal adhesions were detected by confocal microscopy. The localization of active MMPs and actin cytoskeleton were evaluated by in situ zymography.Results: The IGF-1 induced the activation of canonical signaling pathways through the IGF-1R phosphorylation. Culture supernatants showed a relative decrease in the active form of MMP-2, correlating with an increased accumulation of MMP-2 protein in the MGCs´ lysate. The IGF-1 effect on MMP-2 was abolished by an IGF-1R blocking antibody, αIR3, as well as by the PI3-kinase inhibitor, LY294002. The IGF-1 increased the migratory capacity of MGCs, which was blocked by the GM6001 MMP inhibitor, LY294002 and αIR3. Finally, IGF-1 induced the intracellular distribution of MMP-2 toward cellular protrusions and the partial colocalization with β1-integrin and phospo-focal adhesion kinase signals. Gelatinase activity was concentrated along F-actin filaments.Conclusions: Taken together, these data indicate that IGF-1, through its receptor activation, regulates MGCs´ motility by a mechanism that involves the MMP-2 and PI3K signaling pathway.Fil: Lorenc, Valeria Erika. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Jaldín Fincati, Javier Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Luna Pinto, Jose Domingo. Departamento de Vítreo-retina, Centro Privado de Ojos R; ArgentinaFil: Chiabrando, Gustavo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Sanchez, Maria Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentin

Desarrollo de un helado hipocalórico y funcional con adición de acidos grasos Omega 3

Se desarrolló un helado bajo en grasas, sin azúcar, enriquecido con ácidos grasosomega 3, y con características sensoriales de un helado estándar. Para encontraruna mezcla base adecuada se realizó un diseño experimental 23 sin adición deomega 3 ni esencias. A la mezcla base elegida se le realizó un segundo diseñoexperimental 21 para observar el efecto del omega 3 en la calidad del producto.Se encontró que la adición de un 0,33% de omega 3 no afectó (p>0,05) laspropiedades organolépticas del helado. Un estudio de estabilidad del omega 3,determino que este presentó una cinética de deterioro de 2° orden, y que el índicede peróxido se mantuvo bajo 3 meq O2/kg aceite, menor al límite permitido (10meq O2/kg aceite). El análisis proximal indicó que el helado con 0,33% de omega3 tuvo 78 kcal por 100 g de producto.Palabras claves: Helado funcional, omega 3, hipocalóric

Plan de mejoramiento institucional una perspectiva desde la gestión participativa y la calidad educativa, horizontes – retos

Maestría en EducaciónThis investigation Magdalena has as aim Understand the dynamics of the plan of improvement of the educational institutions of the department of so that there are constructed spaces of analysis that they contribute to the transformation in the construction and implementation of the PMI. It developed bearing the methodological approach in mind investigation action in two phases: diagnosis and intervention, informing three educational public establishments of the department of Magdalena.As sources: document PMI card of characterization, interviews with group of discussion workshops of formation and accompaniment. The results were; the members of the educational community know little on the PMI, it is elaborated without the community participation, only as requirement of the secretaries of education. In the phase the II demonstrates clarity on the concept and construction of the PMI, motivation and commitment.Esta investigación tiene como objetivo Comprender la dinámica del plan de mejoramiento de las instituciones educativas del departamento del Magdalena de modo que se construyan espacios de análisis que contribuyan a la transformación en la construcción e implementación de los PMI. Se desarrolló teniendo en cuenta el enfoque metodológico investigación acción en dos fases: diagnóstico e intervención, participando tres establecimientos educativos públicos del departamento del Magdalena. Como fuentes: documento PMI, ficha de caracterización, entrevista con grupo de discusión talleres de formación y acompañamiento. Los resultados fueron; los miembros de la comunidad educativa conocen poco sobre el PMI, es elaborado sin la participación comunitaria, solo como requisito de las secretarias de educación.En la fase II se evidencia claridad sobre el concepto y construcción del PMI, motivación y compromiso

Consideraciones teórico-metodológicas para el abordaje de la paternidad desde una perspectiva historiográfica

Este artigo apresenta algumas considerações teóricas para abordar a paternidade na historiografia. Esta tarefa é assumida desde a pergunta: ¿O que é a paternidade? Realiza-se uma aproximação aos assuntos metodológicos, oferecendo grande importância à legislação como fonte, a partir da idéia de que a paternidade é uma instituição e como tal se institui pelas leis. A legislação é reconhecida como artefato cultural e literário, produto de lutas num campo de poder.Este artículo presenta algunas consideraciones teóricas para abordar la paternidad en la historiografía. Esta tarea es asumida desde la pregunta: ¿Qué es la paternidad? Se efectúa una aproximación a los aspectos metodológicos, colocando en un lugar preponderante la legislación como fuente, a partir de la idea de que la paternidad es una institución y como tal se instituye sobre las leyes. La legislación es presentada como artefacto cultural y literario, producto de luchas en un campo de poder

Using individual tracking data to validate the predictions of species distribution models

The authors would like to thank the College of Life Sciences of Aberdeen University and Marine Scotland Science which funded CP's PhD project. Skate tagging experiments were undertaken as part of Scottish Government project SP004. We thank Ian Burrett for help in catching the fish and the other fishermen and anglers who returned tags. We thank José Manuel Gonzalez-Irusta for extracting and making available the environmental layers used as environmental covariates in the environmental suitability modelling procedure. We also thank Jason Matthiopoulos for insightful suggestions on habitat utilization metrics as well as Stephen C.F. Palmer, and three anonymous reviewers for useful suggestions to improve the clarity and quality of the manuscript.Peer reviewedPostprintPostprintPostprintPostprintPostprin

Nutritional support in cancer patients: A position paper from the Italian Society of Medical Oncology (AIOM) and the Italian Society of Artificial Nutrition and Metabolism (SINPE)

Malnutrition is a frequent problem in cancer patients, which leads to prolonged hospitalization, a higher degree of treatment-related toxicity, reduced response to cancer treatment, impaired quality of life and a worse overall prognosis. The attitude towards this issue varies considerably and many malnourished patients receive inadequate nutritional support. We reviewed available data present in the literature, together with the guidelines issued by scientific societies and health authorities, on the nutritional management of patients with cancer, in order to make suitable and concise practical recommendations for appropriate nutritional support in this patient population. Evidence from the literature suggests that nutritional screening should be performed using validated tools (the Nutritional Risk Screening 2002 [NRS 2002], the Malnutrition Universal Screening Tool [MUST], the Malnutrition Screening Tool [MST] and the Mini Nutritional Assessment [MNA]), both at diagnosis and at regular time points during the course of disease according to tumor type, stage and treatment. Patients at nutritional risk should be promptly referred for comprehensive nutritional assessment and support to clinical nutrition services or medical personnel with documented skills in clinical nutrition, specifically for cancer patients. Nutritional intervention should be actively managed and targeted for each patient; it should comprise personalized dietary counseling and/or artificial nutrition according to spontaneous food intake, tolerance and effectiveness. Nutritional support may be integrated into palliative care programs. "Alternative hypocaloric anti-cancer diets" (e.g. macrobiotic or vegan diets) should not be recommended as they may worsen nutritional status. Well-designed clinical trials are needed to further our knowledge of the nutritional support required in different care settings for cancer patients