Redox Regulation of Ras Proteins in Dictyostelium discoideum

Reactive oxygen species are a normal consequence of life in an aerobic environment. However when they deviate from the narrow permissible range in cells, oxidative damage can occur. Dictyostelium discoideum is a model organism ideal for the study of cell signaling events such as those affected by oxidative stress. It was previously shown that Ras signaling in Dictyostelium is affected by genetic inactivation of the antioxidant enzyme Superoxide dismutase C (SodC) and in vitro data suggests that the NKCD motif of Ras is the redox target of superoxide. The main objective of this project was to determine the mechanism of superoxide mediated Ras regulation in vivo. To accomplish the main objective, we cloned, and in some cases, mutated different Ras proteins and later determined their activity in wild type and sodC- cells. RasC and RasD showed normal activation in sodC- cells, however RasG and RasS displayed high Ras activity. These last two Ras proteins contain the NKC118D motif inside the nucleotide binding region. A mutation of cysteine118 to alanine in RasG rendered the protein less active in sodC- than the wild type RasG protein and a mutation alanine118 to cysteine in RasD conferred redox sensitivity to this small GTPase. Additionally, the propensity of RasG to be targeted by superoxide was evident when the environment of wild type cells was manipulated to induce the internal generation of superoxide through changes in the extracellular ion levels mainly magnesium. Lack of magnesium ions increased the intracellular level of superoxide and severely hampered directional cell migration. Chemotaxis of cells expressing RasG was negatively impacted by the absence of magnesium ions; however rasG- cells did not seem to be affected in their ability to perform chemotaxis. The last experiment implies that RasG is an important mediator of cell signaling during oxidative stress, responsible for preventing cells from continuing their developmental program. Our study suggests that the cysteine residue in the NKCD motif is essential for mediating the redox sensitivity of Ras proteins in Dictyostelium and that RasG is an essential mediator of the response to oxidative stress in this organism.

Intracellular Signaling and Trafficking in Cancer: Role of Rab5-GTPase in Migration and Invasion of Breast Cells

Metastasis is characterized pathologically by uncontrolled cell invasion, proliferation, migration and angiogenesis. Steroid hormones, such as estrogen, and growth factors, which include insulin growth factor I/II (IGF-1/IGF-2) therapy has been associated with most if not all of the features of metastasis. It has been determined that IGF-1 increases cell survival of cancer cells and potentiate the effect of E2 and other ligand growth factors on breast cancer cells. However not much information is available that comprehensively expounds on the roles of insulin growth factor receptor (IGFR) and Rab GTPases may play in breast cancer. The latter, Rab GTPases, are small signaling molecules and critical in the regulation of many cellular processes including cell migration, growth via the endocytic pathway. This research involves the role of Rab GTPases, specifically Rab5 and its guanine exchange factors (GEFs), in the promotion of cancer cell migration and invasion. Two important questions abound: Are IGFR stimulation and downstream effect involved the endocytic pathway in carcinogenesis? What role does Rab5 play in cell migration and invasion of cancer cells? The hypothesis is that growth factor signaling is dependent on Rab5 activity in mediating the aggressiveness of cancer cells. The goal is to demonstrate that IGF-1 signaling is dependent on Rab5 function in breast cancer progression. Here, the results thus far, have shown that while activation of Rab5 may mediate increased cell proliferation, migration and invasion in breast cancer cells, the Rab5 GEF, RIN1 interacts with the IGFR thereby facilitating migration and invasion activities in breast cells. Furthermore, endocytosis of the IGFR in breast cancer cells seems to be caveolin dependent as the data has shown. This taken together, the data shows that IGF-1 signaling in breast cancer cells relies on IGF-1R phosphorylation, caveolae internalization and sequestration to the early endosome RIN1 function and Rab5 activation.

A ROS-based Workspace Control and Trajectory Planner for a seven degrees of freedom Robotic arm

In this Bachelor Thesis I want to provide readers with tools and scripts for the control of a 7DOF manipulator, backed up by some theory of Robotics and Computer Science, in order to better contextualize the work done. In practice, we will see most common software, and developing environments, used to cope with our task: these include ROS, along with visual simulation by VREP and RVIZ, and an almost "stand-alone" ROS extension called MoveIt!, a very complete programming interface for trajectory planning and obstacle avoidance. As we will better appreciate and understand in the introduction chapter, the capability of detecting collision objects through a camera sensor, and re-plan to the desired end-effector pose, are not enough. In fact, this work is implemented in a more complex system, where recognition of particular objects is needed. Through a package of ROS and customized scripts, a detailed procedure will be provided on how to distinguish a particular object, retrieve its reference frame with respect to a known one, and then allow navigation to that target. Together with technical details, the aim is also to report working scripts and a specific appendix (A) you can refer to, if desiring to put things together.

Changes in [18F]Fluoro-2-deoxy-D-glucose incorporation induced by doxorubicin and anti-HER antibodies by breast cancer cells modulated by co-treatment with metformin and its effects on intracellular signalling

Peer reviewed

Interspecific correlation between red blood cell mitochondrial ROS production, cardiolipin content and longevity in birds

This work was supported by the Swiss National Science Foundation (31003A-124988/1 to PB, 31003A-138187 and 31003A-159600/1 to PC). We are grateful to the staff of La Vaux-Lierre for giving us access to the bird care centre, to Sylvie Massemin and Jean-Patrice Robin for their help in data sampling and to Olivier Glaizot and two anonymous reviewers for their comments on the manuscript.

Changes in [18F]Fluoro-2-deoxy-D-glucose incorporation induced by doxorubicin and anti-HER antibodies by breast cancer cells modulated by co-treatment with metformin and its effects on intracellular signalling

Peer reviewed

Rab32 restriction of intracellular bacterial pathogens

Experiments in the authors’ laboratory are supported by the Wellcome Trust (grant no. 109680/Z/15/Z), the Royal Society (grant no. RG150386), and Tenovus Scotland (grant no. G14/19) to SS. VSC is recipient of a European Union’s Horizon 2020 research and innovation programme Marie Sklodowska Curie Fellowship (grant no. 706040).

Changes in [18F]Fluoro-2-deoxy-D-glucose incorporation induced by doxorubicin and anti-HER antibodies by breast cancer cells modulated by co-treatment with metformin and its effects on intracellular signalling

Peer reviewed

Protection of primary cultures of mouse hepatocytes against fas-induced apoptosis : role of EGF receptor intrinsic activity and intracellular redox state

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Skeletal muscle disuse atrophy : implications on intracellular signaling pathways and mitochondrial permeability transition pore function

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

NOX-driven ROS formation in cell transformation of FLT3-ITD positive AML

In different types of myeloid leukemia, increased formation of reactive oxygen species (ROS) has been noted and associated with aspects of cell transformation including the promotion of leukemic cell proliferation and migration, as well as DNA-damage and accumulation of mutations. Work reviewed in this article has shown the involvement of NADPH oxidase (NOX)-derived ROS downstream of oncogenic protein-tyrosine kinases in both processes, and the related pathways have been partially identified. FLT3-ITD, an important oncoprotein in a subset of AML, causes activation of AKT and subsequently stabilization of p22phox, a regulatory subunit for NOX1-4. This process is linked to ROS formation and DNA damage. Moreover, FLT3-ITD signaling through STAT5 enhances expression of NOX4, ROS formation and inactivation of the protein-tyrosine phosphatase DEP-1/PTPRJ, a negative regulator of FLT3 signaling, by reversible oxidation of its catalytic cysteine residue. Genetic inactivation of NOX4 restored DEP-1 activity and attenuated cell transformation by FLT3-ITD in vitro and in vivo. Future work is required to further explore these mechanisms and their causal involvement in leukemic cell transformation, which may result in the identification of novel candidate targets for therapy.

Protection of primary cultures of mouse hepatocytes against fas-induced apoptosis : role of EGF receptor intrinsic activity and intracellular redox state

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Skeletal muscle disuse atrophy : implications on intracellular signaling pathways and mitochondrial permeability transition pore function

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Una propuesta de enseñanza de kayakismo para navegar en aguas abiertas, mar y ríos de montaña

Esta ponencia tiene como objetivo compartir el trabajo realizado durante los últimos cuatro años en un curso de kayak que funciona en la ciudad de Berisso. En él se trabaja para desarrollar en los alumnos las habilidades motrices necesarias para poder navegar en el mar, ríos de montaña, aguas abiertas y saltar cascadas. En la primera parte se describirán a grandes rasgos las características de la propuesta, el contexto donde se desarrolla, los destinatarios, el lugar donde funciona, equipo docente, organización de los grupos y los recursos materiales utilizados. Luego nos introduciremos de lleno en la planificación y su organización. Analizaremos los objetivos, los contenidos y su organización en niveles, las actividades y viajes que se realizan para que los alumnos apliquen las habilidades motrices desarrolladas en el curso. Para finalizar realizaremos un análisis a partir de la propia experiencia sobre la complejidad de la enseñanza y del aprendizaje de la técnica del roll, las estrategias didácticas utilizadas, la importancia de trabajar la tranquilidad debajo del agua y por último cerraremos hablando de la evaluación en el curso de kayak en pileta