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  Microvascular Biology and Inflammation - Research
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Our unit is currently addressing issues related with the mechanisms underlying the activation and the function of blood cells in inflammation by studying the role of fibrinogen (Fib) in erythrocyte and neutrophil activation and the role of neutrophil chemoattractants and blood hemodynamics in their recruitment in inflammatory conditions. Concerning this, our current and future aims are to:

Dissect the Fib-CD47 interaction and its influence on the erythrocyte NO pathway;

Study the players and mechanisms underlying Fib-neutrophil interaction and the Fib-mediated endothelium-neutrophil communication;

Understand the influence of Fib on neutrophil apoptosis and autophagy in sepsis;

Assess circulating miRNA expression profile as a prognostic marker in AMI patients outcome;

Study the interplay between neutrophil deformability and shear stress using numerical application;

Understand the mechanisms underlying the generation and function of gradients of hydrogen peroxide (H2O2) and Cxcl8 chemokines in neutrophil activation/recruitment and how their action is concerted in this regard.

 

For studies on the influence of Fib on erythrocyte and neutrophil biology, cellular samples will be obtained from human healthy volunteers, AMI and sepsis patients. The study of the erythrocyte NO pathway under Fib/CD47 dependence will be assessed by using pharmacological inhibitors, CD47 agonist peptides and antibodies through amperometry, spectrophotometry and fluorescence microscopy. The molecular characterization of Fib-CD47 binding will be investigated using CD47 and Fib antibodies and peptides by flow cytometry and confocal microscopy. 

Contact person: carlotasaldanha at medicina.ulisboa.pt 

 

In AMI patients, we expect to be able to contribute for the miRNAs expression profiles with adverse cardiovascular events. Moreover, we expect to manufacture a micro-hemorheological prototype apparatus to be used in patient hospital bed side. The numerical models developed are expected to constitute useful tools for understanding the influence of biomechanical factors in the onset and progression of vascular diseases. Finally, we expect to understand how distinct chemotactic cues concert their action in neutrophil recruitment in inflammation. Altogether, we hope our studies will enable us to unravel novel potential targets and strategies for the development of therapeutics for inflammatory diseases, cardiovascular diseases as well as sepsis.

Contact person: pnapoleao at medicina.ulisboa.pt

 

Zebrafish_1The function and crosstalk of H2O2 and Cxcl8 gradients in neutrophil recruitment will be investigated in zebrafish viathe use of molecular and cellular biology techniques, signaling studies and in vivo cell tracking and chemotactic gradient visualization by fluorescence microscopy. To directly visualize Cxcl8 gradients in vivo, we will develop transgenic fish expressing mCherry-tagged versions of Cxcl8s under the action of their native promoters. 

Contact person: acalado at medicina.ulisboa.pt

 

 

Fluorescence microscopy images of the recruitment of eGFP-labelled neutrophils in tail transected 3dpf mpx:GFP larvae, pre-injected with the indicated morpholinos (MO), at 6 hours post wounding (hpw). Scale bar = 200µm. by Sofia de Oliveira

 

 

Numerical models will be developed based on the quantification of rolling and adherent leukocytes under blood flow and on the visualization of membrane deformability changes by intravital microscopy studies in mice post-capillary venules. Shear stress variations will be addressed through the use of an Optical Doppler Velocimeter. 

Contact person: anarmsilva at medicina.ulisboa.pt  

 Microcirculation_1

 

 

 Post-capillary venule in mouse cremaster muscle superfused with bicarbonate-bufered saline and observed by intravital microscopy (200x). by Ana Silva-Herdade and Vanda Vitorino de Almeida

 

Numerical_simulation_of_rolling_leukocyte_1

Numerical simulation of the influence of a free flowing leukocyte on the endothelial wall shear stress. Movement towards the wall disturbes the flow at longer distances.

 

 

Ongoing projects

2014/2016 Perfis da Agregação Eritrocitária e da IL-8 em sobreviventes de enfarte agudo de miocárdio. Dr. Daniel Ferreira (Hospital da Luz), Coordinator: C. Saldanha

2013/2015 Evaluation of inflammatory and adaptative immunity imbalance to improve prognosis of ischemic cardiovascular diseases, Liga dos Amigos do Hospital de Santa Marta/2012. Coordinator: Patrícia Napoleão.

2013/2015 Contribution of circulating microRNA to improve prognosis of ischemic cardiovascular diseases, Liga dos Amigos do Hospital de Santa Marta/2012. Coordinator: Patricia Napoleão

2012/2015 Mathematical and computational modeling of human physiology EXCL/Mat-NAN/0114/2012 A. Sequeira (CEMAT/IST).

2012/2015 Evaluation of inflammation profiles in septic patients – contribution to multiple cross prognostic biomarkers. Coordinator: Carlota Saldanha.

2011/2013 Functional characterization of the neutrophil in alcoholic and non-alcoholic steatohepatiti. Coordinator: Ângelo Calado.

2011/2013 Functional characterization of the neutrophil in alcoholic and non-alcoholic steatohepatitis. Coordinator: A. Calado.

2009/2013 In vivo imaging of inflammation in zebrafish: the role of chemokine system in the neutrophil and macrophage migration, Sofia de Oliveira, PhD Fellowship SFRH/BD/62674/2009, FCT. Coordinator: A. Calado.

 

Previous projects 

2009/2011 Development and validation of vaso-occlusion early predictors in a a mendelian model of vascular disease, FCT. Coordinator: João Lavinha, INSA. PIC/IC/83084/2007

2009/2011 Cardiovascular risk and inflammation- contribution to the etiopathogenesis of atherosclerosis and cardiovascular disease, FCT. Coordinator: João Eurico, IMM, FML, UL. PIC/IC/82920/2007.

2007/ 2012 The relationship of circulating biomarkers of apoptosis and endothelial function with the plaque composition using VH IVUS, FCT. Coordinator: Teresa Pinheiro, Grupo de Estudos Biomédicos, Instituto Tecnológico e Nuclear (ITN). FCT project PIC/IC/82734/2007

2007/2011 Fibrinogen-dependent signalling in microvascular blood cell function – implications of inflammation and cardiovascular disease, FCT. Coordinator: Carlota Saldanha. PTDC/SAU-OSM/73449/2006

Fibrinogen-dependent signalling in microvascular blood cell function – Implications of Inflammation and Cardiovascular disease. Foundation for Science and Technology, Portugal – Project PTDC/SAU-OSM/73449/2006 (PI).

Multiscale Mathematical Modelling in Biomedicine. Foundation for Science and Technology, Portugal – Project PTDC/ MAT/ 68166/2006

Molecular Medicine – from genome to clinical practice (Team member; responsible for selection of a high-speed intravital confocal microscope). Foundation for Science and Technology, Portugal – Project REEQ/650/SAU/2005

(2002-2006) - Mathematical and Numerical Modelling in Haemodynamics – HaeModel. EU-RTN European Research Training Network - HPRN-CT 2002-00270

Mathematical and Numerical Modelling of the Human Cardiovascular System. Foundation for Science and Technology, Portugal - Project POCTI/MAT/41898/ 2001

 

Main collaborations

Adélia Sequeira (Center of Mathematics and its Applications, Technical University of Lisbon, Portugal)

Victoriano Mulero (Faculty of Biology, University of Murcia, Murcia, Espanha)

Stephen Andrew Renshaw (MRC Centre for Development and Biomedical Genetics, University of Sheffield, Sheffield, Reino Unido)

Nadia Antonova, Institute of Mechanics and Biomechanics of the Bulgarian Academy of Sciences, Bulgary

Caterina Faggio, Departamento de Ciência Biológica e Ambiental, Universidade de Messina, Sicily, Italy, ERASMUS Program

Sarka Necasova, Institute of Mathematics of the Academy of Sciences of the Czech Republic, Czech Republic

Narla Mohandas, New York Blood Center, USA

Hisayuki Nomiyama (Graduate School of Medical Sciences - Department of Molecular Enzymology, Kumamoto University, Kumamoto, Japão)

Naoki Osada (National Institute of Biomedical Innovation, Osaka, Japão)

Rui L. Reis (3Bs Research Group, Department of Polymer Engineering, University of Minho, Guimarães, Portugal)

Luís L. Cardoso (Universidade de Trás-os-Montes e Alto Douro, Vila Real, Portugal)

Manuela Rodeia (Faculdade de Medicina Veterinária, Universidade Técnica de Lisboa, Lisboa, Portugal)

João Garcia da Fonseca, Nuno Reis, Ricardo Cabeça (Biosurfit SA, Lisboa, Portugal)

Geert Schmid-Schönbein (Microcirculation Lab/ Departament of Bioengineering, University of California, San Diego, EUA)

Gerard Nash (Center of Cardiovascular Sciences, University of Birmighan, Reino Unido)

Hermona Soreq (Faculty of Science, Hebrew University, Jerusalem, Israel)

Felix M. Gõni (Biophysics Unit, University of Basque Country, Espanha)


 



 

Av. Professor Egas Moniz
Edifício Egas Moniz,
1649-028 Lisboa
Portugal
 

Tel: (+351)217985136
Fax: (+351)217999477


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