Manuel E Patarroyo

Fundación instituto de Inmunología de Colombia, Μπογκοτά, Bogota D.C., Colombia

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Publications (343)1257.97 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: Developing novel generations of subunit-based antimalarial vaccines in the form of chemically-defined macromolecule systems for multiple antigen presentation represents a classical problem in the field of vaccine development. Many efforts involving synthesis strategies leading to macromolecule constructs have been based on dendrimer-like systems, the condensation of large building blocks and conventional asymmetric double dimer constructs, all based on lysine cores. This work describes novel symmetric double dimer and condensed linear constructs for presenting selected peptide multi-copies from the apical sushi protein expressed in Plasmodium falciparum. These molecules have been proved to be safe and innocuous, highly antigenic and have shown strong protective efficacy in rodents challenged with two Plasmodium species. Insights into systematic design, synthesis and characterisation have led to such novel antigen systems being used as potential platforms for developing new anti-malarial vaccine candidates.
    Biochemical and Biophysical Research Communications 07/2014; · 2.41 Impact Factor
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    ABSTRACT: The design of new healthcare schemes which involve using molecular HPV screening means that both persistence and clearance data regarding the most prevalent types of HR-HPV occurring in cities in Colombia must be ascertained.
    BMC Infectious Diseases 07/2014; 14(1):395. · 3.03 Impact Factor
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    ABSTRACT: Tuberculosis (TB) continues being one of the diseases having the greatest mortality rates around the world, 8.7 million cases having been reported in 2011. An efficient vaccine against TB having a great impact on public health is an urgent need. Usually, selecting antigens for vaccines has been based on proteins having immunogenic properties for patients suffering TB and having had promising results in mice and non-human primates. Our approach has been based on a functional approach involving the pathogen–host interaction in the search for antigens to be included in designing an efficient, minimal, subunit-based anti-TB vaccine. This means that Mycobacterium tuberculosis has mainly been involved in studies and that lipoproteins represent an important kind of protein on the cell envelope which can also contribute towards this pathogen's virulence. This study has assessed the expression of four lipoproteins from M. tuberculosis H37Rv, that is, Rv1411c (LprG), Rv1911c (LppC), Rv2270 (LppN) and Rv3763 (LpqH), and the possible biological activity of peptides derived from these. Five peptides were found for these proteins which had high specific binding to both alveolar A549 epithelial cells and U937 monocyte-derived macrophages which were able to significantly inhibit mycobacterial entry to these cells in vitro.
    Chemical Biology &amp Drug Design 07/2014; · 2.47 Impact Factor
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    ABSTRACT: HIV infection leads to a decreasing immune response, thereby facilitating the appearance of other infections, one of the most important ones being HPV. However, studies are needed for determining associations between immunodeficiency caused by HIV and/or the presence of HPV during the course of cervical lesions and their degree of malignancy. This study describes the cytological findings revealed by the Papanicolaou test, laboratory characteristics and HPV molecular profile in women with and without HIV infection.
    BMC Cancer 06/2014; 14(1):451. · 3.33 Impact Factor
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    ABSTRACT: Topological and stereo-electron characteristics are essential in major histocompability class II-peptide-T-cell receptor (MHC-p-TCR) complex formation for inducing an appropriate immune response. Modified high activity binding peptides (mHABPs) were synthesised for complete full protection antimalarial vaccine development producing a large panel of individually fully protection-inducing protein structures (FPIPS) and very high long-lasting antibody-inducing (VHLLAI) mHABPs. Most of those which did not interfere, compete, inhibit or suppress their individual VHLLAI or FPIPS activity contained or displayed a polyproline II-like (PPIIL) structure when mixed. Here we show that amino acid side-chains located in peptide binding region (PBR) positions p3 and p7 displayed specific electron charges and side-chain gauche(+) orientation for interacting with the TCR. Based on the above, and previously described physicochemical principles, non-interfering, long-lasting, full protection-inducing, multi-epitope, multistage, minimal subunit-based chemically synthesised mHABP mixtures can be designed for developing vaccines against diseases scourging humankind, malaria being one of them.
    Vaccine 02/2014; · 3.77 Impact Factor
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    ABSTRACT: Plasmodium falciparum (Pf) malaria causes 200 million cases worldwide, 8 million being severe and complicated leading to ∼1 million deaths and ∼100,000 abortions annually. Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) has been implicated in cytoadherence and infected erythrocyte rosette formation, associated with cerebral malaria; chondroitin sulphate-A attachment and infected erythrocyte sequestration related to pregnancy-associated malaria and other severe forms of disease. An endothelial cell high activity binding peptide is described in several of this ∼300 kDa hypervariable protein's domains displaying a conserved motif (GACxPxRRxxLC); it established H-bonds with other binding peptides to mediate red blood cell group A and chondroitin sulphate attachment. This motif (when properly modified) induced PfEMP1-specific strain-transcending, fully-protective immunity for the first time in experimental challenge in Aotus monkeys, opening the way forward for a long sought-after vaccine against severe malaria.
    PLoS ONE 01/2014; 9(2):e88420. · 3.53 Impact Factor
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    ABSTRACT: Non-human primates belonging to the Aotus genus have been shown to be excellent experimental models for evaluating drugs and vaccine candidates against malaria and other human diseases. The immune system of this animal model must be characterised to assess whether the results obtained here can be extrapolated to humans. Class I and II major histocompatibility complex (MHC) proteins are amongst the most important molecules involved in response to pathogens; in spite of this, the techniques available for genotyping these molecules are usually expensive and/or time-consuming. Previous studies have reported MHC-DRB class II gene typing by microsatellite in Old World primates and humans, showing that such technique provides a fast, reliable and effective alternative to the commonly used ones. Based on this information, a microsatellite present in MHC-DRB intron 2 and its evolutionary patterns were identified in two Aotus species (A. vociferans and A. nancymaae), as well as its potential for genotyping class II MHC-DRB in these primates.
    PLoS ONE 01/2014; 9(5):e96973. · 3.53 Impact Factor
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    ABSTRACT: Human papillomavirus (HPV) is the main risk factor associated with the development of cervical cancer (CC); however, there are other factors that favour the progression of the illness such as immunosuppression caused by the human immunodeficiency virus (HIV). This study was thus aimed at evaluating the functionality of classical polymerase chain reaction (PCR)-based molecular tests for the generic identification of HPV-DNA (GP5+/6+, MY09/11, pU1M/2R individually or in combination) using cervical and urine samples from 194 HIV-positive women. Infected samples were tested with type-specific primers for six high-risk types (HPV-16, -18, -31, -33, -45 and -58) and two low risk ones (HPV-6/11). HPV infection prevalence in cervical samples was 70.1% and 63.9% in urine samples. HPV-16 was the most prevalent viral type in both cervical and urine samples, having high multiple infection frequency compared to single infections detected in such samples. HPV-DNA detection by PCR (mainly pU1M/2R primer set) in urine samples was positively associated with abnormal cytological findings (ASCUS/SIL). It was determined that the operative characteristics for detecting cytological abnormalities were similar for cervical and urine samples. This suggested using PCR for detecting HPV-DNA in urine samples as a potential screening strategy for CC prevention in future prevention and control programmes along with currently implemented strategies for reducing the impact of the disease, i.e. they are economic, easy to collect, have wide acceptability amongst women and have similar operative characteristics to those of the cervical samples.
    Journal of clinical microbiology 08/2013; · 4.16 Impact Factor
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    ABSTRACT: Tuberculosis (TB) remains one of the most worrying infectious diseases affecting public health around the world; 8.7 million new TB cases were reported in 2011. The search for an Mycobacterium tuberculosis H37Rv protein sequence which is functionally important in host-pathogen interaction has been proposed for developing a new vaccine which will allow efficient and safe control of the spread of this disease. The present study thus reports the results obtained for the Rv1268c protein described in the M. tuberculosis H37Rv genome as a hypothetical unknown, probably secreted, protein based on a highly robust, specific, sensitive and functional approach to the search for potential epitopes to be included in an anti-tuberculosis vaccine. Rv1268c presence was determined by immunoblotting after obtaining polyclonal sera against mycobacterial total sonicate or subcellular fractions. Such sera were used in electron immunomicroscopy (EIM) for confirming protein localisation on the M. tuberculosis envelop by recognising colloidal gold-labelled immunoglobulin. Screening assays revealed the presence of two sequences having high binding activity: one binding A549 alveolar epithelial cells ((141)TGMAALEQYLGSGHAVIVSI(160)) and other binding U937 monocyte-derived macrophages ((21)AVALGLASPADAAAGTMYGD(40)). Such sequences' ability to inhibit mycobacterial entry during in vitro assays was analysed. The structure of synthetic peptides binding to target cells was also determined, bearing in mind the structure-function relationship. These results, together with those obtained for other proteins, have been involved in selecting peptides which might be included in a subunit-based anti-tuberculosis vaccine.
    Bioorganic & medicinal chemistry 08/2013; · 2.82 Impact Factor
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    ABSTRACT: Plasmodium falciparum malaria parasite invasion of erythrocytes is an essential step in host infection and the proteins involved in such invasion are the main target in developing an antimalarial vaccine. Secretory organelle-derived proteins (micronemal AMA1 protein and the RON2, 4, and 5 rhoptry neck proteins) have been recently described as components of moving junction complex formation allowing merozoites to move into a newly-created parasitophorous vacuole. This study led to identifying RON5 regions involved in binding to human erythrocytes by using a highly robust, sensitive and specific receptor-ligand interaction assay; it is further shown that the RON5 protein remains highly conserved throughout different parasite strains. It is shown that the binding peptide-erythrocyte interaction is saturable and sensitive to chymotrypsin and trypsin. Invasion inhibition assays using erythrocyte binding peptides showed that the RON5-erythrocyte interaction could be critical for merozoite invasion of erythrocytes. This work provides evidence (for the first time) suggesting a fundamental role for RON5 in erythrocyte invasion.
    Peptides 08/2013; · 2.52 Impact Factor
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    ABSTRACT: The aim of obtaining novel vaccine candidates against malaria and other transmissible diseases can be partly based on selecting non-polymorphic peptides from relevant antigens of pathogens, which have to be then precisely modified for inducing a protective immunity against the disease. Bearing in mind the high degree of the MSA-2(21-40) peptide primary structure's genetic conservation among malaria species, and its crucial role in the high RBC binding ability of Plasmodium falciparum (the main agent causing malaria), structurally defined probes based on non-natural peptide-bond isosteres were thus designed. Thus, two peptide mimetics were obtained (so-called reduced amide pseudopeptides), in which naturally made amide bonds of the (30)FIN(32)-binding motif of MSA-2 were replaced with ψ-[CH2-NH] methylene amide isostere bonds, one between the F-I and the second between I-N amino acid pairs, respectively, coded as ψ-128 ψ-130. These peptide mimetics were used to produce poly- and monoclonal antibodies in Aotus monkeys and BALB/c mice. Parent reactive mice-derived IgM isotype cell clones were induced to Ig isotype switching to IgG sub-classes by controlled in vitro immunization experiments. These mature isotype immunoglobulins revealed a novel epitope in the MSA-2(25-32) antigen and two polypeptides of rodent malaria species. Also, these antibodies' functional activity against malaria was tested by in vitro assays, demonstrating high efficacy in controlling infection and evidencing neutralizing capacity for the rodent in vivo malaria infection. The neutralizing effect of antibodies induced by site-directed designed peptide mimetics on Plasmodium's biological development make these pseudopeptides a valuable tool for future development of immunoprophylactic strategies for controlling malarial infection.
    Amino Acids 07/2013; · 3.91 Impact Factor
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    ABSTRACT: Abstract Tuberculosis (TB) is an air-born, transmissible disease, having an estimated 9.4 million new TB cases worldwide in 2009. Eventual control of this disease by developing a safe and efficient new vaccine able to detain its spread will have an enormous impact on public health policy. Selecting potential antigens to be included in a multi-epitope, minimal subunit-based, chemically-synthesized vaccine containing the minimum sequences needed for blocking mycobacterial interaction with host cells is a complex task due to the multiple mechanisms involved in M. tuberculosis infection and the mycobacterium's immune evasion mechanisms. Our methodology, described here takes into account a highly robust, specific, sensitive and functional approach to the search for potential epitopes to be included in an anti-TB vaccine; it has been based on identifying short mycobacterial protein fragments using synthetic peptides having high affinity interaction with alveolar epithelial cells (A549) and monocyte-derived macrophages (U937) which are able to block the microorganism's entry to target cells in in vitro assays. This manuscript presents a review of the results obtained with some of the MTB H37Rv proteins studied to date, aimed at using these high activity binding peptides (HABPs) as platforms to be included in a minimal subunit-based, multiepitope, chemically-synthesized, antituberculosis vaccine.
    Critical Reviews in Microbiology 02/2013; · 5.07 Impact Factor
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    ABSTRACT: Identifying the minimal functional regions of the proteins which the malaria parasite uses when invading its host cells constitutes the first and most important approach in an effective design for a chemically synthesised, multi-antigen, multi-stage, subunit-based vaccine. This work has been aimed at identifying the PfRh1 protein binding regions (residues 1-2580) belonging to the reticulocyte binding-like (RBL or P. falciparum Rh [PfRh]) family implicated in the parasite's alternative target cell invasion routes. Eighteen peptide regions (called high activity binding peptides - HABPs) binding to red blood cells (RBC) were identified in peptides mapped in a highly robust, specific and sensitive receptor-ligand assay. These HABPs were saturable in the experimental conditions assayed here and most had an alpha helix structure. Polymorphism studies revealed that only six of the eighteen HABPs identified had changes at amino acid level amongst the seven P. falciparum strains evaluated. Most HABPs' specific binding became altered when RBC were treated with neuraminidase, chymotrypsin and trypsin, suggesting differing sensitivity for RBC membrane receptors. After ascertaining that the Rh1 gene was transcribed and expressed in late-stage schizonts of the FCB-2 strain, invasion inhibition assays were carried out. When most of these HABPs were assayed in P. falciparum in vitro culture they were able to inhibit high percentages of FVO strain invasion compared to low inhibition percentages observed with the FCB-2 strain. This data shows small Rh1 regions' participation during invasion and suggests that these units should be included in further immunological and structural studies.
    Vaccine 02/2013; · 3.77 Impact Factor
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    ABSTRACT: Infection, coinfection and type-specific human papillomavirus (HPV) distribution was evaluated in human immunodeficiency virus (HIV)-positive women from paired cervical and urine samples. Paired cervical and urine samples (n = 204) were taken from HIV-positive women for identifying HPV-DNA presence by using polymerase chain reaction (PCR) with three generic primer sets (GP5+/6+, MY09/11 and pU1M/2R). HPV-positive samples were typed for six high-risk HPV (HR-HPV) (HPV-16, -18, -31, -33, -45 and -58) and two low-risk (LR-HPV) (HPV-6/11) types. Agreement between paired sample results and diagnostic performance was evaluated. HPV infection prevalence was 70.6% in cervical and 63.2% in urine samples. HPV-16 was the most prevalent HPV type in both types of sample (66.7% in cervical samples and 62.0% in urine) followed by HPV-31(47.2%) in cervical samples and HPV-58 (35.7%) in urine samples. There was 55.4% coinfection (infection by more than one type of HPV) in cervical samples and 40.2% in urine samples. Abnormal Papanicolau smears were observed in 25.3% of the women, presenting significant association with HPV-DNA being identified in urine samples. There was poor agreement of cervical and urine sample results in generic and type-specific detection of HPV. Urine samples provided the best diagnosis when taking cytological findings as reference. In conclusion including urine samples could be a good strategy for ensuring adherence to screening programs aimed at reducing the impact of cervical cancer, since this sample is easy to obtain and showed good diagnostic performance.
    PLoS ONE 01/2013; 8(2):e56509. · 3.53 Impact Factor
  • Jose Manuel Lozano, Manuel Elkin Patarroyo
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    ABSTRACT: The molecular basis for obtaining novel anti-malarial vaccine candidates depends on a considered selection of antigenic peptides, mainly derived from Plasmodium antigens’ non-polymorphic regions. Since such targeted-molecules are poorly immunogenic when tested as vaccine components, they usually have to be modified to overcome their immunological phenotype. Transition state theory, explaining how peptidases catalyse a given peptide bond breakage, thus led to reduced amide pseudopeptides being proposed as possible mimetics for a transition-state. Stabilising such high-energy molecular stages could become a strategy for inducing antibodies potentially harbouring catalytic properties. Hence, isostere-bond peptido-mimetics represented a rational choice as potential abzyme-inducers and site-directed designed reduced amide pseudopeptides for obtaining peptide-analogues from selected malarial high-binding motifs. This novel family of vaccine candidates has proved to be an efficient functional antibody-inducer, the latter acting as efficient blockers of Plasmodium infection of human and mouse RBCs.
    Current Immunology Reviews 01/2013; 9(4).
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    ABSTRACT: Previous BAC clone analysis of the Platyrrhini owl monkey KIRs have shown an unusual genetic structure in some loci. Therefore, cDNAs encoding KIR molecules from eleven Aotus vociferans monkeys were characterized here; ten putative KIR loci were found, some of which encoded atypical proteins such as KIR4DL and transcripts predicted to encode a D0+D1 configuration (AOTVOKIR2DL1*01v1) which appear to be unique in the Aotus genus. Furthermore, alternative splicing was found as a likely mechanism for producing activator receptors in A. vociferans species. KIR proteins from New World monkeys may be split into three new lineages according to domain by domain phylogenetic analysis. Although the A. vociferans KIR family displayed a high divergence among paralogous genes, individual loci were limited in their genetic polymorphism. Selection analysis showed that both constrained and rapid evolution may operate within the AvKIR family. The frequent alternative splicing (as a likely mechanism generating activator receptors), the presence of KIR4DL and KIR2DL1 (D0+D1) molecules and other data reported here suggest that the KIR family in Aotus has had a rapid evolution, independent from its Catarrhini counterparts.
    PLoS ONE 01/2013; 8(11):e79731. · 3.53 Impact Factor
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    ABSTRACT: Modified HABP (mHABP) regions interacting with HLA-DRβ1∗ molecules have a more restricted conformation and/or sequence than other mHABPs which do not fit perfectly into their peptide binding regions (PBR) and do not induce an acceptable immune response due to the critical role of their ϕ and ψ torsion angles. These angles' critical role was determined in such highly immunogenic, protection-inducing response against experimental malaria using the conformers (mHABPs) obtained by (1)H-NMR and superimposed into HLA-DRβ1∗-like Aotus monkey molecules; their phi (Φ) and psi (Ψ) angles were measured and the H-bond formation between these molecules was evaluated. The aforementioned mHABP propensity to assume a regular conformation similar to a left-handed polyproline type II helix (PPII(L)) led to suggesting that favouring these conformations according to their amino acid sequence would lead to high antibody titre production and sterile protective immunity induction against malaria, thereby adding new principles or rules for vaccine development, malaria being one of them.
    Biochemical and Biophysical Research Communications 11/2012; · 2.41 Impact Factor
  • Manuel E Patarroyo, Adriana Bermudez, Martha P Alba
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    ABSTRACT: The importance of CSP- and STARP-derived Φ and Ψ dihedral angles in mHABP structure was analysed by (1)H-NMR in the search for molecules which can be included as components of a first-line-of-defence P. falciparum sporozoite multi-epitope vaccine against the most lethal form of human malaria. Most of the aforementioned dihedral angles were left-hand-like polyproline type II (PPII(L)) structures whilst others had right-hand-like α-helix (α(R)), thus allowing mHABPS to fit better into MHCII molecules and thereby form an appropriate pMHCII complex and also establish the H-bonds which stabilise such complex and by this means induce an appropriate immune response. This information has great implications for vaccine development, malaria being one of them.
    Biochemical and Biophysical Research Communications 11/2012; · 2.41 Impact Factor
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    ABSTRACT: If ever there were a truism then it would be that a completely protective Plasmodium falciparum malaria vaccine is desperately needed. Our institute has devoted all its efforts during the last 30 years to developing a fully protective, minimal subunit-based, multiepitope, multistage (targeting sporozoite and merozoite proteins), chemically synthesized antimalarial vaccine, given that peptides with high binding activity to their corresponding host cells (liver cells or red blood cells) form the springboard for vaccine design. However, such conserved high activity binding peptides have to be specifically modified to render them into highly immunogenic and protection-inducing peptides since they are immunologically silent. These modifications, analyzed at the 3D structural level by (1)H-NMR, allow them a better fit into the MHC II-peptide-T-cell receptor complex to induce an appropriate immune response, providing a rational and logical approach (analyzed at the single atom level) for vaccine development, particularly in the field of malaria.
    Expert Review of Vaccines 09/2012; 11(9):1057-1070. · 4.22 Impact Factor
  • Armando Moreno-Vranich, Manuel E Patarroyo
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    ABSTRACT: Conserved Plasmodium falciparum high activity binding peptides' (HABPs) most relevant proteins involved in malaria parasite invasion are immunologically silent; critical binding residues must therefore be specifically replaced to render them highly immunogenic and protection-inducing. Such changes have a tremendous impact on these peptides' steric-electronic effects, such as modifications to peptide length peptide bonds and electronic orbitals' disposition, to allow a better fit into immune system MHCII molecules and better interaction with the TCR which might account for the final immunological outcome.
    Biochemical and Biophysical Research Communications 06/2012; 423(4):857-62. · 2.41 Impact Factor

Publication Stats

5k Citations
1,257.97 Total Impact Points

Institutions

  • 2001–2014
    • Fundación instituto de Inmunología de Colombia
      Μπογκοτά, Bogota D.C., Colombia
  • 2010–2013
    • Universidad del Rosario
      • School of Medicine and Health Sciences
      Bogotá, Bogota D.C., Colombia
  • 1983–2013
    • National University of Colombia
      • Facultad de Medicina
      Μπογκοτά, Bogota D.C., Colombia
  • 2011
    • Johns Hopkins University
      • Department of Biophysics and Biophysical Chemistry
      Baltimore, MD, United States
  • 2001–2010
    • University of São Paulo
      • Faculty of Medicine (FM)
      San Paulo, São Paulo, Brazil
  • 1998–2005
    • Swiss Tropical and Public Health Institute
      Bâle, Basel-City, Switzerland
  • 2001–2004
    • Universidad del País Vasco / Euskal Herriko Unibertsitatea
      • Faculty of Pharmacy
      Bilbao, Basque Country, Spain
  • 1992–2003
    • Universidad Autónoma de Madrid
      • • Departamento de Farmacología
      • • Centro de Biología Molecular "Severo Ochoa" (CSIC-UAM)
      Madrid, Madrid, Spain
  • 1988–2002
    • Hospital San Juan de Dios Pamplona
      Quilichao, Cauca, Colombia
  • 2000
    • University of Minnesota Duluth
      Duluth, Minnesota, United States
    • Hospital Clínic de Barcelona
      Barcino, Catalonia, Spain
    • Universidad de Cartagena
      Cartagena de Indias, Bolívar, Colombia
  • 1995
    • Instituto do Coração
      San Paulo, São Paulo, Brazil
  • 1987
    • Stockholm University
      • Department of Mathematics
      Tukholma, Stockholm, Sweden