Ricardo Santos Simões

University of São Paulo, San Paulo, São Paulo, Brazil

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Publications (119)150.26 Total impact

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    ABSTRACT: Introduction: osteoprotegerin has emerged as a new candidate for the treatment of osteoporosis. However, high levels of osteoprotegerin have been linked to vascular calcification, an independent and well-defined risk factor for cardiovascular disease (CVD) and mortality. Thus, the action of osteoprotegerin in these situations has been questioned. Objective: to evaluate the effect of osteoprotegerin (OPG) on the human body, especially in bone tissue and in vascular diseases. Methods: the scientific databases consulted were PubMed-Medline and Cochrane, using keywords (MeSH terms) grouped into the following syntaxes: (Osteoprotegerin OR Osteoclastogenesis Inhibitory Factor OR Receptors, Tumor Necrosis Factor, Member 11b OR Tumor Necrosis Factor Receptor Superfamily, Member 11b OR FDCR-1 Protein OR FDCR 1 Protein OR OCIF Protein OR Follicular Dendritic Cell-Derived Receptor-1) AND (Bones AND Bone OR Bones AND Bone Tissue OR Bones OR Bone Tissue OR Cardiovascular Diseases). Results: Osteoprotegerin is present in various organs and binds to two ligands: nuclear factor kB (RANKL) related to the differentiation of osteoclasts, and tumor necrosis factor related to the apoptosis-inducing ligand (TRAIL). OPG inhibits the regulation effects of nuclear factor kB on inflammation and on the skeletal and vascular systems, preventing the apoptosis induced by TRAIL, being related to the preservation of bone tissue. Conclusion: a deeper knowledge of the mechanisms involved in the association between OPG serum levels, bone integrity and cardiovascular disease can provide important data for future therapeutic interventions.
    Full-text · Article · Dec 2015
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    ABSTRACT: Objective: To quantify the collagen fibers in the lacrimal gland of female mice with hyperprolactinemia. Methods: Forty adult female mice were randomly divided into two groups with 20 animals each: nonpregnant control (CTR1, control group, 0.2 mL of saline solution) and nonpregnant experimental (HPRL1, experimental group, 200 µg/day metoclopramide). Treatments lasted for 50 consecutive days. On day 50, 10 females from each group (control and experimental) were euthanized in the proestrus phase; then, the blood was collected and the lacrimal glands were removed. Thereafter, the remaining females were placed with the mates and continued to receive treatment with saline solution or metoclopramide. On the 6th post-coital day, 10 pregnant females from the control group (CTR2) and 10 pregnant females from the experimental group (HPRL2) were euthanized, after which blood was collected and the lacrimal glands removed. The lacrimal glands were processed for morphological analyses and collagen quantification, and prolactin and sex steroid levels were measured in the blood samples. Data were statistically analyzed using an unpaired Student t test (p<0.05). Results: Morphological analysis revealed greater structural tissue disorganization of the lacrimal glands in the metoclopramide-treated groups. The total collagen content was significantly higher in the HPRL1 group than in the CTR1 group (p<0.05), whereas the difference between the CTR2 and HPRL2 groups was not significant. Conclusion: Our data suggest an impairment in the functioning of the lacrimal gland as a consequence of increased prolactin levels and decreased serum levels of estrogen and progesterone.
    Full-text · Article · Sep 2015 · Clinics (São Paulo, Brazil)
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    ABSTRACT: The aim of this study was to evaluate the effects of metoclopramide-induced hyperprolactinemia on the tibial epiphyseal plate of hormone-treated oophorectomized mice. For this purpose, 18 animals with intact ovaries were allocated to two groups, M (metoclopramide) and V (vehicle). One hundred and eight oophorectomized animals were allocated to 12 subgroups: Oophx/V (vehicle); Ooph/M (metoclopramide); Oophx/V + E (vehicle + estradiol); Oophx/M + E (metoclopramide + estradiol); Oophx/V + P (vehicle + progesterone); Oophx/M + P (metoclopramide + progesterone); Oophx/V + T (vehicle + testosterone); Oophx/M + T (metoclopramide + testosterone); Oophx/V + E + P (Vehicle + estradiol + progesterone); Oophx/M + E + P (metoclopramide + estradiol + progesterone); Oophx/V + E + P + T (vehicle + estradiol + progesterone + testosterone); Oophx/M + E + P + T (metoclopramide + estradiol + progesterone + testosterone). After a 50-day treatment was performed histomorphometric and immunohistochemical cell death analysis. In the epiphyseal plate of the hyperprolactinemic and/or oophorectomized animals, cell proliferation and bone formation decreased, inducing intensified cell death. In the sex steroid-treated animals, estrogen boosted cell proliferation; progesterone, bone formation and testosterone, both cell proliferation and bone formation. These findings suggest that oophorectomy and hyperprolactinemia changed epiphyseal plate morphology causing cartilage degeneration. Treatment with combined sex steroids may diminish such deleterious effects.
    No preview · Article · Jul 2015 · Gynecological Endocrinology
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    ABSTRACT: To evaluate the sex steroid profile and histomorphometry of the adrenal cortical zones of androgenized rats (wistar) with polycystic ovary syndrome treated with metformin. Thirty animals were divided into three groups: GC (regular estrous cycle), GPE (permanent estrus), and GPEM (permanent estrus + metformin 28 mg/kg for 50 days). At the end of this period, blood was collected for hormone measurement. The width of the adrenal cortical zones and the nuclear volumes were analyzed by histomorphometry. The ANOVA test was used in the statistical analysis. The adrenal glands of the androgenized animals were larger and more intensely vascularized than those of the other groups. The concentration of androstenedione in GPE was higher than that in the other groups (0.4 ± 0.1*>= 0.2 ± 0.1 = 0.2 ± 01, *p < 0.05). The width of the zona glomerulosa and of the zona reticularis and their nuclear volumes were greater in GPE compared to those of the other groups (GPE* > GPEM = GC, *p < 0.05). Metformin treatment may decrease the serum levels of androstenedione as well as the width and the nuclear volumes of the zona glomerulosa and of the zona reticularis in androgenized animals.
    No preview · Article · Jun 2015 · Gynecological Endocrinology
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    ABSTRACT: To evaluate the effects of angico bark extract (Anadenanthera colubrina var. cebil) in the healing process of the skin of rats. Twenty adult male rats were divided into four groups of five animals each, according to the respective postoperative days, as follow: G4, G7, G14 and G21. Each group received two incisions on skin and subcutaneous tissue in the right and left antimere of the thoracic region, separated by a distance of 2 cm. The right lesion was treated daily with saline and the left with the angico alcoholic extract (5%). At the end of each experimental period, the animals were euthanized and fragments of the wound area with the edges were removed, fixed in 10% formaldehyde solution and processed for paraffin embedding. Histological sections (5 μm of thickness) were stained with hematoxylin and eosin (HE), Gomori trichromic and picrosisirus red for morphological and morphometric analyses. Statistical analysis was done by ANOVA and Tukey-Kramer test (p<0.05). Morphological analysis showed larger fibroblasts and a higher concentration of collagen fibers in skyn wounds treated with the angico extract. Morphometric analysis demonstrated a significant increase in the number of fibroblasts at 7th and collagen in 7th and 14th days (p<0.01) in wounds treated with the angico extract. The angico alcoholic extract (Anadenanthera colubrina var. cebil) induces the acceleration of wound healing in skin wounds of rats.
    Full-text · Article · May 2015 · Acta cirurgica brasileira / Sociedade Brasileira para Desenvolvimento Pesquisa em Cirurgia
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    ABSTRACT: Soybeans as a whole or their food isolates are found in many foods, such as tofu, soy milk and yogurt. In women, the consumption of these foods is especially associated with beneficial health effects, such as menopause vasomotor symptoms reduction, osteoporosis prevention, genital atrophy mitigation, and breast cancer attenuation. Among them, some elements have nonsteroidal structure and can bind to estrogen receptors, which are classified as phytoestrogens. Isoflavones are phytoestrogens with high biological activity, especially those named genistein and daidzein. Both bind to estrogen receptors with greater affinity to β receptors. Genistein (4', 5,7 trihydroxyisoflavone) is considered a potent anticancer agent due to its capacity of inhibiting tumor cell growth factors in several tissues and diseases, such as breast, prostate, and neck cancers as well as melanoma and leukemia. Moreover, they block the cell cycle and induce cell death by apoptosis, inactivating NF-kB and activating caspase-3 in prostate cancer cells. In addition, they act as potent angiogenic factors and tumor cell proliferation inhibitors. Some studies suggest that genistein inhibited the growth of tumor cells due to inhibition of gene expression of protein tyrosine kinase of topoisomerases I and II in different cell types. Other studies also showed that genistein was capable to prevent apoptosis of normal cells through their antioxidant properties. Daidzein (7,4-dihydroxy isoflavone), another isoflavone, when ingested is metabolized in equol by intestinal bacterias. It has been hypothesized that production of equol is the key to the effectiveness of isoflavone, which brings health benefits. Taking together, clinical, experimental and cell cultures studies suggest that isoflavones, mainly genistein and daidzein, play an important role in preventing chronic diseases, such as osteoporosis, cancer and diabetes.
    Full-text · Chapter · Apr 2015
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    ABSTRACT: To evaluate wound contraction and the concentration of mast cells in skin wounds treated with 5% BPT essential oil-based ointment in rats. Twenty rats, male, of adult age, were submitted to skin surgery on the right (RA) and left antimeres (LA) of the thoracic region. They were divided into two groups: control (RA - wounds receiving daily topical application of vaseline and lanolin) and treated (LA - wounds treated daily with the topical ointment). The skin region with wounds were collected at days 4, 7, 14 and 21 after surgery. Those were fixed in 10% formaldehyde and later processed for paraffin embedding. Sections were obtained and stained by H.E for histopathology analysis. The degree of epithelial contraction was measured and mast cell concentration were also evaluated. The treated group showed higher mast cell concentrations (p<0.05) associated with increased contraction at day 7 and 14 respectively. Ointment containing 5% Brazilian pepper tree oil increases mast cell concentration and promotes skin wound contraction in rats.
    Full-text · Article · Apr 2015 · Acta cirurgica brasileira / Sociedade Brasileira para Desenvolvimento Pesquisa em Cirurgia
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    ABSTRACT: To evaluate the morphology, necrotic area and collagen content in skin flaps of rats subjected to hyperbaric oxygenation (HBO). Forty adult rats were divided into four groups: GEC - epilated; GE/HBO - epilated subjected to HBO; GER - epilated submitted to skin flap (2 cm in width /8 cm length in the dorsal area) and GER/HBO - epilated, subjected to skin flap and HBO. HBO (2.4 ATA) was performed for two hours during seven consecutive days. In the eighth day, the rats were anesthetized and the skin flaps were removed and separated into three portions, relative to pedicle fixation. The material fixed in 10% formalin was processed for paraffin embedding; sections were stained by H.E and subjected to picrosirius-red method. The slides examined under light microscopy for evaluation of the collagen content in polarized light microscope and ImageLab(r) software (Bio-Rad). The data showed larger area of necrosis and lower levels of collagen in the three regions of the GER group, whereas in the GER/HBO group the collagen content was similar to the GEC and GE/HBO groups. Hyperbaric oxygenation reduced the area of necrosis and preserved the morphology and collagen content in skin flaps of rats.
    Full-text · Article · Apr 2015 · Acta cirurgica brasileira / Sociedade Brasileira para Desenvolvimento Pesquisa em Cirurgia
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    ABSTRACT: To analyze steroidogenesis-related gene expression in the rat ovary exposed to melatonin supplementation. Thirty-two virgin adult female rats were randomized to two groups as follows: the control group GI received vehicle and the experimental group GII received melatonin supplementation (10 µg/night per animal) for 60 consecutive days. After the treatment, animals were anesthetized and the collected ovaries were immediately placed in liquid nitrogen for complementary deoxyribonucleic acid microarray analyses. A GeneChip¯ Kit Rat Genome 230 2.0 Affymetrix Array was used for gene analysis and the experiment was repeated three times for each group. The results were normalized with the GeneChip¯ Operating Software program and confirmed through analysis with the secondary deoxyribonucleic acid-Chip Analyzer (dChip) software. The data were confirmed by real-time reverse transcription polymerase chain reaction analysis. Genes related to ovarian function were further confirmed by immunohistochemistry. We found the upregulation of the type 9 adenylate cyclase and inhibin beta B genes and the downregulation of the cyclic adenosine monophosphate response element modulator and cytochrome P450 family 17a1 genes in the ovarian tissue of GII compared to those of the control group. Our data suggest that melatonin supplementation decreases gene expression of cyclic adenosine monophosphate, which changes ovarian steroidogenesis.
    Full-text · Article · Feb 2015 · Clinics (São Paulo, Brazil)
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    ABSTRACT: Background Infertility associated with polycystic ovary syndrome (PCOS) could be related to many mechanisms including endometrial factors. Objectives To review cell adhesion molecule and estrogen receptor expression in the endometrium. Search strategy A systematic review was performed of the Medline and Cochrane databases for papers published in any language between 2004 and 2014. The search term was "'polycystic ovary syndrome' OR 'Stein Leventhal syndrome' OR 'anovulation' AND 'endometrium' OR 'endometria.'" Selection criteria Research studies on endometrial cell adhesion molecules and estrogen receptor expression among women with PCOS diagnosed according to the Rotterdam criteria were included. Data collection and analysis Data were extracted from identified studies and the quality of assessment was analyzed. Main results Six studies were included. Data were controversial with respect to MUC1 and αVβ3 integrin expression with significantly higher and lower levels, respectively, in women with PCOS. Estrogen receptor expression was enhanced among patients with PCOS as compared with healthy women. Conclusions Endometrial factors influence embryo receptivity as indicated by the molecular mediators identified in the studies, including cell adhesion molecules and the estrogen receptor. © 2014 International Federation of Gynecology and Obstetrics. Published by Elsevier Ireland Ltd. All rights reserved.
    No preview · Article · Dec 2014 · International Journal of Gynecology & Obstetrics

  • No preview · Conference Paper · Dec 2014
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    ABSTRACT: Abstract The aim of this study was to quantify the sulfated glycosaminoglycans in the endometria of women with polycystic ovary syndrome (PCOS). Of the 18 patients recruited for this study, 10 patients with PCOS comprised the PCOS group (PCOSG), and eight patients with regular and ovulatory menstrual cycles comprised the control group (CG). The clinical, biochemical, morphological and endometrial data from both groups were analyzed. Biopsies were performed during the proliferative phase of the menstrual cycle for the CG and during the persistent proliferative phase for the PCOSG (all women were amenorrheic). In the PCOSG, there was a significant increase in the endometrial concentration levels of heparan sulfate (p = 0.03), but no difference in the concentrations of chondroitin sulfate was determined between the two groups (p = 0.77). Period of time without menstruation (p = 0.001) and body mass index (BMI) (p = 0.04) correlated directly and positively with heparan sulfate concentration. There was no association between heparan sulfate levels and basal insulin values (p = 0.08). High levels of endometrial heparan sulfate in women with PCOS indicate an interference with maternal-fetal recognition, which contributes to infertility; thus, endometrial heparan sulfate may be a predictive marker of future neoplasia risk.
    No preview · Article · Dec 2014 · Gynecological Endocrinology
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    ABSTRACT: Objective: We evaluated whether genistein or estrogen treatment has the same effect when administered immediately or late to rats induced with menopause using ovariectomy. Methods: Sixty adult female rats were divided into six treatment groups: GI = vehicle immediately after ovariectomy; GII = vehicle 30 days after ovariectomy; GIII = genistein immediately after ovariectomy; GIV = genistein 30 days after ovariectomy; GV = estrogen immediately after ovariectomy; and GVI = estrogen 30 days after ovariectomy. All animals were treated for 30 consecutive days. At the end of the treatment, part of the uteri was removed for subsequent histological studies and another part was used to evaluate estrogen receptors 1 and 2, cell proliferation (cyclin A1 and A2, cyclin D1, cyclin-dependent kinase inhibitors 1, 1B and 2, antigen identified by the monoclonal antibody Ki67) and angiogenesis (vascular endothelial growth factor, VEGF-A) gene expression. Results: Late treatment after castration in rats resulted in more developed endometrium, enhanced cell proliferation and estrogen-signalling pathways, particularly the cyclin-related genes Ki67 and VEGF-A, compared to early treatment. Interestingly, these same effects were less intense with genistein compared to those induced by estrogen, especially when genistein was administered late. Conclusion: Our data show that isoflavone renders a lower risk of cancer when compared to estrogen in treatments.
    Full-text · Article · Sep 2014 · Climacteric

  • No preview · Article · Aug 2014 · Microscopy and Microanalysis
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    ABSTRACT: Objective: To evaluate the effects of soy isoflavone extract in the pro-oxidant/antioxidant balance in the uterus of ovariectomized rats. Methods: Twenty 3-month-old adult female Wistar rats were divided into four equal groups: GI: sham-operated (estrous phase); GII: control ovariectomized rats; GIII: ovariectomized rats treated with genistein (50 μg/kg/day) by gavage; GIV: ovariectomized rats subcutaneously treated with estrogen (10 μg/kg/day). After 30 consecutive days of treatment, the rats were euthanized and the uterus removed. The distal thirds of the uterine horns were processed for histomorphometric analyses of endometrial and myometrial thicknesses and glandular area. Other regions of the uteri were kept in liquid nitrogen and subsequently processed for analysis of reactive species quantification (DCF), total antioxidant capacity (TAC) and lipid oxidation status (TBARS). Data were statistically analyzed by one-way ANOVA, complemented by the Tukey-Kramer test (p < 0.05). Results: GII and GIII exhibited lower endometrial thickness, glandular area and myometrial thickness than GI and GIV, while a higher myometrial thickness was observed in GIV compared with the other groups. Moreover, the isoflavone-treated group showed lower DCF and TBARS compared to GII, and also an improvement of TAC compared to GI and GIV. Despite the significant decrease in TBARS, no significant difference in DCF nor a decrease in TAC were observed in GIV when compared to GII. Conclusion: Our data show that isoflavones improve antioxidant status and counteract oxidative stress, without promoting a trophic effect in the uterus of rats.
    Full-text · Article · Aug 2014 · Climacteric
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    ABSTRACT: Objective: to evaluate the immunohistochemical expression of proliferative, apoptotic and steroidogenic enzyme markers in the ovaries of rats with polycystic ovary syndrome (PCOS). Methods: twenty rats were divided into two groups: GCtrl - estrous phase, and PCOS - with polycystic ovaries. The GCtrl animals were subjected to a lighting period from 7 am to 7 pm, while the animals with PCOS group remained with continuous lighting for 60 days. Subsequently, the animals were anesthetized, the ovaries were removed and fixed in 10% formaldehyde, prior to paraffin embedding. Sections were stained using H.E. or subjected to immunohistochemical methods for the detection of Ki-67, cleaved caspase-3, CYP11A1, CYP17A1 and CYP19A1. The results were analyzed using Student's t-test (p < 0,05). Results: morphological results showed evidence of interstitial cells originating from the inner theca cells of degenerating ovarian cysts in PCOS. Immunoexpression of Ki-67 was higher in the granulosa cells in GCtrl, and the theca interna cells in PCOS, while cleaved caspase-3 was higher in granulosa cells of ovarian cysts from PCOS and in the theca interna cells of GCtrl. Immunoreactivity of CYP11A1 in the theca interna, granulosa and interstitial cells was similar between the two groups, while CYP17A1 and CYP19A1 were higher in the granulosa and interstitial cells in the PCOS group. Conclusion: the results indicate that the interstitial cells are derived from the theca interna and that enzymatic changes occur in the theca interna and interstitial cells in ovaries of rats with PCOS, responsible for the high levels of androgens and estradiol.
    Full-text · Article · Jul 2014 · Revista da Associação Médica Brasileira
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    ABSTRACT: To analyze the expression of genes related to steroidogenesis in the ovary of pinealectomized rats. Experimental research. University research laboratory. Thirty female adult rats. Administration of vehicle (GI), pinealectomy with vehicle (GII), or pinealectomy with melatonin replacement (10 μg/night) for 60 consecutive days (GIII), then euthanasia after 2 months of treatment, ovary collection complementary DNA microarray analyses, confirmatory quantitative reverse-transcriptase polymerase chain reaction analyses, and immunohistochemical analyses for localizing steroidogenesis changes in the ovary. Biologic molecular study followed by immunohistochemical analysis. The changes in the expression of CYP11A1, CYP17A1, and CYP19A1 after pinealectomy (GII) compared with control (GI) showed the Cyp17a1 expression level increased in the theca interna and interstitial cells in the GII rats compared with the other groups. Melatonin deprivation (pinealectomy) or administration may influence the ovarian CYP17A1 expression and steroidogenesis.
    Full-text · Article · May 2014 · Fertility and sterility
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    ABSTRACT: Cryopreservation of the ovarian tissue has shown promising results. However, there remain controversial issues such as the short half-life of grafts. In this aspect, there are some evidences that preconditioning the ovarian tissue before transplantation is beneficial. To determine the effect of hypoxic preconditioning in vitro on ovarian tissue prior to transplantation. Eighteen female adult Wistar rats, were sorted into three experimental groups. Ovaries were maintained in DMEM low glucose serum free at 37[degree sign]C with 5% CO2, at atmospheric oxigen concentration (normoxia) or 1% O2 (hypoxia) for 16 hours. Oxigen concentration was determined by injection of nitrogen in the incubator. Animals submitted to ovarian transplantation immediately after oophorectomy were the Control Group (C). After this, the ovaries were implanted in the retroperitoneum with nonabsorbable suture and animals evaluated for thirty days after transplantation. Beginning on postoperative (PO) day 11, a daily collection of vaginal smear was carried out. Analyses comprised morphological, morphometric (counting ovarian follicles and corpora lutea) and immunohistochemistry for cleaved caspase-3 (apoptosis). In normoxia and control groups all animals recovered their estrous cycles, while in the hypoxia group, two animals did not ovulate but, among those which did, resumption took longer than in the other groups (p < 0.05). The number of ovarian follicles and corpora lutea decreased significantly in the hypoxia group when compared to the other two groups (p < 0.001) and apoptosis was increased in the few ovarian follicles which remained viable (p < 0.001). The hypoxic preconditioning in vitro was not beneficial to the graft and worsened their viability, compromising its functionality or delaying the return of this.
    Full-text · Article · Mar 2014 · Journal of Ovarian Research
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    ABSTRACT: Literature data indicate that the infertility in women with polycystic ovary syndrome (PCOS) is not only attributed to anovulation but also to endometrial dysfunction. Endometrial biopsies were performed in the proliferative and secretory phases of women with normal cycle and in women with PCOS before and after oral treatment with micronized progesterone. After the treatment, the endometrium of the women with PCOS exhibited a lower number of glands and thicker luminal epithelium compared to the normal women in the secretory phase. In addition, the PCOS group exhibited reduced integrin and MECA-79 immunoexpression during the secretory phase. The expression of E-cadherin was higher in the PCOS and the expression of intercellular adhesion molecule 1 was lower in PCOS, during the secretory and proliferative phases, respectively. Also, there is a negative correlation with MECA-79 and integrin expression and body mass index. Conventional doses of progesterone may not be enough to correct the changes of endometrial histomorphology and the receptive markers of PCOS-bearing women. The obesity may be a factor that interferes with this response.
    No preview · Article · Jan 2014 · Reproductive sciences (Thousand Oaks, Calif.)

  • No preview · Conference Paper · Dec 2013

Publication Stats

382 Citations
150.26 Total Impact Points

Institutions

  • 2008-2015
    • University of São Paulo
      • • Department of Obstetrics and Gynecology
      • • Faculty of Medicine (FM)
      San Paulo, São Paulo, Brazil
  • 2004-2015
    • Universidade Federal de São Paulo
      • • Departamento de Morfologia e Genética
      • • Departamento de Ginecologia
      San Paulo, São Paulo, Brazil
  • 2012-2014
    • Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo
      San Paulo, São Paulo, Brazil
    • Federal University of Pernambuco
      Arrecife, Pernambuco, Brazil
  • 2013
    • Federal University of Minas Gerais
      Cidade de Minas, Minas Gerais, Brazil
  • 2011
    • Senac São Paulo
      San Paulo, São Paulo, Brazil