Inscrição votiva em língua lusitana (Arronches Portalegre)

Propõe-se leitura, interpretação e integração histórica da epígrafe redi- gida em língua lusitana, proveniente de uma herdade dos arredores de Arronches. Documenta o sacrifício de animais, designadamente de dez ovelhas, a divindades indígenas – Banda, Reva, Munis, Broeneia... – cujos nomes se fazem acompanhar de epítetos, um dos quais repetido com grafias diferentes (em dativo, Haracui, Aharacui, Harase), passí- vel de relacionar-se com o topónimo actual, Arronches. Na segunda parte, os três dedicantes, que poderão identificar-se como criadores de ovelhas, suplicam às divindades que lhes aceitem os sacrifícios. Considera-se muito viável a hipótese de relacionar esta e as outras epígrafes em língua lusitana – de Lamas de Moledo e Cabeço das Frá- guas – com as rotas da transumância logo nos primórdios da domina- ção romana

Mechanistic Insights into the Anti-angiogenic Activity of Trypanosoma cruzi Protein 21 and its Potential Impact on the Onset of Chagasic Cardiomyopathy

Chronic chagasic cardiomyopathy (CCC) is arguably the most important form of the Chagas Disease, caused by the intracellular protozoan Trypanosoma cruziit is estimated that 10-30% of chronic patients develop this clinical manifestation. The most common and severe form of CCC can be related to ventricular abnormalities, such as heart failure, arrhythmias, heart blocks, thromboembolic events and sudden death. Therefore, in this study, we proposed to evaluate the anti-angiogenic activity of a recombinant protein from T. cruzi named P21 (rP21) and the potential impact of the native protein on CCC. Our data suggest that the anti-angiogenic activity of rP21 depends on the protein's direct interaction with the CXCR4 receptor. This capacity is likely related to the modulation of the expression of actin and angiogenesis-associated genes. Thus, our results indicate that T. cruzi P21 is an attractive target for the development of innovative therapeutic agents against CCC.Univ Fed Sao Paulo, Escola Paulista Med, Departamento Microbiol Imunol Parasitol, BR-05508 Sao Paulo, SP, BrazilUniv Fed Uberlandia, Inst Ciencias Biomed, Dept Imunol, Lab Tripanosomatideos, Uberlandia, MG, BrazilUniv Fed Uberlandia, Inst Genet & Bioquim, Lab Bioquim & Toxinas Animais, Uberlandia, MG, BrazilCeTICS, Inst Butantan, Sao Paulo, BrazilUniv Fed Uberlandia, Fac Med, Centro Referencia Nacl Dermatol Sanitaria Hanseni, Lab Patol Mol & Biotecnol, Uberlandia, MG, BrazilUniv Fed Uberlandia, Inst Ciencias Biomed, Dept Immunol, Lab Osteoimunol & Imunol Tumores, Uberlandia, MG, BrazilUniv Fed Sao Paulo, Escola Paulista Med, Departamento Microbiol Imunol Parasitol, BR-05508 Sao Paulo, SP, BrazilUniv Fed Sao Paulo, Escola Paulista Med, Departamento Microbiol Imunol Parasitol, BR-05508 Sao Paulo, SP, BrazilWeb of Scienc

Trypanosoma cruzi

Trypanosoma cruzi is the etiological agent of American trypanosomiasis, or Chagas disease, and is transmitted mainly by blood-sucking reduviid insects in endemic countries. Metacyclic trypomastigotes released in the feces during the insect blood meal enter a mammalian host through skin wounds or mucosal membranes and invade sur- rounding cells. After cell invasion, metacyclic trypomastigotes are restrained within a parasitophorous vacuole (PV), from where they escape, transform into amastigotes, and multiply in the cytosol. Later, following binary division, amastigotes differentiate back into highly motile trypomastigotes that are released upon cell lysis. They can infect neighboring cells, migrate to different tissues, or be ingested by an insect vector. The parasites in the tissues, as- sociated with an immune response, contribute to the chronic symptoms of the disease. Reactive oxygen species (ROS), among other factors, play an important role during parasite multiplication and interstage transformation.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)CNPq: 424729/2018FAPESP: 2018/09948-

Trypanosoma cruzi cell invasion and traffic: Influence of Coxiella burnetii and pH in a comparative study between distinct infective forms

Previous studies have shown that Coxiella burnetii, an intracellular bacterium that resides within acidified vacuoles with secondary lysosomal characteristics, is an effective modulator of the intracellular traffic of trypomastigote forms of Trypanosoma cruzi.. in addition, vacuolar and cellular pH are related to fusion events that result in doubly infected phagosomes. T cruzi, the etiological agent of Chagas' disease, occurs as different strains grouped in two major phylogenetic lineages: T cruzi I, associated with the sylvatic cycle, and T cruzi II, linked to the human disease. in this work we compared extracellular amastigotes (EA), metacyclic trypomastigotes (NIT) and tissue Culture derived trypomastigotes (TCT) belonging to T. cruzi I or T. cruzi II for their ability to invade and escape from their parasitophorous vacuole (PV), in Vero cells or Vero cells harboring the bacterium, C burnetti. Distinct invasion patterns were observed between different infective stages and between infective forms of different strains. Studies on the transference kinetics revealed that pH modulates the intracellular traffic of each infective stage, but this influence is not exclusive for each phylogenetic group. Endosomal to lysosomal sequential labeling with EEA-1 and LAMP-1 of the PV formed during the entry of each infective form revealed that the phagosome maturation processes are distinct but not strain-dependent. Due to their low hemolysin and trans-sialidase activities, MTs are retained for longer periods in LAMP-1 positive vacuoles. Our results thus suggest that despite the contrasting invasion capabilities, parasites of distinct phylogenetic group behave in similar fashion once inside the host cell. (c) 2007 Elsevier B.V. All rights reserved.Universidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Immunol & Parasitol, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Immunol & Parasitol, BR-04023062 São Paulo, BrazilWeb of Scienc

Comparative histopathology of endomyocardial biopsies in chagasic and non-chagasic heart transplant recipients

Background: Heart transplantation has been an option for the treatment of chagasic (C) cardiomyopathy despite difficulties concerning the control of rejection and reactivation. The parasite-host interaction under the influence of immunosuppressive therapy may affect the immunological response to the graft in a pattern different from that in non-chagasic (NC) patients. The aim of this study was to compare the major histopathological features in heart: transplantation in C and NC patients.Methods: We studied 293 endomyocardial biopsies from two groups of heart transplanted patients, including 18 C and 15 NC. Both groups had identical surgical and clinical procedure except immunosuppressive therapy was lower in C patients. The histopathological parameters evaluated were the Quilty effect, rejection, C myocarditis reactivation, fibrosis, hypertrophy, and ischemia. In addition, lymphocytic cellular infiltration of myocarditis due to rejection or reactivation was immunophenotyped in the biopsies of both groups with rejection grades 3 to 4, in biopsies with signs of reactivation, and in fragments of the receptor heart with chronic C myocarditis. A search for Trypanosoma cruzi was performed in all biopsies in the C group in which lymphocyte immunophenotyping was done. We used immunofluorescence and confocal microscopy.Results: The Quilty effect was present in 23% of the biopsies, involving 69.7% of the patients without a significant difference between groups (p = 0.509). Rejection was frequently observed in biopsies with the Quilty effect and the effect often recurred in the same patient. Rejection grades 3 to 4 was more frequent in the C group (p = 0,023). There were 5 episodes of Chagas' disease reactivation with myocarditis in 2 cases. The mean numbers of CD8+ and CD4+ T cells, and the CD4+-to-CD8+ ratio were similar for rejection in both groups (p > 0.05), while the CD4+-to-CD8+ ratio was significantly lower in chronic C myocarditis compared to rejection in the C group (p = 0.043). There was no significant difference in ischemic damage or interstitial fibrosis in the groups but there was a higher frequency of hypertrophy in the NC group (p = 0.007).Conclusions: The histopathological features of heart transplantation in C patients did not differ from that in NC patients in regard to the Quilty effect, development of myocardial fibrosis and ischemia. However, the higher involvement of the C group for rejection grades 3 to 4 suggested higher susceptibility to this event. The similarity of the lymphocytic cellular composition for rejection in both groups indicates that C patients respond to immunological stimulus in a similar pattern as NC patients.UNIFESP, Escola Paulista Med, Dept Patol, Sao Paulo, BrazilUNIFESP, Escola Paulista Med, Disciplina Cardiol, Sao Paulo, BrazilUNIFESP, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, Sao Paulo, BrazilUNIFESP, Escola Paulista Med, Dept Patol, Sao Paulo, BrazilUNIFESP, Escola Paulista Med, Disciplina Cardiol, Sao Paulo, BrazilUNIFESP, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, Sao Paulo, BrazilWeb of Scienc

Role of the second disulfide bridge (Cys(18)-Cys(274)) in stabilizing the inactive AT(1) receptor

Previous research showed that disruption of the Cys(18)-Cys(274) bond in the angiotensin II (AngII) AT(1) receptor mutant (C18S), expressed in CHO cells, causes an increase in the basal activity and attenuation of the maximum response to AngII. in addition, this mutant was mostly intracellularly distributed. Our aim was to investigate whether the intracellular presence of the mutant was due to a constitutive internalization or to a defective maturation of the receptor. the first hypothesis was assessed by pretreating the cells with losartan or [Sar(1)Leu(8)]-AngII, specific AT(1) receptor antagonists, a maneuver to revert the receptor internalization. the second hypothesis was tested using calnexin, an endoplasmic reticulum marker. We found that treatment with AT(1) receptor antagonists causes an increase in the binding ability of the mutant to AngII. Furthermore, whereas the maximum effect is increased, it reduces the enhanced basal levels of IP3. the hypothesis for a lack of maturation of the mutant receptor was ruled out because calnexin was poorly colocalized with the intracellular C18S receptor. Our results suggest that the mutation of the AT(1) receptor leads to a conformational structure similar to that of the active mode of the AT(1) receptor, favoring its internalization in the absence of the agonist.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, Dept Biophys, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biophys, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, BR-04023062 São Paulo, BrazilFAPESP: 07/01910-0Web of Scienc

beta-Actin-binding Complementarity-determining Region 2 of Variable Heavy Chain from Monoclonal Antibody C7 Induces Apoptosis in Several Human Tumor Cells and Is Protective against Metastatic Melanoma

Complementarity-determining regions (CDRs) from monoclonal antibodies tested as synthetic peptides display anti-infective and antitumor activities, independent of the specificity of the native antibody. Previously, we have shown that the synthetic peptide C7H2, based on the heavy chain CDR 2 from monoclonal antibody C7, a mAb directed to a mannoprotein of Candida albicans, significantly reduced B16F10 melanoma growth and lung colony formation by triggering tumor apoptosis. the mechanism, however, by which C7H2 induced apoptosis in tumor cells remained unknown. Here, we demonstrate that C7H2 interacts with components of the tumor cells cytoskeleton, being rapidly internalized after binding to the tumor cell surface. Mass spectrometry analysis and in vitro validation revealed that beta-actin is the receptor of C7H2 in the tumor cells. C7H2 induces beta-actin polymerization and F-actin stabilization, linked with abundant generation of superoxide anions and apoptosis. Major phenotypes following peptide binding were chromatin condensation, DNA fragmentation, annexin V binding, lamin disruption, caspase 8 and 3 activation, and organelle alterations. Finally, we evaluated the cytotoxic efficacy of C7H2 in a panel of human tumor cell lines. All tumor cell lines studied were equally susceptible to C7H2 in vitro. the C7H2 amide without further derivatization significantly reduced lung metastasis of mice endovenously challenged with B16F10-Nex2 melanoma cells. No significant cytotoxicity was observed toward nontumorigenic cell lines on short incubation in vitro or in naive mice injected with a high dose of the peptide. We believe that C7H2 is a promising peptide to be developed as an anticancer drug

Expression of angiotensin I-converting enzymes and bradykinin B-2 receptors in mouse inner medullary-collecting duct cells

We described in mouse inner medullary-collecting duct cells (m1MCD-3) the somatic and the N-domain ACE synthesis and its interaction with the kallikrein-kinin system co-localized in the same cells. We purified two ACE forms from culture medium, M1 (130 kDa) and M2 (N-domain, 60 kDa), and cellular lysate, C1 (130 kDa) and C2 (N-domain, 60 kDa). Captopril. and enalaprilat inhibited the purified enzymes. the immunofluorescence studies indicated that ACE is present in the membrane, cytoplasm and in the cell, nucleus. Kinin B-1 and B-2 receptors were detected by immunofluorescence and showed to be activated by BK and DesR(9) BK, increasing the acidification rate which was enhanced in the presence of enalaprilat. the presence of secreted and intracellular ACE in mIMCD-3 confirmed the hypothesis previously proposed by our group for a new site of ACE secretion in the collecting duct. (c) 2007 Elsevier B.V. All rights reserved.Universidade Federal de São Paulo, Escola Paulista Med, Depto Med, Disciplina Nefrol, BR-04023900 São Paulo, BrazilDept Biofis, BR-04023900 São Paulo, BrazilDept Microbiol Imunol & Parasitol, Disciplina Parasitol, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Depto Med, Disciplina Nefrol, BR-04023900 São Paulo, BrazilWeb of Scienc