Ludmila R. P. Ferreira

Macrophage signaling by glycosylphosphatidylinositol-anchored mucin-like glycoproteins derived from Trypanosoma cruzi trypomastigotes

Activation of cells from the innate immune system has an important role in host resistance to early infection with the intracellular protozoan parasite, Trypanosoma cruzi. Here we review the studies that have identified and structurally characterized the glycosylphosphatidylinositol (GPI) anchors, as parasite molecules responsible for the activation of cells from the macrophage lineage. We also cover the studies that have identified the receptor, signaling pathways as well as the array of genes expressed in macrophages that are activated by these glycoconjugates. We discuss the possible implications of such response on the host resistance to T. cruzi infection and the pathogenesis of Chagas disease.

Photosynthetic efficiency and water use in eucalyptus plants sprayed with glyphosate

Objetivou-se com este trabalho avaliar a eficiencia fotossintetica e o uso da agua por plantas de clones de eucalipto submetidas ao herbicida glyphosate. O experimento foi realizado em esquema fatorial 4 x 5, com quatro clones de eucalipto (57, 386, 1203 e 1213) e quatro doses de glyphosate (43,2; 86,2; 129,6; e 172,8 g ha-1) e uma testemunha sem herbicida, considerada dose zero, com quatro repeticoes. Aos 7, 14, 21 e 28 dias apos aplicacao do herbicida (DAA) foi avaliada a intoxicacao das plantas, e aos 7 e 21 DAA, o fluxo de gases pelos estomatos (U - mmol s-1), a atividade fotossintetica (A - mmol m-2 s-1), a condutância estomatica (Gs - mol m-1 s-1), a transpiracao (E - mol H2O m-2 s-1) e a eficiencia do uso da agua (QUE - mol CO2 mol H2O-1). Aos 50 DAA, as plantas de eucalipto foram coletadas e colocadas em estufa de ventilacao forcada a 70 oC ate atingirem massa constante. Aos 21 DAA, o clone 1203 comportou-se como mais sensivel ao herbicida. Nao houve diferenca entre clones para as variaveis fisiologicas avaliadas. Aos 21 DAA constatou-se que, com o incremento da dose de glyphosate, houve reducao na condutância estomatica, na taxa de fluxo de gases pelos estomatos, na taxa fotossintetica e na eficiencia do uso da agua. Plantas dos clones 1213 e 1203 apresentaram maior acumulo de massa seca. O aumento da dose do glyphosate promoveu menor acumulo de massa seca das plantas de eucalipto. O glyphosate afetou negativamente o crescimento e a eficiencia fotossintetica e de uso da agua dos clones estudados.

Controle de Digitaria horizontalis pelos herbicidas glyphosate, sulfosate e glifosate potássico submetidos a diferentes intervalos de chuva após a aplicação

The effectiveness of sulfosate, potassium glyphosate and different glyphosate formulations on Digitaria horizontalis was evaluated when the products were applied in post emergence conditions and the plants were submitted to 20 mm simulated rainfall during 50 minutes, at intervals of 1, 2, 4 and 6 hr after herbicide application. The experiment was carried out under greenhouse conditions from January to April 2000, in Vicosa, Minas Gerais, Brazil, arranged in a split split-plot randomized block design with four replications. The phytotoxic effect of the herbicides on D. horizontalis was evaluated at 3, 7, 14, 21 and 28 DAA (days after application). At 45 DAA, the shoot dry biomass was compared to the control (without herbicide). No differences were observed among potassium glyphosate, sulfosate and glyphosate formulations (Roundup Transorb®) for similar application conditions. These treatments provided lower regrowth of D. horizontalis, when the interval without rain following application was of 46 hr. Glyphosate SL and WG formulations were the most affected by rainfall in all the intervals evaluated, when compared to other herbicide treatments.

Expression of serum amyloid A3 mRNA by inflammatory macrophages exposed to membrane glycoconjugates from Trypanosoma cruzi

Differential display reverse transcriptase‐polymerase chain reaction (PCR) was used to identify genes expressed by murine macrophages exposed to glycosylphosphatidylinositol‐anchored mucin‐like glycoproteins isolated from Trypanosoma cruzi trypomastigotes. Among the different PCR product bands identified in the differential display gel, one showed high homology with the serum amyloid A3 protein (SAA3). Northern blot assays showed augmentation of SAA3 mRNA expression by inflammatory macrophages exposed to live trypomastigotes or parasite glycolipids as compared to unstimulated macrophages. Our results also showed the expression of SAA3 mRNA, in liver and heart from animals in the acute phase of Chagas disease. It is important that expression of SAA3 mRNA was closely associated with tissue parasitism and presence of inflammatory cells. Together, our findings indicate the possible involvement of SAA3 protein on immunopathology of Chagas disease and establish a new infectious disease model to study the pathophysiological role of this acute‐phase protein. J. Leukoc. Biol. 66: 593–600; 1999.

Identification and characterization of a novel mouse gene encoding a Ras-associated guanine nucleotide exchange factor: expression in macrophages and myocarditis elicited by Trypanosoma cruzi parasites.

The ability of Trypanosoma cruzi to activate macrophages is, at least in part, attributed to the glycosylphosphatidylinositol‐anchored mucin‐like glycoproteins (GPI‐mucins) expressed in the surface of the trypomastigote stage of the parasite. The differential display reverse transcriptase‐polymerase chain reaction and the reverse Northern blot were used to study modulation of gene expression in murine macrophages exposed to GPI‐mucins and in cardiac tissues from mice infected with T. cruzi. Among several cDNAs that were more abundant in lanes corresponding to macrophages stimulated with GPI‐mucins as compared with resting cells, we confirmed the differential expression of A1, interleukin‐18, and GPIγ4. Some of these genes were also shown to have enhanced expression in the cardiac tissue (DAP‐12, A1, and GPIγ4) from infected animals. The expression of GPIγ4 was also enhanced in human monocytes stimulated with GPI‐mucins or bacterial lipopolysaccharides. The complete sequence of the GPIγ4 transcript and its gene including the 5′ upstream region was defined. GPIγ4 was encoded by a novel, single copy gene present in mouse as well as human genomes and showed conserved homology to different members of the guanine nucleotide exchange factor family.

Myocardial Infarction–Associated Transcript, a Long Noncoding RNA, Is Overexpressed During Dilated Cardiomyopathy Due to Chronic Chagas Disease

Long noncoding RNAs (lncRNAs) modulate gene expression at the epigenetic, transcriptional, and posttranscriptional levels. Dysregulation of the lncRNA known as myocardial infarction-associated transcript (MIAT) has been associated with myocardial infarction. Chagas disease causes a severe inflammatory dilated chronic cardiomyopathy (CCC). We investigated the role of MIAT in CCC. A whole-transcriptome analysis of heart biopsy specimens and formalin-fixed, paraffin-embedded samples revealed that MIAT was overexpressed in patients with CCC, compared with subjects with noninflammatory dilated cardiomyopathy and controls. These results were confirmed in a mouse model. Results suggest that MIAT is a specific biomarker of CCC.

Induction of IL-12 Production in Human Peripheral Monocytes by Trypanosoma cruzi Is Mediated by Glycosylphosphatidylinositol-Anchored Mucin-Like Glycoproteins and Potentiated by IFN-γ and CD40-CD40L Interactions

Chagas disease, caused by the protozoan parasite Trypanosoma cruzi (T. cruzi), is characterized by immunopathology driven by IFN-γ secreting Th1-like T cells. T. cruzi has a thick coat of mucin-like glycoproteins covering its surface, which plays an important role in parasite invasion and host immunomodulation. It has been extensively described that T. cruzi or its products—like GPI anchors isolated from GPI-anchored mucins from the trypomastigote life cycle stage (tGPI-mucins)—are potent inducers of proinflammatory responses (i.e., cytokines and NO production) by IFN-γ primed murine macrophages. However, little is known about whether T. cruzi or GPI-mucins exert a similar action in human cells. We therefore decided to further investigate the in vitro cytokine production profile from human mononuclear cells from uninfected donors exposed to T. cruzi as well as tGPI-mucins. We observed that both living T. cruzi trypomastigotes and tGPI-mucins are potent inducers of IL-12 by human peripheral blood monocytes and this effect depends on CD40-CD40L interaction and IFN-γ. Our findings suggest that the polarized T1-type cytokine profile seen in T. cruzi infected patients might be a long-term effect of IL-12 production induced by lifelong exposure to T. cruzi tGPI-mucins.

Exsudato radicular de imazapyr aplicado sobre mudas de diferentes clones de eucalipto

In the renewal of eucalypt crops, after tree harvesting, it is necessary to eliminate the sprouts that interfere with the initial development of the new stands. This elimination can be made by applying some herbicides on the shoots. One of the herbicides used is imazapyr, which, according to preliminary works carried out at the Universidade Federal de Vicosa, is capable to exsudate through the root system. In this case, questions arise concerning the exsudate ability to interfere in the development of the new plants; whether this capacity is the same for all eucalypt clones; at which imazapyr dose this problem is more serious; and the intensity at which the herbicide exsudate will interfere with the growth of the new stands. This work evaluated whether the root systems of different clones present different imazapyr exsudation intensities and the effect of the applied doses on herbicide concentration in the soil profile and its influence in subsequent crops. A dry mater decrease was verified in the sorghum plants when compared to the checks, in both 0-15 and 15-30 cm soil layers, evidencing imazapyr exsudation through the root system of the four clones analyzed. The toxic effect of the exsudates was observed in the entire soil volume reached by the eucalypt root system. Increased imazapyr doses increased the toxicity symptoms in sorghum imazapyr exsudation by all eucalypt clones was similar at the two depths analyzed.

Efeitos da chuva na eficiencia de formulacoes e doses de Glyphosate no controle de Brachiaria decumbens

The objective of this study was to evaluate the dose effects the glyphosate formulations - ammonium, isopropylamine and potassic - on Brachiaria decumbens plant control, under controlled conditions, when submitted to 20 mm of rainfall intensity for 30 minutes, at intervals of 1, 2, 4, 6, 12 and 24 hours after application of the herbicide treatments. Rainfall events after glyphosate application reduced the control of B. decumbens and this reduction was higher as the rainfall event was closer to the herbicide application. Potassic and isopropylamine glyphosate formulations showed a higher control of B. decumbens than ammonium glyphosate formulation. This effect was also observed on the dry biomass accumulation of B. decumbens regrowth, which was evaluated at 19 days after excising the plantshoot. At least 8, 11 and 12 hours after glyphosate application without rainfall were required, respectively, for potassic, isopropylamine and ammonium glyphosate at the dose of 1,440 g ha-1, to obtain a control over 80%; however, for the rate of 2,160 g ha-1, intervals required were 5, 7, and 9 hours, at least, respectively, to obtain the same control of B. decumbens. Dry biomass accumulation oduring weed regrowth was lower than 5%, providing an excellent control of the weed when four to five hours without rainfall occurred after herbicide application at the highest two doses (2,160 and 1,440 g ha-1). For isopropylamine glyphosate at the doses of 2,160 and 1,440 g ha-1, the same control was obtained from 7 and 12 hours without rainfall, respectively. It was concluded that both the potassic and isopropylamine glyphosate formulations are less affected by rainfall events after herbicide application, tolerating rainfall in a shorter interval after herbicide application than ammonium glyphosate.

Circulating miR-1 as a potential biomarker of doxorubicin-induced cardiotoxicity in breast cancer patients

Cardiotoxicity is associated with the chronic use of doxorubicin leading to cardiomyopathy and heart failure. Identification of cardiotoxicity-specific miRNA biomarkers could provide clinicians with a valuable prognostic tool. The aim of the study was to evaluate circulating levels of miRNAs in breast cancer patients receiving doxorubicin treatment and to correlate with cardiac function. This is an ancillary study from “Carvedilol Effect on Chemotherapy-induced Cardiotoxicity” (CECCY trial), which included 56 female patients (49.9±3.3 years of age) from the placebo arm. Enrolled patients were treated with doxorubicin followed by taxanes. cTnI, LVEF, and miRNAs were measured periodically. Circulating levels of miR-1, -133b, -146a, and -423-5p increased during the treatment whereas miR-208a and -208b were undetectable. cTnI increased from 6.6±0.3 to 46.7±5.5 pg/mL (p<0.001), while overall LVEF tended to decrease from 65.3±0.5 to 63.8±0.9 (p=0.053) over 12 months. Ten patients (17.9%) developed cardiotoxicity showing a decrease in LVEF from 67.2±1.0 to 58.8±2.7 (p=0.005). miR-1 was associated with changes in LVEF (r=-0.531, p<0.001). In a ROC curve analysis miR-1 showed an AUC greater than cTnI to discriminate between patients who did and did not develop cardiotoxicity (AUC = 0.851 and 0.544, p= 0.0016). Our data suggest that circulating miR-1 might be a potential new biomarker of doxorubicin-induced cardiotoxicity in breast cancer patients.