Annexin A1 Mimetic Peptide Ac2-26 Modulates the Function of Murine Colonic and Human Mast Cells.

Mast cells (MCs) are main effector cells in allergic inflammation and after activation, they release stored (histamine, heparin, proteases) and newly synthesized (lipid mediators and cytokines) substances. In the gastrointestinal tract the largest MC population is located in the lamina propria and submucosa whereas several signals such as the cytokine IL-4, seem to increase the granule content and to stimulate a remarkable expansion of intestinal MCs. The broad range of MC-derived bioactive molecules may explain their involvement in many different allergic disorders of the gastrointestinal tract. Annexin A1 (AnxA1) is a 37 KDa glucocorticoid induced monomeric protein selectively distributed in certain tissues. Its activity can be reproduced by mimetic peptides of the N-terminal portion, such as Ac2-26, that share the same receptor FPR-L1. Although previous reports demonstrated that AnxA1 inhibits MC degranulation in murine models, the effects of exogenous peptide Ac2-26 on intestinal MCs or the biological functions of the Ac2-26/FPR2 system in human MCs have been poorly studied. To determine the effects of Ac2-26 on the function of MCs toward the possibility of AnxA1-based therapeutics, we treated WT and IL-4 knockout mice with peptide Ac2-26, and we examined the spontaneous and compound 48/80 stimulated colonic MC degranulation and cytokine production. Moreover, in vitro, using human mast cell line HMC-1 we demonstrated that exogenous AnxA1 peptide is capable of interfering with the HMC-1 degranulation in a direct pathway through formyl peptide receptors (FPRs). We envisage that our results can provide therapeutic strategies to reduce the release of MC mediators in inflammatory allergic processes.

Leishmania infection in blood donors: A new challenge in leishmaniasis transmission?

Transfusion-transmitted leishmaniasis has been a concern in regions endemic for the disease. Whether immediate or delayed, the risks posed by this mode of transmission call for careful assessment. The purpose of this study was to detect Leishmania infection in blood donors living in an endemic area and to investigate progression to the disease in these individuals. Immunofluorescent antibody test, enzyme-linked immunosorbent assay, leishmaniasis rapid test, and the polymerase chain reaction were applied to 430 donors in an initial evaluation. Of those donors with at least one positive test, 50 were reevaluated four years later by the same methods, as were 25 controls who had been negative on the same tests. In the first evaluation, Leishmania infection was detected in 41.4% (95% CI: 36.7-46.1) of donors (n = 430). None of the 75 reevaluated individuals had developed the disease, but retesting revealed positivity in at least one test in 36.0% (95% CI: 25.1-46.9) of donors. Of the 50 initially testing positive, 50% remained so on retesting. Of the 25 initially negative controls, two tested positive in the subsequent evaluation. The severity of the parasitosis and the risk of transfusion transmission warrant investigation of the potential inclusion of methods for Leishmania detection into blood banks for effective screening of infected donors.

Intestinal parasitism among waste pickers in Mato Grosso do Sul, Midwest Brazil.

The purpose of this study was to estimate the prevalence of intestinal parasites in both cooperative-affiliated and independent waste pickers operating at the municipal sanitary landfill in Campo Grande, Mato Grosso do Sul, Brazil, and associate these findings with hemoglobin, eosinophils, vitamin A and C levels and interleukin 5 and 10 (IL-5 and IL-10) production. Biological samples were collected, in addition to clinical, epidemiological, and sociodemographic data. Stool analyzes were based on sedimentation by centrifugation and on spontaneous sedimentation. High-performance liquid chromatography was used to determine vitamin A and C levels. ELISA was employed to quantify interleukins. Intestinal parasites were found in 29 of the 66 subjects assessed (43.9%). Endolimax nana (22.7%), Entamoeba coli (21.1%), Giardia lamblia (6.1%), Entamoeba histolytica/E. dispar (4.5%), and Ascaris lumbricoides (4.5%) were the most prevalent species. Pathogenic parasites were detected in 11 individuals (16.7%). Hypovitaminoses A and C were detected in 19.6% (13/66) and 98.4% (65/66) of subjects, respectively. IL-5 and IL-10 production was observed in 21 (31.8%) and 32 (48.4%) subjects, respectively. Infection with pathogenic intestinal parasites was not a cause of vitamin A and C deficiency or IL-5 and IL-10 production among these workers.

Is Leishmania (Viannia) braziliensis parasite load associated with disease pathogenesis?

BACKGROUND: Leishmania (Viannia) braziliensis is the main etiological agent of tegumentary leishmaniasis in the Americas. Parasite molecular diversity and host immune status contribute to extensive variations in its clinical presentation within endemic areas of Brazil. Pentavalent antimonials have been used for more than 60 years as the first-line drug for all cases, despite the potential for severe side effects and refractoriness. In Rio de Janeiro, Brazil, most L. (V.) braziliensis infections are benign with a scarcity of parasites, although metastasis and refractory infections can arise. In this scenario, the use of novel molecular tools can be useful for diagnosis and to assess tissue parasitism, and is of benefit to clinical and therapeutic management. METHODS: In this study, parasite load was assessed by real-time PCR based on the leishmanial small subunit ribosomal RNA gene. RESULTS AND CONCLUSION: The data revealed a tendency to higher tissue parasitism in the skin compared to mucous lesion sites and a reduction with disease progression. Parasite load was lower in poor compared to good responders to antimonials, and was also reduced in recurrent lesions compared to primary ones. However, parasite load became higher with sequential relapses, pointing to an immune system inability to control the infection. Therefore the parasite burden does not seem to be a good predictor of disease progression.

Severity of tegumentary leishmaniasis is not exclusively associated with Leishmania RNA virus 1 infection in Brazil.

Leishmania RNA virus (LRV) has been shown to be a symbiotic component of Leishmania parasites in South America. Nested retro-transcription polymerase chain reaction was employed to investigate LRV1 presence in leishmaniasis lesions from Brazil. In endemic areas of Rio de Janeiro (RJ), no LRV1 infection was observed even with mucosal involvement. LRV1 was only detected in Leishmania (V.) guyanensis cutaneous lesions from the northern region, which were obtained from patients presenting with disease reactivation after clinical cure of their primary lesions. Our results indicated that the severity of leishmaniasis in some areas of RJ, where Leishmania (V.) brazi-liensis is the primary etiological agent, was not associated with Leishmania LRV1 infection.

Severity of tegumentary leishmaniasis is not exclusively associated with Leishmania RNA virus 1 infection in Brazil

Leishmania RNA virus (LRV) has been shown to be a symbiotic component of Leishmania parasites in South America. Nested retro-transcription polymerase chain reaction was employed to investigate LRV1 presence in leishmaniasis lesions from Brazil. In endemic areas of Rio de Janeiro (RJ), no LRV1 infection was observed even with mucosal involvement. LRV1 was only detected in Leishmania (V.) guyanensis cutaneous lesions from the northern region, which were obtained from patients presenting with disease reactivation after clinical cure of their primary lesions. Our results indicated that the severity of leishmaniasis in some areas of RJ, where Leishmania (V.) brazi-liensis is the primary etiological agent, was not associated with Leishmania LRV1 infection.

Clinical features of cutaneous and disseminated cutaneous leishmaniasis caused by Leishmania (Viannia) braziliensis in Paraty, Rio de Janeiro.

BACKGROUND: American tegumentary leishmaniasis (ATL) caused by Leishmania (Viannia) braziliensis is endemic in Rio de Janeiro State (RJ), where the disease shows epidemiologic and clinical characteristics distinct from those of ATL in other Brazilian regions. Paraty is the second most important endemic area in RJ; however, reports on leishmaniasis in this region refer to the occurrence of the disease without describing its characteristics. METHODS: The clinical features of 71 cases of ATL reported between 1991 and 1997 in Paraty are presented. Thirty patients were re-evaluated 10 years later. RESULTS: Males and females were affected in similar proportions, and the disease was more prevalent in patients aged between 10 and 49 years (63.4%). Cutaneous leishmaniasis was the most prevalent clinical form observed. Unique lesions were present in 69% of cases, 91.6% of which displayed an ulcerated aspect. Although mucosal leishmaniasis was not observed, severe clinical manifestations, such as disseminated cutaneous lesions caused by L. braziliensis, were diagnosed in two patients. These patients presented skin lesions with different clinical aspects spread throughout the body, as well as low cellular immune responses. Montenegro skin test (92% positivity) and serology (8% IgM and 56% IgG anti-Leishmania positive results) were the most utilized tests for supporting the diagnosis of leishmaniasis. Parasites, detected in 27 of the 33 cases analyzed, were characterized as L. braziliensis. CONCLUSION: ATL in Paraty shares the clinical and laboratory characteristics reported for ATL in other regions of RJ, probably because of the similar epidemiologic context related to the Atlantic rainforest region.

Leishmania (Viannia) braziliensis: human mast cell line activation induced by logarithmic and stationary promastigote derived-lysates.

Herein we investigate the ability of live promastigotes and total lysate of Leishmania (Viannia) braziliensis, derived from parasites in the logarithmic (L-Lb) or stationary phase (S-Lb), to induce human mast cell line (HMC-1) activation. In comparison with medium-treated cells, a significant histamine release was observed in HMC-1 cultures stimulated with S-Lb. Lipophosphoglycan also induced histamine release by HMC-1 cells. In immunocytochemical assays, we found a marked staining for tryptase in medium-treated HMC-1 cells, however, stimulation with L-Lb or S-Lb caused a marked decrease in the color reaction as well as in the number of tryptase-positive cells. L-Lb and S-Lb induced an evident decrease in the intracellular expression of IL-4 but not IL-12. Live stationary promastigotes were able to induce high levels of IL-4 release in HMC-1 cultures. Furthermore, these cells released significant amounts of IL-12 when incubated with both types of live promastigotes. These results indicate that L. (V.) braziliensis promastigotes differ in their ability to induce direct human mast cells activation, according to the growth phase of the parasite. Furthermore, the release of pro-inflammatory mediators and cytokines could represent an important phenomenon that might favor the initial establishment of the infection.

Mastócitos e a Leishmaniose Tegumentar Americana Humana / Mast cells and Human american cutaneous leishmaniasis

Os mastócitos exercem um papel fundamental em reações alérgicas, mas também em resposta imune contra agentes infecciosos e parasitários. A meta principal deste estudo foi avaliar a densidade e a distribuição de diferentes populações de mastócitos em lesões cutâneas e em mucosas ativas de pacientes com leishmaniose tegumentar americana (LTA). Além disso, objetivou-se investigar a reatividade de mastócitos após a estimulação com diferentes preparações antigênicas derivadas de promastigotas de Leishmania tanto in vitro como in vivo. No infiltrado inflamatório crônico de lesões cutâneas e mucosas, os mastócitos mostraram-se desgranulados, com a presença de histamina difusa no tecido. Nas lesões cutâneas, foi observada uma maior densidade de mastócitos, quando comparada às amostras de pele normal, contrastando com as lesões mucosas onde os mastócitos foram encontrados em menor número. Em pele normal, foi observada uma predominância da subpopulação MCTC. Entretanto, nas lesões cutâneas, especialmente aquelas com evolução recente, verificou-se uma modificação no padrão fenotípico de mastócitos, sendo a subpopulação MCCT encontrada em maior proporção. Nenhuma alteração no padrão fenotípico de mastócitos foi observada em lesões mucosas. Utilizando modelo in vitro, uma liberação significativa de histamina por mastócitos humanos da linhagem HMC-1 foi observada em culturas estimuladas com antígeno total de L. braziliensis. As análises de citometria de fluxo mostraram que a estimulação antigênica induziu à liberação de IL-4, mas não de IL-12. Injeções intradérmicas de antígeno total e lipofosfoglicana purificada, uma molécula expressa em grande concentração na superfície de formas promastigotas, induziram rápida desgranulação dos mastócitos seguida de um aumento significativo na exsudação plasmática em ratos normais Wistar. A modificação do padrão fenotípico de mastócitos em lesões cutâneas, associado à liberação de IL-4 e histamina sugere o envolvimento de mastócitos na patogênese da LTA, possivelmente participando do estabelecimento inicial da infecção.

Análise do processo de ensino e aprendizagem de leitura e escrita: o comportamento de atentar ao som das palavras

Que comportamentos podem facilitar a aprendizagem de leitura e escrita? O objetivo deste trabalho foi iniciar desenvolvimento de procedimentos específicos para simplificar o processo de ensino-aprendizagem de leitura e escrita. Neste trabalho 20 alfabetizandos adultos foram submetidos a um pré-teste com questões envolvendo a discriminação de características sonoras de palavras. Em seguida, foram feitos exercícios de atenção à dimensão sonora da palavra falada e de registro escrito simplificado dessa dimensão, seguidos de pós-teste. Os exercícios produziram mudanças na atenção à dimensão sonora das palavras. O ensino, após o pré-teste, envolveu a apresentação visual e sonora de uma palavra de quatro sílabas, do universo vocabular dos alfabetizandos, seguida da apresentação das suas sílabas e das respectivas famílias silábicas e das respectivas famílias silábicas, em exercícios de leitura e escrita. Cada nova sílaba foi ensinada com auxílio do procedimento de exclusão. A discriminação anterior de características sonoras das palavras ajudou no processo de aquisição de leitura e escrita. Outras características de procedimentos foram utilizadas, como atendimento individualizado, exercícios específicos para cada relação envolvida no processo de leitura, e reforçamento imediato. A contribuição específica de cada procedimento ao procedimento de aprendizagem não foi verificada neste trabalho (AU)

Tumor necrosis factor-æ in human American tegumentary leishmaniasis

Tumor necrosis factor-alpha (TNF-alpha) is a cytokine produced by activated macrophages and other cells. In order to verify whether the serum levels of TNF-alpha in American tegumentary leishmaniasis patients are associated with the process of cure or aggravation of the disease, 41 patients were studied: 26 cases of cutaneous leishmaniasis (CL) and 15 of mucocutaneous leishmaniasis (MCL). During active disease the serum levels of TNF-alpha of MCL patients were significantly higher than those of CL patients and control subjects (healthy individuals and cutaneous lesions from other etiologies). The MCL patients had serum titers of TNF-alpha significantly lower at the end of antimonial therapy than before therapy. After a six-month follow-up, the MCL patients had serum levels of TNF-alpha similar to those observed at the end of the therapy as well as to those of Cl patients and control subjects. No significant variation in the serum levels of TNF-alpha was observed in CL patients throughout the study period (before, at the end of therapy and after a six-month follow-up). The possible relationship between the high TNF-alpha serum levels and severity of the disease is discussed.