Effective anti-leishmanial activity of minimalist squaramide-based compounds.

In order to evaluate the in vitro leishmanicidal activity of N,N'-Squaramides derivatives, compounds that feature both hydrogen bond donor and acceptor groups and are capable of multiple interactions with complementary sites, against Leishmania infantum, Leishmania braziliensis and Leishmania donovani a series of 18compounds was prepared and assayed on extracellular and intracellular parasite forms. Infectivity and cytotoxicity tests were performed on J774.2 macrophage cells using meglumine antimoniate (Glucantime) as the reference drug. Changes in metabolite excretion by 1H-NMR and the ultrastructural alterations occurring in the parasites treated using transmission electron microscopy (TEM), was analyzed. Compounds 1, 7, 11, 14 and 17 were the more active and less toxic. Infection rates showed that the order of effectiveness was 17 > 11 > 14 > 7 for both L. infantum and L. braziliensis and in the same way, the compound 1 for L. donovani. All these compounds have altered the typical structure of the promastigotes, glycosomes and mitochondria. These severe modifications by the compounds are the ultimate reasons for the alterations observed in the excretion products. The Squaramide 17 (3-(butylamino)-4-((3-(dimetilamino)propyl)(methyl)amino)cyclobut-3-en-1,2-dione) was clearly the most efficient of all compounds. The data appear to confirm that the severe modifications generated in organelles such as glycosomes or mitochondria by the compounds are the ultimate reasons for the alterations observed in the excretion products of all species. The activity, stability, low cost of starting materials, and straightforward synthesis make amino squaramides appropriate molecules for the development of an affordable anti-leishmanial agent.

Diagnosis of Congenital Chagas Disease Using an Iron Superoxide Dismutase Excreted as Antigen, in Mothers and Their Children During the First Year of Life.

BACKGROUND: Chagas disease caused by Trypanosoma cruzi is endemic in Latin America. Human infection is mainly spread by Triatominae insects. Other forms of transmission are congenital, blood transfusion and organ transplantation. METHODS: Anti-T. cruzi antibodies were determined by enzyme-linked immunosorbent assay (ELISA) and Western blot (WB) in 155 serum samples from mothers and their babies. Indirect immunofluorescence (IFA) and a commercial test were used to validate efficacy of a specific ELISA-iron-excreted superoxide dismutase assay. Sera from babies were collected at 6 and 12 months, whereas maternal samples were obtained after delivery. Calostrum and umbilical cord samples were simultaneously obtained. RESULTS: Anti-T. cruzi antibodies were detected in 8 (5.16%) mothers by ELISA-WB, in 7 (4.51%) using IFA and in 1 (0.64%) by a commercial kit. Nine (5.80%) 6-month-old children were positive by ELISA-WB and 7 (4.51%) by IFA; negative results were obtained when the commercial kit was used. At 12 month of age, 15 (9.67%) children were positive by ELISA-WB, 13 (8.38%) by IFA and 1 (0.64%) by the commercial test. Antibodies were detected in 4 mothers whose children were serologically negative. Four other mothers and their children were positive, but only one of them had detected antibodies in umbilical cord up to 12 months, thus assuming vertical transmission. CONCLUSIONS: The use of iron-excreted superoxide dismutase as antigen in serologic tests for detection of T. cruzi yielded promising results as diagnostic procedure.

Anti-Trypanosoma cruzi antibody detection in eastern Andalusia (Spain).

BACKGROUND: Chagas disease caused by the protozoan haemoflagellate Trypanosoma cruzi is no longer found exclusively in Latin America; the disease is occurring in Europe, and Spain is the country with the highest prevalence. METHODS: Our aim was to detect anti-T. cruzi antibodies in blood donors from southeast Spain, and we performed eight serological diagnostic assays on each of 550 blood samples collected in March-June 2010. Two in-house ELISA methods were used to test against a parasite lysate (ELISA-H) and the semi-purified superoxide dismutase excreted by T. cruzi (ELISA-SODe); we also used the Western blot technique against the same antigen (WB-SODe), indirect immunofluorescence (IFA) and four commercial tests. RESULTS: The serological test results showed a range of seroprevalence values, the lowest being 1.1%, determined by IFA and two commercial tests (Ab rapid and Chagascreen); other values were: 1.3% (commercial ELISA [Chagas ELISA IgG+IgM]); 2.1% (immunochromatographic test [Stick Chagas]); 2.7% (ELISA-H); 4.0% (WB-SODe); and 4.2%, the highest value (ELISA-SODe). CONCLUSIONS: The excellent specificity of SODe antigen for the detection of antibodies to T. cruzi in donors lead us to affirm that the serological test performed with this biomarker could provide a useful screening and confirmatory test method for cases of Chagas disease.