We evaluated the toxicity of a semipurified extract (EPA fraction, containing caffeine and several flavan-3-ols and proanthocyanidins) of seeds of the native Amazon plant Paullinia cupana (guaraná) in rodents. Acute toxicity was tested in male Swiss mice, which received different doses orally (OR) and intraperitoneally (ip); control groups received water. These tests produced acute mortality, with LD(50) of 1.825 g/kg (OR) and 0.827 g/kg (ip), and a significant weight decrease in lungs of mice receiving a dose of 0.1g/kg. In the repeated-dose toxicity test, the EPA was administered OR daily to male and female Wistar rats at doses of 30, 150, and 300 mg/kg/day/90 days. Their behavior, mortality, weight changes, laboratory tests, and the weights and histopathology of organs were evaluated. No rats died during the tests. Males dosed at 150 or 300 mg/kg gained weight more slowly and lost kidney weight (absolute and relative weights, compared to the control group). Hematological and biochemical tests showed few changes, differing somewhat between males and females; the histopathological evaluation indicated no significant changes. These results indicate that the EPA fraction of guaraná caused no toxicity in rats at the smallest dose evaluated (30 mg/kg). No other species was evaluated.

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There is a difference of high diluted medication effect in four and eight-week-old mice. In the group of animals 8 weeks the immunomodulatory effect seems to have been higher. Hence, treatment with the medicine produced from T. cruzi modulates the inflammatory response with increased apoptosis and decreased serum levels of TGF-β.

The main results of the current study will be published when all patients have completed three years from the start of chemotherapy. Radical anterior resection and bone grafting was done on all patients and a comparison was made between six-and nine-month durations of chemotherapy with daily isoniazid and rifampicin plus twice-weekly streptomycin. The operation was usually undertaken within 14 days of the start of chemotherapy. Specimens taken at operation were examined fresh at the laboratory of the Ruttonjee Sanatorium in Hong Kong and were also sent at -78°C in solid CO2 by air to London. Further specimens were examined histologically by Professor J Ball, University of Manchester. CULTURE MEDIAThe media used were slopes of L6wenstein-Jensen medium (LJ), Lowenstein-Jensen medium containing 0-5% sodium pyruvate (LJ + P), Middlebrook's 7H11 oleic acid-albumin agar medium4 (Difco) (7H11) and liquid Kirchner medium.5 Kirchner medium was prepared without penicillin and was 662

Introduction: about 10 million people worldwide suffer from Chagas’ disease [1]. The World Health Organization (WHO) has explicitly acknowledged the significance of this condition and supports the use of Complementary and Alternative Medicine by health systems integrated with conventional treatments. Even so, one century after its discovery it still represents a global challenge [1,2]. Biotherapics are ultradiluted medicines and the infection of mice by Trypanosoma cruzi is an excellent model to understand their effect [3,4]. At 8 weeks, mice are physiologically more developed than at age 4 weeks, including a more competent immune system [5]. Aim: the aim of this study was to assess the effect of biotherapic of T. cruzi in dilution 17x on liver and spleen tissue of mice of different ages infected by this protozoon. Method: in a blind, randomized controlled trial 12 male Swiss mice aged 4 and 8 weeks, infected by 1,400 blood trypomastigotes T. cruzi Y strain were divided into groups control – treated with 7% hydroalcoholic solution (CI-4=3 animals or CI-8=3 animals) and treated with biotherapic 17x (BIOT-4=3 animals or BIOT-8=8 animals). Treatment (0.2 ml biotherapic/day/animal, per gavage) started after infection was verified (4th day) and animals were sacrificed on the 3rd day of treatment. For histopathological exam, the liver and spleen were removed and fixated in 4% paraformaldehyde for 24 hours and then processed for inclusion in paraffin. Semi-serial 7mm cuts were made and subjected to hematoxylin-eosin stain. It was performed a quantitative analysis of the number of nests of amastigotes and inflammatory foci in the liver. Slides were observed under microscope Olympus BX41 (Tokyo, Japan) and images captured with camera Qcolor3 (Olympus) coupled to the microscope. In the spleen it was counted the number of nests of amastigotes and the number of foreign-body giant cells. In each organ, 20 microscopic fields/cut were counted under power 40x totaling 120 fields/animal with microscope Olympus CBA (Tokyo, Japan). To analyze data it was used software Statistica 8.0. For data not exhibiting normal distribution it was used Kruskal-Wallis’ test at 5% significance and ANOVA for the ones with normal distribution. Chi-square test was used to compare percentages. Biotherapic 17d was prepared by adding 0.9 ml of blood with T. cruzi (10E+7 trypomastigotes/ml) to 9.1 ml of distilled water in laminar flow. Following dilutions were prepared in 86% hydroalcoholic solutions up to 16x. Dilution 17x was prepared with 7% hydroalcoholic solution [6]. It was performed microbiological control and in vivo biological risk of the biotherapic. Results showed a number of colony forming units proper for use (>1CFU/ml). Intraperitoenal inoculation of the biotherapic did not cause infection in animals. This study was approved by the Ethics Committee for Animal Experimentation/UEM protocol 030/2008. Results and Discussion: in the liver, animals of group BIOT-8 exhibited less nests of amastigotes (p