Highly diluted medication reduces tissue parasitism and inflammation in mice infected by Trypanosoma cruzi.

AIM: To evaluate the effects of Kalium causticum, Conium maculatum, and Lycopodium clavatum 13cH in mice infected by Trypanosoma cruzi. MATERIALS AND METHODS: In a blind, controlled, randomized study, 102 male Swiss mice, 8 weeks old, were inoculated with 1400 trypomastigotes of the Y strain of T. cruzi and distributed into the following groups: CI (treated with 7% hydroalcoholic solution), Ca (treated with Kalium causticum 13cH), Co (treated with Conium maculatum 13cH), and Ly (treated with Lycopodium clavatum 13cH). The treatments were performed 48 h before and 48, 96, and 144 h after infection. The medication was repertorized and prepared in 13cH, according to Brazilian Homeopathic Pharmacopoeia. The following parameters were evaluated: infectivity, prepatent period, parasitemia peak, total parasitemia, tissue tropism, inflammatory infiltrate, and survival. Statistical analysis was conduced considering 5% of significance. RESULTS: The prepatent period was greater in the Ly group than in the CI group (p = 0.02). The number of trypomastigotes on the 8th day after infection was lower in the Ca group than in the CI group (p < 0.05). Total parasitemia was significantly lower in the Ca, Co, and Ly groups than in the CI group. On the 12th day after infection, the Ca, Co, and Ly groups had fewer nests and amastigotes/nest in the heart than the CI group (p < 0.05). Decreases in the number of nests and amastigotes in the intestine were observed in the Ly group compared with the CI group (p < 0.05). In the liver (day 12), Ly significantly prevented the formation of inflammatory foci compared with the other groups. In skeletal muscle, Co and Ly decreased the formation of inflammatory foci compared with CI (p < 0.05). Ly afforded greater animal survival compared with CI, Ca, and Co (p < 0.05). The animals in the Co group died prematurely compared with the CI group (p = 0.03). CONCLUSIONS: Ly with 13cH potency had significantly more benefits in the treatment of mice infected with T. cruzi, reducing the number of blood parasites, amastigote nests in tissue, and the number of amastigotes per nest and increasing animal survival.

Trypanosoma cruzi: biotherapy made from trypomastigote modulates the inflammatory response.

UNLABELLED: This study evaluates the effect of Trypanosoma cruzi biotherapy 17dH (BIOT) on mice of different ages, infected with the protozoa concerned. METHOD: Performing a blind, controlled, randomized by drawing experiment, 110 animals four or eight-week-old, Swiss, male mice were divided into infected control treated hydroalcoholic 7% (CI-4 = 34 or CI-8 = 21 animals) and infected control treated with biotherapy 17dH-0.2 mL/animal/20 consecutive days/oral regimen (BIOT-4 = 33 or BIOT-8 = 21 animals). Animals were inoculated intraperitoneally with 1400 trypomastigote, T. cruzi Y-strain. Parasitological, immunological and histopathologic parameters were evaluated statistically, using Statistica-8.0 and R 3.0.2 program to analysis of survival. The study was approved by the Ethics Committee for Animal Experimentation/UEM. RESULTS: Four-week-old mice showed no statistical difference in parasitemia (P = 0.5718) between the treated and control group. Eight-week-old mice from the treated group had a higher parasite peak (P = 0.0424) and higher parasitemia (P < 0.005) than the control. To both groups of 4 and 8 weeks of age, treated or untreated, survival of mice was higher in the treated group than in the control, although it was not statistically significant (p-value = 0.32, 0.55 respectively). Four-week-old mice displayed a spleen section with a number of amastigote nests significantly higher in BIOT-4 than CI-4 (P = 0.01). In eight-week-old mice the number of amastigote nests (P < 0.001) and inflammatory foci (P < 0.06-10% significance) in the liver section were smaller in BIOT-8 than CI-8. Spleen giant cells were significantly higher in CI-8 than in BIOT-8 (P < 0.01). Eight-week-old animals treated with biotherapy showed higher parasitemia and lower tissue parasitism. Opposite pattern was observed in four-week-old animals. CONCLUSION: 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-ß.

Different forms of administration of biotherapy 7dH in mice experimentally infected by Trypanosoma cruzi produce different effects.

OBJECTIVE: To evaluate the effects of different forms of administration of the blood trypomastigotes biotherapy 7dH in mice experimentally infected with Trypanosoma cruzi. MATERIAL AND METHODS: Male swiss mice were inoculated with 1400 blood trypomastigotes of the Y strain of T. cruzi and allocated into 5 treatment groups: IC (distilled water); TCBZ (benznidazole); TBA(7dH) (biotherapy 7dH 20 days after infection); TBB(7dH)7 (biotherapy 7dH seven days before infection); TBB(7dH)30 (biotherapy 7dH 30 days before infection). Parasitological parameters assessed included pre-patent and patent periods, parasitemia peak, total parasitemia, mortality and survival rates. Cure index was obtained by fresh blood examination, hemoculture and polymerase chain reaction (PCR). RESULTS: The TBB(7dH)7 group showed a reduction in parasitemia peak, parasitemia area under the curve and total parasitemia. TBB(7dH)30 showed a tendency to increased pre-patent and survival periods, peak parasitemia was increased without increased total parasitemia. TBA(7dH) did not present significant alterations in the parasitological parameters analyzed. CONCLUSIONS: Biotherapy 7dH given before infection (7 or 30 days) produces different effects suggesting modulation of the host's immune system. The effects range from reduced parasitemia to its effective increase. The use of biotherapy to treat T. cruzi infection including dose, potency and schedule deserves further investigation.

Canova medication modifies parasitological parameters in mice infected with Trypanosoma cruzi.

The goals of this study were to evaluate the effect of the Canova medication, a homeopathic immune-system modulator, on the evolution of infection induced by the Trypanosoma cruzi Y strain in mice. The animals were divided into five groups: (i) untreated infected controls (I), (ii) infected animals treated with benznidazole (Bz), (iii) infected animals treated with the Canova medication (CM), (iv) infected animals treated with benznidazole and the Canova medication (Bz+CM), and (v) uninfected controls that received only the vehicle (grain alcohol) (C). The parameters evaluated were: parasitemia, mortality, control of cure, and tissue parasitism analysis. Our results showed that the evolution of the experimental infection was modified by treatment with CM, and that daily and consecutive doses were harmful to the animals, causing death in 100% of the infected animals in a brief period. The analysis of parasitism performed on the organs on the 12th day postinfection showed that in infected animals treated with CM, the number of amastigote/nests in the spleen was significantly reduced, while in cardiac tissue, intestine, and liver the number was significantly increased compared with infected control animals. These results indicate that CM has a negative influence on the host-parasite relationship, modifying the tropism of the parasite for tissues, and increasing the parasitemia peak in this experimental model.

Changes of RAPD profile of Trypanosoma cruzi II with Canova and Benznidazole.

Chagas disease, caused by the protozoan Trypanosoma cruzi, involves immunomediated processes. Canova (CA) is a homeopathic treatment indicated in the diseases in which the immune system is depressed. This study evaluated the Random Amplification of Polymorphic DNA (RAPD) profile of T. cruzi under the influence of CA and Benznidazole (BZ). Mice infected with the genetic lineage of T. cruzi II (Y strain) were divided into 4 groups: Infected animals treated with saline solution (control group); treated with CA; treated with BZ; treated with CA and BZ combined. Treatment was given at the 5th-25th days of infection (D5-25). The parasites were isolated by haemoculture in Liver Infusion Tryptose (LIT) medium: at D5 (before treatment), D13, 15 and 25 (during treatment) and D55 and 295 (after treatment). DNA was extracted from the mass of parasites. RAPD was done with the primers lambdagt11-F, M13F-40 and L15996, the amplified products were eletrophoresed through a 4% polyacrylamide gel. Data were analyzed by the coefficient of similarity using the DNA-POP program. 163 markers were identified, 5 of them monomorphic. CA did not act against the parasites when used alone. The RAPD profiles of parasites treated with BZ and CA+BZ were different from those in the control group and in the group treated with CA. The actions of the CA and BZ were different and the action of BZ was different from the action of CA+BZ. These data suggest that CA may interact with BZ. The differences in the RAPD profile of the Y strain of T. cruzi produced by BZ, CA+BZ and the natural course of the infection suggest selection/suppression of populations.

Highly diluted medication reduces parasitemia and improves experimental infection evolution by Trypanosoma cruzi.

BACKGROUND: There is no published information about the use of different protocols to administer a highly diluted medication.Evaluate the effect of different protocols for treatment with biotherapic T. cruzi 17 dH (BIOT(Tc17dH)) on clinical/parasitological evolution of mice infected with T. cruzi-Y strain. METHODS: A blind, randomized controlled trial was performed twice, using 60 28-day-old male Swiss mice infected with T. cruzi-Y strain, in five treatment groups: CI - treated with a 7% ethanol-water solution, diluted in water (10 µL/mL) ad libitum; BIOT(PI) - treated with BIOT(Tc17dH) in water (10 µL/mL) ad libitum during a period that started on the day of infection; BIOT(4DI) - treated with BIOT(Tc17dH) in water (10 µL/mL) ad libitum beginning on the 4th day of infection; BIOT(4-5-6) - treated with BIOT(Tc17dH) by gavage (0.2 mL/ animal/day) on the 4th, 5th and 6th days after infection; BIOT(7-8-9) - treated with BIOT(Tc17dH) by gavage (0.2 mL/ animal/day) on the 7th, 8th and 9th days after infection. We evaluated: parasitemia; total parasitemia (P(total)); maximum peak of parasites; prepatent period (PPP) - time from infection to detection of the parasite in blood; patent period (PP) - period when the parasitemia can be detected in blood; clinical aspects; and mortality. RESULTS: Parasitological parameters in the BIOT(PI) and mainly in the BIOT(4PI) group showed better evolution of the infection compared to the control group (CI), with lower P(total), lower maximum peak of parasites, higher PPP, lower PP and longer survival times. These animals showed stable body temperature and higher weight gain and water consumption, with more animals having normal-appearing fur for longer periods. In contrast, groups BIOT(4-5-6) and BIOT(7-8-9) showed worse evolution of the infection compared to the control group, considering both parasitological and clinical parameters. The correlation analysis combined with the other data from this study indicated that the prepatent period is the best parameter to evaluate the effect of a medication in this model. CONCLUSIONS: The BIOT(4DI) group showed the best clinical and parasitological evolution, with lower parasitemia and a trend toward lower mortality and a longer survival period. The prepatent period was the best parameter to evaluate the effect of a medication in this model.

Serologic testing for Trypanosoma cruzi: comparison of radioimmunoprecipitation assay with commercially available indirect immunofluorescence assay, indirect hemagglutination assay, and enzyme-linked immunosorbent assay kits.

The radioimmunoprecipitation assay (RIPA) has been used as a confirmatory test in several ongoing and published studies of Trypanosoma cruzi in blood donors in the United States. Despite its use as a confirmatory test, few studies are available comparing RIPA to commercially available serologic test methods. Thus, we compared RIPA with two indirect hemagglutination assays (Biolab Diagnostica SA, São Paulo, Brazil; Hemagen Diagnostics, Inc., Waltham, Mass.) and four different enzyme-linked immunosorbent assays (Abbott Laboratories, Abbott Park, Ill.; Embrabio, São Paulo, Brazil; Organon Teknika, São Paulo, Brazil; and Gull Laboratories, Salt Lake City, Utah) using a panel of 220 serum specimens from Brazilian blood donors with a range of T. cruzi antibody titers as determined by indirect immunofluorescence assay (IFA). A titer of 1:20 was used as the baseline for seropositivity. All IFA-negative serum specimens (n = 19) were nonreactive on all tests. At a titer of 1:20 (n = 9), reactivity rates varied considerably among the tests, with only the RIPA and the Organon and Gull assays identifying reactive specimens. For specimens at a 1:40 titer (n = 35), most assays identified at least 32 of 35 (91%) specimens as reactive, but the Biolab assay only identified 24 (69%). At higher titers (1:80, n = 56; 1:160, n = 101) the assays were comparable, with the exception of the Biolab assay, demonstrating rates of agreement with IFA of >/=98%. Overall, when compared with several other test formats, RIPA demonstrated equivalent or superior rates of agreement with IFA-positive specimens across all titers examined. In particular, at titers of >1:40, the RIPA compared favorably with other test methods currently in use, supporting its application as a confirmatory test, particularly in a research setting.

Estudo hispatologico de alteracoes do figado e coracao de camundongos inoculados com T. cruzi e tratados com bioterapico D30 / Histopathologil study in liver and heart of mices infected with T. cruzi and treated with biotherapic D30

The authors studied the action of Biotherapic D30 on liver and heart alterations of mice infected with T.cruzi. Was the liver showed increase cell and nucleus volume, sinusoids with increase size. A diminution of tissue parasitemia were observed on both organs