Denise Lessa Aleixo

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 hosts 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.

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

BackgroundThere 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 (BIOTTc 17dH) on clinical/parasitological evolution of mice infected with T. cruzi-Y strain.MethodsA 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; BIOTPI - treated with BIOTTc 17dH in water (10 μL/mL) ad libitum during a period that started on the day of infection; BIOT4DI - treated with BIOTTc 17dH in water (10 μL/mL) ad libitum beginning on the 4th day of infection; BIOT4-5–6 - treated with BIOTTc 17dH by gavage (0.2 mL/ animal/day) on the 4th, 5th and 6th days after infection; BIOT7-8–9 - treated with BIOTTc 17dH by gavage (0.2 mL/ animal/day) on the 7th, 8th and 9th days after infection. We evaluated: parasitemia; total parasitemia (Ptotal); 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.ResultsParasitological parameters in the BIOTPI and mainly in the BIOT4PI group showed better evolution of the infection compared to the control group (CI), with lower Ptotal, 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 BIOT4-5–6 and BIOT7-8–9 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.ConclusionsThe BIOT4DI 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.

Predominance of Th1 response, increase of megakaryocytes and Kupffer cells are related to survival in Trypanosoma cruzi infected mice treated with Lycopodium clavatum.

We investigated the number of megakaryocytes, Kupffer cells and ratios of Th1/Th2 and Th1/Th17 cytokines in survival of mice infected with Y strain of Trypanosoma cruzi and treated with Lycopodium clavatum. In a blind, randomized and controlled assay, Swiss male mice, 8weeks-old, infected with 1400 trypomastigotes (Y strain) were divided into groups and treated with: GLy - Lycopodium clavatum dynamization13c and GCI - alcohol solution 7° GL (vehicle medicine). The treatment was offered two days before infection and on the 2nd, 4th and 6th days after infection, overnight (1mL/100mL) and ad libitum. Parameters assessed were: survival rate, number of megakaryocytes and Kupffer cells, cytokines dosage (TNF-α, IFN-γ, IL-2, IL-4, IL-6, IL-10, IL-17), Th1/Th2 and Th1/Th17 ratios. The increase in megakaryocytes, Kupffer cells, predominance of Th1 response, with increased TNF-α, IL-10, TNF-α/IL-4, TNF-α/IL-17 and decreased IL-6 IL-6/IL-4, are related to increased survival in mice infected with T. cruzi and treated with Lycopodium clavatum 13c. This result demonstrates the possibility of an alternative approach for the treatment of Chagas disease with dynamized drugs.

Histopathological lesions in encephalon and heart of mice infected with Toxoplasma gondii increase after Lycopodium clavatum 200dH treatment

In many cases, symptoms of toxoplasmosis are mistaken for the ones of other infectious diseases. Clinical signs are rare in immunocompetent people. However, when they arise, in the acute phase of infection, several organs are affected due to the rapid spread of tachyzoites through the bloodstream. In the present study, the reduction of tachyzoites in peripheral blood of mice of G72 (infected 72h after treatment) and G48 (infected 48h after treatment and treated three more times), when compared with IC (infected and non-treated), suggests protective effect exerted by Lycopodium clavatum. If on the one hand L. clavatum brought benefits, reducing parasitemia, on the other hand, the parasitism became exacerbated. Histopathological analysis demonstrated focal, multifocal and diffuse inflammatory infiltrates, ranging from absent, discreet, moderate to intense, in heart and encephalon of mice of NIC (non-infected and non-treated), IC, G48 and G72 groups, respectively. In the perivascular region and meninges, the injuries were enlarged. The presence of tachyzoites was demonstrated through immunohistochemical (IHC) assay in myocardium. Toxoplasma gondii induced increase of collagen fibers in myocardium of mice of G72 and G48 groups, compared with IC (p<0.05) and NIC (p<0.001). The presence of inflammatory infiltrates, as well as the progressive fibrosis, caused myocardial remodeling in animals treated with L. clavatum. Counterstaining with H&E suggests TGF-β expression by mononuclear cells in the inflammatory infiltrate. Based on our results, we can conclude that the adopted regimen and potency exerted a protective effect, reducing parasitemia. However, it intensified the histopathological lesions in encephalon and heart of mice infected with T. gondii.

Phosphorus protects cardiac tissue by modifying the immune response in rats infected by Trypanosoma cruzi

HighlightsPhosphorus 13cH promotes beneficial effects in murine infection by T. cruzi.The Treatment beneficially modulates as inflammatory cytokines IFN‐&ggr; e TNF‐&agr;.The drug reduces inflammation of cardiac tissue in Wister rats infected with T. cruzi. Abstract Aim: This study evaluates and correlates the number of myocarditis focuses and production of cytokines in Rattus norvegicus (Wistar lineage), experimentally infected with T. Cruzi and treated with Phosphorus. Methods: In two blind, controlled and randomized trials, 53 45‐day‐old, male animals were allocated into groups Control (n = 24): Control group infected and treated with 7% hydroalcoholic solution, the preparation vehicle of the test medication; and Phosphorus (n = 24 on days 0, 5, 10 and 24 after infection): group infected and treated with Phosphorus 13cH, diluted 10−26 and dynamized (test medication). The animals were inoculated intraperitoneally with 5 × 106 blood trypomastigotes of T. cruzi‐Y strain. The medication was administered overnight (16 consecutive hours), diluted in water (1 mL/100 mL) in amber water bottles. The animals were treated 2 days before and 2, 4, and 6 days after infection. Enumeration of inflammatory foci in cardiac tissue (Hematoxylin‐Eosin) and dosage of cytokines TNF‐&agr; and IFN‐&ggr; in the serum were performed on days 0, 5, 10 and 24 after infection, using three animals/group. Mann‐Whitney, Friedman ANOVA, Spearman correlation (p < 0.05), and Statistica Single User Software version 13.2 were used for data analysis. Results: The animals treated with Phosphorus 13cH had high concentration of INF‐&ggr; on the 5th day of infection with significant decrease on the 10th and 24th days (p < 0.05), and high concentration of TNF‐&agr; on the 5th and 10th days of infection with decrease on the 24th day (p < 0.05). The treatment with Phosphorus caused a significant increase of INF‐&ggr; and TNF‐&agr; on the 5th day of infection compared with the Control (p < 0.05), with reestablishment on the 24th day, as well as in the Control group. The group treated with Phosphorus had 52.5% less number of myocarditis focuses in heart than Control group (p < 0.05) on the 10th day of infection. The significant increase in cytokines on the5th day of infection in the Phosphorus group is related to a significant decrease in the number of inflammatory foci in cardiac tissue on the 10th day of infection in this group. Discussion and conclusion: Treatment with Phosphorus 13cH promotes beneficial effects in T. cruzi infection in Wistar rats by modulating the secretion of IFN‐&ggr; and TNF‐&agr; with decreased inflammation in cardiac tissue. These results reinforce the importance of considering the use of homeopathy for establishing new therapeutic approaches in the management of patients with Chagas disease.

Dynamized ethyl alcohol improves immune response and behavior in murine infection with Trypanosoma cruzi

Aim To evaluate the effects of dynamized ethyl alcohol (Ethylicum) 6cH and 30cH in mice infected with T. cruzi. Methods In a blind, randomized and controlled assay, 63 eight‐week‐old, Swiss, male mice, infected with IP (1400 trypomastigotes, T. cruzi‐Y‐strain), were allocated into groups: CNI‐non‐infected (n = 12), CI‐infected and non‐treated (n = 17), Et6cH‐infected, treated with Ethylicum 6cH (dilution 1:1012) (n = 17), Et30cH‐infected, treated with Ethylicum 30cH (dilution 1:1060) (n = 17). Treatment was administered 48 h before and after infection, followed by 56 h/56 h periods, until the 9th day after infection (a.i), for 16 h. Survival and mortality were assessed until the 82nd day after infection (a.i.). TNF‐&agr;, IL‐6, IL‐10, IL‐5 and IL‐17A cytokines were assessed in serum (3–4 animals/group), at time T0 (before infection), T8 and T12 (8th and 12th a.i), using the Mouse Cytokine 20‐Plex Panel Magnetic Kit (Invitrogen, USA). Inflammation was determined in heart sections (eosin‐hematoxylin staining) and behavior was analyzed with ANY‐maze® software. The study was approved by the Animal Ethics Committee/UEM. Statistica 8.0 and R 3.0.2 software were used for statistical analyses. Results The greater survival observed in the Et6cH group was related to decreased inflammation in heart tissue and increased IL‐5 at T0 (p < 0.05) and IL‐10 at T8 (p < 0.05), characterizing the Th2 response. It was also related to shorter periods of immobility, observed on day 12 a.i. The higher mortality in the Et30cH group was related to increased inflammation in the heart and a higher concentration of IL‐6 and TNF‐&agr; cytokines, characterizing the Th1 response. Conclusion The results demonstrate the beneficial effect of Ethylicum 6cH in acute murine infection by T. cruzi. HighlightsBeneficial effect of Ethylicum 6cH in acute murine infection by T. cruzi.Ethylicum 6cH presented pronounced Th2 response.Ethylicum 6cH decreased inflammation in heart and increased hosts survival.Ethylicum 30cH exhibited pronounced Th1 response. Graphical abstract Figure. No Caption available.

Randomized study using biotherapeutic and #8220;T.cruzi 3dH and #8221; impairs experimental infection by Trypanosoma cruzi

Objective: The aim of this study was to evaluate the effect of a biotherapeutic in very low dynamization (3dH) in murine infection by Trypanosoma cruzi. Methods: Swiss male mice (Mus musculus) infected by T. cruzi Y strain were divided into three groups according to their treatment: Control infection - 7% ethanolwater solution; 3dH each day (ED) - daily biotherapeutic T. cruzi 3dH, from the day of infection until the end of experiment; 3dH single day (SD) - biotherapeutic T. cruzi 3dH, on the day of infection for 12 h. Parasitological (total parasitemia, peak of parasites, day of the peaks, prepatent period, patent period, and survival) and clinical (body temperature and body weight) parameters were evaluated. Results: No significant differences were observed among groups for parasitological evaluation. 3dH ED group presented an earlier mortality compared to control. Clinical parameters showed that animals treated with biotherapeutic had worse outcome when compared with control animals. Conclusion: The low dynamization of T. cruzi biotherapeutic worsened its murine infection by means of clinical evaluation. Considering other studies using biotherapeutic of T. cruzi, suggests the possibility for an efficacy of different dynamizations regarding oscillatory potency-effect-curves.