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.

Effect of an homeopathic complex on fatty acids in muscle and performance of the Nile tilapia (Oreochromis niloticus).

BACKGROUND: The homeopathic complex HomeoAqua Mega 3(®) was designed to stimulate fish liver function resulting in enhanced lipid metabolism and improved overall performance. The effect of the complex in fatty acid compounds in the muscle tissues and the performance of Nile tilapia (O. niloticus) were evaluated. METHODS: A control diet with 40 mL of alcohol solution (30° Gay Lussac (GL) alcohol) per kg of feed and another diet with 40 mL/kg of the homeopathic complex were analysed in sex-reversed juvenile male Nile tilapia with mean initial weights of 89.54 g (±7.97) and 89.74 g (±8.83) and initial total mean lengths of 16.93 cm (±0.56) and 16.85 cm (±0.56) for the control and homeopathy-treated fish, respectively. Overall, 200 fish were distributed into 10 water tanks, with 20 specimens in each 600 L tank, they were kept for 63 days. Monitoring of the water's physical and chemical parameters was performed. Additionally, the percentage chemical composition and the composition of the muscle tissue fatty acids were determined, and fish performance was evaluated. RESULTS: No significant differences (p > 0.05) were found between treatments with regard to the water's physical and chemical parameters, moisture, ashes, proteins of the Nile tilapia muscular tissue and mean rates of total weight and length of the animals in the final period of the assay. The total lipid (TL) rates of the tilapia muscle tissue were 1.133 ± 0.2% at the start (control and homeopathy) and 0.908 ± 0.14% (control) and 0.688 ± 0.14% (homeopathy) at the end of the experiment (p < 0.06). The sum total of the SFAs (saturated fatty acids) was 338.50 ± 0.15 at the start (control and homeopathy) and 271.49 ± 0.34 mg/g of the TLs for the control and 226.12 ± 0.77 mg/g of TL for the homeopathy at the end of the experiment (p < 0.05). The n-6/n-3 ratio was also lower for the homeopathy-treated fish (8.45 ± 0.40) compared with the control fish (9.60 ± 0.14), (p < 0.05) at the end of the 63-day period. CONCLUSION: Nile tilapia juveniles that received the HomeoAqua Mega-3(®) in their diets had a decrease in the TL rates, mainly for the SFAs, and the n-6/n-3 ratio, compared with the control group, without any changes in the animal performance during the experimental period.

Morpho-functional response of Nile tilapia (Oreochromis niloticus) to a homeopathic complex.

BACKGROUND: This study evaluated the performance, prevalence of ectoparasites and morpho-functional response of the liver and the branchiae of Nile tilapia (Oreochromis niloticus) raised on fish meal with added of the homeopathic complex Homeopatila 100(®) at different concentrations. METHODS: Post-reversed juvenile Nile tilapia (O. niloticus) of the GIFT (Genetic Improvement of Farmed Tilapia) strain were used in this study. The performance, ectoparasite prevalence and parasite load in the branchiae and skin as well as the liver and branchial histology. Fish were randomly assigned to receive one of four treatments: control, 20 mL hydroalcoholic solution (alcohol 30° GL); 20 mL Homeopatila 100(®) per kg of meal; 40 mL Homeopatila 100(®) per kg of meal; or 60 mL of Homeopatila 100(®) per kg of meal, compared to control with out the addition of the complex. There were four replications per treatment type (16 experimental units total) at a density of 40 fish per m(3) over a period of 57 days. The Kruskal-Wallis H test (p < 0.05) was employed to analyse the physical and chemical parameters of water as well as for parasite prevalence; whereas analysis of variance was used for liver performance. If the values were significant (p < 0.05), they were compared by Tukey's test. Multiple comparisons of averages were performed using Student's t test (p < 0.05). RESULTS: There were no significant between the physical and chemical parameters of the water between the different groups at the end of the experiment. Significant differences (p < 0.05) in the mixed parasite conditions were found within the different Homeopatila 100(®) treatments. The hepatosomatic ratio of fish treated with Homeopatila 100(®) was significantly lower than that of fish from the control group. The best results in the liver and branchiae occurred in fish receiving Homeopatila 100(®) at 40 mL/kg in terms of the number of hepatocytes/mm(2), the intercellular glycogenic behaviour, the rates of histological changes (hyperplasia, lamella fusion and telangiectasia) and the percentage of neutral and acidic mucin-producing cells. CONCLUSION: The addition of Homeopatila 100(®) at a concentration 40 mL per kg/meal to the diet of juvenile Nile tilapias resulted in improved hepatocytes and intracellular glycogen levels as well as the lowest mean rate of branchial histological changes with an increase in acidic mucin-producing cells compared to neutral mucin-producing cells, compared to control.

Antimalarial potential of Nosode 30 and 200 against Plasmodium berghei infection in BALB/c mice.

BACKGROUND & OBJECTIVES: Homeopathy is considered as an emerging area of alternative medicine which could be established for the global health care. One of the greatest objections to this science lies in its inability to explain the mechanism of action of the micro doses based on scientific experiments and proofs. The present study has been undertaken to screen in vivo antimalarial activity of Malaria Co Nosode 30 and Nosode 200 against Plasmodium berghei infection in BALB/c mice. METHODS: Peter's 4-day test was used to evaluate the in vivo schizontocidal effect of Nosode 30 and Nosode 200. One month follow-up study was done to calculate the mean survival time of mice in each group. Biochemical analysis was carried out to assess the liver and kidney function tests using diagnostic kits. RESULTS: Nosode 30 and 200 exhibited 87.02 and 37.97% chemosuppression on Day 7 and mean survival time (MST) of 18.5 ± 2.16 and 16.5 ± 1.37 days respectively, which were extremely statistically significant when compared to MST of infected control (8.55 ± 0.83 days). The safety of Nosode 30 was also confirmed by the comparable levels of ALP, SGOT, SGPT activities, concentration of bilirubin, urea and creatinine to CQ treated group. CONCLUSION: Nosode 30 possesses considerable in vivo antiplasmodial activity against P. berghei infection as compared to Nosode 200 as evident from the chemosuppression obtained using Peter's 4-day test. Further, studies on the drug can be carried out to establish its antimalarial potential in monotherapy or in combination with other homeopathic drug formulations.

[Cardiac involvement in adults with echinococcosis].

Experiments have established that the first target for echinococcus is the liver and lung and that for pathogenic fungi and protozoa is the heart. Adult patients with hepatic hydatid disease complicated by paecilomycosis have been found to have atypical paecilomycosis-associated myocarditis, the treatment of which was developed by the authors, by using antibiotics, fungicides, and homeopathic remedies.

[Effect of centipede extract on cervical tumor of mice and its mechanism].

UNLABELLED: To study the anti-tumor activity of centipede extract on cervical tumor of mice and its mechanism. METHODS: The tumor-bearing mice were treated with centipede extract from two solvents [ether (CE) and alcohol (CA)] at different comcentration. The mice' life span, tumor inhibition rate and immune function were estimated. RESULTS: No mice died in CE and CA treatment groups and the tumor inhibition rate was 52.85% and 33.65% respectively. Observed the tumor tissue slices with light microscope and found infiltration of tumor cells in striated muscle in the control group but centipede treatment groups had massive necrosis and apoptosis. Karyopyknosis and apoptotic tumor cells were observed in the treatment groups under transmission electron microscopy. Compared with control group, the expression of Bax increased, the expressions of Bcl-2 and Survivin decreased, but the content of VEGF, the indexes of thymus and spleen had no significant change in treatment groups. The number of CD3+ T lymphocytes had no significant change while the ratio of CD4+ and CD8+, the number of CD19+ B lymphocytes decreaed in the CE group. The numbers of CD3+ and CD4+ lymphocytes decreased in the CA group. The pathological examine indicated no obvious change in the tissue slices of mice's liver and kidney, manifested the concentrations of CE and CA between the article's had no visible side effect. CONNCLUSION: The two extracts (CE and CA) can suppress the growth of cervical tumor and its mechanism may be related to Bax and Caspase-3 medicated the mitochondrial signal transit pathway.

[Amino acid exchange in paeci lomycosis-complicated echinococcosis].

The authors have detected atypical paecilomycosis-associated myocarditis with impaired amino acid exchange and pain syndrome for the first time. At first, pain occurs in the chest and radiates into the axilla, to the left arm to the finger tips, by paralyzing the arm. In some patients, pain manifests itself in both arms with radiation to the belly, by accompanying by fainting. The skin is wet, cold; the pulse is frequent and of poor volume and difficult-to-count. Heart pain spreads into the armpit and down the arm, by making the fingers numb. Attempts to use current analgesics (movalis, sirdalud, nimesil, morphine) in combination with fungicides (diflucan, mycosist, orungal) have failed to yield positive results. The homeopathic drug Latrodectus mactans, prepared from caracurt venom, in combination with the authors' designed diet and other homeopathic agents have relieved pain syndrome and normalized amino acid exchange, which offered possibilities for successful surgical treatment for echinococcosis with later recovery.

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.

Hepatic ethoxy-, methoxy- and pentoxyresorufin O-dealkylase activities in Landrace and Duroc pigs stimulated with HCG.

The effect of human chorionic gonadotropin (hCG) stimulation on the activities of ethoxyresorufin O-deethylase (EROD), methoxyresorufin O-demethylase (MROD) and pentoxyresorufin O-depentylase (PROD) was studied in intact male pigs of purebred Landrace and Duroc breeds. Pigs were divided into four groups: two control groups of each breed, without hCG stimulation (n = 20 for each breed), and two experimental groups (n = 18 for each breed), with hCG stimulation (Pregnyl(®); N.V. Organon, Oss, The Netherlands, 30 IU/kg live weight). Pigs were slaughtered 3 days after hCG stimulation and enzyme activities were measured in hepatic microsomes using two approaches. First, only one substrate concentration was used for the analysis of each enzyme activity. We found that EROD activity was suppressed by hCG-stimulation in Landrace (p = 0.004), but not Duroc pigs (p > 0.05). Generally, EROD activity was higher in Duroc pigs compared with Landrace (p = 0.017). Methoxyresorufin O-demethylase and PROD activities did not differ between groups (p > 0.05). To further characterize EROD, MROD and PROD, enzyme kinetic studies were performed. V(max) values for EROD and MROD in both breeds were lower after hCG stimulation (p < 0.001 for Landrace and p < 0.05 for Duroc). Additionally, V(max) values for EROD significantly differed between Landrace and Duroc pigs being higher in Duroc pigs (p < 0.05). We concluded that both hCG stimulation and breed differences may be important in the regulation of EROD and MROD activities. This study provides the first data on the effect of hCG stimulation and thus high testicular steroids, on EROD, MROD and PROD activities. Further studies are needed to investigate individual CYP450 enzymes and their regulation in porcine tissues.

[Therapeutic effect of Haobieyangyinruanjianfang on the mouse liver fibrosis caused by schistosomiasis].

OBJECTIVE: To explore therapeutic effect of Haobieyangyinruanjianfang (HBYYRJ) on mouse liver fibrosis by schistosomiasis. METHODS: Mice except for normal control were infected with Japanese schistosome cercarias, after 12 weeks, infected mice were divided into 7 groups: low HBYYRJ group, middle HBYYRJ group, high HBYYRJ group, Fufangbiejiaruangan tablet (FFBJRG) group, colchicine group, 3 months infection group and 6 months infection group. Hepatic fibrosis was found in 3 months infection group. Liver hydroxyproline (Hyp) was determined, matrix metalloproteinase-2 and 9 (MMP-2, MMP-9) were detected with gelatin zymography, serum hyaluronic acid (HA) and precollagen III (PC-III) were detected using RIA. RESULTS: HBYYRJ obviously reduced hepatic fibrosis (probability value less than 0.01). Collagen and HA in 3 months infection group and 6 months infection group were higher than that in normal group (probability value less than 0.01), collagen in high and middle HBYYRJ groups and HA in middle and low HBYYRJ groups were lower than that in 6 months infection group (P less than 0.01, probability value less than 0.05). The expression of MMP-9 and MMP-2 in 3 months infection group and 6 months infection group was higher than that in normal group (probability value less than 0.01), The expression of MMP-9 in three HBYYRJ groups and the expression of MMP-2 in high HBYYRJ group were lower than that in 6 months infection group (probability value less than 0.05). CONCLUSION: HBYYRJ can reduce liver fibrosis caused by schistosomiasis.

[Effects of centipede extracts on normal mouse and S180, H22 bearing mouse].

OBJECTIVE: To study the effects of centipede extracts on H22 tumor-bearing mouse, sarcoma S180 mouse and normal mouse. METHODS: Normal and tumor-bearing mouse were orally administrated by centipede extracts. Rate of restraining tumor, index of thymus and spleen were calculated after 12 days treatment. Acute toxicity testing tried to figure out In LD50 of centipede extracts. RESULTS: The restraining tumor rates of centipede ethanol extracts at low and medium doses were 22.2% and 17. 88%. There was no tumor restraining effect by the high dose treatment. The tumor growth of the H22 model mouse was not restrained by the centipede water extracts. There were no significant differences among the treatments in their spleen weight and spleen index. In LD50 test, the administrating dosages of centipede extracts given to the mouse were 48 times those given to patients on clinic. The result showed no mouse dead in centipede group. CONCLUSION: Centipede water extracts had no anti-tumor effect on tumor-bearing mouse. There is certain toxicity in ethanol extracts of centipede, suggesting that centipede alone for treatment of cancer needs further study.

Hepatoprotective activity of six polyherbal formulations in paracetamol induced liver toxicity in mice.

BACKGROUND & OBJECTIVE: Polyherbal formulations available with a wide range of indications like protective to liver, appetite and growth promoters, gastrointestinal and hepatic regulator, as treatment for hepatic dysfunction, for hepatic regeneration as well as liver stimulant and tonic. Despite the widespread use, there is a lack of scientific evidence on their efficacy and safety. This study was undertaken to evaluate the hepatoprotective activity of six commercially available formulations, namely Liv 52, Livergen, Livokin, Octogen, Stimuliv and Tefroliv in acute liver toxicity in mice model induced by paracetamol (PCM). METHODS: Swiss albino mice of either sex were used, divided in 28 groups with six in each group. The dose of the polyherbal formulations was calculated from human dose (20 ml/day) using a standard conversion table. They were given as pretreatment (2.60 ml/kg/day) for 7 days by oral route twice a day prior to PCM administration. Hepatotoxicity was induced by administering a single oral dose of PCM (500 mg/kg bw) on day 8. The study parameters were conducted on day 9. The biochemical parameters included liver enzyme levels alanine tranaminases (ALT), aspartate transaminases (AST) and alkaline phosphatase (ALP). The pharmacological and pathological parameters were phenobarbitone sleeping time and macroscopic and microscopic changes of liver tissues respectively. RESULTS: PCM toxicity significantly increased ALT, AST and ALP (321.00 +/- 87.93, 273.17 +/- 45.68, 257.50 +/- 17.64 IU/l vs normal control, 33.33 +/- 0.61, 89.33 +/- 9.50, 152.17 +/- 11.40 IU/l respectively, P<0.05), prolonged phenobarbitone induced sleeping time (from 277.50 +/- 8.04 min to 335.83 +/- 7.00 min, P<0.05). When PCM higher dose (1g/kg p.o. single dose) was used, the liver tissue, in macroscopic appearance, showed extensive necrosis associated with haemorrhages. Low dose (500 mg/kg p.o. single dose) showed punctate haemorrhagic necrosis of liver tissue. In the microscopic studies, PCM induced toxicity showed haemorrhages, fatty changes and necrosis. The pretreatment in low doses (2.6 ml/kg/day) with liquid formulations of Liv 52 and Livergen reversed the PCM induced liver toxicity. At higher doses (5.2 ml/ kg/day), all the six herbal formulations conclusively showed marked beneficial effects in the studied pharmacological, biochemical and histological parameters. INTERPRETATION & CONCLUSION: The present findings demonstrated the efficacy of polyherbal liquid formulations at two dose levels in PCM induced hepatotoxicity in mice. However, it suggests that a dose adjustment may be necessary to optimize the effects in clinical settings.

Effects of a homeopathic complex in Nile tilapia (Oreochromis niloticus L.) on performance, sexual proportion and histology.

This study aimed to evaluate the effects of the homeopathic complex Homeopatila RS (REAL Homeopathy, Brazil), in the performance, sexual proportion and gills and liver histology of the Nile tilapia fish (Oreochromis niloticus L.). 4,800 post-larvae were treated for 45 days, in 12 tanks (500 L), in a closed environment, with density of 400 larvae per tank. Three treatments were given: alcohol 30%, negative control (C); hormonal, positive control, 17 alpha-methyltestosterone (H); homeopathic, Homeopatila RS, in ethanol (HH), with four replications. Mean values for length, weight, liver somatic index, condition factor, survival rate, average values of histological alterations and sexual proportion were determined. Analysis revealed significant differences (p<0.05) in growth, survival rate, liver somatic index and average values of hepatic histological alterations between treatments. It was concluded that the addition of Homeopatila RS to the diet of Nile tilapias, during the phase of gonadal differentiation, did not induce any alteration in the sexual proportion. Homeopathically treated fish were significantly smaller, but had significantly greater survival than the other two groups, there was no significant difference in final total biomass. The homeopathically treated fish had a lower liver/somatic index with less hepatic lipid inclusions than the other groups.

The contraceptive implant.

Contraceptive implants provide long-acting, highly effective reversible contraception. Currently, the only subdermal implant available to women in the United States is the single rod etonogestrel implant, Implanon (N.V. Organon, Oss, the Netherlands) approved by the Food and Drug Administration in July 2006. Implanon is currently approved for 3 years of use, provides excellent efficacy throughout its use, and is easy to insert and remove. Similar to other progestin-only contraceptives, Implanon can cause irregular vaginal bleeding. Implanon has been shown to be safe to use during lactation, may improve dysmenorrhea, and does not significantly affect bone mineral density, lipid profile, or liver enzymes.

Inhibition of chemically induced carcinogenesis by drugs used in homeopathic medicine.

Homeopathy is considered as one modality for cancer therapy. However, there are only very few clinical reports on the activity of the drugs, as well as in experimental animals. Presently we have evaluated the inhibitory effects of potentized homeopathic preparations against N'-nitrosodiethylamine (NDEA) induced hepatocellular carcinoma in rats as well as 3-methylcholanthrene-induced sarcomas in mice. We have used Ruta, Hydrastis, Lycopodium and Thuja, which are commonly employed in homeopathy for treating cancer. Administration of NDEA in rats resulted in tumor induction in the liver and elevated marker enzymes such as gamma-glutamyl transpeptidase, glutamate pyruvate transaminase, glutamate oxaloacetate transaminase and alkaline phosphatase in the serum and in liver. Concomitant administration of homeopathic drugs retarded the tumor growth and significantly reduced the elevated marker enzymes level as revealed by morphological, biochemical and histopathological evaluation. Out of the four drugs studied, Ruta 200c showed maximum inhibition of liver tumor development. Ruta 200c and phosphorus 1M were found to reduce the incidence of 3-methylcholanthrene-induced sarcomas and also increase the life span of mice harboring the tumours. These studies demonstrate that homeopathic drugs, at ultra low doses, may be able to decrease tumor induction by carcinogen administration. At present we do not know the mechanisms of action of these drugs useful against carcinogenesis.

[Pharmacokinetic study on baicalin of Qingkailing injection in rats].

OBJECTIVE: Studying the metabolic pharmacokinetic of baicalin of Qingkailing injection in rat, to search for effector substance of Qingkailing injection in vivo. METHOD: Qingkailing sterile injection powder was given by caudal vein, then blood, liver and lung were collected in various time, the concentration of baicalin from samples were determined by HPLC-MS. Pharmacokinetic evaluation was carried out using the 3P87. RESULT: After Qingkailing injection, Baicalin was consistent with two-compartment model in rat. 45 min, the concentration of baicalin in hepatic tissue reached maximum, followeded by decrease sharply, 120 min began to rise slowly, present double hump phenomenon. In lung, baicalin concentration was far more than in liver, was eliminated more slowly, but they have the same t(max). CONCLUSION: After Qingkailing injection, baicalin distributed quickly to liver and lung, baicalin is one of effector substance of qingkailing injection in vivo. Baicalin might have hepatoenteral circulation.