A morphometric and molecular study of the apoptosis observed on tadpoles' tail explants under the exposition of triiodothyronine in different homeopathic dilutions.

BACKGROUND: As a therapeutic system, homeopathy is supported by: i) similitude and experimentation in healthy individuals, ii) potentization. A challenge for researchers consists in looking for signals in water (or vehicle) to explain the storage of information in extremely high dilutions and the transfer of such information to the living systems. Anuran amphibian metamorphosis is controlled by thyroid hormones (TH), including the resorption of the tadpole tail. Apoptosis is a genetically regulated form of cell death that can be triggered by various extracellular and intracellular stimuli resulting in coordinated activation of a family of cysteine proteases called caspases. METHODS: This study was blind and randomized. It performed in three stages: I) the identification of the most effective T3 homeopathic dilution to induce apoptotic reactions in Rana (Lithobates) catesbeianus tadpole tail explants stimulated by T3 in substantial, II) study of different controls and III) detection in explants under the action of the most effective dilution of T3, as established in Stage I. RESULTS: There was no statistically significant difference between tail macroscopic dimensions between the groups. T3 10cH decreased the expression of caspase 3/7 mRNA, in explants treated with T3 20 nM. CONCLUSION: The present experiment is in agreement with the hypothesis that T3, at a 10cH homeopathic dilution, changes the metamorphosis molecular network.

Ultra high dilution of triiodothyronine modifies cellular apoptosis in Rana catesbeiana tadpole tail in vitro.

BACKGROUND: Ultra High Dilutions (UHD) are diluted beyond the Avogadro limit with dynamization (dilution with succussion). The process of anuran amphibian metamorphosis is controlled by thyroid hormones, including the resorption of the tadpole tail. METHODS: A randomized and blinded study was performed to investigate the influence of triiodothyronine (T3) 5·10(-24)M (10cH) on apoptosis induced by T3 100 nM in Rana catesbeiana tadpoles' tail tips, in vitro. Explants were randomized to three groups: control: no T3 in pharmacological or UHD dose; test: T3 100 nM and challenged with T3 10cH (UHD); positive control: T3 100 nM, treated with unsuccussed ethanol. The apoptotic index and the area of explants of test and control groups at the first and final day of the experiment were compared by t-test. RESULTS: There was no difference in tail tip area between test and control groups, but a significantly higher (p<0.01) index of apoptosis in explants of the test group. CONCLUSION: This data suggest that T3 10cH modifies the effect of T3 at pharmacological dose, opening new perspectives for further studies and investigation of the dose-effect curve.

[Effect of Wulong Dan on treatment of acute cerebral infarction and change of hormones in hypothalamus-pituitary-thyroid axis].

OBJECTIVE: To explore the effect of Wulong Dan (WLD) on treatment of acute cerebral infarction and change of hormones in hypothalamus-pituitary-thyroid axis. METHODS: Thirty five cases of the treated group treated with WLD by taken orally and 31 cases of the control group treated with nimoton taken orally. And dextran was given intravenously to both groups. Blood level of triiodothyronine (T3), free triiodothyronine (FT3), thyroxine (T4), free thyroxine (FT4) and thyroid stimulating hormone (TSH) of all patients were determined before and after treatment by radioimmunoassay. RESULTS: After 4 weeks of treatment, there were significant difference between the treated group and the control group in cure rate, markedly effective rate, cure score and disappearance of clinical symptoms (P < 0.01). The levels of T3, FT3, TSH of all patients were lower than those of the normal control before treatment. After treatment with WLD and dextran, the above-mentioned indexes turned to normal. CONCLUSIONS: WLD has a significant effect on patients with cerebral infarction on the basis of improved microcirculation induced by dextran. It also has regulatory effect on the hypothalamus-pituitary-thyroid axis disorder so as to be helpful in maintaining the homeostasis.

Triiodothyronine, diluted according homeopathic techniques, modifies the programmed cell death of tadpole tail?s explants / Triiodothyronine, diluted according homeopathic techniques, modifies the programmed cell death of tadpole tail?s explants

High Dilution is a solution beyond the Avogadro limits that, in the dependence of the applied succussion elicits a suppressive or a stimulant effect on a living cell, with a consequent generation of an oscillatory doseeffect curve. According to Bonamin et al. [1], ?Perhaps, the most enigmatic feature regarding the properties of high dilutions is the non-linearity of their effects. In several studies employing in vivo and ex vivo models, especially involving iso-endopathy, an oscillatory potency-effect curve has appeared. The first observations were initially considered as artifacts, but the repetition of this pattern in different studies involving completely different experimental models, in times and places equally different, points out to the existence of a property intrinsic to dynamized systems.? The entire process of anuran amphibian metamorphosis is under thyroid hormones control, included the complete resorption of the tadpole tail. In the present study, we had successfully established a protocol model to culture Rana catesbeiana tadpoles? tail tips in vitro. A random and blind study was performed, with the intent to prove that T3 5.10-24 M (10 cH) modifies the apoptosis induction of T3 100 nM in explants of Rana catesbeiana tadpoles? tail. 60 explants were distributed in three ways: Group A: without T3 action, at pharmacological and HD dose; Group B (test): under the action of T3 100 nM and treated with T3 10 cH (HD); Group C (control): under the action of T3 100 nM and treated with ethanol 70% unsuccussed. After 96 hours of tissue culture, the mean of initial and final area (1.05 vs. 0.98 cm2) and apoptotic index of the explants from Group A were with minimal difference range and for this reason it wasn?t included in the statistical study. In order to identify significant differences in the area and in the apoptotic index of the remainder explants of the 2 groups, B (test) and C (control), we used a student t-test. However, the mean initial and final explants? area from test and control groups were respectively 1.09 vs. 0.22 cm and 1.00 cm vs.0.24 cm, with a mean reduction of 0.87 cm2 and 0.76 cm2, but this difference didn?t achieve statistical significance (p>0.05). In contrast, apoptosis index was significantly higher in test than in control group 11.7 vs. 7.9 (p<0.05), with is confirmed at the table 1.(AU)

Triiodothyronine, diluted according homeopathic techniques, modifies the programmed cell death of tadpole tail?s explants / Triiodothyronine, diluted according homeopathic techniques, modifies the programmed cell death of tadpole tail?s explants

High Dilution is a solution beyond the Avogadro limits that, in the dependence of the applied succussion elicits a suppressive or a stimulant effect on a living cell, with a consequent generation of an oscillatory doseeffect curve. According to Bonamin et al. [1], ?Perhaps, the most enigmatic feature regarding the properties of high dilutions is the non-linearity of their effects. In several studies employing in vivo and ex vivo models, especially involving iso-endopathy, an oscillatory potency-effect curve has appeared. The first observations were initially considered as artifacts, but the repetition of this pattern in different studies involving completely different experimental models, in times and places equally different, points out to the existence of a property intrinsic to dynamized systems.? The entire process of anuran amphibian metamorphosis is under thyroid hormones control, included the complete resorption of the tadpole tail. In the present study, we had successfully established a protocol model to culture Rana catesbeiana tadpoles? tail tips in vitro. A random and blind study was performed, with the intent to prove that T3 5.10-24 M (10 cH) modifies the apoptosis induction of T3 100 nM in explants of Rana catesbeiana tadpoles? tail. 60 explants were distributed in three ways: Group A: without T3 action, at pharmacological and HD dose; Group B (test): under the action of T3 100 nM and treated with T3 10 cH (HD); Group C (control): under the action of T3 100 nM and treated with ethanol 70% unsuccussed. After 96 hours of tissue culture, the mean of initial and final area (1.05 vs. 0.98 cm2) and apoptotic index of the explants from Group A were with minimal difference range and for this reason it wasn?t included in the statistical study. In order to identify significant differences in the area and in the apoptotic index of the remainder explants of the 2 groups, B (test) and C (control), we used a student t-test. However, the mean initial and final explants? area from test and control groups were respectively 1.09 vs. 0.22 cm and 1.00 cm vs.0.24 cm, with a mean reduction of 0.87 cm2 and 0.76 cm2, but this difference didn?t achieve statistical significance (p>0.05). In contrast, apoptosis index was significantly higher in test than in control group 11.7 vs. 7.9 (p<0.05), with is confirmed at the table 1.

Ultradiluição homeopática de triiodotironina altera a apoptose celular da cauda de girinos de Rana catesbeiana: in vitro / Ultra high dilution of triiodothyronine modifies the cellular apoptosis of Rana catesbeiana tadpole tail: in vitro

Ultradiluição (UHD) é o efeito de uma solução, diluída acima do número de Avogrado, que na dependência da sua dinamização (diluição com sucussão) induz um efeito celular supressivo ou estimulante, com conseqüente obtenção de uma curva dose-efeito oscilatória. Por outro lado, a 3,3,5 Triiodo-L-Tironina (T3) é o hormônio mais importante na indução e manutenção das mudanças metamórficas dos girinos, nelas incluídas a absorção da cauda. O presente estudo, cego e randomizado, tem como objetivo comprovar que o T3 5.10-24M (10ª cH) altera a apoptose induzida pelo T3 100 nM na cauda de girinos de Rana catesbeiana, in vitro. Foram distribuídos 60 explantes em três grupos: Grupo A: sem o estímulo do T3 em dose farmacológica e em UHD; Grupo B (teste): sob a ação de T3 100 nM e T3 10ª cH (5.10-24 M); Grupo C (controle): sob a ação do T3 100 nM e etanol 70% sem sucussão. A análise estatística da área dos explantes, no primeiro e ultimo dia do experimento, e do índice apoptótico foi realizado através do teste t Student e foi considerado estatisticamente significante quando p<0,05. Embora sem diferenças significativas na área dos explantes do grupo teste e no grupo controle, um maior e significante índice apoptótico foi identificado nos explantes do grupo teste. Este resultado confirma que o T3 na 10ª cH altera a ação do T3 em dose farmacológica. Futuros experimentos serão realizados, com diferentes dinamizações, com o objetivo da parametrização da curva dose-efeito.

Ultra High Dilution (UHD) is the effect of a solution, beyond the Avogadro limits, that in the dependence of the applied dinamization (dilution with succussion) elicits a suppressive or a stimulant effect on a living cell, with a consequent generation of an oscillatory dose-effect curve. The entire process of anuran amphibian metamorphosis is under thyroid hormones control, included the complete resorption of the tadpole tail. A random and blind study was performed, with the intent to prove that T3 5.10-24 M (10ª cH) modifies the apoptosis induction of T3 100 nM in Rana catesbeiana tadpoles tail tips, in vitro. 60 Explants were distributed in three ways: Group A: without T3 action, at pharmacological and UHD dose; Group B (test): under the action of T3 100 nM and treated with T3 10ª cH (UHD); Group C (control): under the action of T3 100 nM and treated with ethanol 70% unsuccussed. In order to identify significant differences in the area of the remainder explants, at the first and final day of the experiment, and in the apoptotic index we used a student t-test. Although we didnt find statistical difference in macroscopic tadpoles tail tips area from test and control groups, a high and significant (p<0,05) index of apoptosis in histology was found in explants of test group. This data confirms that T3 10 cH modifies the effect of T3 at pharmacological dose. More studies will be necessary, using different dinamizations, to the parameterization of the dose-effect curve proceeding from these experiments.