Ultra High Dilution of triiodothyronine modifies cellular apoptosis in Rana catesbeiana tadpole tail in vitro
Received14 June 2010
revised27 April 2011
accepted06 May 2011
20 December 2017 (online)
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.
- 1 Tata J.R., Kawahara A., Baker B.S. Prolactin inhibits both thyroid hormone-induced morphogenesis and cell death in cultured amphibian larval tissues. Dev Biol 1991; 146: 72-80.
- 2 Ji L., Domanski D., Skirrow R.C., Helbing C.C. Genistein prevents thyroid hormone-dependent tail regression of Rana catesbeiana tadpoles by targetting protein kinase C and thyroid hormone receptor α. Dev Dyn 2007; 236: 777-790.
- 3 Endler P.C., Pongratz W., Smith C.W., Schulte J. Non-molecular information transfer from thyroxin to frogs with regard to homeopathic toxicology. Vet Hum Toxicol 1995; 37 (03) 259-260.
- 4 Endler P.C., Pongratz W., Smith C.W., Schulte J., Senekowitsch F., Citro M. Non-molecular information transfer from thyroxin to frogs; by means of homeopathic preparation and electronic processing. In: Bastide M. Signals and Images . 1997. Dordrecht: Kluwer; 149-159.
- 5 Guedes J.R.P., Ferreira C.M., Guimarães H.M.B., Saldiva P.H.N., Capelozzi V.L. Homeopathically prepared dilution of Rana catesbeiana thyroid glands modifies its rate of metamorphosis. Homeopathy 2004; 93: 132-137.
- 6 Brazilian homeopathic pharmacopoeia. 2nd edn São Paulo: Andrei; 1997.
- 7 Rasband W.S. ImageJ. Bethesda, Maryland, USA: U.S. National Institutes of Health; 1997– 2005. http://rsb.info.nih.gov/ij/.
- 8 Wilcox C.D., Dove S.B., McDavid W.D., Greer D.B. UTHSCSA image tool: help on-line. San Antonio: Universidade de San Antonio; 1997.
- 9 Bodian D. The staining of paraffin sections of nervous tissue with activated protargol. The roles of fixatives. Anat Rec 1937; 69: 153-162.
- 10 Gavrieli Y., Sherman Y., Bem-Sasson S.A. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol 1992; 119 (03) 493-501.
- 11 Wijsman J.H., Jonker R.R., Keijzer R. A new method to detect apoptosis in paraffin sections: in situ end labeling of fragmented DNA. J Histochem Cytochem 1993; 41 (01) 7-12.
- 12 Weibel E.R. Morphometry: stereological theory and practical methods. In: Gil J. Models of Lung Disease-microscopy and Structural Methods. 1990. New York: Marcel Dekker; 199-247.
- 13 Bonamin L.V., Martinho K.S., Nina A.L., Caviglia F., Do Rio R.G. Very high dilutions of dexamethasone inhibit its pharmacological effects in vivo. Br Hom J 2001; 90: 198-203.
- 14 Ruiz-Vega G., Pérez-Ordaz L., Cortés-Galván L., Juárez-G F.M. A kinetic approach to caffeine–Coffea cruda interaction. Homeopathy 2003; 92: 19-29.
- 15 Welles S.U., Endler P.C., Scherer-Pongratz W. et al. Pretreatment with thyroxin 10−8 and the effect of homeopathically prepared thyroxin 10−30 on highland frogs – a multi-researcher study. Forsch Komplementärmed 2007; 14 (06) 353-357.
- 16 Weber S., Endler P.C., Welles S.U. et al. The effect of homeopathically prepared thyroxine on highland frogs: influence of electromagnetic fields. Homeopathy 2008; 97: 3-9.
- 17 Chirumbolo S., Brizzi M., Ortolani R., Vella A., Bellavite P. Inhibition of CD203c membrane up-regulation in human basopils by high dilutions of histamine: a controlled replication study. Inflamm Res 2009; 58 (11) 755-764.
- 18 Bonamin L.V., Endler P.C. Animal models for studying homeopathy and high dilutions: conceptual critical review. Homeopathy 2010; 99: 37-50.
- 19 Derby A. An in vitro quantitative analysis of the response of tadpole tissue to thyroxine. J Exp Zool 1968; 168: 147-156.
- 20 Smith K.B., Tata J.R. Are new proteins synthesized during hormone-induced tadpole tail regression?. Exp Cell Res 1976; 100: 129-146.
- 21 Bastide M. Is the classical experimental method necessary to unconventional medicine basic research? Example of homeopathy. Texte publié dans le rapport du Programme Europíen COST B4 (Co-Operation in Science and Technology) 1998. Online: http://www.webstore.fr/eim/costmad.htm.
- 22 Carmine T.C. Effects of high potencies of tumor necrosis factor alpha on H2O2 production in cultured neuroblastoma cells by enhanced luminol-dependent chemiluminescence (ECL). Br Hom J 1997; 86: 67-72.
- 23 Siqueira CM. Alterações celulares induzidas por um novo bioterápico do tipo nosodio vivo sobre linhagens MDCK e J774.G8. Dissertation for Master of Science Graduation, Universidade Federal do Rio de Janeiro, Brazil, 2009.
- 24 Tata J.R. Amphibian metamorphosis as a model for studying the developmental actions of thyroid hormone. Biochimie 1999; 81: 359-366.
- 25 Hofmann K. The modular nature of the apoptotic signaling proteins. Cell Mol Life Sci 1999; 55: 1113-1118.
- 26 Liu J., Burkin D.J., Kaufman S.J. Increasing α7β1-integrin promotes muscle cell proliferation, adhesion, and resistance to apoptosis without changing gene expression. Am J Physiol Cell Physiol 2008; 294 (02) 627-640.
- 27 Sachs L.M., Abdallah B., Hassan A. et al. Apoptosis in Xenopus tadpole tail muscles involves Bax-dependent pathways. FASEB J 1997; 11: 801-808.
- 28 Calabrese E.J., Baldwin L.A. Toxicology rethinks its central belief. Nature 2003; 421 (6924): 691-692 [Commentary].
- 29 Rot-Nikcevic I., Wassersug R.J. Tissue sensitivity to thyroid hormone in athyroid Xenopus levis larvae. Dev Growth Differ 2003; 45 (04) 321-325.
- 30 Bonamin L.V., Lagache A., Bastide M. Research on ultra-dilutions and the Theory of Corporeal Signifiers: the follow up. In: Bonamin L.V. Signals and Images: Contributions and Contradictions of High Dilution Research. 2008. Dordrecht: Springer; 9-43.