">NON-MOLECULAR INFORMATION TRANSFER
FROM THYROXINE TO FROGS

By Means of Homoeopathic Preparation and Electronic Processing

P.C. ENDLER¹, W. PONGRATZ¹, C.W. SMITH², J. SCHULTE³, F. SENEKOWITSCH⁴, M. CITRO⁵

¹Ludwig Boltzman Institut für Homöopathie, Durerg.4, 8010 Graz, Austria

²Department of Electric and Electronic Engineering, University of Salford, 827221 Salford, UK.

³National Superconducting Cyclotron Laboratory, Michigan State University, East Lansing, USA

⁴Institute of Bioinformatics, Graz, Austria

⁵Research Institute Alberto Sorti (IDRAS), Turin, Italy

1 - Introduction
2 - Methods
3 - Results
4 - Discussion
5 - References

1. Introduction

Research on the scientific basis of homoeopathy can generally proceed on two lines: (a) with regard to the principles underlying inversion effects of specially prepared dilutions (van Wijk and Wiegant, 1994; Bastide, 1994; Endler et al., 1994a) and (b)with regard to the biophysical properties of the information transfer process. This paper is dedicated to (b).

Substances so much diluted that no original molecule is present still exert biological effects, i.e. molecular bio-information may be transduced via water (Bastide et al., 1987; Hadji et al., 1991; Benveniste et al., 1992; Youbicier-Simo et al., 1993; Smith, 1994; Endler et al., 1994b; Pongratz et al., 1995; for a survey, see Endler and Schulte (1994), and hence inhibit climbing activity of juvenile frogs (Endler et al., 1994b). In the work presented here, we have studied the thyroxine-controlled metamorphosis of the amphibian Rana temporaria from the two- to the four-legged stage and further on to the juvenile stage.

Thyroxine, added to the water of a basin (final concentration 10^-9 or 10^-8 part by weight, 2 or 20 nM), induces or accelerates respectively, the development of tadpoles (Pitt-Rivers and Trotter, 1964; Endler and Schulte, 1994). It was postulated that perimolecular energetic fields are involved in the interaction of biologically active substances and the organism (Endler and Schulte, 1994).

The blind experiments on non-molecular information from thyroxine presented in this paper were carried out between 1990 and 1994. In experiments A, the influence of a high dilution of thyroxine was tested. In experiments B, it was attempted to electronically digitize information from this high dilution. In experiments C, it was attempted to transfer information from molecular thyroxine by means of a high gain amplifier. In experiments D, the high dilution of thyroxine was sealed in glass vials hung into the basin water.

2. Methods

In previous experiments, it was observed that a homoeopathically prepared dilution such as 10^-30M of thyroxine can exert an inhibiting effect on tadpole metamorphosis (Endler and Schulte, 1994). However, the relation between the seasonal time the experiment is performed at and the frequency of application of the dilutions was found to be crucial. In spring, more frequent stimulation is needed than in autumn (Table 1).

TABLE 1. Relation of seasonal time and frequency of application with regard to the effect of homoeopathically prepared thyroxine (10^-30) on amphibian metamorphosis. strong, weak, none = strong, weak, no inhibiting effect

Application at spring summer autumn

intervals of

______________________________________________________________

0h/8h strong weak none /opposite

48 h weak weak strong

This relation between seasonal time and frequency of application (as was derived from protocol A) was also taken into consideration in the further experiments B, C and D

2.1. PROTOCOL A: USE OF AN AGITATED DILUTION

2.1.1. Animals and Staging

The first experiments were performed in summer with Rana temporaria from an Austrian pool about 1500 m above sea level. For the main study, Rana temporaria were taken with the transition from the two-legged stage onwards in autumn. We chose only those tadpoles where the hindlegs slightly started to be straddled (stage 31, Gosner's staging table - Gosner, 1960 -). The two-legged tadpoles were observed at regular intervals for 1-2 weeks until (a) the forelegs, preformed under the skin, broke through. After a developmental period of 1 - 2 weeks, the forelegs broke through within a few minutes. Thus, this parameter seems adequate to define a final stage. Furthermore (b), the animals were observed until they reached the juvenile stage (stage of tail reduction).

2.1.2. Further conditions

The transitions were observed in disposable basins (8l). The room temperature was 19^oC. Indirect natural light was used. The tadpoles were fed with a standardized aquarium product ad libidum. Two sets of basins were always used for the treatment with the two test solutions (see below). The number of animals per basin was alike (10 - 20) in each experiment. The basins were rotated in the course of the experiment.

2.1.3. Preparation of testing solutions and exposition to probes

The tadpoles were observed under the influence of tetra-iodo-thyronine sodium pentahydrate (Sigma) specially prepared in an aqueous solution 1 : 10³⁰ part by weight. The stock solution had a concentration of 1 : 10⁴ part tetra-iodo-thyronine sodium pentahydrate of weight; it was diluted in steps of 1 : 10. The diluted solution (10^-5, 10^-6 etc.) was agitated according to standardized instructions (Homöopathisches Arzneibuch, 1991): at every step, a sterile bottle was filled with the dilution, and was pushed down at short regular intervals (e.g. against a rubber impediment) to create mechanical shocks. Disposable pipettes were used. Water was similarly "diluted" (water 10^-30) as control. Check for hormone-active contamination was described previously (Endler and Schulte, 1994). Always 3 drops of the test solutions were added to the basin water at intervals of 48 h.

2.1.4. Evaluation of the data

The cumulative frequencies (fa, fb) of (a) four-legged tadpoles and (b) juveniles were compared by a chi-square test to those of two- or three-legged animals at the respective time intervals. In order to give a further survey on the results, from the graphs the delay was extrapolated with which the test animals reached the fa- and fb- level of the control animals. Further statistical evaluation is discussed elsewhere (Endler and Schulte, 1994; Endler and Pongratz, 1994).

2.1.5. Laboratories/researchers

For the first study (summer 1990), experiments were performed in three independent laboratories (P.C.E., W.P., R.V.W. see List of Authors and Acknowledgements). For the main study (autumn 1993), experiments were performed by W.P.

2.2. PROTOCOL B: COMPACT DISC RECORD /REPLAY

2.2.1. Animals, staging and further conditions are described in protocol (A).

2.2.2. Preparation of testing solutions and exposition to probes

Vials of thyroxine (10^-30 M, or water were placed on an input coil linked to a filter and to an amplifier with a gain of E6. Frequencies below 80 kHz were digitized at the Nyquist frequency, buffered in a RAM and multiplexed on to a CD. After noise reduction and filtering, the signal was attenuated by E6 to restore the original analog level.

Water vials were placed on an output coil for 4 min (device: Fa. Hoefler, Austria). At intervals of 48 h, always 3 drops of the test solutions were added to the basin water. For further details, see experiments (A).

2.2.3. Laboratory /researcher

Experiments were performed in one laboratory by W.P.

2.2.4.Evaluation was done as described in protocol (A).

2.3. PROTOCOL C: USE OF AN ELECTRONIC CIRCUITRY

2.3.1. Animals

Rana temporaria were taken from Austrian pools about 400 m above sea level (spring). Staging and further conditions are described in protocol (A).

2.3.2. Preparation of testing solutions and exposition to probes

Vials of thyroxine (1 mM) or water were placed on an input coil, one end of which was connected to a special amplifier (linear from DC to HF) by a single insulated wire (devices used: Fa. Brügemann, FRG - main study - ; Fa. Medtronik, FRG). Water vials were placed on a similar output coil for 4 min. (Brügemann - device) or 15 min. (Medtronik - device) respectively. At intervals of 8 h always 8 ml of basin water were replaced by the propes (test or control) produced at the output coil. For standardisation of this method, see (Citro, 1991; 1992: Citro et al., 1994; Aissa et al., 1993; Benveniste et al,. 1994).

2.3.3. Laboratory /researcher

The experiments were performed in different laboratories by two independent researchers (W.P., C.V.).

2.3.4. Evaluation was done as described in protocol (A).

2.4. PROTOCOL D: USE OF AN AGITATED DILUTION IN SEALED VIALS

2.4.1. Animals

The first experiments were performed both in summer with Rana temporaria and with Bufo bufo as well as in autumn with Rana temporaria. For the main experiments with Rana temporaria, animals were taken from the pool 400 m above sea level with the transition from the two-legged stage onwards in spring as described in (C). Staging and further conditions are described in protocol (A).

2.4.2. Preparation of testing solutions

It was done as described in protocol (A). Exposition to probes: Eight ml of the thyroxine or water dilutions were sealed in hardglass vials with an optical transmission spectrum > 350 nm. The coded vials were first placed in a common water bath to make sure that they were not individually contaminated outside and then hung into the corresponding basins. The method of application of test substances in sealed ampoules had been used in therapeutical practice and by different researchers before (Endler and Schulte, 1994)).

2.4.3. Laboratory /researcher

Experiments were performed in different laboratories by three independent researchers (P.C.E., W.P., K.W.). The evaluation was done as described in protocol (A).

3. Results

3.1. PROTOCOL A: USE OF AN AGITATED DILUTION

First experiments were performed in summer by P.C.E. (with a total of 545 animals), by W.P. (230 animals) and by R.v.W. (360 animals). At some, but not all measuring points the f(a,b)-values were significantly below those for reference. This is true for the data from the tree laboratories both when pooled (P<0.01) as well as when treated separately (P<0.01).

Figure 1. The influence of the high dilution of thyroxine, (a) on the transition from the two-legged to the four-legged stage and (b) on the transition to the juvenile stage of Rana temporaria tadpoles. "100%" refers to 140 (fa) or 50 animals (fb), respectively, in each group. Time categories 1 - 4, depending on the experiment: normally 2-3 days (see methods). 1 SD < 10%; * , P < 0.01. For absolute numbers of animals, see Table 2. For further explanation, see the text.

TABLE 2. Absolute numbers of animals in Figure 1

Time category 1 2 3                    4

N_fa (control) 048 077 099 12

N_fa (test probe) 021 060 085 113

N_fb (control) 022 027 031 039

N_fb (test probe) 011 018 025 026

The main experiments (optimized protocol) were performed in autumn by W.P. (with a total of 280 animals). As is shown in Figure 1, at all measuring points in time, the f(a,b)-values for the animals exposed to the test dilution (black squares) were below those for reference (white squares) (P<0.01).

From these results it can be extrapolated that the tadpoles exposed to the test liquid reached the f(a,b)-levels of control animals after a delay of about 1 - 2 d.

In other words, the chance of passing metamorphosis is generally smaller for the group treated with the high dilution of thyroxine, added to the basin water at intervals of 48 h than for the water control group.

3.2. PROTOCOL B: COMPACT DISK RECORD /REPLAY

Several experiments were performed in autumn by W.P. with a total of 468 animals.

As is shown in Figure 2, at all measuring points in time, the f(a,b)-values for animals exposed to the test dilution (black circles) were below those for reference (white circles) (P<0.01).

Figure 2. The influence of the liquid exposed to digitised and CD - stored thyroxine information. "100%" refers to 234 animals in each group. 1 - 4, time categories, depending on the experiment: normally 24 h. 1 SD > 10%; *, P < 0.05; **, P < 0.01. For absolute numbers of animals, see Table 3. For further details, see the text.

TABLE 3. Absolute numbers of animals in Figure 2

Time category 1 2 3 4

N_fa (control) 126 152 171 201

N_fa (test probe) 094 124 151 175

N_fb (control) 070 091 131 161

N_fb (test probe) 057 066 109 137

From these results it can be extrapolated that the tadpoles exposed to the test liquid reached the f(a,b)-level of control animals after a delay of about 1 d. In other words, the chance of passing metamorphosis is generally smaller for the group treated with the liquid exposed to the digitized and CD - stored thyroxine information.

3.3. PROTOCOL C: USE OF AN ELECTRONIC CIRCUITRY

Several experiments were performed in spring by W.P. with a total of 756 animals and one experiment by C.V. (180 animals). As is shown in Figure 3 (see next page), at all measuring points in time, the f(a,b)-values for animals exposed to the test dilution (black triangles) were below those for reference (white triangles) (P<0.01). This is true for the data from the two laboratories both when they were pooled as well as when they were treated separately.

Figure 3. The influence of the liquid exposed to electronically transferred thyroxine information. "100%" refers to 468 animals in each group. Time categories 1 - 4, depending on the experiment: normally 24 h. 1 SD < 10%. For absolute number of animals, see Table 5. For further information, see text and the Figure 1

TABLE 4. Absolute numbers of animals in Figure 3

Time category 1 2 3 4

N_fa (control) 206 309 374 417

N_fa (test probe) 145 248 304 360

N_fb (control) 201 243 295 342

N_fb (test probe) 126 173 229 290

From these results it can be extrapolated that the tadpoles exposed to the test liquid reached the f(a,b)-levels of control animals after a delay of about 1 - 1,5 d. In other words, the chance of passing metamorphosis is generally smaller for the group treated with the liquid exposed to electronically transferred thyroxine information.

3.4. PROTOCOL D: USE OF AN AGITATED DILUTION IN SEALED VIALS

The first experiments were performed in summer with Bufo bufo (P.C.E., 347 animals and K.W., 104 animals) and with Rana temporaria (H.H., 217 animals). At some, but not all measuring points, the fa-values were significantly below those for reference. This is true for the data from the three researchers both when they were pooled as well as when they were treated separately (P < 0.01 or P < 0.05, respectively). Further experiments with Rana temporaria were performed in autumn (W.P. et al., 1995). In general, the differences showed that the test dilution slowed down development to the four-legged stage. However, this could not be statistically proved (Endler and Schulte, 1994). The main experiments were performed in spring by W.P. (792 animals) and by K.W. (with a total of 432 animals). As is shown in Figure 4, at all measuring points in time, the f(a,b)-values for animals exposed to the test dilution (black squares) were below those for reference (white squares) (P < 0.01). This is true for the data from the two laboratories both when they were pooled as well as when they were treated separately.

From these results it can be extrapolated that the tadpoles exposed to the test liquid sealed in glass vials reached the f(a,b)-levels of control animals after a delay of about 1 - 2 d. In other words, the chance of passing metamorphosis is generally smaller for the group treated with the high dilution of thyroxine, sealed in glass vials and hung into the aquaria than for the water control group.

Figure 4. The influence of the high dilution of thyroxine, sealed in glass vials. "100%" refers to 612 (time categories 1 and 2, f_a,b) and 468 (time categories 3 and 4, f_a) or 216 (time categories 3 and 4, f_b) animals in each group. Time categories 1 - 4, depending on the experiment: normally 1 or 2 d. 1 SD < 10%; * , P < 0.01. For absolute numbers, see Table 5. For further details, see the text and the Figure 1

TABLE 5. Absolute numbers of animals in Figure 4

Time category 1 2 3 4

N_fa (control) 113 295 315 390

N_fa (test probe) 072 168 236 324

N_fb (control) 150 258 118

N_fb (test probe) 156 247 105

4. Discussion

These results refer to frogs during their metamorphosis from the two-legged stage (a) to the four-legged stage and (b) to the juvenile stage, treated with the test water to which information from thyroxine was transferred by different means. This caused differences in the rate of metamorphosis as compared with reference (water to which information from water was analoguely transferred). In the blind experiments the test liquids significantly slowed down metamorphosis.

In experiments A, the influence of a high dilution (10^-30 M) of thyroxine, prepared in a special process of stepwise dilution and agitation, or similarly diluted water was tested. These probes were directly added to the basin water. In experiments B, vials of the thyroxine dilution (10^-30 M) or water were placed on an input coil linked to a filter and to an amplifier with a gain of 10⁶. Frequencies up to 80 kHz were digitized at the Nyquist frequency, buffered in a RAM and multiplexed on to a CD. After noise reduction and filtering, the signal was attenuated by 10⁶ to restore the original analog level. Water vials were placed for 4 min on an output coil. These probes were directly added to the basin water. In experiments C, vials of thyroxine (1 mM) or water were placed on an input coil, one end of which was connected to a special amplifier (linear from DC to HF) by a single insulated wire. Water vials were placed for 4 min on a similar output coil. These probes were directly added to the basin water. In experiments D, a thyroxine dilution (10^-30 M, see protocol A) and similarly prepared water were sealed in hardglass vials hung in the basin water.

During the treatment, the cumulative frequencies of four-legged tadpoles (for fa, see Table 6) and of juvenile frogs dropped in all types of experiments.

From Table 6 it can be seen that the chance of entering the four-legged stage is generally smaller in the groups treated with the information from thyroxine than in the control groups. Information from molecular thyroxine can be transferred by a process of stepwise aqueous dilution and agitation or by means of an electronic circuitry, it can be stored on a compact disk and it can exert its effect through the wall of a sealed glass vial (for references, see Table 6). This leads to the assumption that this bio-information is electromagnetic in nature and can be processed by conventional electronic circuits and devices.

Recent physics research has revealed that water dipoles may develop phase coherent oscillations through radiation coupling (del Giudice et al., 1988; del Giudice, 1994). It is proposed that these could be modulated as a time-ordered pattern of signals (Endler and Schulte, 1994) and induce the coherent wave propagation (electron propagation) in metals (Endler and Schulte, 1994). A further theory suggests that the phase coherent oscillations may originate information pattern formation through isotopicity effects in high dilutions (Endler and Schulte, 1994). The theoretical physical aspect of the experiments A - D referenced here has yet to be evaluated in order to test and develop a theory on bio-information storage and transfer in 'homoeopathically' prepared dilutions.

TABLE 6. Cumulative frequency of four-legged tadpoles

Protocol / drop of N of P researchers

Reference f_a animals (chi² test)

(test probe + control)

A 10% 683 + 682 0.001 PCE, WP,

Endler et al.; 1991 RvW

19% 140 + 140 0.001 WP

B 14% 234 + 234 0.001 WP

Endler et al,. 1994d, Senekowitsch et al,. 1995

C 13% 468 + 468 0.001 WP, CV

Citro et al., 1995

D 09% 303 + 313 0.01 PCE, KW,

Endler et al, 1994c HH

Non-molecular information transfer from thyroxine to frogs. For initials, see List of Authors and Acknowledgements. First experiments (the preliminary protocols); Main experiments (optimized protocols). For further explanation, see the text. Main experiments of protocol A and experiments B were performed when natural metamorphosis proceeds relatively slowly (autumn), whereas experiments C and main experiments of D were performed when natural metamorphosis proceeds relatively fast (spring). A less differentiated table was presented in Endler et al., 1995.

The possibility of storing (biologically /chemically active) information from molecules on data carrier such as a CD may open new ways both for therapeutical as well as for technical use. However, the authors wish to emphasize that they understand their work as a contribution towards a deeper understanding of the theoretical background of homoeopathy.

Acknowledgements

We are obliged to those who stimulated the experiments, especially to Thomas Kenner and Max Moser, Institute of Physiology, University of Graz and Max Haidvogl, Boltzmann Institute of Homeopathy, Graz. Furthermore, we would like to express thanks to those who performed independent experiments (see Table 6): Roeland Van Wijk, Department for Molecular Cell Biology, University of Utrecht, The Netherlands; Christina Vinattieri, Department of Holistic Medicine, University of Urbino & IDRAS, Turin, Italy; Karl Waltl, Boltzmann Institute of Homoeopathy, Graz; Helge Hilgers, Institute of Zoology, University of Vienna, Austria.

5. References

Aissa, J., Litime, M.H., Attis, E., and Benveniste, J. (1993) Molecular signalling at high dilution or by means of electronic circuitry, J.Immunol. 150, A146.

Bastide, M. (1994) Immunological examples on ultra high dilution research, in P.C. Endler, J. Schulte (eds.): Ultra High Dilution. Physiology and Physics, Kluwer Academic Publishers, Dordrecht, pp 27-34.

Bastide, M., Daurat, V., Doucet-Jaboeuf, M., Pélegrin, A., and Dorfman, P. (1987) Immunomodulator activity of very low doses of thymulin in mice, Int J Immunotherapy 3, 191-200.

Benveniste, J., Aissa, J., Litime, M.H., Tsaegaca, G.T., and Thomas, Y. (1994) Transfer of the molecular signal by electronic amplification, FASEB J 8, A398.

Benveniste, J., Arnoux, B., and Hadji, L.(1992) Highly dilute antigen increases coronary flow of isolated heart from immunized guinea-pigs, FASEB J 6, A1610.

Citro, M. (1991) Vom Pharmakon zur Frequenz; elektronischer Medikamententransfer, Proc. II int. Symp. Biokybernetische Medizin, Würzburg.

Citro, M.(1992) TFF dal farmaco alla frequenza. Vivibios II 3, 25-30.

Citro, M., Endler, P.C., Pongratz, W., Vinattieri, C., Smith, and C.W., Schulte, J.(1995) Hormone effects by electronic transmission, FASEB J (Abstract 12161).

Citro, M., Smith, C.W., Scott-Morley, A., Pongratz, W., Endler, P.C.(1994) Transfer of information from molecules by means of electronic amplification, in P.C. Endler, J. Schulte (eds.): Ultra High Dilution. Physiology and Physics, Kluwer Academic Publishers, Dordrecht, pp 209-214.

Del Giudice, E. (1994) Is the 'memory of water' a physical impossibility?, in P.C. Endler, J. Schulte (eds.): Ultra High Dilution. Physiology and Physics, Kluwer Academic Publishers, Dordrecht, pp 117-120.

Del Giudice, E., Preparata, G., and Vitiello, G.(1988) Water as a free electric dipole laser, Phys Rev Lett. 61, 1085-1088.

Endler, P.C., Citro, M., Pongratz, W., Smith, C.W., Vinattieri, C., and Senekowitsch, F.(1994d) Übertragung von Molekül-Information, Proc. Int. Symp. Niederenergetische Bioinformation, Bad Waltersdorf.

Endler, P.C., and Pongratz, W.(1994) On effects of agitated highly diluted thyroxine (E-30), Comprehensive report, available at the Institute for Zoology, University of Graz, Universitätsplatz 2, A-8010 Graz.

Endler, P.C., Pongratz, W., Kastberger, G., Wiegant, F.A.C., and Schulte, J.(1994b) The effect of highly diluted agitated thyroxine on the climbing activity of frogs, J Vet Hum Tox. 36, 56-59.

Endler, P.C., Pongratz, W., Smith, C.W., and Schulte, J.(1995) Non-molecular information transfer from thyroxine to frogs with regard to 'homoeopathic' toxicology, J.Vet.Hum.Tox., 37, 259-260.

Endler, P.C., Pongratz, W., Van Wijk, R., Kastberger, G., and Haidvogl, M. (1991) Effects of highly diluted sucussed thyroxine on metamorphosis of highland frogs, Berlin J Res Hom 1, 151-160.

Endler, P.C., Pongratz, W., Van Wijk, R., Waltl, K., Hilgers, H., and Brandmaier, R.(1994c) Transmission of hormone information by non-molecular means, FASEB J 8, A400.

Endler, P.C., Pongratz, W., Van Wijk, R., Wiegant, F.A.C., Waltl, K., Gehrer, M., and Hilgers, H.(1994a) A zoological example on ultra high dilution research, in P.C. Endler, J. Schulte (eds.): Ultra High Dilution. Physiology and Physics, Kluwer Academic Publishers, Dordrecht, pp 39-68.

Endler, P.C. and Schulte, J. (1994) Ultra High Dilution. Physiology and Physics, Kluwer Academic Publishers, Dordrecht.

Gosner, K.L.(1960) A simplified table for staging anuran embryos and larvae with notes on identification, Herpetologica 16, 183-195.

Hadji, L., Arnoux, B., and Benveniste, J. (1991) Effect of dilute histamine on coronary flow of guinea-pig isolated heart, FASEB J 5, A1583.

Homöopathisches Arzneibuch (1991) Stuttgart: Deutscher Apothekerverlag and Frankfurt Govi Verlag.

Pitt-Rivers, R. and Trotter, W.R.(1954) The Thyroid Gland, Butterworth Publisher, London.

Pongratz, W., Endler, P.C., Poitevin, B., and Kartnig, T.(1995) Effect of extremely diluted plant hormone on cell culture, Proc. 1995 AAAS Ann. Meeting, Atlanta.

Senekowitsch, F., Endler, P.C., Pongratz, W., and Smith, C.W.(1995) Hormone effects by CD record /replay, FASEB J Abstract 12025 .

Smith, C.W.(1994) Coherence in living biological systems. Neural Network World 4 3, 379-388.

Van Wijk, R. and Wiegant, F.A.C.(1994) Cultured mammalian cells in homeopathy research: the similia principle in self-recovery, University Utrecht Publisher, Utrecht.

Youbicier-Simo, B.J., Boudard, F., Meckaouche, M., Bastide, M., and Baylé JD (1993) The effects of embryonic bursectomy and in ovo administration of highly diluted bursin on adrenocorticotropic and immune response of chicken, Int. J. Immunother. 9, 169-190.

Home page