Belladonna 110 10-222 6.023 x 10-201
Belladonna 120 10-242 6.023 x 10-221
Belladonna 125 10-252 6.023 x 10-231
Belladonna 130 10-262 6.023 x 10-241
Belladonna 140 10-282 6.023 x 10-261
Belladonna 150 10-302 6.023 x 10-281
Belladonna 160 10-322 6.023 x 10-301
Belladonna 170 10-342 6.023 x 10-321
Belladonna 200 10-402 6.023 x 10-381
Atropine 2 2.87 x 10-3 6.023 x 1018
Acetylcholine 2 5.57 x 10-3 6.023 x 1018
________________________________________________________________________
The dilutions were prepared
in two ways: a) by simple homogeneization, without succussion (N dilutions)
and b) by "dynamization"
with manual succussion (knocking slightly
100 times the vial against a firm substrate, rhythmically, at a frequency
of 1 succussion/second for each dilution), according to Hahnemann's homeopathic
technique (D dilutions).
Likewise, D dilutions 1 C -
5 C were prepared in water, starting from distilled water, as well as from
alcohol 70°, the solvent with which the Belladonna
tincture was prepared. These dilutions were used as control.
All the D and N solutions were
prepared in the Homeopathic pharmacy in Bucharest, especially for this research.
The reference substances, atropine sulphate (ATR) and acetylcholine chlorhydrate
(ACH), were used in standard N aqueous
solutions, 10-2, with a content of 17.33 x 1018 and respectively 33.55 x 1018 molecules/100 ml.
1.2. ANIMALS
Male white Wistar rats, with
an average weight of 150 g, fasting, with access only to water the day of
the experience, were used. They were sacrificed by instantaneous decapitation.
The experiences were carried
out in spring and autumn, between 10 h and 16 h.
1.3. TECHNIQUE
The "in vitro" classical
technique on the isolated rat duodenum, mounted in the isolated organ bath (Turner, 1965), with special precautions for
high dilutions, was used.
Duodenum fragments 2 cm long
in situ were isolated and immediately mounted in the isolated organ bath containing
Tyrode type physiologic salt solution, treated with glucose, regulated thermostatically
at 37 °C and oxygenated by air bubbling at a constant output. The motor activity
(motility) of the isolated organ, both the normal tone and isotonic contraction
was recorded using a classical kimograph.
The traction (stress) constantly exerted upon the duodenum
during the test was of 2 g and the inscription was permanently 10 times amplified.
The accommodation to the in
vitro conditions occurred within 10 minutes. The rest interval between two
tests was of 5 minutes. On a duodenum fragment was performed two tests. After
each test followed the washing of the duodenum (by emptying and refilling
the bath with Tyrode's solution), repeated four times. The volume of the solution
making the object of the research, dropped with a pipette into the bath, was
always of 0.2 ml / 20 ml of bath.
Separate baths were used for the tests with : Belladonna, 1 C - 9 C dilutions, 10 C -
200 C dilutions, N dilutions, D dilutions and atropine. During a working day,
in the same bath, the low solutions were tested in an increasing order of
concentrations and the high dilutions were tested at random.
1.4. TESTS
In each test, each dilution
of Belladonna (B), as well as the
solution of each reference substance, was tested on eight duodenum fragments
(n = 8).
1.4.1. Testing the effect on the normal tone of the rat duodenum
The action of the test dilution
of B, on the normal tone of the isolated rat duodenum, was recorded during
5 minutes. The values of the effects exerted by the B dilutions (tone decrease
or increase) were expressed in percentage against the effects (relaxation
or contraction) produced by the reference substances atropine sulphate (ATR)
and acetylcholine chlorhydrate (ACH) respectively , at the standard dose of
10-6 g/ml of bath.
1.4.2. Testing the effect on the ACH
induced spasm
A spasm of the isolated rat
duodenum was induced by acethylcholine chlorhydrate (ACH) at the standard
dose of 1 mg / ml of bath (10-6 g/ml of bath). Two minutes after the administration
of ACH, during the maintenance "en plateau" of the maximum spasm,
the test dilution of B was administered and its effect on the spasm was observed
for 3 minutes.
The intensity of the effect exerted by B dilutions (decrease
or increase of ACH spasm) was expressed in percentage against the contraction
produced by ACH at the standard dose of 10-6 g/ml bath, on the same duodenum .
1.4.3. Testing the identity principle ("Aequalia aequalibus curentur")
The identity principle, on
which isopathy is based, was tested.
The therapeutic effect of B
dilutions, on the relaxation or contraction induced on normal tone by B itself
at the dilutions which had an significantly effect on normal tone, namely
1 C - 5 C and 45 C respectively (according to the test described at point 1.4.1.), was studied. The number (%) of appearances
of therapeutic effect (determined as the return to the value of normal tone
± 25 %) was established.
1.5. STATISTICAL ANALYSIS
The statistical significance
of the results was calculated by using the Student's "t" and Wilcoxon
tests for 1.4.1. and 1.4.2. tests and Chi square test for 1.4.3. test.
From the point of view of scientific
requests of the method on the isolated rat duodenum and of statistical tests
used, in correlation with the bioethical regulations of research on animals,
we established the number of the determinations for each dilution of B, at
eight duodenum fragments, from eight
rats (n = 8).
3.1. THE EFFECT OF BELLADONNA
(B) DILUTIONS ON THE NORMAL TONE OF THE RAT DUODENUM
The effect of D water was a statistically insignificant rise
in the tone. The D aqueous solutions of alcohol 70° had no effect.
The N aqueous solutions of B had the effect of
lowering the normal tone of the isolated rat duodenum, only up to 6 C (unidirectional,
monophasic effect) (Figure 1).
The D dilutions of B exerted an effect up to 45 C, i.e. relaxation between 1 C-20 C and contraction between 30 C-45 C (bidirectional, biphasic effect)
(Figure 1).
At the same degree of dilution,
the D dilutions had a more intense
effect than the N solutions (Figure 1). The statistically significant values
(p<0.05, at n = 8) are shown in Table 2.
Figure 1. Statistically significant Belladonna (B) effects % (p < 0.05),
on the normal tone of the isolated
rat duodenum
TABLE 2. Statistically significant
Belladonna (B) effects % (p < 0.05),
on the normal tone of the isolated
rat duodenum
___________________________________________________________
Dilution ± Effects
% (Means ±SEM)
___________________________________________________________
C D
N
1 -34.88 ±
0.20 -25.62 ± 0.10
5 -22.48 ±
0.50 -14.22 ± 0.30
30 +28.77 ± 0.40
0
45 +44.38 ± 0.40
0
___________________________________________________________
C = centesimal dilutions
at the 1:100 ratio
D = dynamized dilutions, according to Hahnemann's technique
N = non-dynamized dilutions, prepared by sample homogenization
p = probability by Student's t test (n = 8)
3.2. THE EFFECT OF BELLADONNA
(B) DILUTIONS ON THE SPASM INDUCED BY ACETYLCHOLINE (ACH)
The effect of D water consisted
in a slight stimulation and was statistically non-significant. The
D aqueous dilutions of alcohol 70° had no effect.
The N solutions of B had a
spasmolytic effect only up to the 9 C dilution, with a content of 6.023 x
102 ATR molecules/100 ml (unidirectional,
monophasic effect) (Figure 2).
Figure 2. Statistically significant Belladonna
(B) effects % (P < 0.05)
on the spasm induced by acetylcholin
(ACH)
D= dynalized solutions
N= non-dynamized solutions
TABLE 3. Statistically significant
Belladonna (B) effects % (p < 0.05),
on the spasm induced by acetylcholine
(ACH)
_________________________________________________________
DILUTION ± EFFECTS % (MEANS ± SEM)
_________________________________________________________
C D
N
_________________________________________________________
1 -53.30±1.10 -44.54±0.10
5 -44.30±1.20 -35.79±1.50
10 -36.55±1.30 0.00
20 -31.95±1.30 0.00
30 -25.40±0.50 0.00
40 +42.02±1.20 0.00
45 +67.60±0.10 0.00
55 -27.80±0.30 -
75 +95.45±1.20 -
80 +82.39±0.60 -
90 +68.74±1.30 -
95 +29.81±0.20 -
100 +48.92±1.10 -
120 +34.99±0.70 -
170 -41.15±1.10 -
175 -29.73±1.60 -
200 -71.12±2.00 -
________________________________________________________
C= centesimal dilutions
at the 1:100 ratio
D= dynamized dilutions, according to Hahnemann's technique
N= non-dynamized dilutions, prepared by sample homogenization
p= probability by Student's t test (n = 8)
The D dilutions of B had an
effect up to the 200 C, with a content of 6.023 x 10-381 ATR molecules/100 ml. On the scale of D dilutions 1 C - 200 C, the
effect had a sinusoidal course, i.e. the decrease of the contraction at the
1 C - 25 C, 55 C - 60 C and 160 C - 200 C dilutions and
the increase of the spasm at the 30
C - 50 C and 70 C - 150 C dilutions (bidirectional, multiphasic effect) (Figure
2).
The higher D dilutions were
more efficacious. Thus, among the D dilutions which have antagonized the ACH
spasm, the highest dilution studied, 200 C, had the most intense effect of
71.12 %. Among the dilutions which have amplified the ACH spasm, the 75 C
had the highest effect of 95.45 % (Table 3).
Conversely, at the N dilutions, the efficacy decreased with
the increase of the dilution, hence with the lowering of the concentration.
The statistically significant
values (p<0.05, at n = 8) are listed in Table 3.
Figure 3. Demonstration of the identity principle, with dynamized (D) dilutions of Belladonna (B). The therapeutic effect
of D dilutions of B, on the contraction induced by the D 45 CH dilution of
B.
n = 8, p<0.05 (Chi square
test)
3.3. THE EFFECT OF BELLADONNA
(B) DILUTIONS ACCORDING TO THE IDENTITY PRINCIPLE ("AEQUALIA AEQUALIBUS
CURENTUR") USED IN ISOPATHY
The N solutions of B had no
curative effect, according to the identity principle, on the relaxation induced
by the 1 C solution of B.
All the studied D dilutions
of B had a therapeutic effect (in all the n = 8 cases), according to the identity
principle, both on the relaxation induced by the N or D dilutions 1 C and
on the contraction induced by the D dilution 45 C of B . The prerequisite was to use for the treatment,
higher dilutions than the dilution used to induce the imbalance of tonicity.
Thus, B in the D dilutions 5, 45,
75 and 200 C reduced the hypotonia induced by B in the 1 C dilution, both
N and D. On the other hand, B in the D dilutions 55, 75 and 200 C diminished
the contraction induced by B in the D
dilution 45 C (Figure 3).
The statistically significant
results disclosed quantitative and especially qualitative differences in the
pharmacodynamics of the non-dynamized (N) and dynamized (D) preparations according
to Hahnemann's technique.
Quantitative differences :
-At the same degree of dilution,
the D dilutions had a more intense biological effect than the N dilutions.
Qualitative differences :
-The N dilutions exerted a pharmacodynamic action
only up to the limit of the molecular specificity, calculated on the basis
of Avogadro's number at the 9 C dilution, with a content of 6.023 x 101 molecules/100 ml.
In contrast with the above,
the D dilutions exerted a statistically significant biological effect also
at high dilutions, over 10 C, which mathematically and according to Avogadro's
number, do no more contain any molecule of the dissolved active substance.
-The N dilutions had unidirectional
monophasic pharmacodynamic action on the 1 C-9 C scale.
The action of D dilutions,
on the spasm, was bidirectional (double - way) multiphasic, with a sinusoidal
course,on the 1 C - 200 C scale (Figure
2).
-Belladonna (B) in N dilutions
exerted only a relaxant pharmacodynamic action on the duodenal smooth
muscle, on the 1 C - 9 C scale, an effect known in the allopathic pharmacology.
In D dilutions, on the 1 C
- 200 C scale, B exerted also a contracting pharmacodynamic action on the
duodenal smooth muscle, action which in the allopathic pharmacology is unknown
for B.
-In N solutions the effect
decreases with the increase of the dilution degree and rises with the increase
of the concentration and dose.
In D dilutions, the efficacy
rises with the increase of the dilution degree.
-Only dynamized (D) dilutions
(prepared according to Hahnemann's homeopathic technique) acted according
to the identity principle ("Aequalia
aequalibus curentur") used in isopathy.
The N solutions acted only
according to the principle of contraries ("Contraria contrariis curentur") used in allopathy.
All these qualitative differences, detected between the N and
D preparations, with respect to their biological effect, can be explained only by admitting that another form of
information, qualitatively different from that existing in the N preparations,
is also present in the D preparations (Cristea, 1991). The appearance of this
new form of information in D dilutions can be due only to dynamization by
rhythmical succussions according to Hahnemann's preparation technique.
Since in high D dilutions,
exceeding 10 C, the molecule of dissolved active substance is no more present
as an informational substrate, according to Avogadro's number, the information
in D dilutions should be looked for in another informational substrate : a
submolecular structure, a certain form of energy or, merely, a specific molecular
organization of the solvent.
The clearly superior potency and efficacy of the D preparations
demonstrate that the information conveyed by these preparations is much more
specific to the living matter. Starting from the hypothesis that according
to the laws of information processing and storage, the structural information
carried by the active atropine molecule can be conveyed, under certain condition,
by the solvent, we performed - concomitantly with the pharmacological study-,
a research by physical methods, in order to detect in the solvent, the informational
substrate of high B dilutions.
For this purpose, a calorimetric methodology was used, i.e.
the high resolution mixing calorimetry (HRMC) and the differential scanning
calorimetry (DSC), the sensitivity and reproductiveness of which in the study
of aqueous solutions has been demonstrated on a wide series of cases (Dragan,
1987). The HRMC and DSC experiments disclosed a definite organisation of water
in D dilutions, in comparison with N dilutions. Two distinct structures, with
a sinusoidal multiphasic course, in accordance with the bidirectional multiphasic
pharmacodynamic effects, were observed in D dilutions of B (Cristea et al., 1987; Dragan, 1991).
-A systematic study on bidirectional
rhythmic effect phenomenon was made (Husemann, 1992).
-The results, which we have
obtained in calorimetric studies performed on aqueous Belladonna (B) solutions, highly diluted in a centesimal geometric
progression on the 1 C - 200 C scale, are consistent also with those of other
researchers which were carried out on high dilutions and lead to the same
conclusions (Cabaner and Bastide, 1992; Endler and Schulte, 1994).
-The homeopathic drug, characterized
by infinitesimal doses and by dynamization according to Hahnemann's preparation
technique, has a marked informational character, acting in conformity with
one of the laws of information action : "Minimum quantity, maximum significance,
maximum effect" (Restian, 1971).
Cabaner, C. and
Bastide, M. (1992) Isothermal microcalorimetric analysis of homeopathic dilutions,
6th GIRI Meeting, Paris, France
Cristea, A. (1985)
Farmacologia cibernetica (Cybernetical Pharmacology), Nat.Conf. of Cybernetics, Bucharest, Romania,
3.
Cristea, A.,
Dragan, G. and Darie, V. (1987) Potency of pharmacodynamic action and structure
of highly diluted aqueous solutions of Belladonna, Internat. Conf. on Water
and Ions in Biological Systems, Bucharest, Romania, 4.
Cristea, A.,
Nicula, S. and Darie, V. (1991) Pharmacodynamic effects of very high dilutions
of Belladonna on the isolated rat duodenum, 5th
GIRI Meeting, Paris, France.
Cristea, A. (1991)
Information - A hidden parameter of the homeopathic preparation, Nat Conf. of Homeopathy, Bucharest, Romania,
2.
Cristea, A. (1991)
Pour une pharmacologie cybernétique
et informationnelle, Cybernetica 2, 85-93.
Cristea, A. (1994)
Homeopatia in lumina unei teorii informationale a dozelor (The homeopathy - in the light of the informational
theory of doses), Abstr. Nat. Conf.
of Homeopathy, Sibiu, Romania.
Cristea, A. (1994)
Farmacologia informationala a dozelor, 10-th Nat. Congress of Pharmacy, Cluj-Napoca, Romania.
Cristea, A. (1995)
Conceptia informationala in farmacologie, Farmacia, 3-4, 1-14.
Cristea, A. (1996)
For an informational theory of dosage. Illustration from cell and molecular
biology and pharmacology, Cybernetica
, (in press).
Dragan, G. (1987)
Comparative study on molecular associations in solid and liquid phases of
aqueous solutions. Presentation of high resolution mixing calorimetry (HRMC)
and the differential scanning calorimetry (DSC), Acta Polymerica 4, 211-220.
Dragan, G. and
Darie, V. (1991) Composite structure of homeopathic solutions of Belladonna
as revealed by calorimetric measurements, Abstr.
5-th GIRI Meeting, Paris, France.
Endler, P.C.
and Schulte, J. (Eds), (1994) Ultra high dilution. Physiology and physics,
Kluwer Acad. Publ., Dordrecht.
Husemann, F.
(1992) Rhythmusphanomene beim wirksamkeitsnachweiss potenzierter heilmittel
- Nachgewiesen von Kolisko (1923) bis Cristea (1991), Der Merkurstab 2, 73-90.
Restian, A. (1971)
Informatie minima, semnificatie maxima, Rev. de Filosofie 9, 1229-1235.
Restian, A. (1981)
Homo Ciberneticus, St. Enc.Publisher,
Bucharest, Romania.
Restian, A. (1989)
Integronica, Ed. St., Bucharest,
Romania.
Turner, R.A.
(1965) Screening methods in pharmacology,
Acad. Press, New York.