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Effectiveness of ultra high diluted arsenic is a function of succussion number as evidenced by wheat germination test and droplet evaporation method

Authors:
Lucietta Betti at University of Bologna
Lucietta Betti
Grazia Trebbi at University of Bologna
Grazia Trebbi
Maria Olga Kokornaczyk at Society for Cancer Research, Arlesheim, Switzerland
Maria Olga Kokornaczyk
  • Society for Cancer Research, Arlesheim, Switzerland
Daniele Nani at IMAGO PROXIMA CENTAURI Medical Association
Daniele Nani
  • IMAGO PROXIMA CENTAURI Medical Association
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Int J High Dilution Res 2013; 12(44):127-128
Proceedings of the XXVII GIRI Symposium; 2013 Sep03-04; Bern (Switzerland)
127
Effectiveness of ultra high diluted arsenic is a
function of succussion number as evidenced by wheat
germination test and droplet evaporation method
Lucietta Betti1, Grazia Trebbi1, Maria Olga Kokornaczyk1, Daniele Nani2,
Maurizio Peruzzi3, Maurizio Brizzi4
1 Department of Agricultural Sciences, University of Bologna
2 Italian Society of Anthroposophical Medicine
3 Association for Sensitive Crystallization
4 Department of Statistical Sciences, University of Bologna
Background: In the preparation of ultra high dilutions (UHD) each dilution step is followed by a succussion.
Whereas the physical basis of succussion have been well studied [1], there are only few papers regarding the
effects of succussion number (NS) [2]. In previous studies [3, 4] we showed that arsenic at UHD applied on
arsenic stressed wheat seeds stimulates the germination rate.
Aims: Investigate whether NS applied between the dilution-steps influences the UHD effectiveness.
Methodology:
Plant material and stress treatment: Wheat seeds (Triticum aestivum L.) of cv. Pandas were stressed by 30min
of poisoning with 5mM As2O3 aqueous solution [3, 4].
Classes of treatment: Undiluted and unsuccussed H2O (C), H2O and As2O3 both at the 45th decimal
dilution/succussion (W45x and As45x, respectively). The succussion was handmade and NS differed for each
treatment (NS= 4, 8, 16, 32, 40, 70, 100).
Biological model: In each Petri dish 36 seeds were placed and watered with 20ml of treatment. After 96 hours
non-germinated seeds were counted. The experiment was repeated twice and each one consisted in 6 Petri
dishes per treatment and 6 trials. The results were analyzed by Poisson test [3].
Droplet evaporation method: The droplet patterns [5] were prepared out of the stressed wheat samples treated
either with As45x, W45x (NS=8, 32, 70) or C. The experiment was performed in a 3 day repetition, 3 replicates
per day, 5 droplets per replicate. The patterns were evaluated by means of the ImageJ software for their local
connected fractal dimension (LCFD) values. The data was analyzed by means of the analysis of variance.
Multiple mean comparison was carried out by Turkey’s HSD test.
Int J High Dilution Res 2013; 12(44):127-128
Proceedings of the XXVII GIRI Symposium; 2013 Sep03-04; Bern (Switzerland)
128
Results: NS strongly influenced both the biological effectiveness of the treatment (i) and the LCFD of the
DEM patterns (ii). In particular: (i) the rate of germinated seeds significantly increased vs. C following
treatments with NS≥32 for both As45x and W45x; (ii) all As45x (NS=8, 32, 70) and W45x with NS=70
significantly increased the LCFD of DEM patterns vs. C. Both approaches showed that UHD effects increase
with the growing NS, even if there was a non complete correspondence of the significance levels.
Conclusions: Our results show that NS is an important parameter influencing the UHD effectiveness.
Keywords: Succussion number, ultra high dilutions, arsenic, wheat germination, droplet evaporation method
References
1. Molski M. Quasi-quantum model of potentization. Homeopathy 2011; 100: 259-263.
2. Anick DJ. The octave potencies convention: a mathematical model of dilution and succussion. Homeopathy
2007; 96(3): 202-208.
3. Brizzi M, Elia V, Trebbi G, Nani D, Peruzzi M, Betti L The efficacy of ultramolecular aqueous dilutions on a
wheat germination model as a function of heat and aging-time. eCAM. doi:10.1093/ecam/nep217. 2011
4. Betti L, Elia W, Napoli, E, Trebbi G, Zurla M, Nani D, Peruzzi M, Brizzi M. Biological effects and physico-
chemical properties of extremely diluted aqueous solution as a function of aging-time. Front Life Sci. 2012: 1-
10.
5. Kokornaczyk MO, Dinelli G, Marotti I, Benedettelli S, Nani D, Betti L. Self-organized crystallization
patterns from evaporating droplets of common wheat grain leakages as a potential tool for quality analysis.
The Scientific World Journal 2011; 11:1712–1725.
Licensed to GIRI
Support: Laboratoires Boiron
Conflict of interest: authors declare there is no conflict of interest
Received: 16 June 2013; Revised: 12 August 2013; Published: 30 September 2013.
Correspondence author: Lucietta Betti, lucietta.betti@unibo.it
How to cite this article: Betti L, Trebbi G, Kokornaczyk MO, Peruzzi DNM, Brizzi M. Effectiveness of ultra high diluted
arsenic is a function of succussion number as evidenced by wheat germination test and droplet evaporation method. Int J
High Dilution Res [online]. 2013 [cited YYYY Month DD]; 12(44):127-128. Proceedings of the XXVII GIRI Symposium;
2013 Sep 03-04; Bern (Switzerland). GIRI; 2013; Available from:
http://www.feg.unesp.br/~ojs/index.php/ijhdr/article/view/669/664
... It has been shown in two different plant models that UHD treatments have modified the polycrystalline structures obtained from evaporating solutions prepared from treated plants vs. controls. 15e17 In particular, in 15,16 wheat seeds, previously stressed with a ponderal concentration of arsenic trioxide (As 2 O 3 ), were watered with arsenic trioxide 45th decimal dilution (As45x) or water as control; subsequently, the wheat seed leakage were analyzed by means of the droplet evaporation method (DEM). 18,19 During evaporation polycrystalline structures are formed, which complexity degree, measured by means of the local connected fractal dimension (LCFD), differed significantly between the As45x treated seeds and controls. ...
... As reported in, 16 the LCFD of the polycrystalline structures showed to be also sensitive towards the number of strokes applied following each dilution step during the As45x preparation: As 2 O 3 pretreated seeds watered with As45x treatments prepared with growing stroke numbers created polycrystalline structures characterized by significantly higher LCFD values. Furthermore, positive correlations found between the LCFD values and germination tests indicate that LCFD might be sensitive to the seeds vitality. ...
... In so far this result conforms to our previous findings where polycrystalline structures obtained by means of DEM showed to be sensitive towards As45x applied on a wheat seed model. 15,16 In the present study we did not compare the DEM results with any other analysis e.g. germination test, and so we can only state that Zm induced a change to the model, which lowered the LCFD and the area of the polycrystalline structures. ...
Polycrystalline structures formed in evaporating droplets as a parameter to test the action of Zincum metallicum 30c action in a wheat seed model
Article
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Background: Polycrystalline structures formed inside evaporating droplets of different biological fluids have been shown sensitive towards various influences, including ultra high dilutions (UHDs), representing so a new approach potentially useful for basic research in homeopathy. In the present study we tested on a wheat seed model Zincum metallicum 30c efficacy versus lactose 30c and water. Materials and methods: Stressed and non-stressed wheat seeds were watered with the three treatments. Seed-leakage droplets were evaporated and the polycrystalline structures formed inside the droplet residues were analyzed for their local connected fractal dimensions (LCFDs) (measure of complexity) using the software ImageJ. Results: We have found significant differences in LCFD values of polycrystalline structures obtained from stressed seeds following the treatments (p<0.0001); Zincum metallicum 30c lowered the structures' complexity compared to lactose 30c and water. In non-stressed seeds no significant differences were found. Conclusions: The droplet evaporation method (DEM) might represent a potentially useful tool in basic research in homeopathy. Furthermore our results suggest a sensitization of the stressed model towards the treatment action, which is conforming to previous findings.
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... Succussions are rapid, repetitive up-and-down agitations via vertical strokes of the borosilicate glassware containing the solution against a hard elastic surface or vortexing. Skeptics have focused on the dilutions beyond Avogadro's number for bulk form source materials and ignored the trituration and succussion steps as well as their potential contributions to the final product [18]. Nanotechnology researchers have begun to recognize the overlaps between the traditional crude mechanical attrition manufacturing techniques of homeopathy and top down approaches for making nanostructures from insoluble source materials in modern nanotechnology. ...
Nanoparticle Characterization of Traditional Homeopathically-Manufactured Silver (Argentum Metallicum) Medicines and Placebo Controls
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This exploratory study characterized nanoparticles (NPs) in homeopathically-prepared silver medicine (Argentum Metallicum), succussed and unsuccussed control solutions. Methods included nanoparticle tracking analysis (NTA), zeta potentials, inductively-coupled plasma mass spectroscopy (ICP-MS) for silver, and ultraviolet visible spectroscopy (UV-vis). Homeopathy is a 200 year-old traditional medical system using mineral, plant, and animal sources to treat patients. Debate over the efficacy of homeopathic medicines (HMs) stems from its unique manufacturing methods involving dry milling in lactose and/or serial dilutions and succussions (intense agitation) in water or water-ethanol solvent. The present study examined commercially-made, randomized and blinded verum Argentum Metallicum HMs and succussed solvent controls at 6C, 30C, and 200C potencies, and unsuccussed solvent (95% v/v ethanol) in corkstoppered glass vials. NTA revealed >2 × 108 NPs/ml in 20 of 21 samples, with extensive polydispersity. Verum NP characteristics differed significantly from controls: (a) Although Verum 200C exhibited the highest NP concentrations and exemplar transmission electron microscopy of oneVerum 30C sample showed scattered silver-like dark NPs embedded in an organic matrix, ICP-MS could not reliably detect elemental silver at 1 ppb (lower limits of instrument detection); (b) verum NPs were significantly larger sized across potencies (91 nm) versus unsuccussed controls (53 nm); (c) zeta potentials of verum NPs (mean -44 mV) were significantly more negative than the unsuccussed controls (mean -20 mV); (d) succussed controls were intermediate in NP concentrations and sizes between verums and unsuccussed controls; (e) All samples showed UV-vis absorbance peaks near 200-350 nm wavelengths, consistent with organic cork extract. Taken together, the findings suggest polydisperse heterogeneous nanohybrid mixtures of silver, silica shell/ silver core, silica NPs, and perhaps cork exosomes, self-assembled and surface-stabilized by silica coatings, organic plant extract (cork), and/or lactose in verums. Glassware-derived silica from succussions may contribute NPs and coatings for nonlinear source signal amplification to initiate clinical effects.
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The usage of ultra dilutions in agriculture
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Effectiveness of ultra high diluted arsenic is a function of succussion number as evidenced by wheat germination test and droplet evaporation method
Article
Full-text available
  • Sep 2013
Background: In the preparation of ultra high dilutions (UHD) each dilution step is followed by a succussion. Whereas the physical basis of succussion have been well studied [1], there are only few papers regarding the effects of succussion number (NS) [2]. In previous studies [3, 4] we showed that arsenic at UHD applied on arsenic stressed wheat seeds stimulates the germination rate. Aims: Investigate whether NS applied between the dilution-steps influences the UHD effectiveness. Methodology: Plant material and stress treatment: Wheat seeds (Triticum aestivum L.) of cv. Pandas were stressed by 30min of poisoning with 5mM As2O3 aqueous solution [3, 4]. Classes of treatment: Undiluted and unsuccussed H2O (C), H2O and As2O3 both at the 45th decimal dilution/succussion (W45x and As45x, respectively). The succussion was handmade and NS differed for each treatment (NS= 4, 8, 16, 32, 40, 70, 100). Biological model: In each Petri dish 36 seeds were placed and watered with 20ml of treatment. After 96 hours non-germinated seeds were counted. The experiment was repeated twice and each one consisted in 6 Petri dishes per treatment and 6 trials. The results were analyzed by Poisson test [3]. Droplet evaporation method: The droplet patterns [5] were prepared out of the stressed wheat samples treated either with As45x, W45x (NS=8, 32, 70) or C. The experiment was performed in a 3 day repetition, 3 replicates per day, 5 droplets per replicate. The patterns were evaluated by means of the ImageJ software for their local connected fractal dimension (LCFD) values. The data was analyzed by means of the analysis of variance. Multiple mean comparison was carried out by Turkey’s HSD test. Results: NS strongly influenced both the biological effectiveness of the treatment (i) and the LCFD of the DEM patterns (ii). In particular: (i) the rate of germinated seeds significantly increased vs. C following treatments with NS≥32 for both As45x and W45x; (ii) all As45x (NS=8, 32, 70) and W45x with NS=70 significantly increased the LCFD of DEM patterns vs. C. Both approaches showed that UHD effects increase with the growing NS, even if there was a non complete correspondence of the significance levels. Conclusions: Our results show that NS is an important parameter influencing the UHD effectiveness.
View
Droplet evaporation method as a new potential approach for highlighting the effectiveness of ultra high dilutions
Article
Full-text available
  • Apr 2014
Objective This study sought to verify whether the droplet evaporation method (DEM) can be applied to assess the effectiveness of ultra-high dilutions (UHDs). We studied the shape characteristics of the polycrystalline structures formed during droplet evaporation of wheat seed leakages. Methods The experimental protocol tested both unstressed seeds and seeds stressed with arsenic trioxide 5 mM, treated with either ultra-high dilutions of the same stressor substance, or with water as a control. The experimental groups were analyzed by DEM and in vitro growth tests. DEM patterns were evaluated for their local connected fractal dimension (measure of complexity) and fluctuating asymmetry (measure of symmetry exactness). Results Treatment with arsenic at UHD of both stressed and non-stressed seeds increased the local connected fractal dimension levels and bilateral symmetry exactness values in the polycrystalline structures, as compared to the water treatment. The results of in vitro growth tests revealed a stimulating effect of arsenic at UHD vs. control, and a correlation between the changes in growth rate and the crystallographic values of the polycrystalline structures was observed. Conclusions The results indicate that polycrystalline structures are sensitive to UHDs, and so for the first time provide grounds for the use of DEM as a new tool for testing UHD effectiveness. DEM could find application as a treatment pre-selection tool, or to monitor sample conditions during treatment. Moreover, when applied to biological liquids (such as saliva, blood, blood serum, etc), DEM might provide information about UHD effectiveness on human and animal health.
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Effectiveness of ultra high diluted arsenic is a function of succussion number as evidenced by wheat germination test and droplet evaporation method
Article
Full-text available
  • Sep 2013
Background: In the preparation of ultra high dilutions (UHD) each dilution step is followed by a succussion. Whereas the physical basis of succussion have been well studied [1], there are only few papers regarding the effects of succussion number (NS) [2]. In previous studies [3, 4] we showed that arsenic at UHD applied on arsenic stressed wheat seeds stimulates the germination rate. Aims: Investigate whether NS applied between the dilution-steps influences the UHD effectiveness. Methodology: Plant material and stress treatment: Wheat seeds (Triticum aestivum L.) of cv. Pandas were stressed by 30min of poisoning with 5mM As2O3 aqueous solution [3, 4]. Classes of treatment: Undiluted and unsuccussed H2O (C), H2O and As2O3 both at the 45th decimal dilution/succussion (W45x and As45x, respectively). The succussion was handmade and NS differed for each treatment (NS= 4, 8, 16, 32, 40, 70, 100). Biological model: In each Petri dish 36 seeds were placed and watered with 20ml of treatment. After 96 hours non-germinated seeds were counted. The experiment was repeated twice and each one consisted in 6 Petri dishes per treatment and 6 trials. The results were analyzed by Poisson test [3]. Droplet evaporation method: The droplet patterns [5] were prepared out of the stressed wheat samples treated either with As45x, W45x (NS=8, 32, 70) or C. The experiment was performed in a 3 day repetition, 3 replicates per day, 5 droplets per replicate. The patterns were evaluated by means of the ImageJ software for their local connected fractal dimension (LCFD) values. The data was analyzed by means of the analysis of variance. Multiple mean comparison was carried out by Turkey’s HSD test. Results: NS strongly influenced both the biological effectiveness of the treatment (i) and the LCFD of the DEM patterns (ii). In particular: (i) the rate of germinated seeds significantly increased vs. C following treatments with NS≥32 for both As45x and W45x; (ii) all As45x (NS=8, 32, 70) and W45x with NS=70 significantly increased the LCFD of DEM patterns vs. C. Both approaches showed that UHD effects increase with the growing NS, even if there was a non complete correspondence of the significance levels. Conclusions: Our results show that NS is an important parameter influencing the UHD effectiveness.
View
Self-Organized Crystallization Patterns from Evaporating Droplets of Common Wheat Grain Leakages as a Potential Tool for Quality Analysis
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Biological effects and physico-chemical properties of extremely diluted aqueous solutions as a function of aging-time
Article
Full-text available
  • Apr 2012
This study concerns the biological effects and physico-chemical variations of extremely diluted solutions (EDS) as a function of aging-time. The biological efficacy of As2O3 at the 45th decimal dilution/succussion (As 45×) was tested in a wheat germination model. Ten trials were carried out from 0 to 12 months after treatment preparation, using wheat seeds (Triticum aestivum L.) of the Pandas variety. Seeds were pre-treated by poisoning with 0.1% As2O3 solution to reduce germination, to allow a better evaluation of treatment effects. The outcome variable was the number of non-germinated seeds after 96 h. The As 45× treatment aged for less than three months did not show a significant effect on wheat germination, whereas when aged for longer (3-12 months) the effect became significant. Concerning the physico-chemical characteristics, specific conductivity of As 45×aged from 0 to 12 months was measured, using nine samples for each date. The results showed a clear increasing time trend of specific conductivity, more evident when considering the last three measurements, which correspond to more than six months of aging. The physico-chemical behavior of EDS strongly supports the significant biological effects observed in a wheat model.
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The Efficacy of Ultramolecular Aqueous Dilutions on a Wheat Germination Model as a Function of Heat and Aging-Time
Article
Full-text available
  • Feb 2011
  • EVID-BASED COMPL ALT
This study evaluates the effects of temperature and aging on the efficacy of As(2)O(3) at the 45th decimal potency in a wheat germination model, compared against a control and potentized H(2)O 45×. Each treatment-temperature combination was tested on seeds (Triticum aestivum L.) of Pandas variety, using six Petri dishes (33 seeds/dish) per trial, performing eight trials. Seeds were pre-treated by poisoning with 0.1% As(2)O(3) solution to reduce germination, to allow a better evaluation of homeopathic treatment effects. The outcome variable was the number of non-germinated seeds after 96 h. Temperature effect was investigated by heating each treatment in a water bath for 30 min (at 20, 40 or 70°C), or for 5 min (at 100°C), and that of aging by dividing experimental data, collected over a period of nearly five months, into two groups: early and late experiments. Results seem to show that the efficacy of As(2)O(3) 45× is unaltered at 20 and 40°C, increases at 70°C and decreases at 100°C. As regards aging, a notable difference was found between early trials, with no significant efficacy, and late trials, where As(2)O(3) 45× exhibits a repeated significant effect versus control, except at 100°C. A reduction in variability was observed for As(2)O(3) 45× at 20°C versus control, confirming the findings of previous work. The main conclusion suggested by this experiment is that the efficacy of As(2)O(3) 45× on wheat germination may be influenced by heating degree and seems to have an increasing trend as a function of aging.
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Analytical time-dependent functions describing the change of the concentration of the solvent S(t) and the homeopathic active substance A(t) during decimal and centesimal dilution are derived. The function S(t) is a special case of the West-Brown-Enquist curve describing ontogenic growth, the increase in concentration of the solvent during potentization resembles the growth of biological systems. It is demonstrated that the macroscopic S(t) function is the ground state solution of the microscopic non-local Horodecki-Feinberg equation for the time-dependent Hulthèn potential at the critical screening. In consequence potentization belongs to the class of quasi-quantum phenomena playing an important role both in biological systems and homeopathy. A comparison of the results predicted by the model proposed with the results of experiments on delayed luminescence of a homeopathic medicine is made.
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The octave potencies convention: a mathematical model of dilution and succussion
Article
  • Aug 2007
Several hypothesized explanations for homeopathy posit that remedies contain a concentration of discrete information-carrying units, such as water clusters, nano-bubbles, or silicates. For any such explanation to be sustainable, dilution must reduce and succussion must restore the concentration of these units. Succussion can be modeled by a logistic equation, which leads to mathematical relationships involving the maximum concentration, the average growth of information-carrying units rate per succussion stroke, the number of succussion strokes, and the dilution factor (x, c, or LM). When multiple species of information-carrying units are present, the fastest-growing species will eventually come to dominate, as the potency is increased. An analogy is explored between iterated cycles dilution and succussion, in making homeopathic remedies, and iterated cycles of reseeding and growth, in bacterial cultures. Drawing on this analogy, the active ingredients in low and medium potency remedies may be present at early dilutions but only gradually come to 'dominate', while high potencies may develop from the occurrence of low-probability but faster-growing 'mutations.' Conclusions from this model include: 'x' and 'c' potencies are best compared by the amount of dilution, not the amount of succussion; the minimum number of succussion strokes needed per cycle is proportional to the logarithm of the dilution factor; and a plausible interpretation of why potencies at approximately regular ratios are traditionally used (the octave potencies convention).
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Available from: http://www.feg.unesp.br/~ojs/index
  • Jan 2013
  • 664
GIRI; 2013; Available from: http://www.feg.unesp.br/~ojs/index.php/ijhdr/article/view/669/664
Biological effects and physicochemical properties of extremely diluted aqueous solution as a function of aging-time
  • 1-10
  • L Betti
  • Elia W Napoli
  • E Trebbi
  • G Zurla
  • M Nani
  • D Peruzzi
  • M Brizzi
Betti L, Elia W, Napoli, E, Trebbi G, Zurla M, Nani D, Peruzzi M, Brizzi M. Biological effects and physicochemical properties of extremely diluted aqueous solution as a function of aging-time. Front Life Sci. 2012: 1-10.