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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:
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; 11:17121725.
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. ...
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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.
... 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. ...
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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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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.
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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
  • 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.