Number of succussion strokes affects effectiveness of ultra-high-diluted arsenic on in vitro wheat germination and polycrystalline structures obtained by droplet evaporation method.

OBJECTIVES: The aim of this study is to investigate whether the number of succussion strokes applied after each dilution step when preparing the homeopathic treatments influences the effectiveness of ultra-high-diluted (UHD) arsenic trioxide at the 45th decimal dilution/dynamization (As2O3 45x). DESIGN: Wheat seeds, previously stressed with ponderal As2O3, were treated with: As2O3 45x, H2O 45x (dynamized control), or pure water (negative control). The succussion was done manually, and various succussion durations (numbers of strokes) were tested for each treatment. Treatment effectiveness was tested blind using the in vitro germination test and the droplet evaporation method (DEM). Data were processed by the Poisson test (germination test) and by two-way analysis of variance (DEM). MAIN OUTCOME MEASURES: We evaluated both the in vitro germination rate, by counting the non-germinated seeds, and the complexity of polycrystalline structures (PCS) (local connected fractal dimension (LCFD)) obtained by evaporating leakage droplets from stressed seeds that had been watered with the different treatments. RESULTS: We observed a highly significant increase in germination rate when the number of strokes (NS) was ≥32 for both As2O3 45x and H2O 45x, and a significant increase in the LCFD of PCS for As2O3 45x when the NS was ≥32 and for H2O 45x when it was 70. CONCLUSIONS: Both experimental approaches showed increased effectiveness for treatments prepared with a higher number of succussion strokes. These results indicate that succussion may have an important influence on treatment effectiveness, and so highlight the need for further research.

Basic research in homeopathy and ultra-high dilutions: what progress is being made?

This report summarises the latest research developments in the field of high dilutions and homeopathy, as presented at the GIRI symposium of the leading international organisation of scientists in this field, in Florence, Italy in September 2012. The scientific community's early scepticism concerning the possible biological and pharmacological activity of highly diluted solutions, is giving way to a more open-minded attitude that no longer obstructs critical and experimental investigations in this emerging field of biomedicine.

The efficacy of ultramolecular aqueous dilutions on a wheat germination model as a function of heat and aging-time.

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.

Use of homeopathic preparations in experimental studies with abiotically stressed plants.

BACKGROUND: Experimental research on the effects of homeopathic treatments on impaired plants was last reviewed in 1990. OBJECTIVES: To compile a systematic review of the existing literature on basic research in homeopathy with abiotically stressed plants using predefined criteria. METHODS: The literature search was carried out on publications that reported experiments on homeopathy using abiotically stressed whole plants, seeds, plant parts and cells from 1920 to 2010. Outcomes had to be measured by established procedures and statistically evaluated. Using of a Manuscript Information Score (MIS) we identified those publications that provided sufficient information for proper interpretation (MIS≥5). A further evaluation was based on the use of adequate controls to investigate specific effects of homeopathic preparations and on the use of systematic negative control experiments. RESULTS: A total of 34 publications with abiotically stressed plants was identified, published between 1965 and 2010. The 34 publications described a total of 37 experimental studies. Twenty-two studies included statistics, 13 had a MIS≥5, 8 were identified with adequate controls and 4 with negative control experiments. Significant and reproducible effects with decimal and centesimal potencies were found, including dilution levels beyond Avogadro's number. One experimental model was independently assessed by another research team and yielded inverted results compared to the original trial. CONCLUSIONS: Abiotically stressed plant models seem to be a useful approach to investigate homeopathic basic research questions, but more experimentation and especially more independent replication trials are needed. Systematic negative control experiments should be implemented on a routine basis to exclude false-positive results.

A review of three simple plant models and corresponding statistical tools for basic research in homeopathy.

In this paper, we review three simple plant models (wheat seed germination, wheat seedling growth, and infected tobacco plants) that we set up during a series of experiments carried out from 1991 to 2009 in order to study the effects of homeopathic treatments. We will also describe the set of statistical tools applied in the different models. The homeopathic treatment used in our experiments was arsenic trioxide (As2O3) diluted in a decimal scale and dynamized. Since the most significant results were achieved with the 45th decimal potency, both for As2O3 (As 45x) and water (W 45x), we here report a brief summary of these results. The statistical analysis was performed by using parametric and nonparametric tests, and Poisson distribution had an essential role when dealing with germination experiments. Finally, we will describe some results related to the changes in variability, which seems to be one of the targets of homeopathic treatment effect.

Use of homeopathic preparations in experimental studies with healthy plants.

BACKGROUND: The last comprehensive review of experimental research on effects of homeopathic treatments on plants was published in 1984, and lacked formal predefined criteria to assess study quality. Since then several new studies with more advanced methods have been published. OBJECTIVES: To compile a review of the literature on basic research in homeopathy with healthy plants with particular reference to studies investigating specific effects of homeopathic remedies. METHODS: The literature search included English, French, German, Italian, Portuguese and Spanish publications from 1920 to April 2009, using predefined selection criteria. We included experiments with healthy whole plants, seeds, plant parts and cells. The outcomes had to be measured by established procedures and statistically evaluated. We developed a Manuscript Information Score (MIS) and included only publications which provided enough information for proper interpretation (MIS>or=5). A formalised Study Methods Evaluation Procedure (SMEP) was used to evaluate these studies, and the subgroup of studies with adequate controls to identify specific effects. RESULTS: A total of 86 studies in 79 publications was identified, 43 studies included statistics, 29 had MIS>or=5, and 15 studies investigated the specificity of homeopathic preparations. Specific effects of decimal, centesimal and fifty millesimal potencies were found including dilution levels far beyond the Avogadro number. In consecutive series of potencies only some of the tested potencies showed effects. There were many individual studies with diverse methods and very few reproduction trials. CONCLUSIONS: Healthy plant models seem an useful approach to investigate basic research questions about the specificity of homeopathic preparations. More investigations with more advanced methods are recommended, especially in the sectors of potentisation techniques, effective potency levels and conditions for reproducibility. Systematic negative control experiments should become a routine procedure to control the stability of the experimental systems.

Use of homeopathic preparations in phytopathological models and in field trials: a critical review.

BACKGROUND: The literature on the applications of homeopathy for controlling plant diseases in both plant pathological models and field trials was first reviewed by Scofield in 1984. No other review on homeopathy in plant pathology has been published since, though much new research has subsequently been carried out using more advanced methods. OBJECTIVES: To conduct an up-to-date review of the existing literature on basic research in homeopathy using phytopathological models and experiments in the field. METHODS: A literature search was carried out on publications from 1969 to 2009, for papers that reported experiments on homeopathy using phytopathological models (in vitro and in planta) and field trials. The selected papers were summarized and analysed on the basis of a Manuscript Information Score (MIS) to identify those that provided sufficient information for proper interpretation (MIS>or=5). These were then evaluated using a Study Methods Evaluation Procedure (SMEP). RESULTS: A total of 44 publications on phytopathological models were identified: 19 papers with statistics, 6 studies with MIS>or=5. Publications on field were 9, 6 with MIS>or=5. In general, significant and reproducible effects with decimal and centesimal potencies were found, including dilution levels beyond the Avogadro's number. CONCLUSIONS: The prospects for homeopathic treatments in agriculture are promising, but much more experimentation is needed, especially at a field level, and on potentisation techniques, effective potency levels and conditions for reproducibility. Phytopathological models may also develop into useful tools to answer pharmaceutical questions.

Formation of Self-organized Structures in Drying Droplets: a Promising Analytical Tool for Homeopathy Basic Research

The droplet evaporation method (DEM) is based on the evaporation-induced pattern formation in droplets and is applied mainly for medical diagnosis[1].Here, we present aseries of experiments performed by our team showing DEMs potential also forhomeopathy basic research, in particular, for the investigation of(i) low potencies, (ii) low potency complexes (physical model), and (iii) the action of high potencies (plant-based model).Methods:(i) DEM differentiated significantly between Luffa, Baptisia, Echinacea, and Spongiauntil 4x[2]. Furthermore, the patterns varied in function of the numberof succussion strokes (0, 10, or 100) applied during potentization[3]. The performance of chaotic succussions vs. laminar flow vs. slight mixing during the potentization of Viscum album quercus3x influenced the DEM patterns; the chaotic succussions reduced, whereas laminar flow enhanced the patterns complexity vs. the unsuccussed control.(ii) The addition of Mercurius bijodatus9x to Luffa4x changed significantly the DEM patterns, even if the material quantity present in the 9x potency lied far beyond that of ultrapure water.(iii) Leakages obtained by placing healthy or arsenic-damaged wheat-seeds into Arsenicum album45x orheat-damaged intoZincum metallicum30c vs. water created significantly different DEM structures [4, 5]. Results:The damaged seeds put into the potency created structures characterized by a higher complexity than those obtained from damaged seeds put into control water. Furthermore, the potency action seemed to increase with rising numbers ofsuccussion strokes applied during potentization,ascould be shown by means of DEM patterns and germination rate using the same wheat-seed model[6].In all our studies, the pattern evaluation was computerized (texture and fractal analysis performed by means of ImageJ) or based on deep-learning algorithms and the robustness of the experimental system was checked by means of systematic control experiments.Conclusion:DEM together with other similarmethods has also been reviewed by our team for what concerns theapplication in homeopathy basic research[7].

Weak extremely high frequency microwaves affect pollen-tube emergence and growth in kiwifruit: pollen grain irradiation and water-mediated effects.

OBJECTIVE: This study was designed to evaluate the effects of weak-intensity extremely high frequency (EHF) microwaves in a model system-the plant organism pollen grain-lacking the placebo effect, available in large populations, to ensure accurate statistical analysis, and whose sensitivity is closely relevant to animal and human biology. DESIGN: This study was blinded using an in vitro pollen germination technique. SUBJECTS AND STUDY INTERVENTIONS: Pollen of kiwifruit (Actinidia deliciosa) was either directly irradiated or grown in a medium prepared with irradiated water, using a CromoStim 2000, (PromoPharma, Republic of San Marino) designed for EHF microwave resonance therapy (MRT). It produces weak intensity EHF radiations (40-78 GHz), either continuous wave (cw) or modulated, at a 10 Hz-frequency, with infrared (IR) carried to 635-950 nm, and with an impedance (IPD) of 10(-21) W/Hz cm(2) and a power supply from 0 to 20 mW. OUTCOME MEASUREMENTS: Pollen-tube emergence was expressed as a percent of grains producing a tube and tube elongation was measured at 4 hours of incubation by a turbidimetric assay (A(500)) of cultures, expressed as the net absorbance increase over time 0. RESULTS: At days 2 and 4 during aging, both percent of germination and tube growth significantly and consistently improved over controls in kiwifruit pollen grains irradiated for 30 minutes at day 0 at 10 Hz frequency with the CromoStim 2000. Highly significant effects, either stimulant or inhibitory, were also observed on kiwifruit pollen (stressed or not) growing in a medium prepared with water previously irradiated either cw or modulated. Irradiated water affected pollen germination immediately and even after several days following EHF treatment. CONCLUSIONS: Either direct or indirect EHF irradiation performed by the CromoStim 2000 is effective on pollen growth processes. In both cases, water seemed to play a primary role. According to the quantum electrodynamical coherence theory, our work could also have implications for homeopathy, suggesting a key to explain the efficacy of high dilutions and succussion procedures.

Transcriptome Profiling of Wheat Seedlings following Treatment with Ultrahigh Diluted Arsenic Trioxide.

Plant systems are useful research tools to address basic questions in homeopathy as they make it possible to overcome some of the drawbacks encountered in clinical trials (placebo effect, ethical issues, duration of the experiment, and high costs). The objective of the present study was to test the hypothesis whether 7-day-old wheat seedlings, grown from seeds either poisoned with a sublethal dose of As2O3 or unpoisoned, showed different significant gene expression profiles after the application of ultrahigh diluted As2O3 (beyond Avogadro's limit) compared to water (control). The results provided evidence for a strong gene modulating effect of ultrahigh diluted As2O3 in seedlings grown from poisoned seeds: a massive reduction of gene expression levels to values comparable to those of the control group was observed for several functional classes of genes. A plausible hypothesis is that ultrahigh diluted As2O3 treatment induced a reequilibration of those genes that were upregulated during the oxidative stress by bringing the expression levels closer to the basal levels normally occurring in the control plants.