Lucietta Betti

Effect of high dilutions of Arsenicum album on wheat seedlings from seed poisoned with the same substance

Abstract A blind laboratory experiment was carried out to show the effect of a 45x potency of Arsenicum album (As2O3) on wheat seedlings whose seeds had been previously poisoned with a material dose of the same substance. The effect of the homoeopathic treatment on stem growth was statistically significant. The experimental results were matched with a previous study concerning homoeopathic treatment in the same species.

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 Avogadros 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.

A Biostatistical Insight into the As2O3 High Dilution Effects on the Rate and Variability of Wheat Seedling Growth

Background: Most criticism of homeopathy concerns the lack of scientific bases and theoretical models. Fundamental research could make important contributions to our understanding of the mechanisms of action of homeopathic treatments. Plant-based bioassays are suitable for basic research - lacking the placebo effect and ensuring large data samples for structured statistical analyses. Objective: The aim of this study was to reproduce a previous experiment on the effects of arsenic trioxide (As2O3) high dilutions on wheat seedling growth in order to verify whether the same significant results could be obtained working in a different place and with a different experimental team. A further goal was to investigate high dilution effects on variability. Material and Methods: A structured experiment was performed blind over 9 weeks, using wheat seeds previously stressed with a sublethal dose of As2O3. The seeds were then treated with either potentized As2O3 (5x, 15x, 25x, 35x, 45x), potentized water (equivalent potencies) or diluted As2O3 (10-5, 10-15, 10-25, 10-35, 10-45). The working variable was the stem length, measured after 4, 5, 6 and 7 days. Results: Some potencies (As2O3 45x and H2O 45x) induced a relevant increase in seedling growth and/or a variability decrease. Diluted As2O3 did not induce any significant results. Conclusions: Confirmation of a significant stimulating effect on seedling growth and a significant decrease of variability was obtained with ultra-high dilutions at the 45x potency. The model of wheat germination and growth has been confirmed to be a good tool for basic research in homeopathy.

Exogenous spermidine, arsenic and β-aminobutyric acid modulate tobacco resistance to tobacco mosaic virus, and affect local and systemic glucosylsalicylic acid levels and arginine decarboxylase gene expression in tobacco leaves.

The polyamine spermidine and the metalloid arsenic increased resistance responses in the well-known pathosystem NN tobacco/tobacco mosaic virus (TMV). Both the hypersensitive response to TMV in a leaf disk model system (inoculated disks floating in the 0.1mM treatments) and systemic acquired resistance (SAR) in whole plants were significantly affected. In the latter case, 1mM foliar sprays of spermidine and arsenic were as effective as TMV and dl-beta-aminobutyric acid (BABA), both taken as positive controls, in improving the plants response to subsequent challenge inoculation with TMV. Moreover, this phenotypic response was correlated with changes in the endogenous concentration of the SAR-related molecule salicylic acid and in transcript levels of some pathogenesis/stress-related genes (pathogenesis-related proteins PR-1a and PR-2 and arginine decarboxylase (ADC)). Concentrations of free salicylic acid and of 2-O-beta-d-glucosylsalicylic acid and mRNA amount of PR-1a, PR-2 and ADC were analyzed in plants treated with either spermidine or arsenic, and compared with those from untreated plants and from positive (TMV-inoculated or BABA-treated) controls. Conjugated salicylic acid content and ADC transcripts were found to significantly increase, at both the local and systemic levels, relative to untreated controls.

A pilot statistical study with homoeopathic potencies of Arsenicum album in wheat germination as a simple model

A blind, randomized laboratory trial to study homoeopathic potencies of Arsenicum album on wheat germination is proposed as a simple model which allows a rigorous statistical analysis. The parametric tests show that the differences between the treatment groups cannot be explained as a mere effect of intrinsic seed variability.

Self-organized crystallization patterns from evaporating droplets of common wheat grain leakages as a potential tool for quality analysis.

We studied the evaporation-induced pattern formation in droplets of common wheat kernel leakages prepared out of ancient and modern wheat cultivars as a possible tool for wheat quality analysis. The experiments showed that the substances which passed into the water during the soaking of the kernels created crystalline structures with different degrees of complexity while the droplets were evaporating. The forms ranged from spots and simple structures with single ramifications, through dendrites, up to highly organized hexagonal shapes and fractal-like structures. The patterns were observed and photographed using dark field microscopy in small magnifications. The evaluation of the patterns was performed both visually and by means of the fractal dimension analysis. From the results, it can be inferred that the wheat cultivars differed in their pattern-forming capacities. Two of the analyzed wheat cultivars showed poor pattern formation, whereas another two created well-formed and complex patterns. Additionally, the wheat cultivars were analyzed for their vigor by means of the germination test and measurement of the electrical conductivity of the grain leakages. The results showed that the more vigorous cultivars also created more complex patterns, whereas the weaker cultivars created predominantly poor forms. This observation suggests a correlation between the wheat seed quality and droplet evaporation patterns.

The Role of Variability in Evaluating Ultra High Dilution Effects: Considerations Based on Plant Model Experiments

A series of experiments, performed on plant models with ultra high dilutions (UHD) of arsenic trioxide at 45th decimal potency has been reviewed with a particular focus on variability. The working variables considered are: the number of germinated seeds out of a fixed set of 33, the stem length of wheat seedlings and the number of necrotic lesions in tobacco leaf disks inoculated with tobacco mosaic virus (TMV). A thorough comparison between treatment and control group has been proposed, considering the two main sources of variability in each series of experiments: variability within and between experiments. In treated groups, a systematic decrease in variability between-experiments, as well as a general decrease, with very few exceptions, in variability within experiments has been observed with respect to control. Variability is traditionally considered as control parameter of model systems. Our hypothesis, based on experimental evidences, proposes a new role of variability as a target of UHD action. This hypothesis may help interpret unanswered questions that keep rising in basic and clinical research in homeopathy.

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 Avogadros 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.

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 As2O3 at the 45th decimal potency in a wheat germination model, compared against a control and potentized H2O 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% As2O3 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 As2O3 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 As2O3 45× exhibits a repeated significant effect versus control, except at 100°C. A reduction in variability was observed for As2O3 45× at 20°C versus control, confirming the findings of previous work. The main conclusion suggested by this experiment is that the efficacy of As2O3 45× on wheat germination may be influenced by heating degree and seems to have an increasing trend as a function of aging.