Wheat and ultra high diluted gibberellic acid--further experiments and re-analysis of data.

BACKGROUND: Following studies (a) on wheat seedlings and ultra high diluted silver nitrate, and (b) on amphibians and an ultra high diluted hormone, (c) a bio-assay on wheat and extremely diluted gibberellic acid was standardized. This assay was intended to combine the easy-to-handle aspect of (a) and biologically interesting aspects of (b). The purpose of the data analysis presented here was to investigate the influence of an extreme dilution of gibberellic acid on wheat stalk length and to determine the influence of external factors on the experimental outcome. METHODS: Grains of winter wheat (Triticum aestivum, Capo variety) were observed under the influence of extremely diluted gibberellic acid (10(-30)) prepared by stepwise dilution and agitation according to a protocol derived from homeopathy ('G30x'). Analogously prepared water was used for control ('W30x'). 16 experiments including 8000+8000 grains were performed by 9 researchers. RESULTS: Experiments that were performed between January and April showed inconsistent results, whereas most of the experiments performed between September and December showed shorter stalks in the G30x group. This was confirmed by correlation analysis (p<0.01). Thus winter/spring experiments and autumn experiments were analysed separately. When all 10 autumn experiments were pooled, mean stalk lengths (mm) were 48.3±21.4 for the verum group and 52.1±20.4 for control (mean±SD) at grain level (N=5000 per group) and ±5.3 and ±5.1 respectively at dish level. In other words, verum stalk length (92.67%) was 7.33% smaller than control stalk length (100%). The effect size is small when calculation is done on the basis of grains (d=0.18) but, due to the smaller SD at dish level, medium when done on the basis of dishes (d=0.73). The inhibiting effect was observed by 6 of the 6 researchers who performed the autumn experiments. CONCLUSION: The model may be useful for further research as there exists a theoretical justification due to previous studies with wheat and extremely diluted silver nitrate, as well as to previous studies with amphibians and diluted hormones, and its methods are well standardized. Data confirm the hypothesis that information can be stored in the test liquid, even at a dilution of the original substance beyond Avogadro's value; and that the wheat bio-assay is sensitive to such information.

Reproducibility of effects of homeopathically potentised gibberellic acid on the growth of Lemna gibba L. in a randomised and blinded bioassay.

BACKGROUND: Reproducibility of basic research investigations in homeopathy is challenging. This study investigated if formerly observed effects of homeopathically potentised gibberellic acid (GA3) on growth of duckweed (Lemna gibba L.) were reproducible. METHODS: Duckweed was grown in potencies (14x-30x) of GA3 and one time succussed and unsuccussed water controls. Outcome parameter area-related growth rate was determined by a computerised image analysis system. Three series including five independent blinded and randomised potency experiments (PE) each were carried out. System stability was controlled by three series of five systematic negative control (SNC) experiments. Gibbosity (a specific growth state of L. gibba) was investigated as possibly essential factor for reactivity of L. gibba towards potentised GA3 in one series of potency and SNC experiments, respectively. RESULTS: Only in the third series with gibbous L. gibba L. we observed a significant effect (p = 0.009, F-test) of the homeopathic treatment. However, growth rate increased in contrast to the former study, and most biologically active potency levels differed. Variability in PE was lower than in SNC experiments. The stability of the experimental system was verified by the SNC experiments. CONCLUSIONS: Gibbosity seems to be a necessary condition for reactivity of L. gibba to potentised GA3. Further still unknown conditions seem to govern effect direction and the pattern of active and inactive potency levels. When designing new reproducibility studies, the physiological state of the test organism must be considered. Variability might be an interesting parameter to investigate effects of homeopathic remedies in basic research.

Seasonal variation of the effect of extremely diluted agitated gibberellic acid (10e-30) on wheat stalk growth: a multiresearcher study.

The influence of a homeopathic high dilution of gibberellic acid on wheat growth was studied at different seasons of the year. Seedlings were allowed to develop under standardized conditions for 7 days; plants were harvested and stalk lengths were measured. The data obtained confirm previous findings, that ultrahigh diluted potentized gibberellic acid affects stalk growth. Furthermore, the outcome of the study suggests that experiments utilizing the bioassay presented should best be performed in autumn season. In winter and spring, respectively, no reliable effects were found.

Effects of homeopathic arsenicum album, nosode, and gibberellic acid preparations on the growth rate of arsenic-impaired duckweed (Lemna gibba L.).

This study evaluated the effects of homeopathically potentized Arsenicum album, nosode, and gibberellic acid in a bioassay with arsenic-stressed duckweed (Lemna gibba L.). The test substances were applied in nine potency levels (17x, 18x, 21x-24x, 28x, 30x, 33x) and compared with controls (unsuccussed and succussed water) regarding their influence on the plant's growth rate. Duckweed was stressed with arsenic(V) for 48 h. Afterwards, plants grew in either potentized substances or water controls for 6 days. Growth rates of frond (leaf) area and frond number were determined with a computerized image analysis system for different time intervals (days 0-2, 2-6, 0-6). Five independent experiments were evaluated for each test substance. Additionally, five water control experiments were analyzed to investigate the stability of the experimental setup (systematic negative control experiments). All experiments were randomized and blinded. The test system exhibited a low coefficient of variation (approximately equal to 1%). Unsuccussed and succussed water did not result in any significant differences in duckweed growth rate. Data from the control and treatment groups were pooled to increase statistical power. Growth rates for days 0-2 were not influenced by any homeopathic preparation. Growth rates for days 2-6 increased after application of potentized Arsenicum album regarding both frond area (p < 0.001) and frond number (p < 0.001), and by application of potentized nosode (frond area growth rate only, p < 0.01). Potencies of gibberellic acid did not influence duckweed growth rate. The systematic negative control experiments did not yield any significant effects. Thus, false-positive results can be excluded with high certainty. To conclude, the test system with L. gibba impaired by arsenic(V) was stable and reliable. It yielded evidence for specific effects of homeopathic Arsenicum album preparations and it will provide a valuable tool for future experiments that aim at revealing the mode of action of homeopathic preparations. It may also be useful to investigate the influence of external factors (e.g., heat, electromagnetic radiation) on the effects of homeopathic preparations.

Reproducibility of dwarf pea shoot growth stimulation by homeopathic potencies of gibberellic acid.

OBJECTIVES: Investigation of the conditions for reproducibility of dwarf pea shoot growth stimulation through homeopathic potencies of gibberellic acid. METHODS: 4 batches of pea seed (Pisum sativum L. cv. Früher Zwerg; harvests from 1997, 1998, 1999, and 2000) were tested regarding their reaction to gibberellic acid 17x and 18x (compared to unsuccussed and succussed water (1x) as controls) in 8 independent randomized and blinded experiments. Pea seed was immersed for 24h in watery solutions of homeopathic potencies or controls, and cultivated under controlled laboratory conditions. Pea shoot length was measured after 14 days. Two systematic negative control experiments assessed the stability of the experimental set-up. RESULTS: The systematic negative control experiments yielded no significant effects and confirmed the stability of the experimental set-up. 2 out of 4 seed batches reacted to the homeopathic treatment (p<0.05). Seed batch 1997 showed a reproducible reaction to gibberellic acid 17x (shoot length stimulation of +11.2%, p=0.007), and seed batch 1998 showed a significant varying response (increase/decrease). Seed batch 1997 differed from the other 3 batches by an increased glucose and fructose content, and reduced 1000kernel weight. Meta-analysis with data of earlier experiments is in accordance with the results of the present experimental series. CONCLUSIONS: We identified 'seed quality' as a possible trigger factor for successful reproducibility in homeopathic basic research. Premature harvesting as a possible key factor for responsiveness of dwarf peas to homeopathic potencies of gibberellic acid is our current working hypothesis to be tested in future experiments.

Growth stimulation of dwarf peas (Pisum sativum L.) through homeopathic potencies of plant growth substances.

BACKGROUND: Efficacy of higher homeopathic potencies is controversial. Universally accepted specific detection assays for homeopathic dilutions do not exist. Basic research has to develop a spectrum of standardized tools to investigate the mode of action and nature of homeopathic potencies. OBJECTIVE: Can the shoot growth reaction of dwarf peas (gibberellin- deficient mutants) be regarded as evidence of treatment with homeopathic potencies of plant growth substances? MATERIALS AND METHODS: Pea seed (Pisum sativum L. cv. Fruher Zwerg) is immersed for 24 hours in homeopathic potency or control solutions for soaking. Plants germinate and grow in a standard cultivation substrate under controlled environmental conditions. Shoot length is measured 14 days after planting. RESULTS: A screening of homeopathic potencies (12x-30x) of four different plant growth substances revealed biological activity of certain potency levels of gibberellin and kinetin (p < 0.05). Growth stimulation through gibberellin 17x (5 x 10(-18 M)) was assessed in six independent replications; results confirmed those of the screening (p < 0.05). The effect of gibberellin 17x seemed to weaken during the course of the experiments. CONCLUSION: The results back the hypothesis that homeopathic potencies of plant growth substances affect pea shoot growth. Dwarf peas might thus be an interesting system model for studying the action of homeopathic potencies. Further work is required to identify all boundary conditions modulating the reactivity of this system.

The effect of extremely diluted agitated gibberellic acid (10e-30) on wheat stalk growth--a two researcher pilot study.

OBJECTIVE: Use of a wheat growth bio assay after 7 days in research on homeopathic dilutions of gibberellic acid. METHODS: Grains of winter wheat (Triticum aestivum, Capo variety) were observed under the influence of extremely diluted gibberellic acid (10(-30)) prepared by stepwise dilution and agitation according to a protocol derived from homeopathy (30×). Analogously prepared water was used for control. In a two centre study, 3 experiments with a total of 4880 grains were performed. RESULTS: Data were found to be rather homogeneous within the control group as well as within the verum group in general. Germination rates were around 95%, with no significant difference between verum and control group (p>0.05). Mean stalk lengths (mm) were 40.63±20.96 for the verum and 44.33±21.11 for the control group (mean±S.D.) at grain level (N=2440 per group) and ±5.33 and ±5.89, respectively at dish level (122 cohorts of 20 grains per treatment group). In other words, verum stalk length (91.65%) was 8.35% smaller than control stalk length (100%). This difference is statistically highly significant (p<0.001) and was found by both researchers involved independently. CONCLUSION: These results suggest that there was an influence of gibberellic acid 30× on wheat seedling development, i.e. the wheat growth bio assay can be a useful tool for further experiments on homeopathic dilutions of gibberellic acid.

Effects of potentised substances on growth rate of the water plant Lemna gibba L.

OBJECTIVES: This study investigated, whether the growth rate of Lemna gibba L. (duckweed) can be influenced by the application of homeopathic potencies of gibberellic acid, kinetin, argentum nitricum, and lemna minor. METHODS: Duckweed was grown in either potencies (14x-30x, decimal steps) or water controls (unsuccussed and succussed) over seven days. Frond (leaf-like structure) growth was measured using a non-destructive image analysis system. Growth rates were calculated for three time intervals (0-7, 0-3, 3-7 days). Five to six independent, randomized and blinded experiments were analysed for each of the four tested substances. Water control experiments were performed repeatedly to test the reliability of the experimental set-up (systematic negative controls). RESULTS: The systematic negative control experiments did not yield any significant effects. Hence, false positive results could be excluded. The test system had a low coefficient of variation (1.5%). Out of the four tested substances gibberellic acid had the most pronounced effect (p=0.0002, F-test) on the main outcome parameter frond growth rate (r(area) day 0-7). Potency levels 15x, 17x, 18x, 23x and 24x reduced growth rate of Lemna gibba (p<0.05 against the pooled water control, LSD test). CONCLUSIONS: Lemna gibba may be considered as a suitable test organism for further studies on the efficacy of homeopathic potencies. Evidence accumulates, that adjacent potency levels may strongly differ in their biological activity. Potential consequences for therapeutical application might be worth investigating.

Homeopathically prepared gibberellic acid and barley seed germination.

The potentisation process by which homeopathic preparations are produced raises the concern that these medicines have placebo effects only, since they theoretically no longer contain active molecules of the diluted substance. Plant models offer a method of examining the efficacy of homeopathically prepared solutions. This study examined the effects of homeopathically prepared gibberellic acid (HGA3) on the germination performance of barley (Hordeum vulgare L.) seeds. The effect of HGA3 (4-200 cH) on seed germination rate and seedling development was compared to that of the most commonly used form of gibberellic acid (GA3), 0.5 g l(-1), and control (distilled water). The extent and type of response was dependent on the vigour level of the seedlot. Treating seeds from three vigour groups in HGA3 consistently resulted in larger seedlings. High-vigour seeds treated with HGA3 4, 30 and 200 cH germinated faster, and roots of medium-vigour seedlots treated in HGA3 15 cH were longer. Biphasic effects of HGA3 were also demonstrated. As a plant model, germinating barley seeds successfully demonstrated the ability of HGA3 to produce a biological response.