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.

Polycrystalline structures formed in evaporating droplets as a parameter to test the action of Zincum metallicum 30c in a wheat seed model.

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.

First evidence of Beauvais' hypothesis in a plant model.

INTRODUCTION: Beauvais presented the application of a so-called 'quantum-like model of homeopathy' by introducing the idea of a type of randomization/unblinding which he called 'in situ'. He predicted that randomized studies based on this type of randomization/unblinding lead to more pronounced effects in placebo controlled randomized homeopathic trials. We designed an experiment regarding wheat germination and stalk length to investigate Beauvais' idea of 'in situ randomization/unblinding' using a homeopathic dilution of sulphur (LM VI) as compared to placebo as well as to water. AIM AND METHOD: The primary aim of this double-blind randomized controlled experiment was to investigate whether there are differences of 'in situ randomization/unblinding' vs 'central randomization/unblinding' with respect to the effect of a homeopathic substance compared to placebo. The secondary aim of our study was to examine possible differences between the sulphur and the placebo group in the 'in situ' arm regarding germination and/or stalk growth of wheat seedlings measured after a seven days exposure. Wheat was treated either with sulphur LM VI, placebo, or water. The wheat grains were placed on glass lids and treatment was performed following the 'in situ randomization/unblinding' as well as 'central randomization/unblinding' method. Germination was measured and classified into three categories. RESULTS: Under 'in situ' randomization/unblinding the odds of a seed not to germinate is 40% lower if treated with sulphur compared to placebo (p=0.004). In contrast, these odds are practically equal in the 'central' meta-group (OR=1.01, p=0.954). Under 'in situ' randomization/unblinding the odds of a seed to germinate with a length ≥1mm is practically equal if treated with sulphur or with placebo (OR=0.96, p=0.717). In contrast, these odds are 21% higher under sulphur compared to placebo in the 'central' meta-group (OR=1.21, p=0.062). In summary, we found a sulphur effect that is significantly different between 'in situ' and 'central' randomization/unblinding relating to all three stages of germination.

Diverse biological effects of electromagnetic-treated water.

The effects of water treated with an electromagnetic field (EMF) were investigated on two biological systems, humans and plants. Purified de-ionised water was treated by (1) boiling, (2) exposure to microwave radiation, and (3) low frequency electromagnetic oscillation molecular resonance effect technology (MRET), before being used to prepare media for culturing human peripheral blood mononuclear cells (PBMC) from three healthy females. Our results indicated that PBMC culture in MRET-activated medium showed significantly less oxidative metabolism when compared to media prepared from other types of water. As for the effects on soybean, our results indicated that both MRET- and microwave-treated water greatly enhanced the length of the root. These results suggested that electromagnetic-treated water can have diverse biological effects on both animal and plant cells. Since these effects are related to the 'Memory of Water', hypothesis which has been suggested as an explanation of the action of high homeopathic dilutions, our finding warrant a further investigation on the mechanisms of various types of physically conditioned water on specific cellular activities.

Stimulative influence of germination and growth of maize seedlings originating from aged seeds by 2,4-D potencies.

BACKGROUND: The 2,4-D (2,4-dichlorophenoxyacetic acid) is using as a growth regulator in tissue culture media. Maize seeds have poor ability to maintain germination rate in the long term. OBJECTIVE: To examine the possible restorative effect of homeopathic 2,4-D potencies on maize seedlings originating from seeds damaged by accelerated aging. METHODS: Seeds of four maize lines were subjected to accelerated aging stress treatment. Seed samples were treated with distilled water (control) and a range of potencies of 2,4-D: 3C, 3.75C, 4.5C, 5.25C and 6C. The germination capacity, fresh substance (FS) and length of root and shoot were determined. Hydrolysis and biosynthesis, GSH/GSSG ratio and redox capacity were calculated. RESULTS: Induced seed aging decreased germination rate and growth of seedlings. 2,4-D potencies did not have a statistically significant effect on germination. However, there were statistically significant effects on FS production, root and shoot length and redox capacity. The 3C potency had the largest effect on the FS accumulation, 4.5C increased root and shoot length, compared to control (statistically significant). The GSH/GSSG ratio and the redox capacity were decreased by aging. The 3C and 4.5C potencies tended to reverse the GSH/GSSG ratio (statistically significant) in the root and shoot, (i.e., shifted the redox balance to the reduced state). CONCLUSION: Homeopathic potencies of 2,4-D appear to have a beneficial effect on artificially aged maize seeds: they stimulate growth through better substance conversion from seed rest, and shift the redox capacity towards a reduced environment. Further work is required to determine if this is an useful means of improving maize seed germination and growth.

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.

Seed longevity and germination characteristics of six fen plant species.

Fens are among the most threatened habitats in Europe as their area has decreased considerably in the last centuries. For successful management and restoration conservationists need detailed knowledge about seed bank formation and seed longevity of plants, as these features are closely related to successional and vegetation dynamical processes. I analysed seed longevity and the germination characteristics of six fen plant species by seed burial experiments. Based on seed weight, seed bank was expected for long-term persistent for the light-seeded Schoenus nigricans, Carex appropinquata, C. pseudocyperus, C. davalliana and Peucedanum palustre and also that for the medium-seeded Cicuta virosa. It was proved that, the latter two species have short-term persistent seed banks, while Carex pseudocyperus has a transient seed bank, therefore these species may only have a limited role in restoration from seed banks. It was found that Schoenus nigricans, Carex appropinquata and C. davalliana have persistent seed banks, because some of their four-year-old seeds have emerged. Fresh seeds had low germination rate in all studied species and majority of seeds emerged after winter, except for Carex pseudocyperus. After the germination peak in spring, the majority of the ungerminated seeds of Schoenus nigricans, Peucedanum palustre, Carex appropinquata, C. davalliana and Cicuta virosa entered a secondary dormancy phase that was broken in autumn. I found the seasonal emergence of the latter three species highly similar.

Isopathic treatment effects of Arsenicum album 45x on wheat seedling growth--further reproduction trials.

BACKGROUND: Two experimental studies on wheat preintoxicated with Arsenic trioxide yielded a significant shoot growth increase after an isopathic application of Ars-alb 45x. One independent reproduction trial however, yielded an effect inversion: wheat shoot growth was significantly decreased after application of Ars-alb 45x. AIMS: In this study we investigated the role of three potential confounding factors on the experimental outcome: geographical location of the experiments, influence of the main experimenter, and seed sensitivity to Arsenic poisoning. Laboratory-internal reproducibility was assessed by meta-analysis. MATERIAL AND METHODS: Wheat poisoned with Arsenic trioxide was cultivated in vitro in either Ars-alb 45x, water 45x, or unpotentised water. Treatments were blinded and randomised. Shoot length was measured after 7 days. The stability of the experimental set-up was assessed by systematic negative control (SNC) experiments. RESULTS: The SNC experiments did not yield significant differences between the three groups treated with unpotentised water. Thus the experimental set-up seemed to be stable. We did not observe any shoot growth increase after a treatment with Ars-alb 45x in any of the newly performed experiments. In contrast, the meta-analysis of all 17 experiments performed (including earlier experiments already published) yielded a statistically significant shoot growth decrease (-3.2%, p=0.017) with isopathic Ars-alb 45x treatment. This effect was quantitatively similar across all five series of experiments. CONCLUSIONS: Ultramolecular Ars-alb 45x led to statistically significant specific effects in arsenic poisoned wheat when investigated by two independent working groups. Effect size and effect direction differ, however. The investigated factors (geographical location, experimenter, seed sensitivity to Arsenic poisoning) did not seem to be responsible for the effect inversion. Laboratory external reproducibility of basic research into homeopathic potentisation remains a difficult issue.

Empirical investigation of preparations produced according to the European Pharmacopoeia monograph 1038.

According to the European Pharmacopoeia monograph 1038 (Praeparationes homoeopathicae), homeopathic preparations are produced by successive dilution and succussion steps. Dilution levels beyond Avogadro's limit, however, render specific effects implausible according to standard scientific knowledge. Accordingly, we were interested in a critical empirical investigation of preparations produced according to this monograph. Within a precursor study we developed a bioassay based on a fingerprint metabolomic analysis of Lepidium sativum seeds germinated in vitro in either homeopathic preparations or controls in a blinded and randomized assignment. Results of the precursor study were not consistent with the hypothesis that the effects of a Stannum metallicum 30x preparation are identical to placebo. In the present study we investigated the reproducibility of these effects after scrutinizing and optimizing experimental procedures. Ten independent experiments were performed in a blinded and randomized assignment in two independent laboratories. Additionally, 10 systematic negative water control experiments were performed in both laboratories to critically assess the stability of the experimental set-up. The effects of the Stannum metallicum 30x treatment were reproduced. The systematic negative control experiments did not yield false-positive results, indicating a stable experimental set-up. We thus repeatedly observed biological effects conflicting with the assumption that Stannum metallicum 30x is identical to placebo. We therefore wish to discuss whether these findings are to be considered a scientific anomaly or whether they might stimulate further investigations to clarify whether application of the European Pharmacopoeia monograph 1038 may result in pharmaceutical preparations with specific effects.

Duckweed (Lemna gibba L.) as a test organism for homeopathic potencies.

OBJECTIVES: A bioassay with duckweed (Lemna gibba L.) was used to study the effects of homeopathic potencies on the plant's growth rate. Screening included 12 substances: argentum nitricum, copper sulfate, gibberellic acid, 3-indole acetic acid, kinetin, lactose, lemna minor, methyl jasmonate, metoxuron, phosphorus, potassium nitrate, and sulfur. Each substance was tested in the potency range 14x-30x. Controls were unsuccussed and succussed water. DESIGN: In randomized and blinded experiments, duckweed was grown in either potentized substances or water controls over 7 days. Frond (leaf) growth was measured regularly with a computerized image analysis system and growth rates were calculated for different time intervals (day 0-7, 0-3, 3-7). Additionally, a water control run with unsuccussed water as the only test substance was performed to determine the variability of the bioassay. RESULTS: For the water control run, the between-group coefficient of variance for groups of five replicates was 0.87% for the frond area-related average specific growth rate r(area) compared to 1.60% for the frond number-related average specific growth rate r(num). Thus, the former is the preferred parameter to be used. Of twelve tested substances, potentized argentum nitricum, phosphorus, and kinetin significantly (p<0.05, analysis of variance F-test) affected the main parameter: frond area-related average specific growth rate (day 0-7). Segmented area growth rates (day 0-3 or 3-7) were affected by potentized argentum nitricum, gibberellic acid, lactose, and phosphorus. CONCLUSIONS: The described experimental set-up with L. gibba as test organism appears to be a promising new model system to investigate effects of potentized substances. Yet larger sets of replication experiments with selected test substances and systematic negative controls are necessary to verify the effects found.

Effect of homeopathy on soybean germination through soil and seed treatment

Homeopathy is a technique approved by the MAPA for organic production systems. Experiments demonstrating the effectiveness of homeopathy in agriculture are essential. Aims: To evaluate the effects of homeopathic treatment on soybean seed germination. Methodology: The experiment was performed in MS-Brazil. An area of 70 m² was used, divided into two comparison plots of 30 m² each. The soil of plot 1 (treated group) was treated with 5.25 kg of powdered Lithothamniummixed with 250 ml of 30% hydroalcoholic solution (HS) containing homeopathic ingredients. The soil of plot 2 (control group) was treated with thesame preparationbutwithout homeopathic ingredients. Additionally, the 400 seeds assigned toplot 1 were treated with 0,3 ml of a solution made up of 10 ml of 30% HS with homeopathic ingredients dilutedin 300 ml of water. The 400 seeds assigned toplot 2 were treated with 0.3 ml of a solutionmade up of 10 ml of HS without homeopathic ingredients diluted in 300 ml of water. The homeopathic ingredients Sulph 9C, Mag-c12C, and Calc-p 9C were used on soil and seed treatment. The selection and prescription of those ingredients followed Materia Medica instructions. The experiment was conducted using randomized design and 400 seeds per group were sown. Results and discussion: In plot 1, anaverage of 11 seeds per m² germinated, adding up to 330 germinated seeds at a percentage of 82.5%. In plot 2, an average of 7 seeds per m² germinated, adding up to 210 germinated seeds at a percentage of 52.5%. The frequency of seed germinationin both groups was assessed by the Chi-square test to check for significant differences(p<0.05). There was a statistical difference (p<0.05) in the seed germination in the treated group (n=330) compared to the control group (n=210). Conclusion:Homeopathic compounds can be used as a viable treatment for soybean germination.

Study of germination efficiency and temperature/drowning resistance in some ornamental plants treated with ultra high dilute compounds

The ultra-high dilutions (UHDs) can be used for decreasing stress conditions causing by climate variations. The present research investigated the effects of ultra-highdilutions (UHDs) on some ornamental plants, germination, and hormonal variations.Methods: In order to study the effect of UHDs (Calendula officinalisCalen. andArnica montanaArn.) on the physiological, primary metabolite, and hormonal variations of theOryza sativa L. (rice), 104 experiments were designed and statistically analyzed using the Design Expert 7.0.1 software over the general factorial design methodology. Two qualitative factors, including the UHDs/placebo usage and the type of plant usage, and two quantitative factors, including temperature and irrigation, were studied. The validated analysis was subjected to more extended studies on the variations in physiological growth, carbohydrate, protein content, and levels of plant hormones, including gibberellic acids, indole acetic acid, abscisic acid, and salicylic acid.Results: The statistical analysis resulted in a prediction model which was more than 75% correlates with experimental results. The results showed that the UHDs increased the carbohydrate and protein content of seedlings. Also, compared to placebo, the levels of hormones GA3 and IAA in all samples increase, and the amount of GA4 decreases. The amount of ABA and SA hormones inS. officinalisincreased under UHDs treatment while decreasing in the other two samples.Conclusion: The use of UHDs leads to an increase in the production of carbohydrate and protein content. Moreover, it causes significant variations in the growth-inducing hormone and increases the tolerance of seeds under higher/lower temperatures and draught/drowning. The results of this study open up a window to reduce germination survival and increase their resistance to sudden climate change.

A biostatistical insight into the As(2)O(3) 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 (As(2)O(3)) 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 As(2)O(3). The seeds were then treated with either potentized As(2)O(3) (5x, 15x, 25x, 35x, 45x), potentized water (equivalent potencies) or diluted As(2)O(3) (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 (As(2)O(3) 45x and H(2)O 45x) induced a relevant increase in seedling growth and/or a variability decrease. Diluted As(2)O(3) 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.

The effects of a 45x potency of arsenicum album on wheat seedling growth -- a reproduction trial.

BACKGROUND: Independent replications of preclinical investigations of homeopathic potencies are rare. However, they are a necessary tool to determine the relevant factors modulating the effects of homeopathic potencies in preclinical systems. OBJECTIVE: The goal of the present study was to reproduce a trial published in 1997. An Italian group of researchers investigated the effect of Arsenicum album 45x on the growth of wheat which had been previously poisoned with a material dose of Arsenicum album. The homeopathic treatment was associated with increased wheat shoot growth significantly different from the control group (+24%, p < 0.001). MATERIALS AND METHODS: Wheat poisoned with a sublethal dose of Arsenicum album was cultivated in either Arsenicum album 45x, water 45x, or unpotentized water. After 7 days, shoot length was measured. Reproducibility was assessed in eight independent experiments. RESULTS: Arsenicum album 45x significantly inhibited wheat shoot growth (-3%) compared to treatment with unpotentized water and water 45x (p = 0.011 and p = 0.037). Within the experimental series performed in this reproduction trial, the effects of Arsenicum album 45x proofed to be reproducible. The wheat seed species used did not seem to have a significant impact on the experimental outcome. CONCLUSION: The result of this replication trial is a reversal of the original study, since Arsenicum album 45x inhibited wheat shoot growth instead of enhancing it. Nevertheless, high homeopathic potencies may induce statistically significant effects in biological systems. However, the magnitude and direction of these effects seem to depend on yet unknown parameters.

The influence of ultra-high diluted compounds on the growth and the metabolites of Oryza sativa L

The ultra-high dilutions (UHDs) have been widely used in the field of human, animal and plant treatment. In the present research, the effects of the potentized ultra-high dilutions (UHDs) on physiological and biochemical variations in Oryza sativaL. (rice) were investigated. Methods: To study the effect of UHDs (Calendula officinalisCalen. and Arnica montanaArn.) on the physiological and biochemical variations of the Oryza sativaL. (rice), 28 experiments were designed and statistically analyzed using the Design Expert 7.0.1 software over the general factorial design methodology. Three qualitative factors were studied including the UHDs/placebo usage, sterile/non-sterile experimental condition and the type and timing of the UHDs usage. The validated analysis was subjected to more extended studies on the variations in physiological growth, carbohydrate, protein content, pigment production, and amino acid patterns. To evaluate the effects of UHDs on rice, a desirable response percentage was formed from a number of healthy seedling productions, and the height percentage of the aerial parts and main roots were studied. Results: The statistical analysis resulted in a prediction model which was more than 97% correlates with experimental results. The results showed that the UHDs increased the pH variations, carbohydrate, protein and pigment levels each by ~2.5, ~1.5, ~1.4, and ~1.4 folds, respectively. Also compared to placebo, the amount and proportionof amino acids has significantly varied, showing a statistical effect on the germination and seedling growth of the rice, as well as the stress conditions caused by the sterilization process, seedlings entrance into the light and their transition into hydroponic culture medium.Conclusion: The use of UHDs leads to an increase in the production of chlorophyll, as well as carbohydrate and protein content. Moreover, it causes significant variations in the amino acid profile and the production of amino acids along with the photosynthesis, germination, and metabolism processes.(AU)

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.

Feasibility in seed germination of Hypericum perforatum L. submitted at different temperatures and treatments with high dilutions

Background Hypericum perforatum is used as a medicinal plant, mostly in antidepressant treatment. With endogenous and exogenous dormancy, more studies about the germination of the seeds are necessary. Aim: To evaluate the effect of temperature and of preparations of high dilutions on the germination of seeds of Hypericum perforatum. Methods Three bioassays were carried out in chambers of germination, using a completely randomized experimental design and in duplicate. In the first bioassay, of temperatures of 20, 25 and 30 ºC was evaluated, with 20 replicates and 50 seeds in each replicate. In the second, the effect of high dilutions (Kali carbonicum, Natrum muriaticum, Phosphorus and Silicea terra in 12cH) using 4 replicates with 50 seeds each. In the third, Kali carbonicum was tested at 6, 12, 20, 30 cH with 50 seeds and 4 replicates. The percentage of germination, germination rate index (GRI), average germination time (AGT) and seedlings with cotyledons. The data were analyzed through the R software at a 5% significance. Results At the temperature of 25 °C, 63% of the seeds germinated while at the temperatures of 20 and 30 °C the percentage was 26% and 18%, respectively. In seeds treated with Kali carbonicum 12cH the germination rate increased significantly in relation to the control group. Kali carbonicum 6cH increased the GRI of the seeds, while at 20cH, it increased the AGT, showing delay in germination. Conclusion The use of the Kali carbonicum assists in the breakage of dormancy and average time of germination in Hypericum perforatum seeds. (AU)

Cantharis 200c counters heat stress in germinating seeds of Vigna unguiculata (L) Walp

Background Thermal stress during early imbibitional phase of germination causes disruption of redox-homeostasis by increasing accumulation of ROS Reactive Oxygen species (assessed in terms of hydrogen peroxide) and significant reduction of antioxidative defense (assessed in terms of catalase and peroxidase) in germinating tissues of cowpea (Vigna ungiculata). It also induces oxidative damage to newly assembled membrane system by aggravating membrane lipid peroxidation [measured in terms of thiobarbituric acid reactive substances (TBARS)]. Cantharis is a homeopathic remedy used for treating burn injuries in humans. The objective is to see whether potentized Cantharis could ameliorate heat stress in germinating seeds of cowpea. Methodology Seeds, imbibed in water overnight, were divided into four groups (n=50/group): Control I (Water 200c), Control II (Ethanol 200c), treated with Cantharis 200c and untreated and unstressed. Water soaked seeds were dipped in control/drug solution for 5 min and then washed. Control II and Cantharis 200c were diluted with water 1:1000 to minimize the ethanol effect. Except the fourth group, all other groups were subjected to heat stress (450C for 8 hours). All the groups were allowed to germinate for 5 days in germination chamber at 25 ± 20C. Groups 1 and 2 (Control I and II) served as systematic negative controls throughout the experiment. Ten independent replications were performed for each group in parallel. All experiments were randomized and blinded. Results The systematic negative controls (I, II) did not produce any significant effect. The results in terms of germination, growth, soluble sugar, protein, accumulation of reactive oxygen species and loss of membrane permeability clearly exhibit that Cantharis 200c could mitigate heat stress significantly (p<0.05) as compared to the controls I, II. Final germination percentage after 120 hrs was found to be 60%, 48%, 86%, 88%, for groups 1, 2, 3, 4, respectively. Total chlorophyll (Chl-a + Chl-b), carotenoids and total chl/carotenoid ratio were found to be highest in groups 3 and 4. Assessment of membrane lipid peroxidation in terms of TBARS and accumulation of reactive oxygen species (H2O2) showed minimum concentration in group 3 and 4. Conclusion Cantharis 200c can counteract the deleterious effects of heat stress on juvenile tissues and membranes and can be used effectively to counter heat stress in tropical countries especially during summer. (AU)

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.

Bioatividade de extratos hidroalcoólicos de Cymbopogon citratus (DC. ) Stapf. sobre picão-preto (Bidens pilosa L.) e alface (Lactuca sativa L.) / Bioactivity of hydroalcoholic extracts of Cymbopogon citratus (DC. ) Stapf. on germination and early growth of beggartick (Bidens pilosa L.) and lettuce (Lactuca sativa L.)

Este trabalho visou avaliar a bioatividade de extratos hidroalcoólicos de capim-limão sobre germinação e crescimento inicial da planta daninha picão-preto (Bidens pilosa L.) e da planta teste alface (Lactuca sativa L.). A partir de maceração hidroalcoólica de folhas secas e rasuradas (127,46 g) de capim-limão em 1L de álcool de cereais (70%), foram preparados os tratamentos como extratos hidroalcoólicos (HA) pela diluição (v/v) do macerado filtrado em água deionizada na proporção 1:1 (HA1:1) e 1:2 (HA1:2); dos quais foram obtidos os respectivos extratos secos (ES), a partir da evaporação da fase líquida de duas alíquotas de 50 mL de cada extrato HA, que após re-suspendidas em igual volume de água, sendo uma autoclavada (1 atm por 15 minutos), resultando nos extratos secos de HA1:1 e HA1:2 autoclavados (ES1:1A e ES1:2A) e nos extratos secos não autoclavados (ES1:1 e ES1:2); e água (AG) como controle. No primeiro dia dos bioensaios, foram aplicados 2 mL dos tratamentos, em DIC, com cinco repetições. Avaliou-se a porcentagem de germinação (G%), Índice de Velocidade de Germinação (IVG), altura da parte aérea (AA) e comprimento de radícula (CR) de aquênios (25) de picão-preto e de alface distribuídos em placas de Petri e mantidos em câmara de germinação (B.O.D.) por duas semanas. Os extratos HA1:1 e HA1:2 inibiram a G%, AA e CR das duas espécies. Todos os extratos secos reduziram a G%, IVG e o CR da alface. Para o picão preto, apenas o extrato ES1:1 reduziu a G% e o IVG foi reduzido por todos os extratos, a exceção do ES1:1A, mas nenhum extrato influenciou o crescimento inicial desta espécie.

This study aimed to evaluate the bioactivity of hydroalcoholic extract of lemongrass on the germination and early growth of the weed plant beggartick (Bidens pilosa L.) and the test plant lettuce (Lactuca sativa L.). From the hydroalcoholic maceration of dried and cut leaves of lemon grass (127.46 g) in 1 L grain alcohol (70%), the following treatments were prepared, hydro-alcoholic (HA) extracts by diluting (v/v) the macerate in deionized water at 1:1 (HA1:1) and 1:2 (HA1:2) proportions, obtaining the respective dried extracts (DE) from the evaporation of the liquid phase of two 50-mL aliquots of each HA extract; after resuspension in the same water volume, one aliquot was autoclaved (1 atm for 15 minutes) resulting in HA1:1 and HA1:2 dried autoclaved extracts (DE1:1A and DE1:2A) and in dried non-autoclaved extracts (ES1:1 and ES1:2), and water as control. On the first day of bioassays, 2mL of treatments were applied, in completely randomized design, with 5 replicates. Germination percentage (G%), Germination Speed Index (GSI), shoot height (SH) and radicle length (RL) of achenes (25) of beggartick and lettuce were evaluated after having been distributed in Petri dishes and kept in a germination chamber (B.O.D.) for two weeks. HA1:1 and HA1:2 extracts inhibited G%, SH and RL of both species. All dried extracts reduced G%, GSI and CR of lettuce. For beggartick, only ES1:1 extract reduced G% while GSI was reduced in all extracts, except ES1:1A, but no extract influenced the early growth of this species.