Genome-wide identification and expression analysis of the kinesin gene superfamily suggests roles in response to abiotic stress and fertility of wheat (Triticum aestivum L.).

BACKGROUND: Kinesin is a motor for microtubule-based motility. It plays a vital role in plant growth and development. The kinesin superfamily members are known mainly from Arabidopsis. Little research about kinesin superfamily has been conducted on hexploid wheat (Triticum aestivum L.). The functions of kinesins in wheat growth and development, regulation of cell division and response to stress are still unclear. RESULTS: In this study, we identified 155 kinesin (TaKIN) genes in wheat, which were divided into 10 families and some orphan genes via phylogenetic analysis. Less gene structural differences showed that TaKIN genes had redundant functions. The conserved domains of different family members were different, and some families might have some special functional domains. We found many cis-acting elements related to hormones (GA, Auxin, SA, MeJA), cell cycle and cell division in homeopathic elements of the TaKIN genes. Collinearity analysis showed that TaKIN genes were more conservative in monocotyledons. Expression level in different tissues at different stages suggested that TaKIN family may function during the whole growth and development process in wheat. It was worth noting there were quite different at gene expression level between physiological and heritable male sterile lines during the different stages of pollen development. The differential expression patterns of some TaKIN genes between male sterile line and maintainer line might be related to wheat male sterility. Furthermore, we also found TaKIN genes were involved in response to plant hormones and abiotic stress by stress assays. CONCLUSIONS: The result is useful for further exploration of the molecular mechanism of kinesin genes in wheat male sterility and provides important information concerning response to plant hormones and abiotic stress caused by kinesin genes.

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.

Wheat and ultra high diluted silver nitrate--further experiments and re-analysis of data.

BACKGROUND: Since 1926, an influence of a dilution of silver nitrate (24x) on the growth of coleoptiles of wheat seedlings was described. The aim of the study discussed here is the critical proof of the reliability of a test system which has been quoted as a basic model for the research on homoeopathy for decades. METHODS: Grains of winter wheat (Triticum aestivum) were observed under the influence of extremely diluted silver nitrate (10(-23)) prepared by stepwise dilution and agitation according to a protocol derived from homeopathy ('24x'). Analogously prepared water and/or inert water was used for control. Thirty experiments including 5000+5000 grains were performed by 5 researchers. RESULTS: Stalk lengths clearly indicate that development is enhanced by the probe silver nitrate 24x as compared to control. When the experiments 1989-1995 were pooled, means and SD for silver nitrate 24x-groups were 42.3±26.9 mm and for water control groups 34.7±22.2 mm. Verum stalk length was 21.9% bigger than control (100%) (p<0.01; d=0.31, i.e. small). For the experiments 1998-2014, means and SD were 73.7±21.7 mm and 60.5±20.9 mm. Verum stalk length was 21.7% bigger than control (100%) (p<0.01; d=0.62, i.e. medium). From the results one may hypothesize that the result is more marked in experiments showing an average mean of stalk length between ca. 50 and 90 mm in contrast to smaller or bigger mean lengths. CONCLUSION: The previous findings were confirmed by the results.

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.

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.

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.

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.

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].

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.

Calcium intakes in individuals on diets for the management of cows' milk allergy: a case control study.

Three hundred and twenty-three individuals with self-reported food allergy were recruited by media advertisements. Questionnaire information was collected on all respondents. Chocolate (57%), milk (47%), wheat (36%) and food additives (35%) were the most frequently implicated foods. The most frequently reported symptoms were itching (43%), skin rash (43%) and tiredness (43%). Food avoidance was the most common form of food allergy diagnosis (33%) with only 8% of respondents reporting food challenge in food allergy diagnosis. Self-diagnosis was reported by 34% of respondents with 29% and 24% reporting diagnosis by a general practitioner or a homeopath, respectively. Twenty-four per cent of respondents 'always' avoided and a further 57% 'nearly always' avoided the implicated food(s). A group of 38 adults with self-reported 'milk allergy' was selected for further study. Dietary assessments, using the dietary history method, were carried out on this subgroup and on age-, sex- and occupation-matched controls. The results of the dietary assessments revealed that the 'milk allergy' group had significantly higher intakes of fibre, beta-carotene, vitamin C, vitamin E, iron and folic acid (P < 0.05) and significantly lower intakes of calcium (P < 0.002) in comparison to controls. Eighteen per cent of the group used milk alternatives. Thirty-four per cent of the 'milk allergy' group took calcium-containing supplements. Even after calcium supplementation, the mean calcium intake of those who completely avoided milk was unacceptably low (441 mg/d).

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.

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.

Prestimulation of wheat seedlings with gibberellic acid followed by application of an agitated high dilution of the same hormone

In previous multicentre studies, the influence of a homeopathic ultra-high dilution of gibberellic acid on wheat growth was scrutinized. Data showed that this test dilution slowed down stalk growth when experiments were performed in the autumn season. The aim of this work was to test the hypothesis that pretreatment of grains with high concentrations of gibberellic acid would enhance the growth-inhibiting effect of the ultra-high dilution of the plant hormone. Grains of winter wheat (Triticum aestivum, 500 or 1000 per group) were pretreated with (non-agitated) gibberellic acid 10-5, 10-4 and 10-3 parts by weight (Ge-5, Ge-4, Ge-3) or with water (?W0?) for control prior to further treatment. Grains were then observed under the influence of extremely diluted gibberellic acid (10-30 parts by weigth) prepared by stepwise dilution and agitation according to a protocol derived from homeopathy (?G30x?). Analogously prepared water was used for control (?W30x?). Seedlings were allowed to develop under standardized conditions for 7 days; plants were harvested and stalk lengths were measured. Of the four pretreatment variants under study, Ge-3 yielded most growth, followed by Ge-4 , Ge-5 and finally W. This outcome was modulated by the application of G30x in that the inhibition obtained with G30x as compared to W30x was the greater the lower the pretreatment concentration of G had been. The hypothesis that pretreatment of grains with high concentrations of gibberellic acid would enhance the growth inhibiting effect of G30x had to be rejected. Rather, G30x slowed down stalk growth most in the W0 group with p < 0.001, only moderately in the Ge-5 and Ge-4 group and not at all in the Ge-3 group.

Prestimulation of wheat seedlings with gibberellic acid followed by application of an agitated high dilution of the same hormone

In previous multicentre studies, the influence of a homeopathic ultra-high dilution of gibberellic acid on wheat growth was scrutinized. Data showed that this test dilution slowed down stalk growth when experiments were performed in the autumn season. The aim of this work was to test the hypothesis that pretreatment of grains with high concentrations of gibberellic acid would enhance the growth-inhibiting effect of the ultra-high dilution of the plant hormone. Grains of winter wheat (Triticum aestivum, 500 or 1000 per group) were pretreated with (non-agitated) gibberellic acid 10-5, 10-4 and 10-3 parts by weight (Ge-5, Ge-4, Ge-3) or with water (?W0?) for control prior to further treatment. Grains were then observed under the influence of extremely diluted gibberellic acid (10-30 parts by weigth) prepared by stepwise dilution and agitation according to a protocol derived from homeopathy (?G30x?). Analogously prepared water was used for control (?W30x?). Seedlings were allowed to develop under standardized conditions for 7 days; plants were harvested and stalk lengths were measured. Of the four pretreatment variants under study, Ge-3 yielded most growth, followed by Ge-4 , Ge-5 and finally W. This outcome was modulated by the application of G30x in that the inhibition obtained with G30x as compared to W30x was the greater the lower the pretreatment concentration of G had been. The hypothesis that pretreatment of grains with high concentrations of gibberellic acid would enhance the growth inhibiting effect of G30x had to be rejected. Rather, G30x slowed down stalk growth most in the W0 group with p < 0.001, only moderately in the Ge-5 and Ge-4 group and not at all in the Ge-3 group.(AU)

Wheat germination and highly diluted agitated gibberellic acid (10-30) ? a multi researcher study / Germinação do trigo e ácido giberélico em alta diluição (10-30) e agitado ? um estudo multicêntrico / Germinación de trigo y ácido giberélico en alta dilución (10-30) y agitado ? estudio multicéntrico

Grains of winter wheat (Triticum aestivum L., Capo variety) were observed under the influence of highly diluted gibberellic acid (10-30) prepared by stepwise dilution and agitation according to a protocol derived from homeopathy (?G30x?). Adequate control was used (water prepared according to the homeopathic protocol ?W30x? and/or untreated water ?W0?). Two sets of multicenter experiments were performed, 4 in 2009-2010 and 4 in 2011, involving altogether 6 researchers, 6 laboratories and 4,000 grains per treatment group. Data were found to be homogeneous within the control groups as well as within the verum groups. When the 2009-2010 experiments were pooled, mean germination rates after 24 hours were (85.9 + 2.6) for the control group and (82.1 + 5.7) for G30x (mean + SD at the level of experiments in %) (N = 2,000 per group). Verum germination rate was 4.4% lower than (i.e. equal to 96.6% of) (4.4 + 96.6 = 101) the control germination rate (100%). The difference is statistically significant (p < 0.001) and the effect size (d) is large (> 0.8). Observations at other points in time between 0 and 40 hours of germination yielded similar results. Practically no difference was found between W30x and W0 groups (p > 0.05). When the 2011 experiments were pooled, the mean germination rates after 24 hours were (73 + 12) for the control group and (73 + 14) for G30x (N = 2,000 per group), i.e. there was practically no difference between the groups (p > 0.05). We interpret the data from 2009-2010 on wheat germination within 40 hours as being in line with our previous findings on wheat stalk growth after one week, i.e. as confirmation that gibberellic acid 30x can influence, i.e. slow down, wheat development. Various possible reasons for the absence of any difference between groups in the 2011 experiments, including seasonal variance, are discussed and it is suggested to perform wheat germination experiments in the very beginning of autumn season only.

Grãos de trigo comum (Triticum sativum L., variedade Capo) foram observados sob a influência de uma alta diluição de ácido giberélico (10-30) preparada através de diluição e agitação seriadas seguindo um protocolo derivado da homeopatia (G30x). Foram utilizados controles adequados (água preparada segundo o protocolo homeopático - W30x - e/ou água sem tratamento - W0 -). Foram realizadas duas séries de experimentos multicêntricos, 4 em 2009-2010 e 4 em 2011, incluindo 6 pesquisadores, 6 laboratórios, e 4.000 grãos em cada grupo de tratamento. Os dados foram homogêneos dentro dos grupos controle e verum. Na análise combinada dos experimentos de 2009-2010, as taxas médias de germinação em 24 h foram (85,9 + 2,6) no grupo controle e (82,1 + 5,7) no grupo G30x (média + DP no nível dos experimentos em %, N = 2.000 por grupo). A taxa de germinação de verum foi 4,4% menor (96,6% de 4,4 + 96,6 = 101) que a do controle (100%). Essa diferença é estatisticamente significativa (p < 0,001) e o tamanho do efeito (d) é grande (> 0,8). Observações realizadas em outros momentos entre 0 e 40 horas de germinação constaram resultados similares. Praticamente, não foi achada diferença entre os grupos W30x e W0 (p > 0,05). Na análise combinada dos experimentos de 2011, as taxas médias de germinação em 24 h foram (73 + 12) no grupo controle e (73 + 14) no grupo G30x (N = 2.000 por grupo), ou seja, praticamente não houve diferença entre os grupos (p > 0,05). Consideramos que os dados de 2009-2010 sobre a germinação do trigo em até 40 h concordam com nos achados prévios no crescimento do caule de trigo em uma semana, ou seja, confirmam que ácido giberélico 30x pode influenciar, isto é, tornar mais lento, o desenvolvimento do trigo. São discutidos vários motivos para a ausência de toda diferença entre os grupos nos experimentos conduzidos em 2011, incluindo variações sazonais, e sugere-se que os experimentos com germinação de trigo sejam realizados exclusivamente no começo do outono.

Wheat germination and highly diluted agitated gibberellic acid (10-30) ? a multi researcher study / Germinação do trigo e ácido giberélico em alta diluição (10-30) e agitado ? um estudo multicêntrico / Germinación de trigo y ácido giberélico en alta dilución (10-30) y agitado ? estudio multicéntrico

Grains of winter wheat (Triticum aestivum L., Capo variety) were observed under the influence of highly diluted gibberellic acid (10-30) prepared by stepwise dilution and agitation according to a protocol derived from homeopathy (?G30x?). Adequate control was used (water prepared according to the homeopathic protocol ?W30x? and/or untreated water ?W0?). Two sets of multicenter experiments were performed, 4 in 2009-2010 and 4 in 2011, involving altogether 6 researchers, 6 laboratories and 4,000 grains per treatment group. Data were found to be homogeneous within the control groups as well as within the verum groups. When the 2009-2010 experiments were pooled, mean germination rates after 24 hours were (85.9 + 2.6) for the control group and (82.1 + 5.7) for G30x (mean + SD at the level of experiments in %) (N = 2,000 per group). Verum germination rate was 4.4% lower than (i.e. equal to 96.6% of) (4.4 + 96.6 = 101) the control germination rate (100%). The difference is statistically significant (p < 0.001) and the effect size (d) is large (> 0.8). Observations at other points in time between 0 and 40 hours of germination yielded similar results. Practically no difference was found between W30x and W0 groups (p > 0.05). When the 2011 experiments were pooled, the mean germination rates after 24 hours were (73 + 12) for the control group and (73 + 14) for G30x (N = 2,000 per group), i.e. there was practically no difference between the groups (p > 0.05). We interpret the data from 2009-2010 on wheat germination within 40 hours as being in line with our previous findings on wheat stalk growth after one week, i.e. as confirmation that gibberellic acid 30x can influence, i.e. slow down, wheat development. Various possible reasons for the absence of any difference between groups in the 2011 experiments, including seasonal variance, are discussed and it is suggested to perform wheat germination experiments in the very beginning of autumn season only.(AU)

Grãos de trigo comum (Triticum sativum L., variedade Capo) foram observados sob a influência de uma alta diluição de ácido giberélico (10-30) preparada através de diluição e agitação seriadas seguindo um protocolo derivado da homeopatia (G30x). Foram utilizados controles adequados (água preparada segundo o protocolo homeopático - W30x - e/ou água sem tratamento - W0 -). Foram realizadas duas séries de experimentos multicêntricos, 4 em 2009-2010 e 4 em 2011, incluindo 6 pesquisadores, 6 laboratórios, e 4.000 grãos em cada grupo de tratamento. Os dados foram homogêneos dentro dos grupos controle e verum. Na análise combinada dos experimentos de 2009-2010, as taxas médias de germinação em 24 h foram (85,9 + 2,6) no grupo controle e (82,1 + 5,7) no grupo G30x (média + DP no nível dos experimentos em %, N = 2.000 por grupo). A taxa de germinação de verum foi 4,4% menor (96,6% de 4,4 + 96,6 = 101) que a do controle (100%). Essa diferença é estatisticamente significativa (p < 0,001) e o tamanho do efeito (d) é grande (> 0,8). Observações realizadas em outros momentos entre 0 e 40 horas de germinação constaram resultados similares. Praticamente, não foi achada diferença entre os grupos W30x e W0 (p > 0,05). Na análise combinada dos experimentos de 2011, as taxas médias de germinação em 24 h foram (73 + 12) no grupo controle e (73 + 14) no grupo G30x (N = 2.000 por grupo), ou seja, praticamente não houve diferença entre os grupos (p > 0,05). Consideramos que os dados de 2009-2010 sobre a germinação do trigo em até 40 h concordam com nos achados prévios no crescimento do caule de trigo em uma semana, ou seja, confirmam que ácido giberélico 30x pode influenciar, isto é, tornar mais lento, o desenvolvimento do trigo. São discutidos vários motivos para a ausência de toda diferença entre os grupos nos experimentos conduzidos em 2011, incluindo variações sazonais, e sugere-se que os experimentos com germinação de trigo sejam realizados exclusivamente no começo do outono.(AU)

Variação sazonal do efeito do ácido giberélico altamente diluído (10-30) sobre o crescimento do trigo - um estudo com vários pesquisadores / Seasonal variation of the effect of extremely diluted agitated gibberellic acid (10-30) on wheat stalk growth - a multi researcher study

Control experiments were performed at different seasons of the year as a follow-up to pilot experiments [1] where a homeopathic high dilution of gibberellic acid had influenced growth in a wheat bio assay (7 days). 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”). Following up on 5 pilot experiments (4 in autumn 2007, 1 in spring 2008), 10 experiments were performed (5 in autumn 2008 or 2009 and 5 in winter 2009 or 2010) with a total of 9 experiments in autumn season (5 researchers, about 9,000 grains), and 6 in winter/spring (4 researchers, about 6,000 grains).(AU)

Experimentos controlados foram realizados em diferentes estações do ano, em continuação a um experimento-piloto [1] onde o ácido giberélico altamente diluído e agitado influenciaram o crescimento de trigo, em um ensaio biológico (7 dias). Grão de trigo de inverno (Triticum aestivum, variedade Capo) foram observados sob a influência do ácido giberélico altamente diluído (10-30) preparado por um processo serial de diluição e agitação, de acordo com um protocolo derivado da homeopatia (“G30x”). Analogamente, o mesmo protocol foi usado para preparar a amostra controle, apenas com água (“W30x”). Em continuidade a realização de 5 experimentos-piloto (4 no outono de 2007, 1 na primavera de 2008), 10 experimentos foram realizados (5 no outono de 2008 ou 2009 e 5 no inverno de 2009 ou 2010) com um total de 9 experimentos na estação do outono (5 pesquisadores, aproximadamente 9.000 grãos) e 6 no inverno/primavera (4 pesquisadores, aproximadamente 6.000 grãos).(AU)