scholarly journals Reproducibility of effects of the homeopathic dilutions 14x - 30x of gibberellic acid on growth of Lemna gibba L.

2021 ◽  
Vol 11 (40) ◽  
pp. 196-197
Author(s):  
Vera Majewsky ◽  
Claudia Scherr ◽  
Sebastian Arlt ◽  
Peter Klocke ◽  
Stephan Baumgartner
Keyword(s):  
Gibberellic Acid ◽  
Test Organism ◽  
Test System ◽  
Negative Control ◽  
Control Area ◽  
System Stability ◽  
Lemna Gibba ◽  
Test Substance ◽  
Image Analysis System ◽  
Water Control

Background: Reproducibility of investigations in homeopathy is still challenging. Duckweed (Lemna gibba L.), a monocotyledonous waterplant which mostly reproduces vegetatively and therefore builds genetically identical clones, may be a suitable test system for standardised trials. Aims: This study investigated if formerly observed effects of gibberellic acid 14x – 30x on growth of Lemna gibba were reproducible. Methododology: Duckweed was grown in dilutions of gibberellic acid (14x–30x) as well as once succussed (c1) and unsuccussed (c0) water control. Area-related growth rate for day 0–7 was determined by a computerised image analysis system. Three series including five independent blinded and randomised experiments each were carried out in the same way as in the original study. Only time and conductor of experiments were modified. System stability was controled by three series of systematic negative control (SNC) experiments with the same set-up, but distilled and autoclaved water was used as the only test substance. According to the series with gibberellic acid, each serie of SNC experiments included five experiments. Full two-way ANOVA (α = 5%) was used for statistical analysis. Independent variables were treatment and experiment number, dependent variable was r(area) for day 0–7. Data of each experiment was normalised to its mean value to allow a better comparison between experiments. Only if the global ANOVA F-test was significant (p < 0.05) we compared the investigated groups with Fisher`s LSD test (protected Fisher`s LSD). Results: No specific effects of agitated dilutions of gibberellic acid were found in the first two replication series (p=0.263 and p=0.062). In the third serie with gibbous Lemna gibba L. we observed a significant effect (p=0.009) of the homeopathic treatment, however growth was increased in contrast to decreasing in the former study. Variability in experiments with gibberellic acid 14x – 30x was lower than in SNC experiments. The stability of the experimental system was verified by the SNC experiments. Conclusions: When designing new studies to investigate reproducibility, different physiological states of the test organism must be considered. Variability might be an interesting parameter to investigate effects of homeopathic remedies in basic research.

scholarly journals Effects of Homeopathic Arsenicum Album, Nosode, and Gibberellic Acid Preparations on the Growth Rate of Arsenic-Impaired Duckweed (Lemna gibbaL.)

2010 ◽  
Vol 10 ◽  
pp. 2112-2129 ◽  
Author(s):  
Tim Jäger ◽  
Claudia Scherr ◽  
Meinhard Simon ◽  
Peter Heusser ◽  
Stephan Baumgartner
Keyword(s):  
Growth Rate ◽  
Gibberellic Acid ◽  
Growth Rates ◽  
Statistical Power ◽  
Test System ◽  
Negative Control ◽  
Test Substance ◽  
Image Analysis System ◽  
Water Control ◽  
Analysis System

This study evaluated the effects of homeopathically potentized Arsenicum album, nosode, and gibberellic acid in a bioassay with arsenic-stressed duckweed (Lemna gibbaL.). 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 (≈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 withL. gibbaimpaired 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 the effects of homeopathically potentised Argentum nitricum on the growth of Lemna gibba L. in a randomised and blinded bioassay

Homeopathy ◽  
2017 ◽  
Vol 106 (03) ◽  
pp. 145-154 ◽  
Author(s):  
Claudia Scherr ◽  
Claudia Schneider ◽  
Sebastian Patrick Arlt ◽  
Stephan Baumgartner ◽  
Vera Majewsky
Keyword(s):  
Growth Rate ◽  
Treatment Effect ◽  
Significant Interaction ◽  
Test System ◽  
Negative Control ◽  
Original Study ◽  
System Stability ◽  
Lemna Gibba ◽  
Growth State ◽  
Successful Reproduction

Background: A previous study reported a significant statistical interaction between experiment date and treatment effect of Argentum nitricum 14x–30x on the growth rate of duckweed (Lemna gibba L.). The aim of the present study was to investigate the stability of the test system and intra-laboratory reproducibility of the effects found. Methods: Duckweed was treated with A. nitricum potencies (14x–30x) as well as succussed and unsuccussed water controls. The outcome parameter area-related growth rate for day 0–7 was determined by a computerised image analysis system in two series of independent randomised and blinded experiments. Systematic negative control (SNC) experiments were carried out to investigate test system stability. Statistical analysis was performed with full two-way analysis of variance (ANOVA) and protected Fisher's Least Significant Difference (LSD) test. Results: In the first repetition series we found a significant treatment effect (p = 0.016), while in the second series no effect was observed. The negative control experiments showed that the experimental system was stable. An a posteriori subgroup analysis concerning gibbosity revealed the importance of this growth state of L. gibba for successful reproduction of the statistically significant interaction in the original study; flat: no interaction (p = 0.762); slight gibbosity: no interaction (p = 0.356); medium gibbosity: significant interaction (p = 0.031), high gibbosity: highly significant interaction (p = 0.005). Conclusions: With the original study design (disregarding gibbosity status of L. gibba) results of the original study could not be reproduced sensu stricto. We conclude that the growth state gibbosity is crucial for successful reproduction of the original study. Different physiological states of the test organisms used for bioassays for homeopathic basic research must carefully be considered.

scholarly journals Effects of Homeopathic Preparations of Mercurius corrosivus on the Growth Rate of Severely Mercury-Stressed Duckweed Lemna gibba L.

Homeopathy ◽  
2019 ◽  
Vol 108 (02) ◽  
pp. 128-138 ◽  
Author(s):  
Tim Jäger ◽  
Sandra Würtenberger ◽  
Stephan Baumgartner
Keyword(s):  
Growth Rate ◽  
Stress Intensity ◽  
Growth Rates ◽  
Statistical Power ◽  
Test System ◽  
Lemna Gibba ◽  
Image Analysis System ◽  
Water Control ◽  
Time Period ◽  
The Stability

Background We developed a bioassay with mercury-stressed duckweed (Lemna gibba L.) to study potential effects of homeopathically potentised mercury(II) chloride (Mercurius corrosivus [Merc-c.]). The response of this bioassay to homeopathic treatments as a function of stress intensity was also of interest. Methods Duckweed was severely stressed with mercury(II) chloride for 48 hours. Afterwards plants grew in either Merc-c. (seven different potency levels, 24x to 30x) or water controls (unsuccussed and succussed water) for 7 days. Growth rates of the frond (leaf) area were determined using a computerised image analysis system for different time intervals between the measurements on days 0, 3 and 7. Three independent experiments with potentised Merc-c. each were evaluated. Additionally, three water control experiments were analysed to investigate the stability of the experimental set-up (systematic negative control [SNC] experiments). All experiments were randomised and blinded. Results Unsuccussed and succussed water did not significantly differ in terms of duckweed growth rate. The SNC experiments did not yield any significant effects, providing evidence for the stability of the experimental system. Data from the two control groups and the seven treatment groups (Merc-c. 24x–30x) were each pooled to increase the statistical power. Duckweed growth rates for day 0 to 3 were reduced (p < 0.05) after application of Merc-c. compared with the controls. Growth rates for day 3 to 7 were not influenced by the homeopathic preparations. Conclusions The present test system with Lemna gibba L. that was severely stressed by mercury yielded evidence for specific effects of Merc-c. 24x to 30x, namely a growth reduction in the first time period (day 0–3). This is in contrast to former experiments with slightly arsenic-stressed duckweed, where a growth increase was observed in the second time period (day 2–6). We hypothesise that the differing results are associated with the level of stress intensity (severe versus slight).

scholarly journals Investigation of Arsenic-Stressed Yeast (Saccharomyces cerevisiae) as a Bioassay in Homeopathic Basic Research

2011 ◽  
Vol 11 ◽  
pp. 568-583 ◽  
Author(s):  
Tim Jäger ◽  
Claudia Scherr ◽  
Ursula Wolf ◽  
Meinhard Simon ◽  
Peter Heusser ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Statistical Power ◽  
Growth Parameters ◽  
Basic Research ◽  
Test System ◽  
Negative Control ◽  
Test Substance ◽  
Experimental Setup ◽  
Water Control ◽  
Yeast Saccharomyces Cerevisiae

This study investigated the response of arsenic-stressed yeast (Saccharomyces cerevisiae) towards homeopathically potentized Arsenicum album, a duckweed nosode, and gibberellic acid. The three test substances were applied in five potency levels (17x, 18x, 24x, 28x, 30x) and compared to controls (unsuccussed and succussed water) with respect to influencing specific growth parameters. 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. Yeast grew in microplates over a period of 38 h in either potentized substances or water controls with 250 mg/l arsenic(V) added over the entire cultivation period. Yeast's growth kinetics (slope, Et50, and yield) were measured photometrically. The test system exhibited a low coefficient of variation (slope 1.2%, Et500.3%, yield 2.7%). Succussed water did not induce any significant differences compared to unsuccussed water. Data from the control and treatment groups were both pooled to increase statistical power. In this study with yeast, no significant effects were found for any outcome parameter or any homeopathic treatment. Since in parallel experiments arsenic-stressed duckweed showed highly significant effects after application of potentized Arsenicum album and duckweed nosode preparations from the same batch as used in the present study, some specific properties of this experimental setup with yeast must be responsible for the lacking response.

scholarly journals Effects of Homeopathic Preparations of Mercurius corrosivus on the Growth Rate of Moderately Mercury-Stressed Duckweed Lemna gibba L

Homeopathy ◽  
2021 ◽  
Author(s):  
Tim Jäger ◽  
Sandra Würtenberger ◽  
Stephan Baumgartner
Keyword(s):  
Growth Rate ◽  
Stress Level ◽  
Growth Rates ◽  
Statistical Power ◽  
Experimental System ◽  
Negative Control ◽  
Lemna Gibba ◽  
Image Analysis System ◽  
Analysis System ◽  
Observation Period

Abstract Background A bioassay with severely mercury-stressed duckweed (Lemna gibba L.) had revealed growth-inhibiting effects of homeopathically potentised mercury(II) chloride (Mercurius corrosivus, Merc-c.). We hypothesised that effects of potentised preparations are dependent on the stress level of the organisms used in the bioassay. The aim of the present investigation was to examine the response of duckweed to potentised Merc-c. at a lower stress level. Methods Duckweed was moderately stressed with 2.5 mg/L mercury(II) chloride for 48 hours. Afterwards plants grew in either Merc-c. (seven different potency levels, 24x–30x) or water controls (unsuccussed or succussed water) for 7 days. Growth rates of the frond (leaf) area were determined using a computerised image-analysis system for day 0–3 and 3–7. Three independent experiments with potentised Merc-c. and three systematic negative control experiments were performed. All experiments were randomised and blinded. Results Unsuccussed and succussed water did not significantly differ in their effects on duckweed growth rate. The systematic negative control experiments did not yield any significant effects, thus providing evidence for the stability of the experimental system. Data from the two control groups and the seven treatment groups (Merc-c. 24x–30x) were each pooled to increase statistical power. Duckweed growth rates for day 3–7 were enhanced (p < 0.05) after application of Merc-c. compared with the controls. Growth rates for day 0–3 were not influenced by the homeopathic preparations. Conclusions Moderately mercury-stressed Lemna gibba L. yielded evidence of growth-enhancing specific effects of Merc-c. 24x–30x in the second observation period (day 3–7). This observation is complementary to previous experiments with severely mercury-stressed duckweed, in which a decrease in growth was observed in the first observation period (day 0–3). We hypothesise that the differing results are associated with the level of stress intensity (moderate vs. severe).

REHBaR: a Publication Guideline for Homeopathic Basic Research

2021 ◽  
Vol 11 (40) ◽  
pp. 157-158
Author(s):  
Stephan Baumgartner ◽  
Beate Stock-Schröer
Keyword(s):  
Experimental Design ◽  
Basic Research ◽  
Test System ◽  
Negative Control ◽  
Delphi Process ◽  
System Stability ◽  
Current Evidence ◽  
Randomized Controlled Clinical Trials ◽  
High Standards

Background: Specific efficacy of homeopathically prepared substances in high dilution levels is controversial due to the comparably low number of randomized controlled clinical trials and due to the low concentrations of the substances potentised. No generally accepted theoretical model is currently available to explain specific effects of such preparations. In order to unequivocally demonstrate any such effects, experimental research has to meet very high standards. Aims: Objective was to compile guidelines for homeopathic basic research regarding experimental design, implementation, execution, evaluation and publication. Methods: A Delphi Process was conducted, involving European researchers who published experimental work within the last five years. The Delphi process included a total of five rounds, three rounds of adjusting and phrasing plus two consensus conferences [1]. Eligible criteria were collected from existing publications concerned with the quality of homeopathic basic research. In advance a short questionnaire was sent to a selection of research institutes in Germany active in non-homeopathic basic research. Results: Regarding experimental design, the most important points to consider are: randomized and coded (blinded) allocation of the treatments, several independent experiments (including independent production lots), potentised or succussed controls, positive controls (to control reactivity of the system) and systematic negative control experiments to document test system stability and adequacy of the statistical evaluation [1–4]. A detailed publication guideline for authors was developed. REHBaR (Reporting experiments in Homeopathic Basic Research) provides a checklist of 23 items, supplemented with detailed examples [4]. Background, objectives and possible hypotheses should be given in the part ‘introduction’. Special emphasis is put on the ‘materials and methods’ section, where a detailed description of chosen controls, object of investigation, experimental setup, replication, parameters, intervention, allocation, blinding, and statistical methods is required. The section ‘results’ should present sufficient details on analyzed data, descriptive as well as inferential. Authors should discuss their results and give an interpretation in the context of current evidence. Conclusions: Guidelines how to prepare detailed and informative publications are very common in clinical research [5]. To the best to our knowledge REHBaR is the first guideline to be applied by authors when preparing their manuscripts and to be used by scientific journals in the reviewing process in the field of homeopathic basic research. Furthermore the REHBaR guideline can be helpful for planning and conducting experiments as it includes fundamental qualitative standards. To which extent REHBAR can be used also as an instrument to evaluate the quality of a publication will be discussed.

scholarly journals Reproducibility of effects of homeopathically potentised Argentum nitricum on the growth of Lemna gibba L. in a randomized and blinded bioassay

2021 ◽  
Vol 14 (2) ◽  
pp. 07-08
Author(s):  
Vera Majewsky
Keyword(s):  
Growth Rate ◽  
Gibberellic Acid ◽  
Treatment Effect ◽  
Significant Interaction ◽  
Basic Research ◽  
Original Study ◽  
System Stability ◽  
Lemna Gibba ◽  
Significant Treatment ◽  
Growth State

Background: Reproducibility of basic research investigations in homeopathy is still a challenging issue. A former study reported a significant interaction between date of experiment and treatment effect of Argentum nitricum 14x–30x on the growth rate of duckweed (Lemna gibba L.). In a reproduction trial with duckweed (Lemna gibba L.) and potencies of gibberellic acid duckweed appeared only then sensitive to treatment with homeopathic potencies of gibberellic acid potencies when the plants were in the growth state of gibbosity. Aims: This study investigated if formerly observed effects of Argentum nitricum 14x – 30x on growth of Lemna gibba were reproducible and if sensitivity to homeopathic treatment was influences by the growth state of duckweed. Methods: Duckweed was grown in potencies of Argentum nitricum (14x–30x) and one time succussed and unsuccussed water control (c0 and c1). Area-related growth rate for day 0–7 was determined by a computerised image analysis system. The original study including six independent blinded and randomised experiments each were repeated one time completely and a second time with 3 experiments. Every repetition experiment was carried out in the same way as in the original study, only time and conductor of experiments were modified. System stability was controled by systematic negative control (SNC) experiments with the same set-up, but water as the only test substance. All water used in potency and SNC experiments was distilled and autoclaved before. Statistical analysis was evaluated with full two-way ANOVA (α = 5%), using treatment and experiment number as independent variables and r(area) for day 0–7 as dependent variable. Data of each experiment was normalised to its mean value to allow a better comparison between experiments. Only if the global ANOVA F-test was significant (p < 0.05) investigated groups were compared with Fisher`s LSD test (protected Fisher`s LSD). Results: The original study observed a significant interaction between treatment and experiment number compared to cpool (c0+c1).Caused to the fact that in the first repetition series a significant treatment difference between c0 and c1 was found, additionally a significant interaction between experiment number and c0 and c1, it was not possible to pool c0 and c1. All data (original study, as well as repetition series 1 and 2) was evaluated comparing potencies to one time succussed control c1. This new statistical evaluation confirmed the significant interaction between treatment and experiment number for data of the original study (p = 0.0055). In the first repetition series a significant treatment effect (p = 0.016) was observed, in the second series no effect.Variability in experiments with Argentum nitricum 14x – 30x was higher than in SNC experiments. The stability of the experimental system was verified by the SNC experiments. An a posteriori subgroup analysis suggests that using a specific growth state of Lemna gibba (gibbosity) seems to be crucial for successful reproduction of the significant interaction of the original study. Conclusions: With the original study design chosen (disregarding gibbosity of Lemna gibba), results of the original study could not be reproduced. Bioassays for use in homeopathic basic research must carefully consider different physiological states of the test organisms. Variability might be an interesting parameter to investigate effects of homeopathic remedies in basic research.

scholarly journals Comparative study of two bioassays with weakened duckweed and yeast treated with homeopathic preparations

2021 ◽  
Vol 11 (40) ◽  
pp. 129-130
Author(s):  
Tim Jäger ◽  
Claudia Scherr ◽  
Meinhard Simon ◽  
Peter Heusser ◽  
Ursula Wolf ◽  
...  
Keyword(s):  
Leaf Area ◽  
Basic Research ◽  
Test System ◽  
Negative Control ◽  
System Stability ◽  
Leaf Number ◽  
Yeast Saccharomyces Cerevisiae ◽  
Adequate Test ◽  
Sigmoid Growth Curve ◽  
Homeopathic Remedies

In homeopathic basic research, the question as to the most adequate test systems and apt methodology is still open. This investigation examined the hypothesis that more complex organisms show stronger reactions to homeopathic remedies than less complex ones. We compared two Arsenic (As5+) stressed bioassays with duckweed (Lemna gibba, a multi-cellular autotrophic organism) and yeast (Saccharomyces cerevisiae, a single-cellular heterotrophic organism) regarding their response to homeopathic preparations [1]. For duckweed, growth rates of leaf area and leaf number were evaluated. For yeast, growth kinetics were determined by measuring slope, yield and Et50 (point in time when yield was half maximum) of the sigmoid growth curve. The experiments with duckweed and yeast were performed in parallel (same day, same location and identical homeopathic preparations). After screening 17 substances, three homeopathic preparations (Arsenicum album, nosode, gibberellic acid) were chosen for repeated experimental series [2]. Five independent experiments were conducted for each remedy with both organisms in parallel. Potency levels used were in the range of 17x–33x for duckweed and 17x–30x for yeast. To control for test system stability, systematic negative control experiments were conducted over the complete experimentation period. All experiments were blinded and randomized. The systematic negative control experiments did not yield any significant effects. Application of potentized Arsenicum album in the duckweed bioassay yielded the largest effects compared to water controls without remedies for the parameters leaf area and leaf number (p

scholarly journals Test System Stability and Natural Variability of a Lemna Gibba L. Bioassay

PLoS ONE ◽  
2008 ◽  
Vol 3 (9) ◽  
pp. e3133 ◽  
Author(s):  
Claudia Scherr ◽  
Meinhard Simon ◽  
Jörg Spranger ◽  
Stephan Baumgartner
Keyword(s):  
Test System ◽  
Natural Variability ◽  
System Stability ◽  
Lemna Gibba

Systematic Review of Plant-Based Homeopathic Basic Research: An Update

Homeopathy ◽  
2018 ◽  
Vol 107 (02) ◽  
pp. 115-129 ◽  
Author(s):  
Stephan Baumgartner ◽  
Anezka Sokol ◽  
Roman Huber ◽  
Paul Doesburg ◽  
Tim Jäger ◽  
...  
Keyword(s):  
Influencing Factors ◽  
Basic Research ◽  
Test System ◽  
Negative Control ◽  
Research Field ◽  
Original Study ◽  
System Stability ◽  
Test Systems ◽  
Specific Effects ◽  
Publication Quality

Background Plant-based test systems have been described as a useful tool for investigating possible effects of homeopathic preparations. The last reviews of this research field were published in 2009/2011. Due to recent developments in the field, an update is warranted. Publications on plant-based test systems were analysed with regard to publication quality, reproducibility and potential for further research. Methods A literature search was conducted in online databases and specific journals, including publications from 2008 to 2017 dealing with plant-based test systems in homeopathic basic research. To be included, they had to contain statistical analysis and fulfil quality criteria according to a pre-defined manuscript information score (MIS). Publications scoring at least 5 points (maximum 10 points) were assumed to be adequate. They were analysed for the use of adequate controls, outcome and reproducibility. Results Seventy-four publications on plant-based test systems were found. Thirty-nine publications were either abstracts or proceedings of conferences and were excluded. From the remaining 35 publications, 26 reached a score of 5 or higher in the MIS. Adequate controls were used in 13 of these publications. All of them described specific effects of homeopathic preparations. The publication quality still varied: a substantial number of publications (23%) did not adequately document the methods used. Four reported on replication trials. One replication trial found effects of homeopathic preparations comparable to the original study. Three replication trials failed to confirm the original study but identified possible external influencing factors. Five publications described novel plant-based test systems. Eight trials used systematic negative control experiments to document test system stability. Conclusions Regarding research design, future trials should implement adequate controls to identify specific effects of homeopathic preparations and include systematic negative control experiments. Further external and internal replication trials, and control of influencing factors, are needed to verify results. Standardised test systems should be developed.

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