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Background: Mainly due to the use of different inclusion criteria and quality assessments, systematic reviews (SRs) and meta-analyses (MAs) with homeopathic intervention studies (HOMIS) have shown inconsistent results. We aimed to build recommendations for "Summarizing evidence from Homeopathic Intervention Studies" (Sum-HomIS recommendations) in order to approach standardization., Methods: Against the background of a framework-project to update the evidence from homeopathic intervention studies, we launched an expert panel on how to assess the quality of HOMIS and how to summarize evidence from HOMIS. The results of a literature review and the expert communications in advance of the panel as well as the consensus from the discussions are presented here. We added specific considerations for homeopathic veterinary research., Results: On top of the general guidelines when planning a review we report five basic Sum-HomIS recommendations. These are: 1) A broad literature search including special archives and consideration of so-called grey-literature; 2) The inclusion of controlled observational studies alongside randomized controlled trials; 3) The choice of a clear clinical research question in the terms that, if possible, the review project includes studies with predominantly homogeneous populations, interventions, comparators and outcomes (PICOs); 4) The use of a global quality assessment including the assessment of external, model and internal validity; 5) A summary of evidence using the GRADE-approach if the body of evidence is sufficiently large and homogenous or a descriptive summary if it is not so., Conclusions: We present recommendations for designing, conducting, and reporting SRs and MAs with HOMIS., Competing Interests: Declaration of Competing Interest Authors declare none., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Homeopathic complex remedies, composed of several homeopathic medicines in the low potency range, are frequently used in the treatment of a number of common disorders. At the same time, they represent an almost unexplored area of research. Are complex remedies just additive mixtures of the components, or are there interactions between the latter leading to new properties of the complex?, Methods: In the present study, we analyzed as an example the simple bi-component complex, Luffa 4x - Mercurius bijodatus 9x, by means of patterns from evaporated droplets and tested what influences the complex's single compounds have upon the patterns and if there are any interactions. For this purpose, we compared in a series of five experiments patterns from evaporated droplets of the complex, Luffa 4x - Mercurius bijodatus 9x, and three comparison samples in which one or both of the complex's compounds were replaced by potentized solute. The patterns were photographed and evaluated for their gray-level distribution and texture using the software ImageJ . The experimental set-up's stability was tested by means of systematic control experiments., Results: We found that Mercurius bijodatus 9x significantly influenced the patterns of Luffa 4x, increasing their homogeneity; at the same time, the patterns of Mercurius bijodatus 9x combined with solvent were more heterogeneous than those obtained from a control consisting of two pure solvents., Conclusion: In this phenomenological assay, the complex Luffa 4x - Mercurius bijodatus 9x does not correspond to a simple addition of the components. The exact nature of the underlying interaction needs to be elucidated in further investigations., Competing Interests: None declared., (The Faculty of Homeopathy. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/).)

Objectives: Homeopathy (HOM) is a therapeutic method, which is widely used by patients and medical professionals. The medical conditions as well as the homeopathic medical products investigated vary strongly. There is an extensive amount of research, and this necessitates a bibliography that comprehensively presents the entire body of clinical evidence grouped according to medical conditions. Design: Thirty-seven online sources as well as print libraries were searched for HOM and related terms in eight languages (1980 to March 2021). We included studies that compared a homeopathic medicine or intervention with a control regarding the therapeutic or preventive outcome of a disease (classified according to International Classification of Diseases-10). The data were extracted independently by two reviewers and analyzed descriptively. Results: A total of 636 investigations met the inclusion criteria, of which 541 had a therapeutic and 95 a preventive purpose. Seventy-three percent were randomized controlled trials ( n  = 463), whereas the rest were non-randomized studies ( n  = 173). The leading comparator was placebo ( n  = 400). The type of homeopathic intervention was classified as multi-constituent or complex ( n  = 272), classical or individualized ( n  = 176), routine or clinical ( n  = 161) and isopathic ( n  = 19), or various ( n  = 8). The potencies ranged from 1X (dilution of -10,000) to 10 M (100 - 10.000 ). The included studies explored the effect of HOM in 223 medical indications. We present the evidence in an online database. Conclusions: This bibliography maps the status quo of clinical research in HOM. The data will serve for future targeted reviews, which may focus on the most studied conditions and/or homeopathic medicines, clinical impact, and the risk of bias of the included studies.

Objectives: In parts I and II of our review of physicochemical research performed on homeopathic preparations, we identified relevant publications and analyzed the data in terms of individual experiments, looking for the most promising techniques that were used in the past. In this third part, we analyze the results of the experiments seeking to extract information about the possible modes of action underpinning homeopathic preparations. Methods: We summarized the results from the 11 experimental areas previously introduced, extracting the general findings and trends. We also summarized the results in terms of specific research topics: aging, medium used for potentization, sample volume, temperature, material of potentization vessel, and, finally, the use of molecules to probe homeopathic samples. Results: We identified a number of effects that appear consistently throughout the data: Differences to controls seem to increase with: time, moderate temperature, small samples volume, and in ionic medium, whereas high temperatures seem to abolish differences to controls. Based on the present analysis, there is no consistent evidence to date for the nanoparticle hypothesis to explain specific homeopathic treatment effects. However, the quantum coherence domain hypothesis, the dynamic water cluster hypothesis, and the weak quantum theory are still contenders and need to be further assessed experimentally. Conclusions: The field requires further targeted experimentation to validate past findings reporting differences between homeopathic dilutions and controls, and to expand these findings by specifically testing the three main working hypotheses that are currently at hand.

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)., Competing Interests: None declared., (The Faculty of Homeopathy.)

Background: Evaporation-induced pattern formation in droplets has been applied to test effects of high potencies. Here we propose for the first time the droplet evaporation method (DEM) as a tool to characterize low potencies on a qualitative and quantitative basis., Materials and Methods: The present investigation consisted of: (1) screening of 18 different substances of mineral, vegetal, and animal origin in the 1x to 6x potency range; choice of four substances with characteristic pattern-forming properties; (2) replication experiments aiming at the differentiation of four homeopathic preparations at the same potency levels (2x-6x); and (3) control experiments performed on three preparations. The DEM experimental protocol consisted of the evaporation of droplets of the analyzed potency per se , placed on microscope slides and in controlled conditions. The resulting patterns were photographed and subjected to computerized image analysis., Results: The screening experiments yielded a wide variety of patterns. Homeopathic preparations of mineral origin showed the largest variety of forms, whereas potencies of vegetal origin mostly created dendritic patterns, probably due to diffusion-limited aggregation. The here-analyzed image analysis variables (gray-level distribution, entropy, and inverse difference moment) allowed a highly significant differentiation of patterns prepared from four substances ( Echinacea , Baptisia , Luffa , and Spongia ) at the same potency levels in the range 2x to 4x, whereas patterns obtained from potencies 5x and 6x could no longer be differentiated and resembled the pattern of pure solute (purified water). The control experiments showed reasonable experimental model stability., Conclusions: DEM seems to be a promising tool for qualitative phenomenological characterization of homeopathic preparations in low potency. We propose the application of the current experimental model for investigating further research topics in this field, such as the comparison of potencies versus simple dilutions or the contribution of component remedies to the patterns formed by homeopathic combination medicines., Competing Interests: None declared., (The Faculty of Homeopathy.)

Background: Methods based on phase-transition-induced pattern formation (PTPF) are increasingly used in medical research. Frequent application fields are medical diagnosis and basic research in homeopathy. Here, we present a systematic review of experimental studies concerning PTPF-based methods applied to homeopathy research. We also aimed at categorizing the PTPF methods included in this review., Materials and Methods: Experimental studies were collected from scientific databases (PubMed, Web of Science, Russian eLibrary) and from experts in the research field in question, following the PRISMA guidelines. The studies were rated according to pre-defined scientific criteria., Results: The review included 15 experimental studies. We identified seven different PTPF methods applied in 12 experimental models. Among these methods, phase-transition was triggered through evaporation, freezing, or solution, and in most cases led to the formation of crystals. First experimental studies concerning the application of PTPF methods in homeopathic research were performed in the first half of the 20th century; however, they were not continued in the following years. Only in the last decade, different research groups re-launched the idea, introducing new experimental approaches and computerized pattern evaluation techniques. The here-identified PTPF methods are for the first time proposed to be classified as one group of methods based on the same basic physical phenomenon., Conclusions: Although the number of experimental studies in the area is still rather limited, the long tradition in the application of PTPF methods and the dynamics of the present developments point out the high potential of these methods and indicate that they might meet the demand for scientific methods to study potentized preparations., Competing Interests: None., (The Faculty of Homeopathy.)

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., Competing Interests: None., (The Faculty of Homeopathy.)

Objectives: The last systematic review of physicochemical research performed on homeopathic preparations was published in 2003. The aim of the study is to update and expand the current state of knowledge in the area of physicochemical properties of homeopathic preparations. In part 1 of the study, we aim to present an overview of the literature with respect to publication quality and methods used. In part 2, we aim to identify the most interesting experimental techniques. With this, we aim to be in a position to generate meaningful hypotheses regarding a possible mode of action of homeopathic preparations., Methods: A two-step procedure was adopted: (1) an extensive literature search, followed by a bibliometric and quality analysis on the level of publications and (2) a thorough qualitative analysis of the individual physicochemical investigations found. In this publication, we report on step (1). We searched major scientific databases to find publications reporting physicochemical investigations of homeopathy from its origin to the end of 2015. Publications were assessed using a scoring scheme, the Manuscript Information Score (MIS). Information regarding country of origin of the research and experimental techniques used was extracted., Results: We identified 183 publications (compared to 44 in the last review), 122 of which had an MIS ≥5. The rate of publication in the field was ∼2 per year from the 1970s until 2000. Afterward, it increased to over 5.5 publications per year. The quality of publications was seen to increase sharply from 2000 onward, whereas before 2000, only 12 (13%) publications were rated as "high quality" (MIS ≥7.5); 44 (48%) publications were rated as "high quality" from 2000 onward. Countries with most publications were Germany (n = 42, 23%), France (n = 29, 16%), India (n = 27, 15%), and Italy (n = 26, 14%). Techniques most frequently used were electrical impedance (26%), analytical methods (20%), spectroscopy (20%), and nuclear magnetic resonance (19%)., Conclusions: Physicochemical research into homeopathic preparations is increasing both in terms of quantity and quality of the publications.

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., (Copyright © 2015 The Faculty of Homeopathy. Published by Elsevier Ltd. All rights reserved.)

Objectives: The objective was to evaluate homeopathic basic research studies that use plant-based bioassays. With this in view, a compilation was made of the findings of three systematic literature reviews covering plant-based bioassays in the three fields of healthy, abiotically, or biotically stressed plants. This compilation focused on investigations using advanced experimental methods and detailed descriptions, also with the aim of supporting the design of future experiments., Methods: Publications included had to report on studies into the effects of homeopathic preparations on whole plants, seeds, plant parts and cells. Outcomes had to be measured by established procedures and statistically evaluated. A Manuscript Information Score (MIS) was applied using predefined criteria to identify publications with sufficient information for adequate interpretation (MIS ≥ 5). Additional evaluation focused on the use of adequate controls to investigate specific effects of homeopathic preparations, and on the use of systematic negative control (SNC) experiments to ensure the stability of the bioassay. Only a fraction of the studies reported here were performed with 'ultra high' dilutions, whereas other studies were performed with moderate or high dilutions., Results: A total of 157 publications were identified, describing a total of 167 experimental studies. 84 studies included statistics and 48 had a MIS ≥ 5, thus allowing adequate interpretation. 29 studies had adequate controls to identify specific effects of homeopathic preparations, and reported significant effects of decimal and centesimal homeopathic potencies, including dilution levels beyond Avogadro's number. 10 studies reported use of SNC experiments, yielding evidence for the stability of the experimental set-up., Conclusion: Plant models appear to be a useful approach for investigating basic research questions relating to homeopathic preparations, but more independent replication trials are needed in order to verify the results found in single experiments. Adequate controls and SNC experiments should be implemented on a routine basis to exclude false-positive results., (Copyright © 2015 The Faculty of Homeopathy. Published by Elsevier Ltd. All rights reserved.)

Background: Reproducibility of basic research investigations in homeopathy is challenging. This study investigated if formerly observed effects of homeopathically potentised gibberellic acid (GA3) on growth of duckweed (Lemna gibba L.) were reproducible., Methods: Duckweed was grown in potencies (14x-30x) of GA3 and one time succussed and unsuccussed water controls. Outcome parameter area-related growth rate was determined by a computerised image analysis system. Three series including five independent blinded and randomised potency experiments (PE) each were carried out. System stability was controlled by three series of five systematic negative control (SNC) experiments. Gibbosity (a specific growth state of L. gibba) was investigated as possibly essential factor for reactivity of L. gibba towards potentised GA3 in one series of potency and SNC experiments, respectively., Results: Only in the third series with gibbous L. gibba L. we observed a significant effect (p = 0.009, F-test) of the homeopathic treatment. However, growth rate increased in contrast to the former study, and most biologically active potency levels differed. Variability in PE was lower than in SNC experiments. The stability of the experimental system was verified by the SNC experiments., Conclusions: Gibbosity seems to be a necessary condition for reactivity of L. gibba to potentised GA3. Further still unknown conditions seem to govern effect direction and the pattern of active and inactive potency levels. When designing new reproducibility studies, the physiological state of the test organism must be considered. Variability might be an interesting parameter to investigate effects of homeopathic remedies in basic research., (Copyright © 2014 The Faculty of Homeopathy. Published by Elsevier Ltd. All rights reserved.)

Objectives: A bioassay with arsenic-stressed duckweed (Lemna gibba L.) was developed to study potentially regulative effects of homeopathic preparations. We compared potentized substances (nine different potency levels between 17 x and 33 x ) with two controls (unsuccussed and succussed water) regarding their influence on number- and area-related growth rate and color of fronds (leaves). Screening included 11 potentized substances: Arsenicum album, gibberellic acid, nosode, arsenic(V), phosphorus, Conchae, Acidum picrinicum, Argentum nitricum, Crotalus horridus, Hepar sulfuris, and Mercurius vivus naturalis., Design: Duckweed was stressed with arsenic(V) for 48 hours. Afterwards, plants grew in either potentized substances or water controls for 6 days. Growth rate and color of fronds were determined with a computerized image analysis system for different time intervals (days 0-2, 2-6, 0-6). A systematic negative control experiment with unsuccussed water was used to investigate the stability of the bioassay. All experiments were randomized and blinded., Results: Arsenicum album and nosode potencies increased frond number-related growth rate compared to controls (succussed water controls or pooled water controls [succussed and unsuccussed], p < 0.05, t test). Regarding color classification, no effects were observed., Conclusions: The experimental setup with L. gibba stressed by arsenic(V) provides a valuable tool to investigate regulative effects of potentized substances. In order to verify the effects of Arsenicum album and nosode potencies, further independent replication experiments are necessary.

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%, Et50 0.3%, 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.

Background: The last comprehensive review of experimental research on effects of homeopathic treatments on plants was published in 1984, and lacked formal predefined criteria to assess study quality. Since then several new studies with more advanced methods have been published., Objectives: To compile a review of the literature on basic research in homeopathy with healthy plants with particular reference to studies investigating specific effects of homeopathic remedies., Methods: The literature search included English, French, German, Italian, Portuguese and Spanish publications from 1920 to April 2009, using predefined selection criteria. We included experiments with healthy whole plants, seeds, plant parts and cells. The outcomes had to be measured by established procedures and statistically evaluated. We developed a Manuscript Information Score (MIS) and included only publications which provided enough information for proper interpretation (MIS>or=5). A formalised Study Methods Evaluation Procedure (SMEP) was used to evaluate these studies, and the subgroup of studies with adequate controls to identify specific effects., Results: A total of 86 studies in 79 publications was identified, 43 studies included statistics, 29 had MIS>or=5, and 15 studies investigated the specificity of homeopathic preparations. Specific effects of decimal, centesimal and fifty millesimal potencies were found including dilution levels far beyond the Avogadro number. In consecutive series of potencies only some of the tested potencies showed effects. There were many individual studies with diverse methods and very few reproduction trials., Conclusions: Healthy plant models seem an useful approach to investigate basic research questions about the specificity of homeopathic preparations. More investigations with more advanced methods are recommended, especially in the sectors of potentisation techniques, effective potency levels and conditions for reproducibility. Systematic negative control experiments should become a routine procedure to control the stability of the experimental systems.

Background: The literature on the applications of homeopathy for controlling plant diseases in both plant pathological models and field trials was first reviewed by Scofield in 1984. No other review on homeopathy in plant pathology has been published since, though much new research has subsequently been carried out using more advanced methods., Objectives: To conduct an up-to-date review of the existing literature on basic research in homeopathy using phytopathological models and experiments in the field., Methods: A literature search was carried out on publications from 1969 to 2009, for papers that reported experiments on homeopathy using phytopathological models (in vitro and in planta) and field trials. The selected papers were summarized and analysed on the basis of a Manuscript Information Score (MIS) to identify those that provided sufficient information for proper interpretation (MIS>or=5). These were then evaluated using a Study Methods Evaluation Procedure (SMEP)., Results: A total of 44 publications on phytopathological models were identified: 19 papers with statistics, 6 studies with MIS>or=5. Publications on field were 9, 6 with MIS>or=5. In general, significant and reproducible effects with decimal and centesimal potencies were found, including dilution levels beyond the Avogadro's number., Conclusions: The prospects for homeopathic treatments in agriculture are promising, but much more experimentation is needed, especially at a field level, and on potentisation techniques, effective potency levels and conditions for reproducibility. Phytopathological models may also develop into useful tools to answer pharmaceutical questions.

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

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.

Objective: Systematic assessment of the in vitro research on high potency effects., Method: Publications of experiments were collected through databases, experts, previous reviews, citation tracking., Inclusion Criteria: stepwise agitated dilutions <10(-23); cells or molecules from human or animal. Experiments were assessed with the modified SAPEH score., Results: From 75 publications, 67 experiments (1/3 of them replications) were evaluated. Nearly 3/4 of them found a high potency effect, and 2/3 of those 18 that scored 6 points or more and controlled contamination. Nearly 3/4 of all replications were positive. Design and experimental models of the reviewed experiments were inhomogenous, most were performed on basophiles., Conclusions: Even experiments with a high methodological standard could demonstrate an effect of high potencies. No positive result was stable enough to be reproduced by all investigators. A general adoption of succussed controls, randomization and blinding would strengthen the evidence of future experiments.

Objectives: Biologic effects of high homeopathic potencies can be studied in cell cultures using cell lines or primary cells. We hypothesized that primary cells would be more apt to respond to high potencies than cell lines, especially cancer cell lines. We set out to investigate the effects of low doses and high homeopathic potencies of cadmium chloride, respectively, in an intoxication model with human primary lymphocytes compared to a human leukemia cell line (Jurkat)., Design: Cells were pretreated with either low concentrations (nM-microM) or high potencies (pool 15-20c) of cadmium for 120 hours, following which they were exposed to a toxic treatment with a range of cadmium concentrations (8-80 microM) during 24 hours. Cell viability was eventually assessed by use of the MTS/PES assay. Controls included a vehicle (NaCl 0.9%) for the low concentrations of cadmium or water 15-20c for cadmium 15-20c. A total of 34 experiments were conducted, 23 with low concentrations and 11 with high potencies of cadmium. Data were analyzed by analysis of variance., Results: Pretreatment with low concentrations or high potencies of cadmium significantly increased cell viability in primary lymphocytes after toxic challenge, compared to control cells (mean effect +/- standard error = 19% +/- 0.9% for low concentrations respectively 8% +/- 0.6% for high potencies of cadmium; p < 0.001 in both cases). The pretreatment effect of low doses was significant also in cancerous lymphocytes (4% +/- 0.5%; p < 0.001), albeit weaker than in normal lymphocytes. However, high homeopathic potencies had no effect on cancerous lymphocytes (1% +/- 1.9%; p = 0.45)., Conclusions: High homeopathic potencies exhibit a biologic effect on cell cultures of normal primary lymphocytes. Cancerous lymphocytes (Jurkat), having lost the ability to respond to regulatory signals, seem to be fairly unresponsive to high homeopathic potencies.

Introduction: The Health Technology Assessment report on effectiveness, cost-effectiveness and appropriateness of homeopathy was compiled on behalf of the Swiss Federal Office for Public Health (BAG) within the framework of the 'Program of Evaluation of Complementary Medicine (PEK)'., Materials and Methods: Databases accessible by Internet were systematically searched, complemented by manual search and contacts with experts, and evaluated according to internal and external validity criteria., Results: Many high-quality investigations of pre-clinical basic research proved homeopathic high-potencies inducing regulative and specific changes in cells or living organisms. 20 of 22 systematic reviews detected at least a trend in favor of homeopathy. In our estimation 5 studies yielded results indicating clear evidence for homeopathic therapy. The evaluation of 29 studies in the domain 'Upper Respiratory Tract Infections/Allergic Reactions' showed a positive overall result in favor of homeopathy. 6 out of 7 controlled studies were at least equivalent to conventional medical interventions. 8 out of 16 placebo-controlled studies were significant in favor of homeopathy. Swiss regulations grant a high degree of safety due to product and training requirements for homeopathic physicians. Applied properly, classical homeopathy has few side-effects and the use of high-potencies is free of toxic effects. A general health-economic statement about homeopathy cannot be made from the available data., Conclusion: Taking internal and external validity criteria into account, effectiveness of homeopathy can be supported by clinical evidence and professional and adequate application be regarded as safe. Reliable statements of cost-effectiveness are not available at the moment. External and model validity will have to be taken more strongly into consideration in future studies.

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.

Background: Various investigators have observed significant effects of highly diluted histamine on human basophil degranulation in vitro, compared to corresponding water controls. However, active and inactive dilution levels differed in most studies., Objective: We aimed to reproduce former studies with flow-cytometry using rigorously controlled experimental conditions to minimise confounding factors., Methods: In seven independent experiments, basophils of the same human donor were incubated with diluted histamine (up to 10(-34)M) or water controls and activated with anti-IgE antibodies. Basophil activation was determined by using bi-colour flow-cytometry. Experiments were blinded and performed with a randomised arrangement of the solutions on microtiter-plates., Results: Histamine at the dilutions 10(-2)M and 10(-22)M was associated with a significant inhibition of basophil degranulation (p=0.018, Wilcoxon signed rank test) of 23.1% and 5.7%, respectively, if compared to "diluted" water treated in an identical manner. However, if all controls were pooled, only histamine 10(-2)M had a significant effect. Significant effects were seen for row numbers of the microtiter plates., Conclusion: We were not able to confirm the previously reported large effects of homeopathic histamine dilutions on basophil function of the examined donor. Seemingly, minor variables of the experimental set up can lead to significant differences of the results if not properly controlled.

Objectives: In Part 1 of the review of physicochemical research performed on homeopathic preparations the authors identified relevant publications of sufficient reporting quality for further in-depth analysis. In this article, the authors analyze these publications to identify any empirical evidence for specific physicochemical properties of homeopathic preparations and to identify most promising experimental techniques for future studies. Methods: After an update of the literature search up to 2018, the authors analyzed all publications in terms of individual experiments. They extracted information regarding methodological criteria such as blinding, randomization, statistics, controls, sample preparation, and replications, as well as regarding experimental design and measurement methods applied. Scores were developed to identify experimental techniques with most reliable outcomes. Results: The publications analyzed described 203 experiments. Less than 25% used blinding and/or randomization, and about one third used adequate controls to identify specific effects of homeopathic preparations. The most promising techniques used so far are nuclear magnetic resonance (NMR) relaxation, optical spectroscopy, and electrical impedance measurements. In these three areas, several sets of replicated high-quality experiments provide evidence for specific physicochemical properties of homeopathic preparations. Conclusions: The authors uncovered a number of promising experimental techniques that warrant replication to assess the reported physicochemical properties of homeopathic preparations compared with controls. They further discuss a range of experimental aspects that highlight the many factors that need to be taken into consideration when performing basic research into homeopathic potentization. For future experiments, the authors generally recommend using succussed (vigorously shaken) controls, or comparing different homeopathic preparations with each other to reliably identify any specific physicochemical properties. [ABSTRACT FROM AUTHOR]

Objectives: From a pharmaceutical point of view, we see a need to develop stable preclinical test systems to identify and investigate effects of potentized remedies as used in Anthroposophic Medicine and Homeopathy. We evaluated a plant bioassay regarding its capacity to distinguish homeopathic remedies from placebo, applied as sucrose pillules.Methods: Pea seed (Pisum sativum L) was soaked for 24 hours in water with dissolved homeopathic or placebo pillules, or in water only. Shoot length was measured 14 days after planting and treatment groups were compared by analysis of variance (ANOVA). The stability of the system was validated by systematic negative control experiments.Results: The system is suitable to test a common application form - sucrose pillules - of a potentized preparation without influence of the pharmaceutical carrier substance. A screening of 13 potentized preparations revealed Calcium carbonicum to affect pea shoot growth (p < 0.05). Three independent series of main experiments were performed with potentized Calcium carbonicum to assess reproducibility. Meta-analysis of all data revealed significant effects of Calcium carbonicum 12c and 30c on pea shoot growth (p < 0.05), which were however dependent on the date of experiment and/or the experimental series.Conclusions: Potentized Calcium carbonicum, applied as sucrose pillules, influenced pea shoot growth in the assay investigated. However, due to the small effect size and due to the modulation of the effects by still unknown external factors, further optimization of this bioassay is necessary to be used in pharmaceutical quality control or in investigating the biological or pharmaceutical mode of action of potentized preparations. [ABSTRACT FROM AUTHOR]

Objectives: There is a need for preclinical test systems to assess specific effects of homeopathic preparations in commonly available pharmaceutical form. We evaluated a plant bioassay regarding its capacity to distinguish homeopathically prepared pillules from placebo. Methods: Pea seed (Pisum sativum L. cv. Früher Zwerg) was soaked for 24 hours in water with dissolved homeopathic or placebo pillules. Plants germinated and grew in a standard cultivation substrate under controlled environmental conditions. Shoot length was measured 14 days after planting, and treatment groups were compared by analysis of variance (ANOVA). After a screening, three independent series of main experiments assessing the effects of Calcium carbonicum (12c, 30c, 200c) were performed with different experimenters in the same laboratory to assess reproducibility. The stability of the system was validated by systematic negative control experiments. Results: No false positive results were observed in the systematic negative control experiments. Placebo sucrose pillules did not influence shoot growth. A screening of 13 homeopathic preparations revealed Calcium carbonicum 12c to affect pea shoot growth (p = 0.02). In the first series of repetition experiments (n = 8), Calcium carbonicum 30c influenced shoot growth (p = 0.04). In series II and III (n = 2 x10), no significant main effect could be observed. A meta-analysis of all data revealed the effect of Calcium carbonicum 12c and 30c to be dependent on the date of experiment and/or the experimental series. Conclusion: The system is suitable to test a common application form--sucrose pillules--of a homeopathic preparation without influence of the pharmaceutical carrier substance. We observed some evidence that Calcium carbonicum 12c and 30c exerted specific effects in this bioassay. Further optimization of this bioassay is necessary to be used in quality control or in investigating the biological or pharmaceutical mode of action of homeopathic preparations. [ABSTRACT FROM AUTHOR]

Background: There are different types of phase transition-based methods (PTMs) applied in medicine. Two of them, the copper chloride crystallization and droplet evaporation method, find more and more often use also in homeopathic research. The results of the studies conducted until now indicate that PTMs seem to be highly interesting for the analysis of homeopathic treatment effects. Moreover, they are time-saving, economic, and flexible (e.g., PTMs may be used on different kinds of experimental models). Here we present a review of PTMs applied in homeopathy (part of the project 'Systematic Review of Crystallization Processes Applied for Medical Purposes SyRCrysMed'). Materials and Methods: Articles, book chapters, and other materials have been collected from scientific databases (using search terms "HOMEOPATHY" or "HIGH DILUTIONS" and "CRYSTALLIZATION" or "PHASE TRANSITION"), Internet, and libraries. The literature was divided into experimental studies and other literature. Experimental studies were summarized in accordance to following criteria: method/methodology/experimental model, homeopathic treatment, application. The publications regarding homeopathy were rated in accordance to predefined criteria concerning the REHBaR guidelines and crystallization methodology. Results and Discussion: Fourteen publications (7 articles, 3 abstracts, 4 book chapters) and two other online materials have been collected describing initially six different PTMs applied in homeopathy. Twelve experimental studies met the quality expectations of the predefined criteria and were considered for the review. All considered studies were basic research studies and concerned four different PTMs, 18 remedies, and potencies ranging from 1xH to 200cH (with lactose, and/or potentized or unpotentized water as controls). It seems that the development of PTMs was not constant over time: collected publications can be divided into old and recent studies, with a gap from 1975 to 2009. Conclusion: The results of this review point at a high potential of PTMs in homeopathic basic research. [ABSTRACT FROM AUTHOR]

Objective: A bioassay with arsenic-stressed duckweed (Lemna gibba L.) had revealed effects of potentized Arsenicum album . Our research question is whether this bioassay can be modified by using mercury for stressing the duckweed plants and by applying potentized mercury for treatment. In addition, by interchanging and combining the two stressors and the two potentized substances in the test system, we aim at developing a test system for the simile principle as well as at testing the basic principle of homeopathic combination remedies. Methods: The experimental setting of Jäger et al 2010 had to be built up at a new location and adopted for the usage of mercury as a stressor for duckweed. Different soluble mercury compounds were screened to identify the most suitable stress factor (stable upon dissolution, low standard deviation, broad effect range, stable ED50 values). In addition, a growth chamber with homogeneous lighting and temperature condition as well as minimal air movement to prevent evaporation differences of the nutrient solution had to be designed and built. Results: The highly standardized conditions, which were reached by a special constructed growth chamber allowed achieving very small coefficients of variance (0.64%). In three systematic negative control experiments, no significant effects were observed (p = 0.580) that confirmed the appropriate applicability of mercury(II)chloride for stressing duckweeds and indicated a stable test system. Conclusion and Outlook: A test system that had revealed significant effects of homeopathic remedies on arsenic-stressed duckweed was built up at a second location with mercury-stressed duckweed. As next step, several mercury compounds (e.g., Mercurius corrosivus and Mercurius bijodatus) will be applied as potentized remedies at higher (24-30x) and lower (6-12x) potentization levels. By interchanging and combining the stressors and the remedies, the impact of homeopathic remedies will be investigated. [ABSTRACT FROM AUTHOR]

Introduction: To direct future fundamental research on homeopathic potencies, it is necessary to have a solid overview over previously used methods and experimental results. For this systematic review, we focused on laboratory experiments that investigated physico-chemical properties of homeopathic potencies and compared them to controls or between several potency levels. Methods: Relevant publications were searched in pertinent databases, article references, and personal collections of literature. All articles found were rated by two reviewers according to a manuscript information score (MIS). Articles could score between 0 and 10 points, as 0 to 2 points were given each for description of experimental procedure, materials, measuring instruments, potentization method, and controls. Articles with an average MIS ≥5 were retained for further review. The further review process included a detailed analysis of the used potencies, controls, materials, and methods, the quality of the study design, the statistics used, and finally the reported results. All information was extracted also by two reviewers. The included experiments were classified into groups regarding the used physico-chemical methods, that is, spectroscopy, luminescence, Raman spectroscopy, nuclear magnetic resonance, chromatography, analytical methods, electrical impedance, electrochemistry, calorimetry, imaging methods, and various physical methods. The results per method will be summarized for publication. Results: One hundred and eighty-three publications were submitted to information scoring. Sixty-one publications were excluded due to low MIS, and 122 publications were included in the further review. The categories with the highest number of experiments were electrical impedance, spectroscopy, analytical methods, and nuclear magnetic resonance. High-quality research was characterized by the use of independent production series of the potencies, adequate controls, blinding, randomization, and statistical analyses of the results. Discussion: A rigorous methodology was necessary to receive meaningful results due to the large number of methods and different study designs. We try to identify promising areas for future research. [ABSTRACT FROM AUTHOR]

Objective: Jäger et al observed effects of Arsenicum album in a plant bioassay using arsenic-stressed duckweed (Lemna gibba L.). By reproducing these experiments, the influence of a different location and researcher will be investigated. Moreover, the test system has been expanded with metabolomic analysis to reveal biochemical pathways that can be affected by homeopathic remedies. Methods: The test system with arsenic-stressed duckweed had to be set up at the University of Freiburg. To follow the experimental settings of Jäger et al, a climate chamber was constructed, which allows keeping conditions of temperature, air humidity, and light intensity constant. To prevent cross-contamination in the experiments, a further shielding surrounding each beaker was included. The highly standardized conditions resulted in very small coefficients of variance (≈ 1%). To investigate the stability of the test system, three systematic negative control experiments using only unsuccussed pure water were conducted. In further experiments, decimal diluted and succussed potencies of Arsenicum album in eight potency levels (17x, 18x, 21-23x, 28x, 30x, and 33x) will be tested. Additionally, metabolomic fingerprints of treated and untreated plants will be compared. This complementary analysis allows for a statistical evaluation of several thousand metabolomes with principal components analysis (PCA). Pathways that are affected by homeopathic remedies could possibly be revealed by further analyses. Results: The experimental settings of Jäger et al were reproducible. No significant effects were observed in the systematic negative control experiments, which indicate a stable test system. Conclusion: A test system that revealed significant effects of homeopathic remedies on arsenic-stressed duckweed was recreated at a second location. In addition to the growth rate of duckweed, metabolomics will be used to reveal differences between treated and untreated plants. [ABSTRACT FROM AUTHOR]

Background: In homeopathy, it is usually assumed that ultra-high dilutions (UHDs) need to come in direct contact with the organism to be treated to exhibit their action. However, it has been observed in some models that UHDs may influence organisms also at a distance. Here, we test whether the droplet evaporation method (DEM) might serve as a tool to study such force-like influences. In a series of three DEM experiments (E1-E3), we studied (1) force-like effects occurring between As 2 O 3 45x treatment (As45x) and undiluted, unsuccussed, ultrapure water (W), (2) As45x and wheat seeds, as also (3) whether force-like effects may be shielded by means of aluminum foil. Materials and Methods: Bilayer recipients (falcon tubes of two different sizes put one into another) were used to place near each other, but without a physical contact in E1 As45x and water (called further waterE1), in E2 As45x and seeds (seedsE2), and in E3 As45x and seeds wrapped or not in aluminum foil (seedsE3). After 1 week, in E1 wheat seeds were added to the waterE1 and in E2-3 fresh water to seedsE2 and seedsE3. The 1-hour leakage droplets were then put on slides and evaporated. The resulting patterns were evaluated for their local connected fractal dimension (LCFD). Results: The homeopathic effects passed from As45x to water E1, seeds E2, and seeds E3 through the recipient polyethylene. Conclusion: Our results confirm that UHDs may pass their effects at a distance on water and seeds. Such effects should be considered in homeopathy research, and preventive measures should be applied to avoid cross-over contamination. [ABSTRACT FROM AUTHOR]

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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. [ABSTRACT FROM AUTHOR]

Background: Experimental research on the effects of homeopathic treatments on impaired plants was last reviewed in 1990. Objectives: To compile a systematic review of the existing literature on basic research in homeopathy with abiotically stressed plants using predefined criteria. Methods: The literature search was carried out on publications that reported experiments on homeopathy using abiotically stressed whole plants, seeds, plant parts and cells from 1920 to 2010. Outcomes had to be measured by established procedures and statistically evaluated. Using of a Manuscript Information Score (MIS) we identified those publications that provided sufficient information for proper interpretation (MIS≥5). A further evaluation was based on the use of adequate controls to investigate specific effects of homeopathic preparations and on the use of systematic negative control experiments. Results: A total of 34 publications with abiotically stressed plants was identified, published between 1965 and 2010. The 34 publications described a total of 37 experimental studies. Twenty-two studies included statistics, 13 had a MIS≥5, 8 were identified with adequate controls and 4 with negative control experiments. Significant and reproducible effects with decimal and centesimal potencies were found, including dilution levels beyond Avogadro’s number. One experimental model was independently assessed by another research team and yielded inverted results compared to the original trial. Conclusions: Abiotically stressed plant models seem to be a useful approach to investigate homeopathic basic research questions, but more experimentation and especially more independent replication trials are needed. Systematic negative control experiments should be implemented on a routine basis to exclude false-positive results. [Copyright &y& Elsevier]

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. [Copyright &y& Elsevier]

Background: Inhibition of human basophil activation by highly diluted histamine was reported to be a reliable experimental model to examine biological effects of high dilutions. However, independent replications did not always yield concordant results. Aims: We aimed at performing an independent replication of a former study [1] using rigorously controlled experimental conditions to minimise confounding factors. Materials and Methods: In 20 independent experiments, human basophils were treated with highly diluted histamine (15cH, 16cH, corresponding to 10-30-10-32 M) prior to activation by fMLP (formyl-methionyl-leucyl-phenylalanine peptide). Controls were treated with analogously diluted water (15cH, 16cH). The dilutions were prepared freshly for each experiment in deionised water by successive steps of centesimal dilution and agitation (10 s vortex at high speed). Highly diluted samples were blinded and randomised. All samples were set in triplicates. Activated basophils were determined by flow cytometry using anti-CD203c. 20 independent systematic negative control (SNC) experiments were carried out to investigate possible systematic errors. Results: No difference in basophil activation was observed between the highly diluted histamine samples and the highly diluted water controls. There was no evidence for a blood donor specificity of the results. The SNC experiments demonstrated the stability of the test system. Experimental variability within and between experiments was slightly reduced for the highly diluted histamine samples. Discussion: This study was designed as an independent reproduction of a former study [1]. Though we strictly adopted the experimental procedure described in [1], our results do not confirm the large inhibitory effects observed for histamine 15cH and 16cH. This lack of reproducibility might be due to minor differences in the experimental design, such as blinding and randomising of the samples, which we chose to perform in order to reduce the possibility of artifacts but was omitted in the former study. Conclusions: Laboratory independent replication of homeopathic basic research experiments is still a challenge. Assuming that the results formerly obtained with this model were not due to systematic errors, the quest identifying the crucial factors for successful reproducibility is open for future research. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Background: Experimental research on the effects of treatments with homeopathic preparations on plants was systematically reviewed in three research areas (unstressed plants, abiotically stressed plants, and phytopathological models) in 2009/2011. Aims: The objective of this study was to compile a synthesis of these three recent systematic literature reviews to obtain a general overview on the use of plant bioassays in homeopathic basic research. Methods: Literature search was carried out on publications that reported experiments with homeopathic preparations on whole plants, seeds, plant parts or cells from 1920 to 2010, in healthy, abiotically or biotically stressed conditions. Outcomes had to be measured by established state-of-the-art procedures and statistically evaluated. Using a Manuscript Information Score (MIS) those publications were identified that provided sufficient information for proper interpretation (MIS > 5). Further evaluation focused on the use of adequate controls to investigate specific effects of homeopathic preparations and on the use of systematic negative control experiments to ensure proper system performance. Results: A total of 157 publications with plants were identified. The 157 publications described a total of 167 experimental studies. 84 studies included statistics and 48 had a MIS > 5 allowing proper interpretation. 29 studies were identified with adequate controls to identify specific effects of homeopathic preparations, reporting significant effects of decimal and centesimal homeopathic potencies, including dilution levels beyond Avogadro's number. Studies that tested series of consecutive potency levels reported a non-linear and discontinuous relation between effect and potency level. There were many individual studies with diverse methods and very few replication trials. 10 studies reported use of systematic negative control experiments, yielding evidence for the stability of the experimental set-up. Conclusions: Plant models appear to be a useful approach to investigate basic research questions on homeopathic preparations, but more independent replication trials and systematic research are needed. Systematic negative control experiments should be implemented on a routine basis to exclude false-positive and false-negative results. [ABSTRACT FROM AUTHOR]

Background: A major challenge of homeopathic basic research is to develop test systems that yield consistent results. Outcome of plant bioassays is usually based on growth parameters (e.g. germination rate, seedling length, leaf area). Aims: We aimed to evaluate the potential of a crystallisation method with additives ("biocrystallisation") as complementary outcome measure. This method used is based on the crystallographic phenomenon that when crystallising watery solutions of dihydrate CuCl2 in the presence of organic additives (juices/extracts), reproducible dendritic crystal structures are observed. The resulting biocrystallograms can be evaluated visually and/or by computerized image analysis. Methods: Cress seeds (Lepidium sativum L.) germinated and grew in vitro in either Stannum met. x or water 30x. Per experiment, six coded (blinded) x preparations were applied in randomized order, representing three independent replicates of the two treatments. Seedlings grew for 96 hours in darkness and were subsequently processed into a watery extract. Biocrystallisation was performed on circular glass plates in 6-fold replication per treatment group, yielding 36 biocrystallograms per experiment. A total of 15 independent experiments were performed at two independent laboratories. Biocrystallograms were scanned and analysed by computerized texture image analysis, using 15 second-order parameters as outcome measure. 3-way-ANOVA with the independent parameters treatment (n=2), internal replicate (n=3), and number of experiment (n=15) was used to analyse the data. Results: All 15 texture analysis (TA) variables yielded significant (p<0.05) or highly significant (p<0.01) results for the homeopathic treatment. 12 out of 15 second-order TA variables yielded significant (p<0.05) or highly significant (p<0.01) statistical interactions between experimental day and potency treatment, meaning that the difference between Stannum met. x and water x seemed to exhibit varying results for different experimental days. Three out of 15 TA variables did not show any such interaction meaning that the effects of Stannum met. x versus water x were quite reproducible over the 15 independent experiments for these three TA variables. Two variables yielded differences between the internal replicates (p<0.05), most probably due to a processing order effect. There were only minor differences between the results of the two laboratories. Conclusions: The texture of biocrystallograms of homeopathically treated cress exhibited specific characteristics, differentiating water x and Stannum met. x. Thus, the biocrystallisation method seems to be a promising complementary outcome measure for plant bioassays investigating effects of homeopathic preparations. [ABSTRACT FROM AUTHOR]

Introduction: Homeopathic preparations in low potencies, containing still measurable quantities of the starting substance, constitute a unique research field in homeopathic basic research. Here a series of experiments is presented carried out by means of the droplet evaporation method (DEM), investigating the specificity of the method, and presumed effects of the succussion procedure applied in the production of homeopathic preparations. Methods: DEM analysis consisted in the evaporation of droplets of the potencies per se placed on microscope slides. Resulting patterns were photographed. Images were evaluated by means of ImageJ software, by measuring grey level distribution, texture, and fractality. The experimentation consisted of four series: (i) screening (1x-6x potencies from 19 substances), (ii) differentiation experiments (2x-6x potencies of Echinacea, Baptisia, Luffa, and Spongia), (iii) differentiation between succussed (100 or 10 times) and unsuccussed samples (Echinacea 2x, Baptisia 3x, Baptisia 4x, Luffa 4x, and Spongia 6x). (iv) investigation of the influence upon the patterns of single compounds present in a remedy complex. The experimental set-up stability was examined by systematic positive control experiments. Results: (i) Homeopathic preparations of mineral origin showed the greatest form variety, whereas those of vegetal origin created fractal patterns in the potency range 2x-4x. (ii) Differentiation of potencies of different origin at the same dilution level was possible from 2x to 4x. (iii) In all potency levels, succussed (100 and 10 times) and unsuccussed variants could be significantly differentiated. Significant differences between all variants were found in some cases in potency levels 4x and higher. In general, application of succussion reduced size, homogeneity, and complexity of the DEM patterns. (iv) Patterns of a remedy complex Luffa 4x - Mercurius bijodatum 9x showed a clear predominance of the Luffa 4x; however also the second component, present in a much lower concentration, influenced significantly the pattern of the remedy complex as also differed significantly from the pattern of succussed water control. Conclusions: The results suggest that DEM is a suitable tool for scientific investigation of homeopathic preparations in the low potency range. DEM might be applied to assess further research questions, such different potentization procedures (vessel shape, overhead volume, material), storing time, and difference between batches. [ABSTRACT FROM AUTHOR]

Background: Evaporation induced pattern formation in droplets has been successfully applied to test the effects of high and ultra-high potencies. Here we propose for the first time the droplet evaporation method (DEM) as a tool to distinguish low potencies prepared from different substances on a qualitative and quantitative basis. Materials and methods: The present experimentation consisted of (i) screening of 1x-6x potencies deriving from 19 different substances of mineral, vegetal, and animal origin; choice of 4 substances characterized by most suitable pattern forming properties, (ii) experiments aiming the differentiation of potencies of same dilution level prepared from 4 pre-chosen substances in the potency range 2x-6x, and (iii) control experiments performed on 3 potencies. DEM experimental protocol consisted in the evaporation of droplets of the analyzed potency per se, placed on microscope slides and in controlled conditions. The resulting patterns were photographed and subjected to a computerized pattern evaluation. Results: Screening experiments delivered a wide variety of patterns formed involving different pattern forming phenomena. Potencies of mineral origin showed the greatest form variety, whereas potencies of vegetal origin created in the potency range 2x-4x fractal patterns probably in course of diffusion limited aggregation. The differentiation of potencies in same dilution levels prepared from 4 pre-chosen substances (Echinacea, Baptisia, Luffa, and Spongia) was possible in potency-range 2x-4x, whereas patterns obtained from potencies 5x and 6x could no longer be differentiated and resembled the pattern of pure solute (water). The control experiments showed reasonable experimental model stability. Conclusions: DEM seems to represent a promising tool for further studies of low potencies. We propose the application of the here presented experimental model for investigating questions, such as, comparison of potencies versus dilutions, qualitative studies on complex homeopathic preparations in regard on the influences their single compounds exhibit on the pattern. [ABSTRACT FROM AUTHOR]

Introduction: 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 a series of experiments performed by our team showing DEMs potential also for homeopathy 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 Spongia until 4x [2]. Furthermore, the patterns varied in function of the number of 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 quercus 3x influenced the DEM patterns; the chaotic succussions reduced, whereas laminar flow enhanced the patterns complexity vs. the unsuccussed control. (ii) The addition of Mercurius bijodatus 9x to Luffa 4x 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 album 45x or heat-damaged into Zincum metallicum 30c 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 of succussion strokes applied during potentization, as could 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 similar methods has also been reviewed by our team for what concerns the application in homeopathy basic research [7]. [ABSTRACT FROM AUTHOR]

Background: Since ancient times, preparations from traditional medicinal plants e.g. Arnica montana, Calendula officinalis or Hypericum perforatum have been used for different wound healing purposes. The aim of this study was to investigate the efficacy of the commercial low dilution homeopathic remedy Similasan® Arnica plus Spray, a preparation of Arnica montana 4x, Calendula officinalis 4x, Hypericum perforatum 4x and Symphytum officinale 6x (0712-2) and medium diluted SIM WuS (Petroleum 15x, Arnica montana 15x, Calcium fluoratum 12x, Calendula officinalis 12x, Hepar sulfuris 12x and Mercurius solubilis 15x; 1101-4), on the wound healing in cultured NIH 3T3 fibroblasts. Both remedies were from Similasan AG (Jonen, Switzerland) and prepared according the German Homoeopathic Pharmacopoeia (GHP) following descriptions 4a for arnica, 3a for marigold and St. John's wort, 2a for comfrey, 5a for petroleum, and 6 for calcium fluoride, hepar sulfuris and mercurius solubilis. Materials and Methods: Cell proliferation, migration and wound closure promoting effect of the preparations (0712-2, 1101- 4) and their succussed solvents (0712-1, 1101-3) were investigated on mouse NIH 3T3 fibroblasts. Cell viability was determined by WST-1 assay, cell growth using BrdU uptake, cell migration by chemotaxis assay and wound closure by CytoSelect ™Wound Healing Assay Kit which generated a defined wound area. All assays were performed in three independent controlled experiments. In some experiments diluted unsuccussed alcohol (0712-3) was also investigated. Results: Preparations (0712-1), (0712-2), (0712-3), (1101-3) and (1101-4) were investigated at decimal dilution steps from 1x to 4x. Cell viabilty was not affected by any of the substances and (0712-1) and (0712-2) showed no stimulating effect on cell proliferation. Preparation (0712-2) exerted a stimulating effect on fibroblast migration (31.7%) vs 15% with succussed solvent (0712-1) at 1:100 dilutions (p<0.001). Unsuccussed solvent (0712-3) had no influence on cell migration (6.3%; p>0.05). Positive control 2 ng/ml EGF increased migratory activity of cells by 49.8%. Preparation (0712-2) at a dilution of 1:100 promoted in vitro wound closure by 59.5% and differed significantly (p<0.001) from succussed solvent (0712-1), which caused 22.1% wound closure. Medium diluted remedy (1101-4) exerted accelerating effect on wound closure after 14h of treatment. Wounded area was closed by 20% with (1101-4) and 13% by (1101-3) compared to untreated control. Succussed solvent (1101-3) caused about 23% and the remedy (1101-4) about 30% wound closure after 24h. Remedy (1101-4) and succussed solvent (1101-3) modestly stimulated cell growth at dilutions 1:100 and 1:1000 by about 25% and 15%, respectively. No statistically significant differences between preparations 1101-3 and 1101-4 could be detected. Conclusions: Our results demonstrate that the Similasan® Arnica plus low dilution homeopathic remedy exerted wound healing potential, which is a result of increased ability of fibroblasts to migrate without affecting cell proliferation. Medium diluted preparation SIM WuS exerted stimulating effect on the wound closure accompanied by a cell proliferating effect. Used in vitro wound closure test was sensitive enough for low dilutions preparation, however for medium diluted preparation despite of a trend, no significant differences could be detected. [ABSTRACT FROM AUTHOR]

Background: Several series of experiments from our research group have shown ultraviolet (UV) light transmission of homeopathic preparations to slightly but significantly differ from controls. We now investigated whether visible and near infrared spectroscopy were also useful for exploring properties of homeopathic preparations. Materials and methods: Homeopathic preparations of copper sulfate (CuSO4), hypericum and sulfur (S8) were produced in 30 sequential steps of 1:100 dilutions (c-preparations). As controls, succussed potentization medium was used. Transmission of the samples from 190-1100nm was measured 4 times on 5 days with a double beam Shimadzu UV PC 1601 spectrophotometer. To correct for the daily variations of the spectrophotometer, transmission of the samples at each nm was divided by the average transmission of the controls. Median transmissions of the samples were calculated for the ranges of 190-340nm (near and middle UV), 340-640nm (visible light without red), and 640-1100nm (red and near infrared). Differences in the median transmission between potency levels from 6c to 30c were determined using Kruskal-Wallis and Jonckheere-Terpstra tests. Results: Differences in transmissions of the various potency levels were more pronounced in the UV range than in the visible or red/near infrared range. The Kruskal-Wallis test revealed significant differences for homeopathic preparations of CuSO4, hypericum and S8 in the UV range (p=0.032, 0.008, 0.009, respectively) and of S8 in the visible range (p=0.026). Jonckheere's test showed a tendency towards ascending medians with ascending potency levels for CuSO4 in the UV range (p=0.080). Significant trends were revealed for hypericum in the visible range (p=0.042, descending medians) and S8 in the UV range (p=0.015, ascending medians). Conclusion: UV spectroscopy seemed to be more suitable for investigating homeopathic preparations than visible or near infrared spectroscopy, since differences in transmission were more pronounced in the UV range. [ABSTRACT FROM AUTHOR]