Self-assembled Patterns Formed in Evaporating Droplets to Analyze Bi-component Homeopathic Preparations in the Low Dilution Range.

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.

Series of experiments performed with the droplet evaporation method on low potencies

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 perse 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 (1x­6x potencies from 19 substances), (ii) differentiation experiments (2x­6x 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 2x­4x. (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.

Formation of Self-organized Structures in Drying Droplets: a Promising Analytical Tool for Homeopathy Basic Research

The droplet evaporation method (DEM) is based on the evaporation-induced pattern formation in droplets and is applied mainly for medical diagnosis[1].Here, we present aseries of experiments performed by our team showing DEMs potential also forhomeopathy basic research, in particular, for the investigation of(i) low potencies, (ii) low potency complexes (physical model), and (iii) the action of high potencies (plant-based model).Methods:(i) DEM differentiated significantly between Luffa, Baptisia, Echinacea, and Spongiauntil 4x[2]. Furthermore, the patterns varied in function of the numberof succussion strokes (0, 10, or 100) applied during potentization[3]. The performance of chaotic succussions vs. laminar flow vs. slight mixing during the potentization of Viscum album quercus3x influenced the DEM patterns; the chaotic succussions reduced, whereas laminar flow enhanced the patterns complexity vs. the unsuccussed control.(ii) The addition of Mercurius bijodatus9x to Luffa4x changed significantly the DEM patterns, even if the material quantity present in the 9x potency lied far beyond that of ultrapure water.(iii) Leakages obtained by placing healthy or arsenic-damaged wheat-seeds into Arsenicum album45x orheat-damaged intoZincum metallicum30c vs. water created significantly different DEM structures [4, 5]. Results:The damaged seeds put into the potency created structures characterized by a higher complexity than those obtained from damaged seeds put into control water. Furthermore, the potency action seemed to increase with rising numbers ofsuccussion strokes applied during potentization,ascould be shown by means of DEM patterns and germination rate using the same wheat-seed model[6].In all our studies, the pattern evaluation was computerized (texture and fractal analysis performed by means of ImageJ) or based on deep-learning algorithms and the robustness of the experimental system was checked by means of systematic control experiments.Conclusion:DEM together with other similarmethods has also been reviewed by our team for what concerns theapplication in homeopathy basic research[7].

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

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).

Physicochemical Investigations of Homeopathic Preparations: A Systematic Review and Bibliometric Analysis-Part 3.

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.

Impact of succussion on pharmaceutical preparations analyzed by means of patterns from evaporated droplets.

The aim of the present study was to investigate if patterns obtained from evaporating droplets of pharmaceutical preparations reveal the impact of succussion on such medicinal products. For this purpose, five pharmaceutical preparations (Echinacea 10-2, Baptisia 10-3, Baptisia 10-4, Luffa 10-4, and Spongia 10-6) were prepared according to the European Pharmacopoeia guidelines for the production of homeopathic remedies, in three variants each: with varying numbers of succussion strokes (i) 100, (ii) 10 (succussed samples), and (iii) zero (gently mixed, unsuccussed sample). System stability was studied by means of systematic positive control experiments. Patterns were evaluated by means of computerized image analysis regarding grey level distribution, texture, and fractality. For all investigated pharmaceutical preparations, significant differences were found between the succussed and gently mixed samples; whereas, all three samples (prepared with 100, 10 and zero succussion strokes) could be significantly differentiated for Luffa 10-4 and Spongia 10-6 for one image evaluation parameter each. Control experiments showed a reasonable stability of the experimental set-up.

Physicochemical Investigations of Homeopathic Preparations: A Systematic Review and Bibliometric Analysis-Part 2.

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.

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

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).

Phenomenological Characterization of Low-Potency Homeopathic Preparations by Means of Pattern Formation in Evaporating Droplets.

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.

Physicochemical Investigations of Homeopathic Preparations: A Systematic Review and Bibliometric Analysis-Part 1.

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.

Pesquisas físico-químicas sobre as preparações homeopáticas: revisão sistemática e análise bibliométrica ­ parte 1 / Physical-chemical research on homeopathic preparations: review systematics and bibliometric analysis

Objetivo: A última revisão sistemática das pesquisas físico-químicas sobre preparações homeopáticas foi publicada em 2003. O objetivo do presente estudo foi atualizar e ampliar o estado atual dos conhecimentos na área das propriedades físico-químicas das preparações homeopáticas. Na parte 1 do estudo objetivamos apresentar um panorama da literatura quanto à qualidade e métodos utilizados nas publicações. Na parte 2 objetivamos identificar as técnicas experimentais mais interessantes. Desta maneira, esperamos estar na posição de gerar hipóteses significativas quanto ao possível modo de ação das preparações homeopáticas. Métodos: Foi adotado um procedimento com 2 etapas: 1) pesquisa ampla da literatura, seguida de análise bibliométrica e da qualidade das publicações e, 2) análise qualitativa minuciosa das pesquisas físico-químicas individuais localizadas. Na presente publicação relatamos a etapa 1). Realizamos uma busca nas principais bases científicas de dados de publicações relatando pesquisas físico-químicas da homeopatia desde a sua origem até o final de 2015. As publicações foram avaliadas através de um sistema de pontuação, o Manuscript Information Score (MIS). Foram coletadas informações sobre o país de origem dos estudos e as técnicas experimentais utilizadas. Resultados: Localizamos 183 publicações (versus 44 na revisão anterior), 122 delas com MIS ≥ 5. A taxa de publicações na área foi ˜2 por ano entre as década de 1970 e 2000. Depois aumentou para mais de 5,5 publicações por ano. [...] Conclusões: As pesquisas físico-químicas sobre as preparações homeopáticas está aumentando tanto na quantidade e na qualidade das publicações. (AU)

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. [...]Conclusions: Physicochemical research into homeopathic preparations is increasing both in terms of quantity and quality of the publications. (AU)

Pesquisas físico-químicas sobre as preparações homeopáticas: revisão sistemática e análise bibliométrica ­ parte 1 / Physical-chemical research on homeopathic preparations: review systematics and bibliometric analysis

Objetivo: A última revisão sistemática das pesquisas físico-químicas sobre preparações homeopáticas foi publicada em 2003. O objetivo do presente estudo foi atualizar e ampliar o estado atual dos conhecimentos na área das propriedades físico-químicas das preparações homeopáticas. Na parte 1 do estudo objetivamos apresentar um panorama da literatura quanto à qualidade e métodos utilizados nas publicações. Na parte 2 objetivamos identificar as técnicas experimentais mais interessantes. Desta maneira, esperamos estar na posição de gerar hipóteses significativas quanto ao possível modo de ação das preparações homeopáticas. Métodos: Foi adotado um procedimento com 2 etapas: 1) pesquisa ampla da literatura, seguida de análise bibliométrica e da qualidade das publicações e, 2) análise qualitativa minuciosa das pesquisas físico-químicas individuais localizadas. Na presente publicação relatamos a etapa 1). Realizamos uma busca nas principais bases científicas de dados de publicações relatando pesquisas físico-químicas da homeopatia desde a sua origem até o final de 2015. As publicações foram avaliadas através de um sistema de pontuação, o Manuscript Information Score (MIS). Foram coletadas informações sobre o país de origem dos estudos e as técnicas experimentais utilizadas. Resultados: Localizamos 183 publicações (versus 44 na revisão anterior), 122 delas com MIS ≥ 5. A taxa de publicações na área foi ˜2 por ano entre as década de 1970 e 2000. Depois aumentou para mais de 5,5 publicações por ano. [...] Conclusões: As pesquisas físico-químicas sobre as preparações homeopáticas está aumentando tanto na quantidade e na qualidade das publicações. (AU)

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. [...]Conclusions: Physicochemical research into homeopathic preparations is increasing both in terms of quantity and quality of the publications. (AU)