Environmental Homeopathy: Homeopathic Potencies Regulate the Toxicity and Growth of Raphidiopsis raciborskii (cyanobacteria) and can be Tracked Physico-Chemically. Part 1: Biological Results.

INTRODUCTION: Cyanobacteria are microorganisms found in many parts of the world and several genera, such as Raphidiopsis raciborskii, are producers of cyanotoxins. Homeopathic potencies have been found to modulate toxicity in different biological models, and the present study endeavors to discover whether this might also be the case with cyanobacteria. OBJECTIVES: Our objective was to investigate the possible effects of homeopathic potencies on the resilience of Artemia franciscana (brine shrimp) embryos to saxitoxin (STX; cyanotoxin) and on controlling the growth of R. raciborskii in vitro. METHOD: A. franciscana cysts were cultivated in seawater in 96-well plates to evaluate the hatching rate and vitality, plus the gene expression of heat shock proteins (HSPs), after being challenged with R. raciborskii extract containing 2.5 µg/L of STX and treated with different homeopathic potencies. Untreated wells were used as controls ("base-line"). Potencies were chosen from a screening process based on seven selected homeopathic preparations according to the similitude of STX symptoms (Sulphur, Zincum metallicum, Nitric acidum, Plumbum metallicum, Mercurius solubilis, Phosphoric acidum, Isotherapic from R. raciborskii extract; all at 6cH, 30cH and 200cH). Cultures of R. raciborskii maintained in an artificial seawater medium were equally treated with screened homeopathic potencies selected from the same list but specifically for their growth control as a function of time. RESULTS: A 15% lower rate of hatching of A. franciscana cysts was observed after treatment with Nitric acidum 6cH in comparison with baseline (p = 0.05). A complete toxicity reversal was seen after treatment with Isotherapic 200cH, with a 23-fold increase of Hsp 26 gene expression (p = 0.023) and a 24-fold increase of p26 gene expression (p ≤ 0.001) in relation to baseline. Nitric acidum 200cH and Mercurius solubilis 30cH limited the exponential growth of cyanobacteria up to 95% and 85% respectively (p ≤ 0.003) in relation to baseline. Succussed water presented only a transitory 50% inhibition effect. CONCLUSION: Isotherapic 200cH improved A. franciscana bioresilience to STX; Nitric acidum 200cH and Mercurius solubilis 30cH showed the optimal performance on limiting R. raciborskii growth. The results point to the potential of homeopathic potencies to mitigate environmental problems related to water quality.

Environmental Homeopathy: Homeopathic Potencies Regulate the Growth and Toxicity of Raphidiopsis raciborskii (cyanobacteria) and Can be Tracked Physico-Chemically. Part 2: Physico-chemical Results.

INTRODUCTION: The control of cyanobacterial toxicity and growth by homeopathic potencies was described in Part 1 of this two-part report. Here, a parallel approach characterized the physico-chemical features of the potencies used and the liquid media treated with them, correlating these results with their respective biological effects. OBJECTIVES: Our objective was to establish if physico-chemical parameters can track homeopathic potencies in seawater or artificial seawater medium (ASM)-1 and to discover whether these parameters correlate with previously described biological effects. METHOD: Artemia franciscana (brine shrimp) cysts were cultivated in seawater challenged with Raphidiopsis raciborskii extract and treated with different homeopathic potencies chosen from a screening process. Cultures of R. raciborskii maintained in ASM-1 were also treated with previously screened homeopathic potencies, and their growth was monitored as a function of time. The physico-chemical properties of the treated media (seawater or ASM-1) were evaluated by their interaction with solvatochromic dyes and changes in pH, conductivity and temperature. RESULTS: Coumarin 7 was found to be a marker for Nitric acidum 6cH and Isotherapic (R. raciborskii extract) 200cH in seawater (analysis of variance [ANOVA], p = 0.0015). Nile red was found to be a marker for Nitric acidum 200cH and Mercurius solubilis 30cH in ASM-1 (ANOVA, p ≤ 0.001). An increase in pH of ASM-1 and endothermic effects were observed after these treatments (two-way ANOVA, p = 0.0001). Seawater and ASM-1 to which potencies had been added were also subjected to a constant unidirectional 2,400 Gauss static magnetic field and found to have enhanced effects on the solvatochromic dyes tested. CONCLUSION: Homeopathic potencies were specifically traceable in aqueous media using solvatochromic dyes, especially when the samples were subjected to a magnetic field. Results from monitoring other physical parameters, such as pH and temperature, were less specific in relation to potency tracking. However, potency-induced endothermic effects might provide valuable thermodynamic data relating to the nature of potencies.

Evidence from the Use of Solvatochromic Dyes Indicates that Bulk Pure Water does not Potentise.

BACKGROUND: Whilst it is assumed that water plays a crucial role in homeopathy, it is unclear what specific functions water has in the generation, propagation and maintenance of homeopathic potencies. Solvatochromic dyes have demonstrated themselves to be reliable indicators of the presence of potencies and so are well placed to begin to address these questions. OBJECTIVE: The aim of the current study was to ascertain whether pure water could be potentised. If pure water can be potentised, this would indicate that water plays a crucial role in the generation of homeopathic potencies. If it cannot be potentised, then this result points to water being an inert carrier and medium for the potentisation of substances placed in it. METHODS: A specially designed potentiser capable of consistently and reliably producing homeopathic potencies to rigorous standards (Pinkus Potentiser) was used to run up potencies of "Water 10M" and Arsenicum 10M in two interchangeable stainless-steel vials. "Water 10M" and Arsenicum 10M, along with unsuccussed control water, were then tested against solutions of the solvatochromic dye phenol blue by following the absorbance of the dye at 658 nm. RESULTS ": Water 10M" gave results no different statistically from unsuccussed water (p > 0.05) with no change over the time course of the experiments, whereas Arsenicum 10M gave statistically different results (p < 0.001) with increasing divergence from both "Water 10M" and unsuccussed water over time. Similar results were obtained when the vials in which Arsenicum 10M and "Water 10M" were produced were exchanged, so that the vial in which Arsenicum 10M had been produced was then used to produce "Water 10M" and vice versa, indicating the results could not be explained by any anomalies in the production vials themselves. CONCLUSION: Results using the solvatochromic dye phenol blue to detect and measure homeopathic potencies indicate that bulk water itself does not potentise and that unsuccussed water and water that has been succussed are no different. This indicates that what is potentised during cycles of dilution and succussion are the substances placed in water and not the water itself. In turn, this indicates that water is simply a carrier for the homeopathic signal. Evidence from the practice of homeopathy supports this conclusion.

Immobilization of Solvatochromic Dyes on Transparent Cellulose Films: an Improved Method for the Study of Homeopathic Potencies.

BACKGROUND: Highly diluted and succussed solutions (homeopathic potencies) have been shown to interact with a wide range of solvatochromic dyes based on changes in their UV-visible spectra. Studies so far have involved free dyes in solution, but there is a pressing need to find ways to investigate the potency-dye interaction using isolated dye molecules to ask more searching physico-chemical questions regarding the fundamental nature of potencies. AIMS AND METHODS: The aims of the present study have been to look for ways to covalently immobilize solvatochromic dyes onto transparent cellulose films and hence be in a position to investigate dye-potency interactions without the complication of dye-dye interactions, including dye aggregation, which can occur with free dyes in solution. RESULTS: To date, a total of nine different dyes have been immobilized on cellulose films using epoxide activation of hydroxyl groups on the cellulose surface. Using this method, studies have begun looking at the time course of potency action on one of these immobilized dyes, Brooker's merocyanine. Results show that the interaction of Arsenicum 10M with this dye consists of three phases-an initial growth phase, a sustained plateau of interaction, and a final decline phase lasting several days. CONCLUSION: A method has been developed that successfully immobilizes solvatochromic dyes onto transparent cellulose film. These films can then be used in a spectrophotometer to study at a much more detailed level how potencies interact with dyes compared with using free dyes in solution. Results indicate that the information gained in this way provides new insights regarding the fundamental nature of potencies. Specifically, studies using immobilized Brooker's merocyanine with Arsenicum 10M reveal that the lifetime of the potency is much longer than expected and that its action consists of three distinct phases, suggesting a resonant interaction with the dye. How resonant interaction might help to explain the clinical action of potencies is discussed.

Characterization of physicochemical markers for homeopathic medicines and biological supernatant samples

Solvatochromic dyes are probes to detect variations on the dipole moment of solvents after the insertion of homeopathic potencies. Recent studies have shown they can be useful tools in laboratory and field studies to detect the activity of homeopathic remedies.Objective: Determine whether solvatochromic dyes can be a diagnostic tool for cells infected by different agents and/or markers to identify the activity of homeopathic medicines. Methods: Ethilicum1cH, Siliceaterra6, 30, 200cH; Zincummetallicum6, 30, 200cH and Phosphorus6, 30 and 200cH were analyzed by pouring the samples (in a 1:60 rate) into a series of seven dyes (rhodamine, ET 33, ET 30, coumarin 7, NN DMIA, Nile red, methylene violet) diluted in absolute ethanol using pre-established working concentrations. Oscillations of dye absorbance were observed at visible light spectrophotometry according to the remedy and potency. Water and succussed water were used as controls. In a second moment, the absorbance profile of the remedies will be compared with those of biological samples (supernatants) and checked with the biological effect previously obtained from each treatment.Supernatants of RAW 264.7 macrophages stimulated by Calmette-Guérin bacilli (BCG) or infected with Encephalitozoon cuniculiwill be analyzed. Results: Preliminary results have shown that Siliceaterra6cH, Phosphorus30 and 200cH and Zincummetallicum6, 30 and 200cH reduced the absorbance of methylene violet (p=0.01). Repetitions and analysis of supernatants are expected to be performed in the next steps of the study. Future perspectives: Establish a pattern of reactivity of the studied medicines with different dyes and the putative relation with the corresponding supernatants, as an attempt to obtain a "physicochemical signature" for each kind of infection and/or treatment.

Immobilisation of solvatochromic dyes on transparent cellulose films; an improved methodology for the study of homeopathic potencies

Highly diluted and succussed solutions (homeopathic potencies) have been shown to interact with a wide range of solvatochromic dyes based on changes in their UV-visible spectra. Studies so far have involved free dyes in solution, but there is a pressing need to find ways to investigate the potency-dye interaction using isolated dye molecules in order to ask more searching physico-chemical questions regarding the fundamental nature of potencies. Aims and Methods:The aims of the present study have been tolook for ways to covalently immobilizesolvatochromic dyes onto transparent cellulose films and hence be in a position to investigate dye-potency interactions without the complication of dye-dye interactions, including dye aggregation, which can occur with free dyes in solution. Results: to date a total of nine different dyes have been immobilised on cellulose films using epoxide activation of hydroxyl groups on the cellulose surface. Using this methodology studies have begun looking at the time course of potency action on one of these immobilised dyes, Brooker's merocyanine. Results show that the interaction ofArsenicum10M with this dye consists of three phases ­an initial growth phase, a sustained plateau of interaction and a final decline phase lasting several days.Conclusions: amethodology has been developed that successfully immobilises solvatochromic dyes onto transparent cellulose film. These films can then be used in a spectrophotometer to study at a much more detailed level how potencies interact with dyes compared with using free dyes in solution. Results indicate that the information gained in this way provides new insights regarding the fundamental nature of potencies. Specifically, studies using immobilised Brooker's merocyanine with Arsenicum10M reveal that the lifetime of the potency is much longer than expected and that its action consists of three distinct phases, suggesting a resonant interaction with the dye.

Interaction between Solvatochromic Dyes and Water Sampled from a Natural Source Treated with High Dilutions of Phosphorus.

BACKGROUND: Highly diluted and succussed solutions interact with solvatochromic dyes, indicating that changes in solvent and solute polarity could be related to their mechanism of action. It is not known, however, how the activity associated with succussed high dilutions is transferred to untreated water and what the limits of this process are. AIMS: The aims of the present study were to ascertain whether a succussed high dilution of phosphorus (1.5 × 1-59 M; Phos 30cH) seeded into a natural water source that fed a fjord and two connected lakes could propagate itself through the lake system (total volume 2200 m3) and, moreover, whether the process could be tracked using solvatochromic dyes. METHODS: Samples of water were collected before and after seeding, at different times and places throughout the lake system. Controls comprised water taken from an untreated and adjacent, but independent, lake (1385 m3). RESULTS: Water samples taken up to 72 hours after the source treatment produced significant increases (p ≤ 0.03) in the absorbance of the solvatochromic dye methylene violet (MV), while samples from the control lake produced no changes. CONCLUSIONS: The study indicates that activity associated with Phos 30c can propagate itself through large volumes of water, causing changes throughout a whole connected lake system, and that these changes can be tracked using the solvatochromic dye MV. This in turn means the use of homeopathic medicines in large volumes of drinking water, in farming and ecological contexts, now has the potential to be assessed with physico-chemical monitoring.

Homeopathic Potencies May Possess an Electric Field(-like) Component: Evidence from the Use of Encapsulated Solvatochromic Dyes.

BACKGROUND: Homeopathic potencies have been shown to interact with a range of solvatochromic dyes to produce spectroscopic changes in the visible region of the electromagnetic spectrum. Furthermore, the nature of the changes observed under different experimental conditions is beginning to limit the number of possible hypotheses that can be put forward regarding the fundamental identity of potencies. AIMS AND METHODS: The present study uses ß-cyclodextrins to encapsulate solvatochromic dyes of widely varying structures. The purpose of this approach is to de-couple the primary dye-potency interaction from any subsequent aggregation effects. RESULTS: Despite large differences in molecular structure between dyes, results show that potencies affect all dyes according to the same fundamental principles. Specifically, positively and negatively solvatochromic dyes collectively respond in opposite and complementary ways to potencies in accordance with the differential stabilisation of their excited and ground electronic states. Under the conditions of encapsulation, positively solvatochromic dyes display a bathochromic shift of, on average, 0.4 nm with a 2% absorbance change, and negatively solvatochromic dyes display a hypsochromic shift of, on average, 0.2 nm with a 1% absorbance change. This behaviour is only ever seen in two situations-where solvent becomes more polar or where an electric field is applied to solutions of dyes. CONCLUSIONS: The conditions used in this and previous studies to investigate the interaction of potencies with solvatochromic dyes preclude increased polarity of solvent as being responsible for the observed effects and that an explanation in which potencies carry an electric field (or electric field-like) component is by far the more likely. From the magnitude of the spectral changes induced in the dye Brooker's merocyanine by Arsenicum 10M, an estimate of the strength of the postulated electric field of 1.16 × 107 V/m can be made, which is comparable with the potential difference across cell membranes.

Characterization of Antimonium crudum Activity Using Solvatochromic Dyes.

BACKGROUND: The mechanism by which highly diluted and agitated solutions have their effect is still unknown, but the development in recent years of new methods identifying changes in water and solute dipole moments is providing insights into potential modes of action. OBJECTIVE: The objective of the current study was to compare the biological effects of Antimonium crudum (AC) previously obtained by our group and already described in the literature with now measurable physico-chemical effects on solvatochromic dyes. METHODS: Different dilutions of AC and succussed water have been characterized with respect to their effect on the visible spectra of the solvatochromic dyes methylene violet (MV), a pyridinium phenolate (ET33), and a dimethylamino naphthalenone (BDN) compared with in-vitro action against Leishmania amazonensis-infected macrophages. RESULTS: Dye responses varied according to the dye used and the level of AC dilution and results were found to corroborate previously published in-vivo and in-vitro effects of AC. In addition, a very significant enhancement in the absorbance increase of MV was seen using the supernatant from AC 200cH-treated cells (15%; p < 0.0001) over that seen with AC 200cH itself (4%; p = 0.034), suggesting the amplification of ultra-high dilution effects by biological systems. Furthermore, supernatants from AC-treated cells increased the range of dilutions of AC that were capable of producing effects on the spectra of MV. The effect of AC dilutions on dye ET33 was eliminated by a weak electric current passed through potency solutions. CONCLUSION: The data confirm a correspondence between the biological effects of dilutions of AC in-vitro and physico-chemical effects on solvatochromic dyes as measured by changes in their visible spectra. Results also indicate high dilutions of AC are sensitive to exposure to electric currents and biological systems.

Degree of Response to Homeopathic Potencies Correlates with Dipole Moment Size in Molecular Detectors: Implications for Understanding the Fundamental Nature of Serially Diluted and Succussed Solutions.

BACKGROUND: The use of solvatochromic dyes to investigate homeopathic potencies holds out the promise of understanding the nature of serially succussed and diluted solutions at a fundamental physicochemical level. Recent studies have shown that a range of different dyes interact with potencies and, moreover, the nature of the interaction is beginning to allow certain specific characteristics of potencies to be delineated. AIMS AND METHODS: The study reported in this article takes previous investigations further and aims to understand more about the nature of the interaction between potencies and solvatochromic dyes. To this end, the UV-visible spectra of a wide range of potential detectors of potencies have been examined using methodologies previously described. RESULTS: Results presented demonstrate that solvatochromic dyes are a sub-group of a larger class of compounds capable of demonstrating interactions with potencies. In particular, amino acids containing an aromatic bridge also show marked optical changes in the presence of potencies. Several specific features of molecular detectors can now be shown to be necessary for significant interactions with homeopathic potencies. These include systems with a large dipole moment, electron delocalisation, polarizability and molecular rigidity. CONCLUSIONS: Analysis of the optical changes occurring on interaction with potencies suggests that in all cases potencies increase the polarity of molecular detectors to a degree that correlates with the size of the compound's permanent or ground dipole moment. These results can be explained by inferring that potencies themselves have polarity. Possible candidates for the identity of potencies, based on these and previously reported results, are discussed.

HRI Malta 2017-Cutting Edge Research in Homeopathy: HRI's Third International Research Conference in Malta.

The third international conference on "Cutting Edge Research in Homeopathy" organised by the Homeopathy Research Institute (HRI) was held on the inspiring and historic island of Malta from 9th to 11th of June, 2017. One hundred and two abstracts underwent peer review by the HRI Scientific Advisory Committee and external experts to produce the programme of 36 oral presentations and 37 posters, presented by researchers from 19 countries. The 2.5-day programme covered a diverse range of topics, including quantitative and qualitative clinical research, basic research, veterinary research, and provings. These intensive plenary and parallel sessions were interspersed with multiple opportunities for delegates to discuss and exchange ideas, in particular through interactive panel discussions and a pre-conference workshop. The continuing commitment of the homeopathy research community to generate high-quality studies in this rapidly evolving field was clear. In this conference report, we present highlights from this memorable event.

Interaction of homeopathic potencies with the water soluble solvatochromic dye bis-dimethylaminofuchsone. Part 1: pH studies.

INTRODUCTION: Previous studies have demonstrated the potential of solvatochromic dyes for investigating the physical chemistry of homeopathic potencies. RESULTS: Following examination of this class of dyes in organic solvents, results obtained using the positively solvatochromic dye Bis-dimethylaminofuchsone (BDF) in aqueous solution are now reported. Spectral changes observed with this dye in the presence of potencies are both substantial and reproducible. Studies across a wide range of pH values reveal an unusual pH dependence for the dye's interaction with homeopathic potencies. Results indicate potency enhances dye protonation at pH values below c. 7.0, whilst protecting the dye from attack by hydroxyl ions above c. pH 7.5. CONCLUSION: A possible explanation for these observations is offered in terms of a potency - induced electron density shift in BDF. The interaction of homeopathic potencies with solvatochromic dyes, particularly BDF, points towards a possible physico-chemical model for the nature of potencies, how they may be interacting with this class of dyes, and moreover how their biological effects may be mediated.

Solvatochromic dyes detect the presence of homeopathic potencies.

A systematic approach to the design of simple, chemical systems for investigating the nature of homeopathic medicines has led to the development of an experimental protocol in which solvatochromic dyes are used as molecular probes of serially diluted and agitated solutions. Electronic spectroscopy has been used to follow changes in the absorbance of this class of dyes across the visible spectrum in the presence of homeopathic potencies. Evidence is presented using six different solvatochromic dyes in three different solvent systems. In all cases homeopathic potencies produce consistent and reproducible changes in the spectra of the dyes. Results suggest that potencies influence the supramolecular chemistry of solvatochromic dyes, enhancing either dye aggregation or disaggregation, depending upon dye structure. Comparable dyes lacking the intramolecular charge transfer feature of solvatochromic dyes are unaffected by homeopathic potencies, suggesting potencies require the oscillating dipole of solvatochromic dyes for effective interaction. The implications of the results presented, both for an eventual understanding of the nature of homeopathic medicines and their mode of action, together with future directions for research in this area, are discussed.