Highly Diluted Glyphosate Mitigates Its Effects on Artemia salina: Physicochemical Implications.

Glyphosate is an herbicide widely used in agriculture but can present chronic toxicity in low concentrations. Artemia salina is a common bio-indicator of ecotoxicity; it was used herein as a model to evaluate the effect of highly diluted-succussed glyphosate (potentized glyphosate) in glyphosate-based herbicide (GBH) exposed living systems. Artemia salina cysts were kept in artificial seawater with 0.02% glyphosate (corresponding to 10% lethal concentration or LC10) under constant oxygenation, luminosity, and controlled temperature, to promote hatching in 48 h. Cysts were treated with 1% (v/v) potentized glyphosate in different dilution levels (Gly 6 cH, 30 cH, 200 cH) prepared the day before according to homeopathic techniques, using GBH from the same batch. Controls were unchallenged cysts, and cysts treated with succussed water or potentized vehicle. After 48 h, the number of born nauplii per 100 µL, nauplii vitality, and morphology were evaluated. The remaining seawater was used for physicochemical analyses using solvatochromic dyes. In a second set of experiments, Gly 6 cH treated cysts were observed under different degrees of salinity (50 to 100% seawater) and GBH concentrations (zero to LC 50); hatching and nauplii activity were recorded and analyzed using the ImageJ 1.52, plug-in Trackmate. The treatments were performed blind, and the codes were revealed after statistical analysis. Gly 6 cH increased nauplii vitality (p = 0.01) and improved the healthy/defective nauplii ratio (p = 0.005) but delayed hatching (p = 0.02). Overall, these results suggest Gly 6cH treatment promotes the emergence of the more GBH-resistant phenotype in the nauplii population. Also, Gly 6cH delays hatching, another useful survival mechanism in the presence of stress. Hatching arrest was most marked in 80% seawater when exposed to glyphosate at LC10. Water samples treated with Gly 6 cH showed specific interactions with solvatochromic dyes, mainly Coumarin 7, such that it appears to be a potential physicochemical marker for Gly 6 cH. In short, Gly 6 cH treatment appears to protect the Artemia salina population exposed to GBH at low concentrations.

Metal nanoparticle induced hormetic activation: a novel mechanism of homeopathic medicines.

BACKGROUND: High-potency homeopathic remedies, 30c and 200c have enormous dilution factors of 1060 and 10400 respectively. Therefore, the presence of physical entities in them is inconceivable. As a result, their efficacy is highly debated and often dismissed as a placebo. Despite several hypotheses postulated to explain the claimed homeopathic efficacy, none have satisfactorily answered the qualms of the sceptics. Against all beliefs and principles of conventional dilution, we have shown that nanoparticles (NPs) of the starting metals are unequivocally found in the 30c and 200c remedies at concentrations of a few pg/ml. In this paper, our aim was to answer the important question of whether such negligible metal concentrations elicit a biological response. METHODS: Metal-based homeopathic medicines (30c and 200c) were analysed at doses between 0.003%v/v and 10%v/v in in-vitro HepG2 cell-line. Upon treatment, cell response was estimated by MTT assay, FACS and total intracellular protein. Experiments were performed to discern whether the hormesis was a cell-activation or a proliferation effect. RESULTS: Remedies at doses containing a few femtograms/ml levels of the starting metals induced a proliferation-independent hormetic activation by increasing the intracellular protein synthesis. The metal concentrations (at fg/ml) were a billion-fold lower than the studies with synthetic NPs (at µg/ml). Further, we also highlight a few plausible mechanisms initiating a hormetic response at a billion-fold lower dose. CONCLUSIONS: Hormetic activation has been shown for the first time with standard homeopathic high-potency remedies. These findings should have a profound effect in understanding these extreme dilutions from a biological perspective.

In vitro effects of Zinc in soluble and homeopathic formulations on macrophages and astrocytes.

Zinc is an important metal in body homeostasis. Zinc in soluble form (Zn2+) and homeopathic Zincum metallicum were tested in macrophages and astrocytes in order to investigate its potential toxic or therapeutic effects. We evaluated cell viability (WST assay), cytokine production such as tumour necrosis factor alpha (TNF-α) and interleukin 10 (IL-10) by enzyme-linked immunosorbent assay (ELISA) and nitric oxide release by Griess reaction. The effect of zinc-depletion and high zinc pre-treatments on the cell adaptation capability was also investigated. In THP-1 macrophage cell line and in human primary macrophages, Zn2+ at sub-toxic doses (30 µM) caused stimulation of TNF-α and IL-10 with different dynamics reaching the maximum peak at the zinc concentration 100 µM, before the cell death. Highest doses (300 µM) impaired dramatically cell vitality. Similar effects on cell viability were obtained also in C6 astrocytes, where Zn2+ slightly increased the nitric oxide release only in cells activated by one of the pro-inflammatory stimuli used in our cellular model (interferon gamma plus TNF-α). Zinc depletion markedly reduced IL-10 production and cell viability. Zincum metallicum did not cause toxicity in any cell type and showed some small stimulation in WST assay that was statistically significant in a few experimental conditions.

Unequal brothers : are homeopathy and hormesis linked?

The debate between those who believe homeopathy and hormesis derive from the same root and those who believe the two are different phenomena is as old as hormesis. It is an emotionally loaded discussion, with both sides fielding arguments which are far from scientific. Careful analysis of the basic paradigms of the two systems questions the claim of the homeopaths, who find similarities between them. The authors discuss these paradigms, indicating the differences between the claims of homeopathy and hormesis. It is time for thorough and serious research to lay this question to rest. One possible approach is to compare the activity of a hormetic agent, prepared in the usual way, with that of the same agent in the same concentration prepared homeopathically by serial dilution and succussion.

Hormesis: principles and applications.

Hormesis has emerged as a central concept in biological and biomedical sciences with significant implications for clinical medicine and environmental risk assessment. This paper assesses the historical foundations of the dose-response including the threshold, linear and hormetic models, the occurrence and frequency of the hormetic dose response in the pharmacological and toxicological literature, its quantitative and temporal features, and underlying mechanistic bases. Based upon this integrative foundation the application of hormesis to the process of risk assessment for non-carcinogens and carcinogens is explored.

Update on hormesis and its relation to homeopathy.

INTRODUCTION: Hormesis is a dose-response relationship characterized by a biphasic dose response to stressors with a low dose stimulation and a high dose inhibition. The first systematic description of hormesis appeared toward the close of the 18th century by the German pharmacology professor Hugo Schulz. The stressor agent can be any agent or factor capable of causing a deleterious effect. The biological systems can be diverse: bacteria, fungi, algae, yeasts, animals, humans, protozoa and plants. The range of endpoints covers longevity, reproduction, cancer, survival, growth, metabolic effects and others. Hormesis is a nonspecific phenomenon, which can occur in any biological system and can be caused by any stressor. It is quantifiable and reproducible. The apparent similarity between the basic principle of hormesis and homeopathy's Similia Principle, together with the homeopathic claim that hormesis validates homeopathy caused its marginalization, and its rejection during the past century by central figures in pharmacology. Recent years have seen a slight renaissance in the conventional scientific attitude towards hormesis. METHOD: We compared hormesis and homeopathy. RESULT: There is no convincing evidence of similarity between these two systems. Moreover, there are several crucial differences between them, which seem to refute any idea that they stem from the same root. This paper discusses these differences. The rejection of hormesis on grounds of its similarity to homeopathy is unjustified. CONCLUSION: The authors suggest exploring the differences between both systems. Such exploration may answer the key question of whether they do indeed share a root or embrace the same principles. Such exploration may also spur research within both systems to answer further open questions.

Historical foundations of hormesis.

The present paper provides an historical assessment of the concept of hormesis and its relationship to homeopathy and modern medicine. It is argued that the dose-response concept was profoundly influenced by the conflict between homeopathy and traditional medicine and that decisions on which dose-response model to adopt were not based on "science" but rater on historical antipathies. While the historical dispute between homeopathy and traditional medicine has long since subsided, their impact upon the field has been enduring and generally unappreciated, profoundly adversely affecting current drug development, therapeutic strategies and environmental risk assessment strategies and policies.

Hormesis within a mechanistic context.

This paper provides an assessment of the mechanistic foundations of hormesis and how such understandings evolved over the course of the past century. Particular emphasis is placed on recent developments particularly with respect to receptor-based and cell signaling-based pathways. Of particular importance is that the quantitative feature of the hormetic dose response are independent of mechanism.

Hormetic effects of extremely diluted solutions on gene expression.

This paper summarizes the results of investigations showing how molecular biological tools, such as DNA-microarrays, can provide useful suggestions about the behaviour of human organisms treated with microamounts of drugs or homeopathic medicines. The results reviewed here suggest firstly that the action of drugs is not quenched by ultra-high dilution and proceeds through modulation of gene expressions. The efficacy of drug solutions seems to be maintained in ultra-highly diluted preparations, a fact which constitutes a challenge to the dogma of quantization of matter. The second and more important result is that the different gene expression profiles of cell systems treated with the same drugs at different dilutions suggest the existence of hormetic mechanisms. The gene expression profiles of cells treated with copper(II) sulfate, Gelsemium sempervirens and Apis mellifica, are characterized by the same common denominator of the concentration-dependent inversion of gene expression, which can justify at a molecular level the concept of simile adopted in homeopathy. The main conclusion we draw from these results is that these procedures provide new kinds of information and a tool for disclosing the mechanisms involved in hormetic effects. The application of these effects to modern medicine may allow researchers to conceive unprecedented therapeutic applications or to optimize the currently used ones in the framework of a low-dose pharmacology based on a reliable experimental platform.

Enhancement of adaptive biological effects by nanotechnology preparation methods in homeopathic medicines.

Multiple studies have demonstrated that traditional homeopathic manufacturing reagents and processes can generate remedy source and silica nanoparticles (NPs). Homeopathically-made NPs would initiate adaptive changes in an organism as a complex adaptive system (CAS) or network. Adaptive changes would emerge from several different endogenous amplification processes that respond to exogenous danger or threat signals that manufactured nanomaterials convey, including (1) stochastic resonance (SR) in sensory neural systems and (2) time-dependent sensitization (TDS)/oscillation. SR is nonlinear coherent amplification of a weak signal by the superposition of a larger magnitude white noise containing within it the same frequencies of the weak signal. TDS is progressive response magnitude amplification and oscillatory reversal in response direction to a given low dose at physiological limits with the passage of time. Hormesis is an overarching adaptive phenomenon that reflects the observed nonlinear adaptive dose-response relationship. Remedies would act as enhanced micro- and nanoscale forms of their source material via direct local ligand-receptor interactions at very low potencies and/or by triggering systemic adaptive network dynamical effects via their NP-based electromagnetic, optical, and quantum mechanical properties at higher potencies. Manufacturing parameters including dilution modify sizes, shapes, and surface charges of nanoparticles, thereby causing differences in physico-chemical properties and biological effects. Based on surface area, size, shape, and charge, nanoparticles adsorb a complex pattern of serum proteins, forming a protein corona on contact that constitutes a unique biological identity. The protein corona may capture individualized dysfunctional biological mediator information of the organism onto the surfaces of the salient, i.e., resonant, remedy nanostructures. SR would amplify this weak signal from the salient remedy NPs with protein corona adsorbed, leading to sensitized nonlinear dynamical modulation of gene expression and associated changes in biological signaling pathways. When the system reaches its physiological limits during a homeopathic aggravation or the natural disease state, the amplified remedy signal triggers a nonlinear reversal in dynamical direction back towards health.

Cell sensitivity, non-linearity and inverse effects.

It has been claimed that the homeopathic principle of 'similarity' (or 'similia') and the use of individualized remedies in extremely low doses conflicts with scientific laws, but this opinion can be disputed on the basis of recent scientific advances. Several mechanisms to explain the responsiveness of cells to ultra-low doses and the similarity as inversion of drug effects, have again been suggested in the framework of hormesis and modern paradoxical pharmacology. Low doses or high dilutions of a drug interact only with the enhanced sensitivities of regulatory systems, functioning as minute harmful stimuli to trigger specific compensatory healing reactions. Here we review hypotheses about homeopathic drug action at cellular and molecular levels, and present a new conceptual model of the principle of similarity based on allosteric drug action. While many common drugs act through orthostatic chemical interactions aimed at blocking undesired activities of enzymes or receptors, allosteric interactions are associated with dynamic conformational changes and functional transitions in target proteins, which enhance or inhibit specific cellular actions in normal or disease states. The concept of allostery and the way it controls physiological activities can be broadened to include diluted/dynamized compounds, and may constitute a working hypothesis for the study of molecular mechanisms underlying the inversion of drug effects.

High-dilution effects revisited. 1. Physicochemical aspects.

Several lines of evidence suggest that homeopathic high dilutions (HDs) can effectively have a pharmacological action, and so cannot be considered merely placebos. However, until now there has been no unified explanation for these observations within the dominant paradigm of the dose-response effect. Here the possible scenarios for the physicochemical nature of HDs are reviewed. A number of theoretical and experimental approaches, including quantum physics, conductometric and spectroscopic measurements, thermoluminescence, and model simulations investigated the peculiar features of diluted/succussed solutions. The heterogeneous composition of water could be affected by interactive phenomena such as coherence, epitaxy and formation of colloidal nanobubbles containing gaseous inclusions of oxygen, nitrogen, carbon dioxide, silica and, possibly, the original material of the remedy. It is likely that the molecules of active substance act as nucleation centres, amplifying the formation of supramolecular structures and imparting order to the solvent. Three major models for how this happens are currently being investigated: the water clusters or clathrates, the coherent domains postulated by quantum electrodynamics, and the formation of nanoparticles from the original solute plus solvent components. Other theoretical approaches based on quantum entanglement and on fractal-type self-organization of water clusters are more speculative and hypothetical. The problem of the physicochemical nature of HDs is still far from to be clarified but current evidence strongly supports the notion that the structuring of water and its solutes at the nanoscale can play a key role.

High-dilution effects revisited. 2. Pharmacodynamic mechanisms.

The pharmacodynamics aspects of homeopathic remedies are appraised by laboratory studies on the biological effects at various levels (cellular, molecular and systemic). The major question is how these medicines may work in the body. The possible answers concern the identification of biological targets, the means of drug-receptor interactions, the mechanisms of signal transmission and amplification, and the models of inversion of effects according to the traditional 'simile' rule. These problems are handled by two experimental and theoretical lines, according to the doses or dilutions considered (low-medium versus high dilutions). Homeopathic formulations in low-medium dilutions, containing molecules in the range of ultra-low doses, exploit the extreme sensitivity of biological systems to exogenous and endogenous signals. Their effects are interpreted in the framework of hormesis theories and paradoxical pharmacology. The hypotheses regarding the action mechanisms of highly diluted/dynamized solutions (beyond Avogadro-Loschmidt limit) variously invoke sensitivity to bioelectromagnetic information, participation of water chains in signalling, and regulation of bifurcation points of systemic networks. High-dilution pharmacology is emerging as a pioneering subject in the domain of nanomedicine and is providing greater plausibility to the puzzling claims of homeopathy.

Caffeic Acid: Numerous Chemoprotective Effects are Mediated via Hormesis.

Caffeic acid is a common phenolic acid found in coffee and numerous fruits and vegetables. Known for its antioxidant properties, it is widely used as a dietary supplement as part of a polyphenol mixture or as an extract in the form of a capsule or powder. It is also available in liquid form as a homeopathic supplement. Caffeic acid phenethyl ester (CAPE) is an active component of propolis produced by honey bees. Propolis extract is used as a supplement and is available in various forms. The present paper is a comprehensive review of the biomedical literature, showing that caffeic acid effects are hormetic and occur in numerous biological models and cell types for a broad range of endpoints including many aging-related processes. Hormesis is a biphasic dose/concentration response displaying a low concentration/dose stimulation and a high concentration/dose inhibition. Complex alternative search strategies for caffeic acid were used since publications rarely used the terms hormesis or hormetic. Evaluation of the data provides the first assessment of caffeic acid-induced hormetic concentration/dose responses and their quantitative features. Their mechanistic foundations, extrapolative strengths/limitations, and their biomedical, clinical, and public health implications are discussed. Suggestions for future research are presented.

Therapeutic effect of Arsenicum album on leukocytes.

The therapeutic effects of homoeopathic Arsenicum album potencies were investigated in-vitro, using a continuous cell line (MT4), pre-intoxicated with arsenic trioxide (As(2)O(3)), and then treated with succussed and unsuccussed homoeopathic potencies, 6CH, 30CH and 200CH. This study aimed to verify the homoeopathic law of similars and to determine whether potencies diluted beyond Avogadro's constant had physiological effects on cells; whether various potencies would cause different effects as suggested by the concept of hormesis; whether succussed and unsuccussed homoeopathic potencies had different effects on the cells; and to establish whether a biotechnological method could be used to evaluate the above. As(2)O(3) was used to pre-intoxicate and the MTT assay was used to measure the percentage cytotoxicity and half maximal inhibitory concentration (IC(50)) of the cells. The homoeopathic potencies of Arsenicum album (6CH, 30CH and 200CH) were prepared by either succussing or allowing to diffuse for 30 s. After pre-intoxication of the MT4 cells with the IC(50) As(2)O(3) and treatment with succussed and unsuccussed Arsenicum album (6CH-200CH), the cell viability increased with increasing potency from 81% to 194% (over 72 h). The treatments and the times of exposure were found to be statistically significant determinants of cell viability, whereas succussion did not cause any significant variation in the results. The study provided evidence that a biotechnological method (namely cell viability) may be used to scientifically evaluate the physiological effects of homoeopathic potencies on human cells; it confirmed that the homoeopathic potencies did have therapeutic effects; and that succussion was not required in the potentization method in order to produce a curative remedy.

Exploring Possible Mechanisms of Hormesis and Homeopathy in the Light of Nanopharmacology and Ultra-High Dilutions.

Serially diluted succussed solutions of a suitable drug/toxic substance can exhibit physicochemical and biological properties even far beyond Avogadro's limit defying conventional wisdom. They can show hormesis, and homeopathy uses them as medicines. Many studies confirm that they can have an impact on gene expression different than controls. Water in the exclusion zone phase can have memory but for a short period. However, the nanoparticle as the physical substrate can hold information. Nanoparticle and exclusion zone duo as nanoparticle-exclusion zone shell can provide a prolonged memory. The Nanoparticle-Exclusion Zone Shell Model may be an important step toward explaining the nature and bioactivity of serially diluted succussed solutions used as homeopathic medicines. This model may also provide insight into the workings of hormesis. Hormesis is the primary phenomenon through which homeopathic phenomenon may have evolved exhibiting the principle of similars. Hahnemann exploited it to establish homeopathy. The nanoparticle-exclusion zone shells present in the remedy, selected on the principle of similars, can be patient-specific nanoparticles in a symptom syndrome-specific manner. They can carry the drug-specific information for safer clinical applications in an amplified form for high yielding. It suggests homeopathy is a type of nanopharmacology.

Scientific Papers and Patents on Substances with Unproven Effects. Part 2.

Several examples are discussed in this review, where substances without proven effects were proposed for practical use within the scope of evidence-based medicines. The following is discussed here: generalizations of the hormesis concept and its use in support of homeopathy; phytoestrogens and soy products potentially having feminizing effects; glycosaminoglycans for the treatment of osteoarthritis and possibilities of their replacement by diet modifications; flavonoids recommended for the treatment of chronic venous insufficiency and varicose veins; acetylcysteine as a mucolytic agent and its questionable efficiency especially by an oral intake; stem cells and cell therapies. In conclusion, placebo therapies can be beneficial and ethically justifiable but it is not a sufficient reason to publish biased information. Importantly, placebo must be devoid of adverse effects, otherwise, it is named pseudo-placebo. Therapeutic methods with unproven effects should be tested in high-quality research shielded from the funding bias. Some issues discussed in this review are not entirely clear, and the arguments provided here can initiate a constructive discussion.

Hormesis and homeopathy: The artificial twins.

Homeopathy claims a curative reaction from small doses of a substance, high doses of which cause symptoms similar to those the patient is suffering from. Hormesis is a concept of biphasic dose-response to different pharmacological and toxicological agents. According to this concept, a small dose of a noxious agent can exert a beneficial action. A hypothesis is defended here that hormesis as a general principle can be assumed only for the factors present in the natural environment thus having induced adaptation of living organisms. Generalizations of the hormesis phenomenon used in support of homeopathy are unfounded. Low-dose impacts may be associated with a higher risk in a state of organ sub-compensation or failure especially in the elderly patients. Practical recommendations should be based neither on the hormesis as a default approach nor on the postulates of homeopathy. All clinically relevant effects, hormetic or not, should be tested by the methods of evidence-based medicine.

Integrative nanomedicine: treating cancer with nanoscale natural products.

Finding safer and more effective treatments for specific cancers remains a significant challenge for integrative clinicians and researchers worldwide. One emerging strategy is the use of nanostructured forms of drugs, vaccines, traditional animal venoms, herbs, and nutraceutical agents in cancer treatment. The recent discovery of nanoparticles in traditional homeopathic medicines adds another point of convergence between modern nanomedicine and alternative interventional strategies. A way in which homeopathic remedies could initiate anticancer effects includes cell-to-cell signaling actions of both exogenous and endogenous (exosome) nanoparticles. The result can be a cascade of modulatory biological events with antiproliferative and pro-apoptotic effects. The Banerji Protocols reflect a multigenerational clinical system developed by homeopathic physicians in India who have treated thousands of patients with cancer. A number of homeopathic remedy sources from the Banerji Protocols (eg, Calcarea phosphorica; Carcinosin-tumor-derived breast cancer tissue prepared homeopathically) overlap those already under study in nonhomeopathic nanoparticle and nanovesicle tumor exosome cancer vaccine research. Past research on antineoplastic effects of nano forms of botanical extracts such as Phytolacca, Gelsemium, Hydrastis, Thuja, and Ruta as well as on homeopathic remedy potencies made from the same types of source materials suggests other important overlaps. The replicated finding of silica, silicon, and nano-silica release from agitation of liquids in glassware adds a proven nonspecific activator and amplifier of immunological effects. Taken together, the nanoparticulate research data and the Banerji Protocols for homeopathic remedies in cancer suggest a way forward for generating advances in cancer treatment with natural product-derived nanomedicines.

Encontrar tratamientos más seguros y más eficaces para cánceres específicos sigue siendo un desafío significativo para los médicos integrales e investigadores en todo el mundo. Una estrategia emergente es el uso de formas nanoestructuradas de fármacos, vacunas, venenos animales tradicionales, hierbas y agentes nutracéuticos en el tratamiento del cáncer. El reciente descubrimiento de las nanopartículas en medicinas homeopáticas tradicionales aporta otro punto de convergencia entre la nanomedicina moderna y las estrategias intervencionistas alternativas. Una manera en la que los remedios homeopáticos podrían iniciar efectos anticancerígenos incluye acciones de señalización entre células de nanopartículas exógenas y endógenas (exosoma). El resultado puede ser una cascada de acontecimientos biológicos moduladores con efectos antiproliferativos y proapoptóticos. Los protocolos de Banerji reflejan un sistema clínico multigeneracional desarrollado por médicos homeopáticos en la India que han tratado a millares de pacientes con cáncer. Un número de fuentes de remedios homeopáticos de los protocolos de Banerji (p. ej., calcárea fosfórica; carcinosina, tejido derivado del tumor de cáncer de mama preparado homeopáticamente) se solapan con aquellos estudiados en la investigación de la vacuna para el cáncer de exosomas tumorales nanovesiculares y nanopartículas no homeopáticas). Anteriores investigaciones sobre los efectos antineoplásicos de nanoformas de extractos botánicos como la Phytolacca, Gelsemium, Hydrastis, Thuja y Ruta así como sobre la potencia de los remedios homeopáticos derivados de las mismas clases de materiales de origen sugieren otras coincidencias importantes. El descubrimiento replicado de la liberación de silicio, silicona y nanosilicio de la agitación de líquidos en cristal añade un activador inespecífico probado y un amplificador de los efectos inmunológicos. En conjunto, los datos de la investigación de nanopartículas y los protocolos de Banerji de remedios homeopáticos en el cáncer sugieren un camino a seguir para avanzar en el tratamiento del cáncer con nanomedicinas derivadas de productos naturales.

Aspirina en dosis ultrabajas y hormesis / Aspirin in ultra-low doses and hormesis

La relación entre hormesis y Homeopatía resulta hoy en día controvertida. Por un lado, sus primeros fundamentos fueron enunciados con el objeto de explicar los efectos de la Homeopatía. Por otro lado, la mayoría de los trabajos realizados que muestran efectos horméticos fueron realizados a concentraciones mucho mayores que las utilizadas en Homeopatía. El presente trabajo muestra varios puntos de contacto entre la Homeopatía y el concepto de hormesis, y muestra la relación que existe entre los trabajos realizados sobre los efectos de la aspirina 15CH y los distintos tipos de hormesis.