Effects of ultra-high dilutions of sodium butyrate on viability and gene expression in HEK 293 cells.

BACKGROUND: Several recent studies reported the capability of high diluted homeopathic medicines to modulate gene expression in cell cultures. In line with these studies, we examined whether ultra-high dilutions (30C and 200C) of sodium butyrate (SB) can affect the expression levels of genes involved in acquisition of a senescence-associated secretory phenotype (SASP) in human embryonic kidney (HEK) 293 cells. METHODS: Cell viability was evaluated using a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. The expression levels of TNF-α, interleukin (IL)-2, IL-4, IL-6 and IL-10 genes were determined by real-time PCR assay. RESULTS: Exposure to both 30C and 200C during 48 h led to a significant decrease of the level of expression of TNF-α gene, while expression of IL-2 gene was increased when exposed to 30C, and expression of IL-10 gene was decreased when exposed to 200C. No changes in expression levels of all genes studied were observed in cells treated with both 30C and 200C remedies of SB during the 24 h. CONCLUSION: Observed changes in gene expression levels after exposure to 30C and 200C remedies of SB during 48 h suggest that extremely low concentrations of this agent can modulate the transcriptome of HEK 293 cells. These results are in line with findings from other studies confirming the ability of homeopathic remedies to modulate gene expression in cell cultures.

Number of succussion strokes affects effectiveness of ultra-high-diluted arsenic on in vitro wheat germination and polycrystalline structures obtained by droplet evaporation method.

OBJECTIVES: The aim of this study is to investigate whether the number of succussion strokes applied after each dilution step when preparing the homeopathic treatments influences the effectiveness of ultra-high-diluted (UHD) arsenic trioxide at the 45th decimal dilution/dynamization (As2O3 45x). DESIGN: Wheat seeds, previously stressed with ponderal As2O3, were treated with: As2O3 45x, H2O 45x (dynamized control), or pure water (negative control). The succussion was done manually, and various succussion durations (numbers of strokes) were tested for each treatment. Treatment effectiveness was tested blind using the in vitro germination test and the droplet evaporation method (DEM). Data were processed by the Poisson test (germination test) and by two-way analysis of variance (DEM). MAIN OUTCOME MEASURES: We evaluated both the in vitro germination rate, by counting the non-germinated seeds, and the complexity of polycrystalline structures (PCS) (local connected fractal dimension (LCFD)) obtained by evaporating leakage droplets from stressed seeds that had been watered with the different treatments. RESULTS: We observed a highly significant increase in germination rate when the number of strokes (NS) was ≥32 for both As2O3 45x and H2O 45x, and a significant increase in the LCFD of PCS for As2O3 45x when the NS was ≥32 and for H2O 45x when it was 70. CONCLUSIONS: Both experimental approaches showed increased effectiveness for treatments prepared with a higher number of succussion strokes. These results indicate that succussion may have an important influence on treatment effectiveness, and so highlight the need for further research.

Polycrystalline structures formed in evaporating droplets as a parameter to test the action of Zincum metallicum 30c in a wheat seed model.

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.

The use of animal models in homeopathic research--a review of 2010-2014 PubMed indexed papers.

BACKGROUND: In the 1990s, a study was performed on the effects of highly diluted thyroxine on frog metamorphosis. This model represented one of the most discussed examples of the biological effects of high dilutions over the next two decades. In 2010, another critical conceptual review of the use of animal models in homeopathy and high-dilution research was published. The main contribution of these studies was the elucidation of the biological features and phenomenology of the effects of high dilutions on living systems, representing an important step forward in our understanding of the mechanisms of action of homeopathic medicines. METHODS: We performed a further review of this line of investigation using the same methods. Fifty-three articles that were indexed in the PubMed database and used 12 different animal species were systematically evaluated. Only a fraction of the studies (29/53) reported herein were performed with "ultra high" dilutions. The other studies were performed with dilutions in ranges below 10(-23) (14/53 articles) or commercial complexes (10/53 articles). RESULTS: Only two articles reported negative results; both used in vivo protocols to test commercial complexes, one in fish and one in bees. The quality of the employed techniques improved in 2010-2014 compared with the studies that were reviewed previously in 2010, with the inclusion of more ethically refined protocols, including in vitro primary cell cultures and ex vivo studies (10/53 articles), often with three or more replicates and analyses of epigenetic mechanisms that were previously unknown in 2010. CONCLUSION: In our updated review of the past 5 years, we found further demonstrations of the biological effects of homeopathy using more refined animal models and in vitro techniques.

From high dilutions to digital biology: the physical nature of the biological signal.

The memory of water was a radical idea that arose in the laboratory of Jacques Benveniste in the late 1980s. Twenty-five years have passed and yet the often angry debate on its merits continues despite the increasing number of scientists who have reported confirmation of the basic results. One working hypothesis was that molecules can communicate with each other, exchanging information without being in physical contact and that at least some biological functions can be mimicked by certain energetic modes characteristics of a given molecule. These considerations informed exploratory research which led to the speculation that biological signaling might be transmissible by electromagnetic means. Around 1991, the transfer of specific molecular signals to sensitive biological systems was achieved using an amplifier and electromagnetic coils. In 1995, a more sophisticated procedure was established to record, digitize and replay these signals using a multimedia computer. From a physical and chemical perspective, these experiments pose a riddle, since it is not clear what mechanism can sustain such 'water memory' of the exposure to molecular signals. From a biological perspective, the puzzle is what nature of imprinted effect (water structure) can impact biological function. A parallel can be drawn between this debate on the memory of water, which presumes that the action of molecules is mediated by an electromagnetic phenomenon, and the often acrimonious debate on the transmission of nerve influxes via synaptic transfer of specific molecules, neurotransmitters. The latter debate began in 1921 with the first experiments by Loewi and was still active in 1949, 28 years later. A strong reluctance to accept research that questions basic aspects of long-accepted biochemical paradigms is to be expected. In this paper we will provide a brief summary of experiments relating to the memory of water: the earlier work on high dilutions (HD) and then the experiments, which followed and continue today, on digital biology.

Update on preliminary elements of a theory of ultra high dilutions.

INTRODUCTION: The different mechanisms: A. of the interaction between the molecular mother substance and the solvent water or ethanol B. of the storage of molecule-specific information in the solvent. C. the physiological basis of the sensitivity of the living organism towards an ultra high dilution (UHD). D. the mechanism of the interaction of the test dilution with the organism are largely unknown. Several ideas have been postulated, and experiments to test them carried out in physics and in biology. METHOD: The authors revisited a 1994 contribution on 'preliminary elements of a theory on UHDs' and updated it with regard to more recent literature and research findings. RESULTS: Although the experimental basis can still be questioned in most cases, remarkable fundamental observations have been made to explain the effects of UHDs. For some topics in question, it appears that information specific properties of the diluted substance to be transferred is stored by means electromagnetic fields. The interaction between the UHD and the organism seems to be electromagnetic in nature. The transmission of information from (bio-)molecules to the UHD is of special interest. Again, electromagnetic actions and vector potential fields appear to be implicated. CONCLUSION: The mechanisms of information storage and transfer in UHDs are far from fully understood, but progress has been made at experimental and theoretical levels.

Scientific proving of ultra high dilutions on humans.

BACKGROUND: Homeopathic drug provings or pathogenetic trials (HPTs) are the pillar of homeopathy. This review summarizes the authors' findings and interpretations derived from a series of homeopathic drug proving between 1994 and 2015. It gives an overview over a series of attempts to use modern scientific experimental methodology to answer the question, whether such HPTs produce symptoms in healthy volunteers that can be distinguished from placebo symptoms. METHODS: Various experimental models were used: repeated crossover trials with categorical data collection, and a single-case, randomised study. Final models use diligent qualitative data-collection in experienced volunteers. In those, raters decide whether symptoms are typical for a remedy delivered or not. The design is triple-blind and placebo-controlled. RESULT: While previous attempts were inconclusive, this new model allowed to separate placebo symptoms from verum symptoms repeatedly in a series of two definitive studies following promising pilot studies. Results were statistically significant. Also, some signs of the purported non-local signature of homeopathic effects were visible, and the consequences for future methodology is discussed. CONCLUSION: Provided some cautionary notes are taken into account, HPTs can be used to separate out true specific symptoms from placebo symptoms. By the same token this is a road to experimental proof that homeopathic remedies are not just placebos. However, this needs to be taken forward by independent groups.

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.

Anti-oxidant Effect of High Dilutions of Arnica montana, Arsenicum Album, and Lachesis Mutus in Microglial Cells in Vitro.

Microglial cells play important roles in inflammatory responses. The level of oxidative stress is a well-known marker of inflammation. Homeopathic medicines are often used clinically to alleviate inflammation. We evaluated the anti-oxidative effect of high dilutions of Arnica montana (Arnica m.), Arsenicum album (Arsenicum a.), and Lachesis mutus (Lachesis m.) on production of reactive oxygen species (ROS) in inflamed microglial cells in vitro. Microglial cells, on exposure to lipopolysaccharide (LPS), have induced production of ROS compared with resting cells. The dilutions significantly reduced the oxidative stress by decreasing the level of ROS produced. Arnica m. 1C, 3C, 5C, 7C, 9C, and 30C dilutions had a range of ROS reduction between 15 and 42.1%; Arsenicum a. 3C, 5C, 7C, 15C, and 30C dilutions had a range of ROS reduction between 17.6 and 35.3%; and Lachesis m. 3C, 5C, 7C, 9C, 15C, and 30C dilutions had a range of ROS reduction between 25 and 41.7%. To summarize, the dilutions with the greatest effect were Arnica m. 1C (42.1%), Arsenicum a. 30C (35.3%), and Lachesis m. 7C (41.7%). Arnica m., Arsenicum a., and Lachesis m. did not have the same effect on ROS production and were not dose-dependent.

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.

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.

In vivo evaluation of antipyretic effects of some homeopathic ultra-high dilutions on Baker's yeast-induced fever on Similia principle.

BACKGROUND: Homeopathy is a controversial alternative system of medicine. The action of homeopathic medicines is considered slow and it is assumed that homeopathic medicines are ineffective in acute conditions such as fever. OBJECTIVE: In the present study, effects of 3 homeopathic medicines on baker's yeast induced fever were investigated. MATERIALS AND METHODS: 42 local strain rabbits were equally divided into 7 groups. Normal saline was orally administered to group 1 (normal control) rabbits without fever induction. Group 2 underwent baker's yeast-induced fever (negative control). Groups 3, 4, 5, 6 and 7 underwent baker's yeast-induced fever and were thereafter treated orally with paracetamol, Nux vomica 200C and 1M, Calcarea phos 200C and Belladonna 200C respectively. Rectal temperature was checked hourly. The abdominal writhing and frequency of loose stools were also monitored. ANOVA was applied for checking statistical significance. p ≤ 0.05 was considered significant. RESULTS: The rectal temperature increased significantly (p < 0.05) in the negative control group when compared to the normal control. Abdominal writhing and loose stools monitoring showed increased writhing and loose stools frequency of group 2, 3, 6 and 7 rabbits. However, treatment of paracetamol significantly reduced rectal temperature. Group 4 & 5 showed significant reduction of rectal temperature together with abatement of abdominal writhing and loose stools. CONCLUSION: N. vomica ultra-high dilutions have normalized rectal temperature and prevented the abdominal writhing and loose stools in baker's yeast-induced fever model of rabbits. It could be due to antidotal activity of N. vomica ultra-high dilutions. Therefore, N. vomica ultra-high dilutions can be useful antipyretic agents and can treat conditions associated with gastrointestinal symptoms. However, fixed conclusion can't be asserted due to caveat of small sample size.

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.

Efectos de las diluciones homeopáticas altas sobre los modelos in vitro: revisión de la literatura / Effects of high homeopathic dilutions on in vitro models: a review of the literature

Los efectos de las diluciones homeopáticas altas (HD, por sus siglas en inglés) son controvertidos debido a que superan el número de Avogadro. Objetivo: Realizar una revisión de la literatura sobre los efectos de las HD sobre los modelos in vitro. Métodos: Se realizó una búsqueda sistemática en la base de datos PubMed de estudios que evaluaran las HD simples sobre modelos in vitro, publicados de 2007 en adelante. Resultados: 28 publicaciones cumplieron con los criterios de inclusión/exclusión. 26 estudios mostraron efectos evidentes de las HD simples sobre modelos in vitro. La mayor parte de tales estudios fueron realizados en países en donde la homeopatía ha alcanzado un alto nivel de institucionalización. Conclusiones: Los modelos in vitro evidencian de forma clara la actividad biológica de las HD superiores al número de Avogadro y dan cuenta de los efectos encontrados en la práctica clínica. La mayor parte de los estudios fueron realizados en países en donde la homeopatía es reconocida oficialmente, lo cual facilita el acceso a recursos para el desarrollo de la investigación.

the effects of homeopathic high dilutions (HDs) are controversial because they exceed Avogadro's number. Aim: to perform a literature review on the effects of HDs on in vitro models. Methods: a systematic search was performed in database PubMed for studies assessing simple HDs on in vitro models published from 2007 onward. Results: 28 publications met the inclusion/exclusion criteria; 26 studies demonstrated patent effects of simple HDs on in vitro models; most such studies were conducted in countries where homeopathy attained a high level of institutionalization. Conclusions: in vitro models patently evidence biological activity of HDs above Avogadro's number and account for effects found in clinical practice. Most studies were conducted in countries where homeopathy is officially recognized, which facilitates access to resources for the development of research.

Efectos de las diluciones homeopáticas altas sobre las plantas: revisión de la literatura / Effects of high homeopathic dilutions on plants: a review of the literature

Entre los supuestos no convencionales de la homeopatía, el uso de medicamentos en diluciones altas (HD, por sus siglas en inglés) es una causa de objeciones y escepticismo entre la comunidad científica, formada dentro del paradigma de la dependencia de la dosis de la farmacología clásica. La investigación que busca evidenciar los efectos de las HD homeopáticas recurre a varios modelos experimentales (in vitro, plantas y animales). Objetivo: Describir los resultados de estudios con alta calidad metodológica que han demostrado los efectos positivos de las HD homeopáticas sobre las plantas. Métodos: Tomando como fuente de referencia las revisiones publicadas hasta 2015, actualizamos la información añadiendo datos de estudios recientes incluidos en la base de datos PubMed. Resultados: De los 167 estudios experimentales analizados, 48 cumplieron los criterios mínimos de calidad metodológica, de los cuales 29 detectaron efectos específicos de las diluciones homeopáticas altas sobre las plantas mediante la comparación con controles adecuados. Conclusiones: A pesar de que la mayor parte de los experimentos presentaba una calidad metodológica por debajo del estándar, los estudios que emplearon sistemáticamente reproducibilidad y controles negativos demostraron indiscutibles efectos significativos de las HD homeopáticas sobre las plantas.

Among the non-conventional assumptions of homeopathy, the use of medicines in high dilutions (HD) is a cause for objections and skepticism among the scientific community, trained within the dose-dependency paradigm of classic pharmacology. Research aiming at evidencing the effects of homeopathic HD has resource to several experimental models (in vitro, plants and animals). Aim: To describe the results of studies with high methodological quality that demonstrated positive effects of homeopathic HD on plants. Methods: Taking reviews published until 2015 as reference source, we updated the information through addition of data from recent studies included in database PubMed. Results: From 167 experimental studies analyzed, 48 met the minimum criteria of methodological quality, from which 29 detected specific effects of homeopathic high dilutions on plants through comparison to adequate controls. Conclusions: Despite the substandard methodological quality of most experiments, studies with systematic use of negative controls and reproducibility demonstrated significant indisputable effects of homeopathic HD on plants.

Lycopodium clavatum as a homeopathic treatment in Magellanic penguins (Sphenicus magellanicus)

Uric acid is the main form of excretion of nitrogenous components in birds. The assessment verification samples, control, check of uric acid concentrations is widely used to detect kidney disease. High concentrations (up to five times) of uric acid in plasma can lead to the precipitation of this acid in the form of crystals, which accumulate in the tissues, mainly in the synovial joints and the viscera surface. Aims: report the evolution of homeopathic treatment in increased uric acid in Magellanic penguins (Spheniscus magellanicus) from Santo André Municipal Zoo. Methodology: 12 animals showed an increase in uric acid (reference values 0,3-16,1mg/dL) and received homeopathic treatment with Lycopodium clavatum 6cH. The protocol established was the administration of two globules of Lycopodium clavatum 6cH, once a day, inside fishes offered in the animal's beak. Treatment started on 25/09/2019. On 30/10/2019 was collected a new blood sample, those who were not discharged were kept on treatment and new collections were made on 27/11/2019 and 17/12/2019. Results: 8 animals had an average uric acid of 20.45mg /dl and responded to the 35-days treatment, normalizing the biochemical values with an average of 12.24mg /dl. Two animals with 18.27 mg /dl were discharged in 62 days with 11.70 mg /dl and another two penguins with 19.85 mg /dl were discharged in 82 days later with 9.57 mg /dl of uric acid. Conclusion: Homeopathy proved to be successful in the treatment of increased uric acid in Magellanic penguins, easy to administer, affordable and without side effects.

Silicea terra200cH evaluated by two different spectroscopy methods: a pilot study

Spectroscopy has been shown to bea useful method to study the physicochemical properties of homeopathic preparations. Aim: In this pilot study, we comparedtwo methods (photon scattering and visible-light spectroscopy) in the physicalevaluation of Silicea terra200cH. Methodology: Italian test: Two sampleshave been examinedand compared: Silicea terra200cH and Sac lac200cH, both preparedin water solution, diluted and succussed according to German Homeopathic Pharmacopoeia(Cemon Lab, Italy).Lactose was includedbecause the first 3 potencies of Silicea terraaremadeby trituration in lactosepowder.Measurements were made using an innovative Charge-Coupled Device (CCD) camera developedby Daniele Gullà, called MIRA/CORA(proprietary name). Slight variations in chrominance and luminance due to micro-vibrational 3D phenomenawere analyzed.Allmeasurements have been performed in a dark room at a constant temperature of 22°C +/-0,2°C usingafilter with very narrow spectral bands (10 nm).The measurements have been repeated three times on both Silicea terra200cH and Sac lac200cHwithin a few minutes aftereach other.Brazilian test: In the second test, performed in Brazil, variations in absorbance were used to identify Silicea terra200cH compared with Sac lac200cH andcompared withanother control solution of non-succussed 30% alcohol, using six solvatochromic dyes, following the method developed by Cartwright [1,2]. Both homeopathic samples were imported from Italy(the same sample bottles used in the Italian test),in Brazil they have beendiluted 1:100 in 30% hydro-alcoholic solution, and submitted to 100 succussions using an automatic mechanical arm (Denise, AUTIC, Brazil) prior to being tested. Samples were inserted into dyes solutions and evaluated by visible spectroscopy (FEMTO Spectrophotometer, Brazil). Dyes were prepared in ethanol P.A., according to previous established methods [3]. Three series in triplicate were performed and the results were analyzed by ANOVA / Tukey, comparing both samples and the unsuccussed 30% hydroalcoholic control solution.Results: Italian test: Measurements of the mean entropy of the signals, statistically elaborated with T Student test,yielded a two tailed p value < 0.05, where the entropy of the signal recorded from the Silicea terra200cHsample was statistically lower than the 200cH Sac lacsample.Brazilian test: Among all tested dyes, only BDN(4-(Bis-(4-(dimethylamino)-phenyl)methylene)-1(4H)-naphthalenone) showed aninteraction with Silicea terra200cH, reproducing the conclusions obtained in [3].Conclusions: Two different spectroscopic methods were able to show signal differences betweenSilicea terra200cH andSac lac200cH, suggesting that changes in solvent organization could be involved in the homeopathic signaling process, along withchanges in dipole moments of solvent and dyes. The results are potentially in line witha recent publishedpaper [4], that supportsthe propositionthatthe lower entropy of the verum signal compared with controls could beexplained by increased coherent vibrations of the verum sample, as theoretically explained in previous literature [5].

In vitro evaluation of the effects of Viscum album homeopathic preparations on murine melanoma cells

Viscum album extract (VA) is a complementary treatment in cancer, with in vitro and in vivo cytotoxic effects on several tumor types when applied in phytochemical doses. However, highly diluted ethanolic homeopathic preparations' effects and mechanisms need further study. Aims:To assess the in vitro effects of highly diluted VA from the subspecies V. album abietis and V. album album at different potency levels in different dilution ratios on murine melanoma cells. Methodology:The VA mother tinctures (MT)from Abies alba (MTA) and Quercus robur (MTQ) were prepared with summer and winter samples, harvested in Switzerland. They were submitted to homeopathic ethanolic maceration and a subsequent dynamization process. MTA, MTQ and the following respective potencies were tested in B16F10 murine cells: 3x, 12x, 30x, 6cH, 12cH, 200cH, 2LM, 3LM, and 5LM. Dynamized water, dynamized and non-dynamized ethanol, and carboplatin were used as control groups. The mitochondrial activity and cell viability analysis were performed at 1, 24, 48, and 72 hours by in vitro incubation. MTA and MTQ harvested in summer, as well as 12x, 200cH and 5LM potencies were also tested to cell apoptosis and necrosis markers, reactive oxygens species (ROS) production, inflammatory cytokines profile, cell morphology, and migratory capacity. Results and discussion: MTA and MTQ induced a decrease in cell metabolism and higher cytotoxicity within 1 hour, with significant morphological changes and increased production of ROS and inflammatory cytokines. Both homeopathic dilutions 12x and 5LM showed an influence on cell metabolism, cell replication, and oxidative stress modulation with inflammatory cytokines, mitosis, and migration pattern changes. On the other hand, Quercus robur and Abies alba 200cH showed increased on cytotoxicity and ROS levels, respectively. Conclusion:The in vitro effects of Viscum album homeopathic solutions in melanoma cells highlight the promising antitumoral potential and reinforce the need for further research to better understanding their mechanisms of action.