Water as a sensor of weak impacts on biological systems.

A characteristic feature of weak impacts is the non-monotonic response of living organisms and model biological systems to monotonically decreasing impacts. The qualitative similarity of the effects caused by the different acting factors makes one think about the common cause of the observed effects, which is water. A comprehensive analysis of the actual composition of water indicates that water under normal conditions is a multicomponent open non-equilibrium system. Nanobubbles that are always present in water play a significant role in the properties of dilute aqueous solutions. When collapsed, they can produce active oxygen and nitrogen species that have a strong effect on biological systems. Significant non-monotonic changes in electrical conductivity found in a series of sequentially diluted solutions subjected to vigorous shaking after each dilution convincingly demonstrate the presence of chemical changes in the composition of aqueous solutions explained by mechanochemical processes. Similar changes were observed in water samples prepared in the same manner with vigorous shaking and dilution without the addition of any chemical compounds. The long-term evolution of the conductivity of such solutions depends on the chemical structure of the solutes.

Modifying Distant Effect of High Dilutions of Inorganic and Biological Substances.

It is known that highly diluted substances can exert a modifying effect on the initial substances without direct contact with them (distant interaction). The capability of high dilutions of IFNγ and Na2SO4 for the distant modifying effect was studied by the method of terahertz spectroscopy. Statistically significant differences were shown between terahertz characteristics of the initial solution of IFNγ protein and solution that had interacted with high dilutions of IFNγ; in case of sodium sulfate, no such differences were detected. Thus, high dilutions exert a distant modifying effect on the initial substances with complex spatial structure typical of biological molecules.

Study of the Effects of Highly Diluted Sodium Sulfate and Interferon-Gamma.

High dilutions of solutions containing extremely small amounts of the initial substance can modify the biological effects of the initial substance molecules. Using terahertz spectroscopy, we studied the possibility of modifying the physicochemical properties of the initial substance by adding high dilutions of high-molecular-weight (IFNγ) and low-molecular-weight (Na2SO4) compounds. In addition, the modifying effect produced by high dilutions of a low-molecular electrolyte (a solution of Na2SO4 salt) on the initial substance was confirmed by conductometry. This method allows measuring electrical conductivity that also depends on the physicochemical properties of the solution, namely, the number of ions and velocity of their movement. Statistically significant differences were shown between terahertz and conductometric characteristics of the initial solution (inorganic salt Na2SO4 or a protein IFNγ) and a solution, where high dilutions of the same substances were added in different concentrations. Interestingly, the differences were more pronounced for the biological molecule. Thus, it has been shown that high dilutions can change the properties of the initial solution; the effect is more pronounced for the protein solution.

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.

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.

Investigation of the influence of clathrate hydrate crystals on the structuring of homeopathic high dilutions

High dilutions (HD) of drugs used in homeopathy are mostly too dilute to contain original drug molecules. But evidences support their specific biological and therapeutic effects. The reason behind this is thought to be water structure characteristic of the original drug. Spectroscopic studies indicate that the specific water structure in HDs can be resolved into free water molecules, hydrogen bonding strength of water hydroxyl, number of hydrogen bonds and clathrate hydrate crystals (CHC). HDs are prepared in EtOH water solution by serial dilution and mechanical agitation, and are called potencies. The objective of the present study is to further confirm the presence of CHCs in the two potencies of three drugs. Electronic spectra of the HDs of the potencies indicate two broad peaks and marked difference in intensities of absorption. Furior Transform Infrared (FT-IR) spectra of the test potencies and their control show difference in intensity shift and contour shape of OH stretching and bending bands. All the experimental data indicate the presence of CHCs in varying amounts in the test potencies.

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.

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.

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].

Antibodies Processed Using High Dilution Technology Distantly Change Structural Properties of IFNγ Aqueous Solution.

Terahertz spectroscopy allows for the analysis of vibrations corresponding to the large-scale structural movements and collective dynamics of hydrogen-bonded water molecules. Previously, differences had been detected in the emission spectra of interferon-gamma (IFNγ) solutions surrounded by extremely diluted solutions of either IFNγ or antibodies to IFNγ without direct contact compared to a control. Here we aimed to analyse the structural properties of water in a sample of an aqueous solution of IFNγ via terahertz time-domain spectroscopy (THz-TDS). Tubes with the IFNγ solution were immersed in fluidised lactose saturated with test samples (dilutions of antibodies to IFNγ or control) and incubated at 37 °C for 1, 1.5-2, 2.5-3, or 3.5-4 h. Fluidised lactose was chosen since it is an excipient in the manufacture of drugs based on diluted antibodies to IFNγ. After incubation, spectra were recorded within a wavenumber range of 10 to 110 cm-1 with a resolution of 4 cm-1. Lactose saturated with dilutions of antibodies to IFNγ (incubated for more than 2.5 h) changed the structural properties of an IFNγ aqueous solution without direct contact compared to the control. Terahertz spectra revealed stronger intermolecular hydrogen bonds and an increase in the relaxation time of free and weakly bound water molecules. The methodology developed on the basis of THz-TDS could potentially be applied to quality control of pharmaceuticals based on extremely diluted antibodies.

Study of germination efficiency and temperature/drowning resistance in some ornamental plants treated with ultra high dilute compounds

The ultra-high dilutions (UHDs) can be used for decreasing stress conditions causing by climate variations. The present research investigated the effects of ultra-highdilutions (UHDs) on some ornamental plants, germination, and hormonal variations.Methods: In order to study the effect of UHDs (Calendula officinalisCalen. andArnica montanaArn.) on the physiological, primary metabolite, and hormonal variations of theOryza sativa L. (rice), 104 experiments were designed and statistically analyzed using the Design Expert 7.0.1 software over the general factorial design methodology. Two qualitative factors, including the UHDs/placebo usage and the type of plant usage, and two quantitative factors, including temperature and irrigation, were studied. The validated analysis was subjected to more extended studies on the variations in physiological growth, carbohydrate, protein content, and levels of plant hormones, including gibberellic acids, indole acetic acid, abscisic acid, and salicylic acid.Results: The statistical analysis resulted in a prediction model which was more than 75% correlates with experimental results. The results showed that the UHDs increased the carbohydrate and protein content of seedlings. Also, compared to placebo, the levels of hormones GA3 and IAA in all samples increase, and the amount of GA4 decreases. The amount of ABA and SA hormones inS. officinalisincreased under UHDs treatment while decreasing in the other two samples.Conclusion: The use of UHDs leads to an increase in the production of carbohydrate and protein content. Moreover, it causes significant variations in the growth-inducing hormone and increases the tolerance of seeds under higher/lower temperatures and draught/drowning. The results of this study open up a window to reduce germination survival and increase their resistance to sudden climate change.

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.

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.

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.

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.

Efeito de ultradiluições homeopáticas em modelos in vitro: revisão da literatura / Effect of homeopathic ultradilution on in vitro models: literature review

Introdução: Existem questionamentos quanto aos efeitos das ultradiluições (UDs) homeopáticas por ultrapassarem o número de Avogadro. Objetivo: Realizar uma revisão da literatura sobre os efeitos das UDs em modelos in vitro. Métodos: Foi realizada uma busca sistemática na base de dados PubMed de estudos sobre UDs simples em modelos in vitro a partir de 2007. Resultados: 28 publicações cumpriram os critérios de inclusão/exclusão; 26 estudos demonstraram efeitos evidentes de UDs simples em modelos in vitro, a maioria originada em países onde a homeopatia tem alto grau de institucionalização. Conclusões: Estudos in vitro demonstram inquestionavelmente a atividade biológica de UDs acima do número de Avogadro, dando conta do efeito das mesmas na prática clínica. A maioria das pesquisas se origina em países onde a homeopatia é uma racionalidade médica oficialmente aceita, o que facilita o acesso a recursos de pesquisa. (AU)

Background: the effects of homeopathic high dilutions (HDs) are debated 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 onwards. 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 the access to research. (AU)