The Experts below are selected from a list of 459 Experts worldwide ranked by ideXlab platform

Paolo Bellavite - One of the best experts on this subject based on the ideXlab platform.

  • Fibronectin Gene Up-regulation by Arnica montana in Human Macrophages: Validation by Real-Time Polymerase Chain Reaction Assay.
    Homeopathy : the journal of the Faculty of Homeopathy, 2020
    Co-Authors: Marta Marzotto, Fabio Arruda-silva, Paolo Bellavite
    Abstract:

    Background and Aim Arnica montana L. (Arnica m.) is a popular traditional medicine, used for its therapeutic properties in healing traumas, but little is known about its biological action on tissue formation and repair. This new work tested the effects of Arnica m. Homeopathic Dilutions on human macrophages, key cells in tissue defence and repair. Materials and Methods Macrophages derived from the THP-1 cell line were differentiated with interleukin-4 to induce a ‘wound-healing’-like phenotype, and treated with various Dilutions of Arnica m. centesimal (100 times) Dilutions (2c, 3c, 5c, 9c, and 15c) or control solvent for 24 hours. RNA samples from cultured cells were analysed by real-time quantitative polymerase chain reaction in five separate experiments. Results Arnica montana at the 2c dilution (final concentration of sesquiterpene lactones in cell culture = 10−8 mol/L) significantly stimulated the expression of three genes which code for regulatory proteins of the extracellular matrix, namely FN1 (fibronectin 1, % increase of 21.8 ± standard error of the mean 4.6), low-density lipoprotein-receptor-related protein 1 (% increase of 33.4 ± 6.1) and heparan sulphate proteoglycan 2 (% increase of 21.6 ± 9.1). Among these genes, the most quantitatively expressed was FN1. In addition, FN1, unlike other candidate genes, was upregulated in cells treated with higher Dilutions/dynamisations (3c, 5c, and 15c) of Arnica m. Conclusion The results support evidence that the extracellular matrix is a potential therapeutic target of Arnica m., with positive effects on cell adhesion and migration during tissue development and healing.

  • Arnica montana experimental studies: confounders and biases?
    Journal of integrative medicine, 2018
    Co-Authors: Paolo Bellavite, Marta Marzotto, Clara Bonafini
    Abstract:

    Arnica montana is a popular traditional remedy widely used in complementary and alternative medicine, in part for its wound-healing properties. The authors recently showed that this plant extract and several of its Homeopathic Dilutions are able to modify the expression of a series of genes involved in inflammation and connective tissue regeneration. Their studies opened a debate, including criticisms to the "errors" in the methods used and the "confounders and biases". Here the authors show that the criticisms raised on methodology and statistics are not consistent and cannot be considered pertinent. The present comment also updates and reviews information concerning the action of A. montana Dilutions in human macrophage cells while summarizing the major experimental advances reported on this interesting medicinal plant.

  • Experimental neuropharmacology of Gelsemium sempervirens: Recent advances and debated issues.
    Journal of Ayurveda and integrative medicine, 2018
    Co-Authors: Paolo Bellavite, Clara Bonafini, Marta Marzotto
    Abstract:

    Abstract Gelsemium sempervirens L. (Gelsemium) is traditionally used for its anxiolytic-like properties and its action mechanism in laboratory models are under scrutiny. Evidence from rodent models was reported suggesting the existence of a high sensitivity of central nervous system to anxiolytic power of Gelsemium extracts and Homeopathic Dilutions. In vitro investigation of extremely low doses of this plant extract showed a modulation of gene expression of human neurocytes. These studies were criticized in a few commentaries, generated a debate in literature and were followed by further experimental studies from various laboratories. Toxic doses of Gelsemium cause neurological signs characterized by marked weakness and convulsions, while ultra-low doses or high Homeopathic Dilutions counteract seizures induced by lithium and pilocarpine, decrease anxiety after stress and increases the anti-stress allopregnanolone hormone, through glycine receptors. Low (non-Homeopathic) doses of this plant or its alkaloids decrease neuropathic pain and c-Fos expression in mice brain and oxidative stress. Due to the complexity of the matter, several aspects deserve interpretation and the main controversial topics, with a focus on the issues of high dilution pharmacology, are discussed and clarified.

  • Experimental neuropharmacology of Gelsemium sempervirens: Recent advances and debated issues
    Elsevier, 2018
    Co-Authors: Paolo Bellavite, Clara Bonafini, Marta Marzotto
    Abstract:

    Gelsemium sempervirens L. (Gelsemium) is traditionally used for its anxiolytic-like properties and its action mechanism in laboratory models are under scrutiny. Evidence from rodent models was reported suggesting the existence of a high sensitivity of central nervous system to anxiolytic power of Gelsemium extracts and Homeopathic Dilutions. In vitro investigation of extremely low doses of this plant extract showed a modulation of gene expression of human neurocytes. These studies were criticized in a few commentaries, generated a debate in literature and were followed by further experimental studies from various laboratories. Toxic doses of Gelsemium cause neurological signs characterized by marked weakness and convulsions, while ultra-low doses or high Homeopathic Dilutions counteract seizures induced by lithium and pilocarpine, decrease anxiety after stress and increases the anti-stress allopregnanolone hormone, through glycine receptors. Low (non-Homeopathic) doses of this plant or its alkaloids decrease neuropathic pain and c-Fos expression in mice brain and oxidative stress. Due to the complexity of the matter, several aspects deserve interpretation and the main controversial topics, with a focus on the issues of high dilution pharmacology, are discussed and clarified. Keywords: Gelsemium sempervirens, Anxiety, Neurocytes, Animal models, Homeopathic medicine, Behavior, Gene expressio

  • Gene expression and highly diluted molecules.
    Frontiers in pharmacology, 2014
    Co-Authors: Marta Marzotto, Debora Olioso, Paolo Bellavite
    Abstract:

    Plants of the genus Gelsemium have measurable effects on anxiety-like symptoms in laboratory models (Magnani et al., 2010; Liu et al., 2013; Meyer et al., 2013; Jin et al., 2014). Searching for a possible mechanism of action, we found that very low doses and Homeopathic Dilutions of Gelsemium sempervirens (Gelsemium) modulate the expression of genes involved in neuronal functions (G-protein coupled receptor signaling pathways, calcium homeostasis, inflammatory response and receptors) (Marzotto et al., 2014; Olioso et al., 2014). The commentary's first criticism is that “the search for an involvement of neural genes related to anxiety/depression or mood disorders is biased by the expression of human genes having no orthologs/homologs in mice, where the authors reported evidence about Gelsemium action on behavioral tests in animal anxiety models.” Frankly, we do not see what type of bias can be found in our line of research: Gelsemium is traditionally used as a remedy for anxiety-related symptoms in humans; we have shown that it works in mice models (Magnani et al., 2010; Bellavite et al., 2012), and we wanted to study its mechanism of action at the molecular level, using human neurocyte cell lines. This type of experimental procedure, which addresses various knowledge gaps using both animal studies and in vitro models, is very common in pharmacological studies. Furthermore, the commentary (Chirumbolo, 2014) states that “some genes indicated to be downregulated by Gelsemium 2c, should not be expressed by neuronal cells (e.g., CD163, MPO, C8B, LST1, TREM2, notoriously expressed in immune cell).” Actually, it is well known that neurons express genes of cellular pathways also involved in cytokine/chemokine and immune responses. Genes related to immune and inflammatory responses are expressed in SH-SY5Y cells, as reported by many researchers (Gatta et al., 2011; Toyama et al., 2011; Cui et al., 2012; Xu et al., 2013). Although Gelsemium contains several different compounds (Dutt et al., 2010; Jin et al., 2014), the major active alkaloid of this plant is gelsemine. In the cited commentary we read that “concentration of gelsemine was not assessed, as it was solely calculated on previous spectrometry investigations and new preparations, from ethanol draw extracts, were not further quantified by analytical chemistry.” This statement is misleading because the concentration of gelsemine which we correctly reported (0.021 g/100 ml, corresponding to 6.5 × 10−4 mol/L) (Marzotto et al., 2014, p. 2) was precisely determined by liquid chromatography (not “spectrometry”) in the mother tincture from which the samples were prepared. The concentration of UV-VIS absorbing substances decreased by a factor of 100 at each centesimal dilution step, to become analytically undetectable after a few passages, as we have shown by means of spectrometry in our paper. What we were interested in was to check the accuracy of the procedures as far as possible, not to determine the gelsemine concentration in all subsequent centesimal Dilutions. Perhaps we need to remind readers that normally, when one performs a study of dose-response and the concentration in the highest dose and the dilution factor are known, there is no need to determine the concentration of substances in all successive Dilutions. Another erroneous criticism which forces us to reply is the statement (Chirumbolo, 2014) that our UV-VIS spectra (Marzotto et al., 2014) “showed a peak at 250 nm caused by contaminating millimolar ethanol in Gelsemium 2c.” This is untrue, and we fail to understand how the writer could have reached such a conclusion, since UV-visible absorption spectra were performed with a double-beam spectrophotometer using drug samples and reference controls, having exactly the same ethanol concentration. Subsequently the commentary claims that “the authors tested a complex mixture of G. sempervirens extract, containing at least about 0.154 mM EtOH at 2c, if Dilutions were conducted exactly” and that “concentration of EtOH, set at 30% v/v, faded out to 0.003% in tested Dilutions but the authors did not clarify how much for each centesimal dilution in the Methods section.” The writer supposes that high ethanol concentration could have caused “latent apoptosis.” This is confusing and misleading. As clearly indicated in our paper (Marzotto et al., 2014), the final ethanol concentration was 0.03% v/v (p. 2), and “no significant differences in cell viability were observed between cells treated with the ethanol control solution 0.03% (v/v) and untreated cells” (p. 5). Furthermore, as other researchers have found (Do et al., 2013), cell viability of neurocytes is unaffected by doses of ethanol up to 10 mmol/L (0.06%). Low doses of ethanol may influence gene expression, but would have acted in the same way in the Gelsemium and in the control samples. To rule out any misinterpretation of our findings, Figure ​Figure11 gives a brief outline of our microarray experiments. Figure 1 Workflow of the main steps of the DNA-microarray experiments aimed to discover differentially expressed genes in Gelsemium-treated neuronal cells compared with control treated cells. Cultured SHSY5Y cells were incubated for 24 h with the Gelsemium Dilutions ... Finally, the commentary (Chirumbolo, 2014) maintains that “Gene array profile of expression following 24 h incubation with Gelsemium 2c, showed down-regulation of 49 genes, namely 87.5% gene array.” It is hard to understand how this 87.5% of genes was calculated, since the 49 genes represent about 0.1% of the genes on the microarray chip. The observation that “many gene products, listed in the expression profile of 56 genes array, such as LOC154872, KIAA0825, LOC150763, C1orf167, have not been identified” is bizarre. The presence of some as yet unidentified sequences in the human genome database is of course well known. We hope that these clarifications will be welcomed in the interests of providing correct and truthful information to readers. In summary, we provided reliable evidence that Gelsemium exerts a prevalently inhibitory effect on a series of neurocyte genes across a wide dose range (Marzotto et al., 2014; Olioso et al., 2014). The effect decreases with decreasing doses, but whole genome expression analysis made it possible to detect statistically significant changes even at extremely low doses and Homeopathic Dilutions (e.g., 5th and 9th centesimal dilution, corresponding to final gelsemine concentrations of 6.5 × 10−15 mol/L and 6.5 × 10−23 mol/L respectively). More robust conclusions about the role of the genes involved will require determination whether proteins encoded by the affected genes are similarly changed, using proteomic and phosphoproteomic approaches, and/or further studies using purified active plant compounds.

Marta Marzotto - One of the best experts on this subject based on the ideXlab platform.

  • Fibronectin Gene Up-regulation by Arnica montana in Human Macrophages: Validation by Real-Time Polymerase Chain Reaction Assay.
    Homeopathy : the journal of the Faculty of Homeopathy, 2020
    Co-Authors: Marta Marzotto, Fabio Arruda-silva, Paolo Bellavite
    Abstract:

    Background and Aim Arnica montana L. (Arnica m.) is a popular traditional medicine, used for its therapeutic properties in healing traumas, but little is known about its biological action on tissue formation and repair. This new work tested the effects of Arnica m. Homeopathic Dilutions on human macrophages, key cells in tissue defence and repair. Materials and Methods Macrophages derived from the THP-1 cell line were differentiated with interleukin-4 to induce a ‘wound-healing’-like phenotype, and treated with various Dilutions of Arnica m. centesimal (100 times) Dilutions (2c, 3c, 5c, 9c, and 15c) or control solvent for 24 hours. RNA samples from cultured cells were analysed by real-time quantitative polymerase chain reaction in five separate experiments. Results Arnica montana at the 2c dilution (final concentration of sesquiterpene lactones in cell culture = 10−8 mol/L) significantly stimulated the expression of three genes which code for regulatory proteins of the extracellular matrix, namely FN1 (fibronectin 1, % increase of 21.8 ± standard error of the mean 4.6), low-density lipoprotein-receptor-related protein 1 (% increase of 33.4 ± 6.1) and heparan sulphate proteoglycan 2 (% increase of 21.6 ± 9.1). Among these genes, the most quantitatively expressed was FN1. In addition, FN1, unlike other candidate genes, was upregulated in cells treated with higher Dilutions/dynamisations (3c, 5c, and 15c) of Arnica m. Conclusion The results support evidence that the extracellular matrix is a potential therapeutic target of Arnica m., with positive effects on cell adhesion and migration during tissue development and healing.

  • Arnica montana experimental studies: confounders and biases?
    Journal of integrative medicine, 2018
    Co-Authors: Paolo Bellavite, Marta Marzotto, Clara Bonafini
    Abstract:

    Arnica montana is a popular traditional remedy widely used in complementary and alternative medicine, in part for its wound-healing properties. The authors recently showed that this plant extract and several of its Homeopathic Dilutions are able to modify the expression of a series of genes involved in inflammation and connective tissue regeneration. Their studies opened a debate, including criticisms to the "errors" in the methods used and the "confounders and biases". Here the authors show that the criticisms raised on methodology and statistics are not consistent and cannot be considered pertinent. The present comment also updates and reviews information concerning the action of A. montana Dilutions in human macrophage cells while summarizing the major experimental advances reported on this interesting medicinal plant.

  • Experimental neuropharmacology of Gelsemium sempervirens: Recent advances and debated issues.
    Journal of Ayurveda and integrative medicine, 2018
    Co-Authors: Paolo Bellavite, Clara Bonafini, Marta Marzotto
    Abstract:

    Abstract Gelsemium sempervirens L. (Gelsemium) is traditionally used for its anxiolytic-like properties and its action mechanism in laboratory models are under scrutiny. Evidence from rodent models was reported suggesting the existence of a high sensitivity of central nervous system to anxiolytic power of Gelsemium extracts and Homeopathic Dilutions. In vitro investigation of extremely low doses of this plant extract showed a modulation of gene expression of human neurocytes. These studies were criticized in a few commentaries, generated a debate in literature and were followed by further experimental studies from various laboratories. Toxic doses of Gelsemium cause neurological signs characterized by marked weakness and convulsions, while ultra-low doses or high Homeopathic Dilutions counteract seizures induced by lithium and pilocarpine, decrease anxiety after stress and increases the anti-stress allopregnanolone hormone, through glycine receptors. Low (non-Homeopathic) doses of this plant or its alkaloids decrease neuropathic pain and c-Fos expression in mice brain and oxidative stress. Due to the complexity of the matter, several aspects deserve interpretation and the main controversial topics, with a focus on the issues of high dilution pharmacology, are discussed and clarified.

  • Experimental neuropharmacology of Gelsemium sempervirens: Recent advances and debated issues
    Elsevier, 2018
    Co-Authors: Paolo Bellavite, Clara Bonafini, Marta Marzotto
    Abstract:

    Gelsemium sempervirens L. (Gelsemium) is traditionally used for its anxiolytic-like properties and its action mechanism in laboratory models are under scrutiny. Evidence from rodent models was reported suggesting the existence of a high sensitivity of central nervous system to anxiolytic power of Gelsemium extracts and Homeopathic Dilutions. In vitro investigation of extremely low doses of this plant extract showed a modulation of gene expression of human neurocytes. These studies were criticized in a few commentaries, generated a debate in literature and were followed by further experimental studies from various laboratories. Toxic doses of Gelsemium cause neurological signs characterized by marked weakness and convulsions, while ultra-low doses or high Homeopathic Dilutions counteract seizures induced by lithium and pilocarpine, decrease anxiety after stress and increases the anti-stress allopregnanolone hormone, through glycine receptors. Low (non-Homeopathic) doses of this plant or its alkaloids decrease neuropathic pain and c-Fos expression in mice brain and oxidative stress. Due to the complexity of the matter, several aspects deserve interpretation and the main controversial topics, with a focus on the issues of high dilution pharmacology, are discussed and clarified. Keywords: Gelsemium sempervirens, Anxiety, Neurocytes, Animal models, Homeopathic medicine, Behavior, Gene expressio

  • Gene expression and highly diluted molecules.
    Frontiers in pharmacology, 2014
    Co-Authors: Marta Marzotto, Debora Olioso, Paolo Bellavite
    Abstract:

    Plants of the genus Gelsemium have measurable effects on anxiety-like symptoms in laboratory models (Magnani et al., 2010; Liu et al., 2013; Meyer et al., 2013; Jin et al., 2014). Searching for a possible mechanism of action, we found that very low doses and Homeopathic Dilutions of Gelsemium sempervirens (Gelsemium) modulate the expression of genes involved in neuronal functions (G-protein coupled receptor signaling pathways, calcium homeostasis, inflammatory response and receptors) (Marzotto et al., 2014; Olioso et al., 2014). The commentary's first criticism is that “the search for an involvement of neural genes related to anxiety/depression or mood disorders is biased by the expression of human genes having no orthologs/homologs in mice, where the authors reported evidence about Gelsemium action on behavioral tests in animal anxiety models.” Frankly, we do not see what type of bias can be found in our line of research: Gelsemium is traditionally used as a remedy for anxiety-related symptoms in humans; we have shown that it works in mice models (Magnani et al., 2010; Bellavite et al., 2012), and we wanted to study its mechanism of action at the molecular level, using human neurocyte cell lines. This type of experimental procedure, which addresses various knowledge gaps using both animal studies and in vitro models, is very common in pharmacological studies. Furthermore, the commentary (Chirumbolo, 2014) states that “some genes indicated to be downregulated by Gelsemium 2c, should not be expressed by neuronal cells (e.g., CD163, MPO, C8B, LST1, TREM2, notoriously expressed in immune cell).” Actually, it is well known that neurons express genes of cellular pathways also involved in cytokine/chemokine and immune responses. Genes related to immune and inflammatory responses are expressed in SH-SY5Y cells, as reported by many researchers (Gatta et al., 2011; Toyama et al., 2011; Cui et al., 2012; Xu et al., 2013). Although Gelsemium contains several different compounds (Dutt et al., 2010; Jin et al., 2014), the major active alkaloid of this plant is gelsemine. In the cited commentary we read that “concentration of gelsemine was not assessed, as it was solely calculated on previous spectrometry investigations and new preparations, from ethanol draw extracts, were not further quantified by analytical chemistry.” This statement is misleading because the concentration of gelsemine which we correctly reported (0.021 g/100 ml, corresponding to 6.5 × 10−4 mol/L) (Marzotto et al., 2014, p. 2) was precisely determined by liquid chromatography (not “spectrometry”) in the mother tincture from which the samples were prepared. The concentration of UV-VIS absorbing substances decreased by a factor of 100 at each centesimal dilution step, to become analytically undetectable after a few passages, as we have shown by means of spectrometry in our paper. What we were interested in was to check the accuracy of the procedures as far as possible, not to determine the gelsemine concentration in all subsequent centesimal Dilutions. Perhaps we need to remind readers that normally, when one performs a study of dose-response and the concentration in the highest dose and the dilution factor are known, there is no need to determine the concentration of substances in all successive Dilutions. Another erroneous criticism which forces us to reply is the statement (Chirumbolo, 2014) that our UV-VIS spectra (Marzotto et al., 2014) “showed a peak at 250 nm caused by contaminating millimolar ethanol in Gelsemium 2c.” This is untrue, and we fail to understand how the writer could have reached such a conclusion, since UV-visible absorption spectra were performed with a double-beam spectrophotometer using drug samples and reference controls, having exactly the same ethanol concentration. Subsequently the commentary claims that “the authors tested a complex mixture of G. sempervirens extract, containing at least about 0.154 mM EtOH at 2c, if Dilutions were conducted exactly” and that “concentration of EtOH, set at 30% v/v, faded out to 0.003% in tested Dilutions but the authors did not clarify how much for each centesimal dilution in the Methods section.” The writer supposes that high ethanol concentration could have caused “latent apoptosis.” This is confusing and misleading. As clearly indicated in our paper (Marzotto et al., 2014), the final ethanol concentration was 0.03% v/v (p. 2), and “no significant differences in cell viability were observed between cells treated with the ethanol control solution 0.03% (v/v) and untreated cells” (p. 5). Furthermore, as other researchers have found (Do et al., 2013), cell viability of neurocytes is unaffected by doses of ethanol up to 10 mmol/L (0.06%). Low doses of ethanol may influence gene expression, but would have acted in the same way in the Gelsemium and in the control samples. To rule out any misinterpretation of our findings, Figure ​Figure11 gives a brief outline of our microarray experiments. Figure 1 Workflow of the main steps of the DNA-microarray experiments aimed to discover differentially expressed genes in Gelsemium-treated neuronal cells compared with control treated cells. Cultured SHSY5Y cells were incubated for 24 h with the Gelsemium Dilutions ... Finally, the commentary (Chirumbolo, 2014) maintains that “Gene array profile of expression following 24 h incubation with Gelsemium 2c, showed down-regulation of 49 genes, namely 87.5% gene array.” It is hard to understand how this 87.5% of genes was calculated, since the 49 genes represent about 0.1% of the genes on the microarray chip. The observation that “many gene products, listed in the expression profile of 56 genes array, such as LOC154872, KIAA0825, LOC150763, C1orf167, have not been identified” is bizarre. The presence of some as yet unidentified sequences in the human genome database is of course well known. We hope that these clarifications will be welcomed in the interests of providing correct and truthful information to readers. In summary, we provided reliable evidence that Gelsemium exerts a prevalently inhibitory effect on a series of neurocyte genes across a wide dose range (Marzotto et al., 2014; Olioso et al., 2014). The effect decreases with decreasing doses, but whole genome expression analysis made it possible to detect statistically significant changes even at extremely low doses and Homeopathic Dilutions (e.g., 5th and 9th centesimal dilution, corresponding to final gelsemine concentrations of 6.5 × 10−15 mol/L and 6.5 × 10−23 mol/L respectively). More robust conclusions about the role of the genes involved will require determination whether proteins encoded by the affected genes are similarly changed, using proteomic and phosphoproteomic approaches, and/or further studies using purified active plant compounds.

Salvatore Chirumbolo - One of the best experts on this subject based on the ideXlab platform.

  • Homeopathic Dilutions, Hahnemann Principles, and the Solvent Issue: Must We Address Ethanol as a "Homeopathic" or a "Chemical" Issue?
    Homeopathy : the journal of the Faculty of Homeopathy, 2017
    Co-Authors: Salvatore Chirumbolo, Geir Bjørklund
    Abstract:

    Introduction Homeopathic remedies usually contain a significant amount of ethanol as a co-solvent with water, a pharmaceutical formulation that may raise some concern when remedies are tested in vitro or in laboratory animals, due to the assessed toxicity of ethanol on cell cultures and organisms. The amount of alcohol in a Homeopathic remedy is adjusted following the different Homeopathic pharmacopoeias but it is rarely below 30% v/v, which is a molar mass established to meet both Hahnemann's traditional heritage and the hypothetical role of ethanol in “imprinting” water, through the formation of nanobubbles, with the Homeopathic activity of the remedy. Aims This article aims at discussing the role of ethanol in Homeopathic Dilutions and how its chemical nature should affect the experimental approach in homeopathy. Issues Under Debate While the content of ethanol in a Homeopathic remedy should be as low as 20% v/v, which is a molar fraction able to catalyze the formation of nanobubbles in a dynamized alcohol–water dilution, this amount raises concern about ethanol toxicology in the experimental research with laboratory animals or in vitro. Several authors diluted 1:100 ethanol 30% v/v from their tested Homeopathic Dilutions with distilled water to prevent the cytotoxic effect of the alcohol, but in doing so, they probably reduced the ability of ethanol (now 0.3% v/v) to induce the formation of nanobubbles, thus probably affecting the Homeopathic property of the same dilution. This may generate concerns about how to manage an experimental setting, to meet both the “chemical” nature of ethanol and its role in “homeopathy,” an issue that is discussed in the article. Conclusion Any author working with Homeopathic Dilutions containing a molar fraction of ethanol higher than 20% should take into account the fact that ethanol is cytotoxic and may be a catalyst to the formation of nanobubbles, and so should adjust the experimental approach accordingly.

  • Homeopathic potencies of Arnica montana L. change gene expression in a Tamm-Horsfall protein-1 cell line in vitro model: the role of ethanol as a possible confounder and statistical bias.
    Journal of integrative medicine, 2017
    Co-Authors: Salvatore Chirumbolo, Geir Bjørklund
    Abstract:

    Abstract Marzotto et al. showed that Homeopathic preparations of Arnica montana L. acted directly on gene expression of Tamm-Horsfall protein-1 (THP-1) monocyte/macrophage cell lines activated with phorbol12-myristate13-acetate and interleukin-4 (IL-4). A. montana Homeopathic Dilutions are used in complementary and alternative medicine to treat inflammation disorders and post-traumatic events as well as for wound repair. The French Pharmacopoeia of these remedies uses 0.3% ethanol in each centesimal dilution. In this paper, we discuss how ethanol-containing A. montana Homeopathic centesimal Dilutions can change gene expression in IL-4-treated monocyte/macrophage THP-1. We assessed the role of ethanol in the Arnica Homeopathic Dilutions containing this alcohol by investigating its action on gene expression of THP-1 cell. Evidence would strongly suggest that the presence of ethanol in these remedies might play a fundamental role in the Dilutions ability to affect gene expression, particularly for doses from 5c to 15c. Where, rather than playing a major role in the mesoscopic structure of water, the ethanol might have a chemical-physical role in the induction of THP-1 gene expression, apoptosis, and deoxyribonucleic acid function. This evidence generates a debate about the suggestion that the use of a binary-mixed solvent in Homeopathic chemistry, used by Hahnemann since 1810, may be fundamental to explain the activity of homeopathy on cell models.

  • Adverse effects and homeopathy: may remedies yet contain noxious or toxic molecules?
    British journal of clinical pharmacology, 2014
    Co-Authors: Salvatore Chirumbolo
    Abstract:

    A recent article published in the latest issue of PLoS ONE by Csupor et al. assessed that, at least for a very special subgroup of Homeopathic medicines, a big concern about their reliability and safety still currently exists [1]. As highlighted by these authors' work, Homeopathic products may contain ingredients in allopathic doses. Low Dilutions and mother tinctures still play a role in Homeopathic prescribing, and are particularly prominent in certain systems of homeopathy focusing on the organotropic effects of Homeopathic medicinal products [2]. This fact raises some fundamental ethical issues. In the European Union, the Directive 2001/83/EC allowed the authorization of Homeopathic medicines containing allopathic doses. Therefore the distinction between low potency Homeopathic medications and allopathic herbal medicines might be based on the intention of the manufacturer, i.e. on the production method and the rationale behind their application and not on the dose. I recently reported this concern in a comment of mine [3], where I tried to expand the debate on the urgent need to amend homeopathy by checking commercial preparations about their molecular content and source components before introducing them to the consumer. While people believe that Homeopathic medicines in high Dilutions, prescribed by trained professionals, are probably safe and unlikely to provoke severe adverse reactions, it is difficult to draw definite conclusions from the reported literature in the field, either because of the low methodological quality of reports claiming possible adverse effects [4] or because the many comments coming to an uproar due to critical controversy [5]. In my opinion, even high diluted Homeopathic medicine could contain noxious components for health and I discussed this issue in ref [3]. but this should be confirmed by high tech chemical-physical analysis before rendering Homeopathic remedies available on the public market. The resolution WHA56.31 on traditional medicine, adopted by the 2009 World Health Assembly, requested WHO to provide technical support for developing methodologies to ensure product quality, efficacy and safety in homeopathy. In this perspective, the evidence that the extreme Homeopathic Dilutions retain starting materials was demonstrated by homeopaths themselves using TEM, electron diffraction and chemical analysis by ICP-AE spectroscopy [6]. Ultrastructural physical analysis of water-ethanol interfaces in glass containers, prompts us to make this consideration. During the high mechanical stress, which every Homeopathic medicine undergoes, at the structured solid (glass)–water interfaces occurs the formation of very small gas bubbles, known as ‘nanobubbles’, as shown by the tapping mode atomic force microscopy (TMAFM) imaging technique. This nanobubble formation can affect significantly the role of water interaction with hydrophobic molecules, especially when ethanol is present [7]. The effect of surface treatment with alcohols on interacting hydrophobic surfaces in water has a dramatic effect on the formation of nanobubbles and thus the long range hydrophobic force. Without alcohol–water exchange the population and size of nanobubbles is reduced resulting in either a ‘true’ short range hydrophobic force or a long range nanobubble bridging hydrophobic force, thus changing dramatically the solvent property of water during a dilution step [7]. This might account for the non-linear dynamics observed when a serially diluted compound was checked by quantitative chemical analysis [3,8]. On the basis of recently published results [8] nanoparticles can be concentrated on the liquid surface in a manner similar to the traditional froth-flotation process used in the metal ore purification of larger particles. Lactose during grinding helped in the formation of nanoclusters, whereas the ensuing processes such as mechanical stress and sparging producing numerous large air bubbles aided nanoparticle levitation to the liquid surface, forming a monolayer that was preserved in the serial Dilutions, so altering the real calculation of the diluted material [8]. This could explain why not only ‘low potency’ Homeopathic remedies but even highest Dilutions may contain molecules with yet potentially adverse effects for the organism. As previuosly suggested [3], further insights are needed to highlight this issue.

  • Advances in homeopathy and immunology: a review of clinical research.
    Frontiers in bioscience (Scholar edition), 2011
    Co-Authors: Paolo Bellavite, Salvatore Chirumbolo, Marta Marzotto, Anita Conforti
    Abstract:

    The present paper reviews the clinical research carried out over the past three decades to evaluate the effectiveness of homeopathy for the treatment of respiratory allergies, common upper respiratory tract infections, otorhinolaryngologic complaints, and rheumatic diseases. We include in the analysis both randomised and non-randomised trials, assigning them different weightings in the final balance of evidence, on the basis of semi-quantitative criteria. Overall, the literature concerning a total of 83 original studies suggests that homeopathy may have significant effects in some conditions, e.g. Galphimia glauca (low Homeopathic Dilutions/dynamizations) in allergic oculorhinitis, Anas barbariae (high Homeopathic dilution/dynamization) in influenza-like syndromes, classical individualised homeopathy in otitis, in allergic complaints and in fibromyalgia, and a few low-potency Homeopathic complexes in sinusitis, rhinoconjunctivitis, arthritis. The evidence for individualised Homeopathic therapy in the field of upper respiratory tract infections and for Homeopathic immunotherapy in respiratory allergies is more conflicting. Pragmatic equivalence trials suggest that, in primary care, Homeopathic treatment is not inferior to conventional treatment. A larger number of observational studies and of clinical trials -- conducted in a methodologically correct manner without altering the treatment setting-- are needed before sure conclusions concerning the application of homeopathy for specific diseases can be drawn.

Ursula Wolf - One of the best experts on this subject based on the ideXlab platform.

  • Spatial allocation effects within a potentization basic research model - evidence for field-like effects of Homeopathic preparations?
    International Journal of High Dilution Research, 2014
    Co-Authors: Stephan Baumgartner, Peter Heusser, Lucietta Betti, Mascha Binder, Ursula Wolf
    Abstract:

    Background: The mode of action of ultramolecular Homeopathic preparations is still unknown. Interactions between objects or entities can be grouped in four main general scientific categories: material, force-/field-like, entanglement-like or informational. Should Homeopathic preparations have a field-like mode of action, there is greater probability of cross-contamination as long as the means to “shield” objects from each other is unknown. A field-like interaction would also lead to treatment at a distance effects that are distance-dependent. Aims: We analysed a set of experiments with Arsenicum album 45x treated wheat seedlings regarding a possible distance-dependent cross-contamination. Materials and Methods: We performed an a posteriori analysis of a set of 17 independent experiments [1,2] with wheat seedlings pre-treated with 1‰ arsenic. Three treatments were applied (Arsenicum album 45x, water 45x, or unpotentized water) with 150 seedlings in each treatment group per experiment. Seedlings were arranged in hanging plastic bags side-by-side in identically treated blocks of 10 seedlings. The 3x15 blocks were coded and randomly allocated to the three treatments. Wheat shoot length was measured after 7 days. Treatment effects were analysed as function of the position (1–10) within the blocks of 10 seedlings. Results: Analyzing all data, Arsenicum album 45x exerted an inhibiting effect (–3.2%, p=0.01) compared to both water and water 45x. When restricting the analysis to the outermost seedlings of all subgroups (pos. 1, 10), the treatment effect vanished (0.3%, p=0.92). In contrast, the innermost seedlings of all subgroups (pos. 5, 6) showed a treatment effect of –5.6% (p=0.02). Intermediate pairs of positions (pos. 2–4, 7–9) showed intermediate effects. Regarding shoot length, dependency on spatial position was observed for the plants of the water control groups, but not for the plants treated with Arsenicum album 45x. Conclusions: Whilst the effect of Arsenicum album 45x on wheat-shoot growth was not dependent on the spatial position within the subgroup, the water-control plants became smaller the closer they were to Arsenicum album 45x-treated seedlings. This observation is compatible with the existence of a field-like effect of Homeopathic Dilutions. Another possible explanation that cannot be ruled out by the present experiments, is contamination through the gas-phase. Future investigations of ultramolecular Homeopathic preparations should control any such effects since they may mask treatment effects, leading to false-negative results. Closer investigation of the nature of this distance-dependent effect might contribute to identification of the mode of action of ultramolecular Homeopathic preparations.

  • Comparison of globules prepared from high Dilutions of various starting materials by ultraviolet light spectroscopy
    2013
    Co-Authors: Sabine D Klein, Ursula Wolf
    Abstract:

    Background: High and ultra-high Dilutions of various starting materials, e.g. copper sulfate, Hypericum perforatum and sulfur, showed significant differences in ultraviolet light (UV) transmission from controls and amongst different dilution levels [1,2]. Verum and placebo globules of Aconitum napellus 30c or calcium carbonate/quercus e cortice 6x from the same packs as used in previous clinical trials and dissolved in water could be distinguished by UV spectroscopy [3]. However, it was unclear whether the differences in UV absorbance originated from specific characteristics of the starting materials, from differences in the production of verum and placebo globules, and/or other unknown interference factors. Aims: The aim of this study was to investigate whether globules produced with high and ultra-high Dilutions (6x, 12x, 30c, 200c, 200CF (centesimal discontinuous fluxion), 10,000CF) of various starting materials (Aconitum napellus, Atropa belladonna, phosphorus, sulfur, Apis mellifica, quartz) could be distinguished by UV spectroscopy. Methodology: The globules were specially produced for this study by Spagyros AG (Gumligen, Switzerland) and differed only in the starting materials of the Dilutions (but not in the batch of globules or ethanol used). Globules were dissolved in water at 10 mg/ml, in quadruplicates, approximately 22 h prior to the measurements. Absorbance of the samples in the UV range (from 190 to 340 nm) was measured in a randomized order with a Shimadzu double beam UV-1800 spectrophotometer equipped with an auto sampler. Samples of each starting material were prepared and measured on 5 independent days. The daily variations of the spectrophotometer as well as the drift during the measurements were corrected for. The average absorbance from 200 to 340 nm was compared among various starting materials within equal dilution levels using a Kruskal-Wallis test. Results: Statistically significant differences were found among 30c (Figure 1), 200c and 200CF Dilutions of the various starting materials. No differences were found among 6x, 12x and 10,000CF Dilutions. Conclusions: Globules prepared from high Dilutions of various starting materials may show significantly different UV absorbance when dissolved in water. References [1] Wolf U, Wolf M, Heusser P, Thurneysen A, Baumgartner S. Homeopathic preparations of quartz, sulfur and copper sulfate assessed by UV-spectroscopy. Evid Based Complement Alternat Med. 2011;2011:692798. [2] Klein SD, Sandig A, Baumgartner S, Wolf U. Differences in median ultraviolet light transmissions of serial Homeopathic Dilutions of copper sulfate, Hypericum perforatum, and sulfur. Evid Based Complement Alternat Med. 2013;2013:370609. [3] Klein SD, Wolf U. Investigating Homeopathic verum and placebo globules with ultraviolet spectroscopy. Forsch Komplementmed. 2013, accepted.

  • Comparison of globules prepared from high Dilutions of various starting materials by ultraviolet light spectroscopy
    2013
    Co-Authors: Sabine D Klein, Ursula Wolf
    Abstract:

    Background: High and ultra-high Dilutions of various starting materials, e.g. copper sulfate, Hypericum perforatum and sulfur, showed significant differences in ultraviolet light (UV) transmission from controls and amongst different dilution levels [1,2]. Verum and placebo globules of Aconitum napellus 30c or calcium carbonate/quercus e cortice 6x from the same packs as used in previous clinical trials and dissolved in water could be distinguished by UV spectroscopy [3]. However, it was unclear whether the differences in UV absorbance originated from specific characteristics of the starting materials, from differences in the production of verum and placebo globules, and/or other unknown interference factors. Aims: The aim of this study was to investigate whether globules produced with high and ultra-high Dilutions (6x, 12x, 30c, 200c, 200CF (centesimal discontinuous fluxion), 10,000CF) of various starting materials (Aconitum napellus, Atropa belladonna, phosphorus, sulfur, Apis mellifica, quartz) could be distinguished by UV spectroscopy. Methodology: The globules were specially produced for this study by Spagyros AG (Gumligen, Switzerland) and differed only in the starting materials of the Dilutions (but not in the batch of globules or ethanol used). Globules were dissolved in water at 10 mg/ml, in quadruplicates, approximately 22 h prior to the measurements. Absorbance of the samples in the UV range (from 190 to 340 nm) was measured in a randomized order with a Shimadzu double beam UV-1800 spectrophotometer equipped with an auto sampler. Samples of each starting material were prepared and measured on 5 independent days. The daily variations of the spectrophotometer as well as the drift during the measurements were corrected for. The average absorbance from 200 to 340 nm was compared among various starting materials within equal dilution levels using a Kruskal-Wallis test. Results: Statistically significant differences were found among 30c (Figure 1), 200c and 200CF Dilutions of the various starting materials. No differences were found among 6x, 12x and 10,000CF Dilutions. Conclusions: Globules prepared from high Dilutions of various starting materials may show significantly different UV absorbance when dissolved in water. References [1] Wolf U, Wolf M, Heusser P, Thurneysen A, Baumgartner S. Homeopathic preparations of quartz, sulfur and copper sulfate assessed by UV-spectroscopy. Evid Based Complement Alternat Med. 2011;2011:692798. [2] Klein SD, Sandig A, Baumgartner S, Wolf U. Differences in median ultraviolet light transmissions of serial Homeopathic Dilutions of copper sulfate, Hypericum perforatum, and sulfur. Evid Based Complement Alternat Med. 2013;2013:370609. [3] Klein SD, Wolf U. Investigating Homeopathic verum and placebo globules with ultraviolet spectroscopy. Forsch Komplementmed. 2013, accepted.

  • Differences in Median Ultraviolet Light Transmissions of Serial Homeopathic Dilutions of Copper Sulfate, Hypericum perforatum, and Sulfur
    Evidence-based complementary and alternative medicine : eCAM, 2013
    Co-Authors: Sabine D Klein, Stephan Baumgartner, Annegret Sandig, Ursula Wolf
    Abstract:

    Homeopathic remedies are produced by potentising, that is, the serial logarithmic dilution and succussion of a mother tincture. Techniques like ultraviolet spectroscopy, nuclear magnetic resonance, calorimetry, or thermoluminescence have been used to investigate their physical properties. In this study, Homeopathic centesimal (c) potencies (6c to 30c) of copper sulfate, Hypericum perforatum, and sulfur as well as succussed water controls were prepared. Samples of these preparations were exposed to external physical factors like heat, pressure, ultraviolet radiation, or electromagnetic fields to mimic possible everyday storage conditions. The median transmissions from 190 nm to 340 nm and 220 nm to 340 nm were determined by ultraviolet light spectroscopy on five measurement days distributed over several months. Transmissions of controls and potencies of sulfur differed significantly on two of five measurement days and after exposure to physical factors. Transmissions of potencies exposed to ultraviolet light and unexposed potencies of copper sulfate and Hypericum perforatum differed significantly. Potency levels 6c to 30c were also compared, and wavelike patterns of higher and lower transmissions were found. The Kruskal-Wallis test yielded significant differences for the potency levels of all three substances. Aiming at understanding the physical properties of Homeopathic preparations, this study confirmed and expanded the findings of previous studies.

  • Potential effects of exposure to external influencing factors on Homeopathic Dilutions as determined by ultraviolet light spectroscopy
    European Journal of Integrative Medicine, 2008
    Co-Authors: Ursula Wolf, B. Marschollek, Peter Heusser, Stephan Baumgartner, Martin Wolf
    Abstract:

    Introduction Statistically significant and clinically relevant effects of Homeopathic Dilutions have been demonstrated in various clinical trials. In contrast to the clinical evidence, the mode of action of Homeopathic Dilutions is not yet identified and may be difficult to explain within a conventional scientific context, in particular, when the dilution levels exceed Avogadro's number. By investigating potential specific physical properties of Homeopathic Dilutions, we aim to understand their modes of action and thus contribute to provide a scientific basis for their use in medicine. In a previous study, we demonstrated that Homeopathic Dilutions have lower UV-light transmissions than controls. The aim of the current study was to explore the effect of external influencing factors, namely UV light and 2 types of temperature on the Homeopathic Dilutions. Methods Homeopathic centesimal (c) Dilutions, 1c to 30c, of CuSO 4 and 10 controls (succussed potentization medium) were prepared in sterile conditions, and after preparation randomized and blinded. The code was only disclosed after data analysis was completed. Three separate sets of samples of each potentization level were filled in test tubes and exposed to three types of external procedures: exposure to UV light of a sterilization lamp for 12 h, incubating at 37 °C for 24 h and heated to 90 °C for 15 min. Each sample and control was measured 4 times with a Shimadzu UV-1601 spectrometer. UV-light transmission values from 220 to 340 nm (in %) were averaged across 4 measurements. The mean values and standard deviations for Homeopathic Dilutions and controls were determined for each condition. Results The mean differences in UV-light transmission±SE (%) between Homeopathic Dilutions and controls and the significances were: UV exposure: −0.0469±0.0264%, p =0.0562; Temperature 37 °C: −0.0685±0.0235%, p =0.0024; Temperature 90 °C:  0.0382±0.0327%, p =0.0787. Conclusion For each form of exposure, the UV-light transmission of the Homeopathic Dilutions compared to the controls was reduced. The difference in the UV-light transmission between Homeopathic Dilutions and controls increased for all exposure forms and heating to 37 °C had the strongest effect.

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  • stimulation of bovine sperm mitochondrial activity by Homeopathic Dilutions of monensin
    Homeopathy, 2005
    Co-Authors: D M Aziz, Heinrich Enbergs
    Abstract:

    Mitochondrial activity is an important viability parameter of spermatozoa and is linked to sperm motility. Monensin is commonly used as an inhibitor for sperm mitochondrial activity in the laboratory. This study was conducted to evaluate the influence of some Homeopathic Dilutions of monensin on sperm mitochondrial activity. Fresh ejaculates from 6 mature bulls were used in the study. Samples of the semen were tested using a flow cytometer for mitochondrial activity and sperm viability using Rhodamine 123 and SYBR-14, respectively. The 9× dilution of monensin resulted in very highly significant (P<0.001) stimulation of mitochondrial activity. Monensin 5×, 7×, 8× and 13× caused highly significant (P<0.01) stimulation of the sperm mitochondrial activity. Other Homeopathic Dilutions of monensin (6×, 10×, 11×, 12× and 14×) also had a significant (P<0.05) stimulatory effect. The use of monensin did not have any negative effect on sperm viability. We conclude that some Homeopathic Dilutions of monensin increase mitochondrial activity of bovine spermatozoa without negative effect on sperm viability, the 9× dilution was the most effective. Further in vivo studies are required to estimate the effect of Homeopathic Dilutions of monensin on semen quality.

  • Stimulation of bovine sperm mitochondrial activity by Homeopathic Dilutions of monensin.
    Homeopathy : the journal of the Faculty of Homeopathy, 2005
    Co-Authors: D M Aziz, Heinrich Enbergs
    Abstract:

    Mitochondrial activity is an important viability parameter of spermatozoa and is linked to sperm motility. Monensin is commonly used as an inhibitor for sperm mitochondrial activity in the laboratory. This study was conducted to evaluate the influence of some Homeopathic Dilutions of monensin on sperm mitochondrial activity. Fresh ejaculates from 6 mature bulls were used in the study. Samples of the semen were tested using a flow cytometer for mitochondrial activity and sperm viability using Rhodamine 123 and SYBR-14, respectively. The 9× dilution of monensin resulted in very highly significant (P