Maurizio Brizzi

Effect of high dilutions of Arsenicum album on wheat seedlings from seed poisoned with the same substance

Abstract A blind laboratory experiment was carried out to show the effect of a 45x potency of Arsenicum album (As2O3) on wheat seedlings whose seeds had been previously poisoned with a material dose of the same substance. The effect of the homoeopathic treatment on stem growth was statistically significant. The experimental results were matched with a previous study concerning homoeopathic treatment in the same species.

A Biostatistical Insight into the As2O3 High Dilution Effects on the Rate and Variability of Wheat Seedling Growth

Background: Most criticism of homeopathy concerns the lack of scientific bases and theoretical models. Fundamental research could make important contributions to our understanding of the mechanisms of action of homeopathic treatments. Plant-based bioassays are suitable for basic research - lacking the placebo effect and ensuring large data samples for structured statistical analyses. Objective: The aim of this study was to reproduce a previous experiment on the effects of arsenic trioxide (As2O3) high dilutions on wheat seedling growth in order to verify whether the same significant results could be obtained working in a different place and with a different experimental team. A further goal was to investigate high dilution effects on variability. Material and Methods: A structured experiment was performed blind over 9 weeks, using wheat seeds previously stressed with a sublethal dose of As2O3. The seeds were then treated with either potentized As2O3 (5x, 15x, 25x, 35x, 45x), potentized water (equivalent potencies) or diluted As2O3 (10-5, 10-15, 10-25, 10-35, 10-45). The working variable was the stem length, measured after 4, 5, 6 and 7 days. Results: Some potencies (As2O3 45x and H2O 45x) induced a relevant increase in seedling growth and/or a variability decrease. Diluted As2O3 did not induce any significant results. Conclusions: Confirmation of a significant stimulating effect on seedling growth and a significant decrease of variability was obtained with ultra-high dilutions at the 45x potency. The model of wheat germination and growth has been confirmed to be a good tool for basic research in homeopathy.

A pilot statistical study with homoeopathic potencies of Arsenicum album in wheat germination as a simple model

A blind, randomized laboratory trial to study homoeopathic potencies of Arsenicum album on wheat germination is proposed as a simple model which allows a rigorous statistical analysis. The parametric tests show that the differences between the treatment groups cannot be explained as a mere effect of intrinsic seed variability.

The Role of Variability in Evaluating Ultra High Dilution Effects: Considerations Based on Plant Model Experiments

A series of experiments, performed on plant models with ultra high dilutions (UHD) of arsenic trioxide at 45th decimal potency has been reviewed with a particular focus on variability. The working variables considered are: the number of germinated seeds out of a fixed set of 33, the stem length of wheat seedlings and the number of necrotic lesions in tobacco leaf disks inoculated with tobacco mosaic virus (TMV). A thorough comparison between treatment and control group has been proposed, considering the two main sources of variability in each series of experiments: variability within and between experiments. In treated groups, a systematic decrease in variability between-experiments, as well as a general decrease, with very few exceptions, in variability within experiments has been observed with respect to control. Variability is traditionally considered as control parameter of model systems. Our hypothesis, based on experimental evidences, proposes a new role of variability as a target of UHD action. This hypothesis may help interpret unanswered questions that keep rising in basic and clinical research in homeopathy.

Effects of Gelsemium sempervirens L. on pathway-focused gene expression profiling in neuronal cells

ETHNOPHARMACOLOGICAL RELEVANCE Gelsemium sempervirens L. is a traditional medicinal plant mainly distributed in the southeastern of the United States, employed in phytotheraphy and homeopathy as nervous system relaxant to treat various types of anxiety, pain, headache and other ailments. Although animal models showed its effectiveness, the mechanisms by which it might operate on the nervous system are largely unknown. This study investigated for the first time by a real-time PCR technique (RT-PCR Array) the gene expression of a panel of human neurotransmitter receptors and regulators, involved in neuronal excitatory signaling, on a neurocyte cell line. MATERIALS AND METHODS Human SH-SY5Y neuroblastoma cells were exposed for 24h to Gelsemium sempervirens at 2c and 9c dilutions (i.e. 2 and 9-fold centesimal dilutions from mother tincture) and the gene expression profile compared to that of cells treated with control vehicle solutions. RESULTS Exposure to the Gelsemium sempervirens 2c dilution, containing a nanomolar concentration of active principle gelsemine, induced a down-regulation of most genes of this array. In particular, the treated cells showed a statistically significant decrease of the prokineticin receptor 2, whose ligand is a neuropeptide involved in nociception, anxiety and depression-like behavior. CONCLUSIONS Overall, the results indicate a negative modulation trend in neuronal excitatory signaling, which can suggest new working hypotheses on the anxiolytic and analgesic action of this plant.

The Efficacy of Ultramolecular Aqueous Dilutions on a Wheat Germination Model as a Function of Heat and Aging-Time

This study evaluates the effects of temperature and aging on the efficacy of As2O3 at the 45th decimal potency in a wheat germination model, compared against a control and potentized H2O 45×. Each treatment-temperature combination was tested on seeds (Triticum aestivum L.) of Pandas variety, using six Petri dishes (33 seeds/dish) per trial, performing eight trials. Seeds were pre-treated by poisoning with 0.1% As2O3 solution to reduce germination, to allow a better evaluation of homeopathic treatment effects. The outcome variable was the number of non-germinated seeds after 96 h. Temperature effect was investigated by heating each treatment in a water bath for 30 min (at 20, 40 or 70°C), or for 5 min (at 100°C), and that of aging by dividing experimental data, collected over a period of nearly five months, into two groups: early and late experiments. Results seem to show that the efficacy of As2O3 45× is unaltered at 20 and 40°C, increases at 70°C and decreases at 100°C. As regards aging, a notable difference was found between early trials, with no significant efficacy, and late trials, where As2O3 45× exhibits a repeated significant effect versus control, except at 100°C. A reduction in variability was observed for As2O3 45× at 20°C versus control, confirming the findings of previous work. The main conclusion suggested by this experiment is that the efficacy of As2O3 45× on wheat germination may be influenced by heating degree and seems to have an increasing trend as a function of aging.

Arnica montana effects on gene expression in a human macrophage cell line. Evaluation by quantitative Real-Time PCR

BACKGROUND Arnica montana is a popular traditional remedy widely used in complementary medicine, also for its wound healing properties. Despite its acknowledged action in clinical settings at various doses, the molecular aspects relating to how A. montana promotes wound healing remain to be elucidated. To fill this gap, we evaluated the whole plant extract, in a wide range of dilutions, in THP-1 human cells, differentiated into mature macrophages and into an alternative IL-4-activated phenotype involved in tissue remodelling and healing. METHODS Real-time quantitative Reverse Transcription Polymerase Chain Reaction (PCR) analysis was used to study the changes in the expression of a customized panel of key genes, mainly cytokines, receptors and transcription factors. RESULTS On macrophages differentiated towards the wound healing phenotype, A. montana affected the expression of several genes. In particular CXC chemokine ligand 1 (CXCL1), coding for an chief chemokine, exhibited the most consistent increase of expression, while also CXC chemokine ligand 2 (CXCL2), Interleukin8 (IL8) and bone morphogenetic protein (BMP2) were slightly up-regulated, suggesting a positive influence of A. montana on neutrophil recruitment and on angiogenesis. MMP1, coding for a metalloproteinase capable of cleaving extracellular matrix substrates, was down-regulated. Most results showed non-linearity of the dose-effect relationship. CONCLUSIONS This exploratory study provides new insights into the cellular and molecular mechanisms of action of A. montana as a promoter of healing, since some of the genes it modifies are key regulators of tissue remodelling, inflammation and chemotaxis.

A Review of Three Simple Plant Models and Corresponding Statistical Tools for Basic Research in Homeopathy

In this paper, we review three simple plant models (wheat seed germination, wheat seedling growth, and infected tobacco plants) that we set up during a series of experiments carried out from 1991 to 2009 in order to study the effects of homeopathic treatments. We will also describe the set of statistical tools applied in the different models. The homeopathic treatment used in our experiments was arsenic trioxide (As2O3) diluted in a decimal scale and dynamized. Since the most significant results were achieved with the 45th decimal potency, both for As2O3 (As 45x) and water (W 45x), we here report a brief summary of these results. The statistical analysis was performed by using parametric and nonparametric tests, and Poisson distribution had an essential role when dealing with germination experiments. Finally, we will describe some results related to the changes in variability, which seems to be one of the targets of homeopathic treatment effect.

Biological effects and physico-chemical properties of extremely diluted aqueous solutions as a function of aging-time

This study concerns the biological effects and physico-chemical variations of extremely diluted solutions (EDS) as a function of aging-time. The biological efficacy of As2O3 at the 45th decimal dilution/succussion (As 45×) was tested in a wheat germination model. Ten trials were carried out from 0 to 12 months after treatment preparation, using wheat seeds (Triticum aestivum L.) of the Pandas variety. Seeds were pre-treated by poisoning with 0.1% As2O3 solution to reduce germination, to allow a better evaluation of treatment effects. The outcome variable was the number of non-germinated seeds after 96 h. The As 45×treatment aged for less than three months did not show a significant effect on wheat germination, whereas when aged for longer (3–12 months) the effect became significant. Concerning the physico-chemical characteristics, specific conductivity of As 45×aged from 0 to 12 months was measured, using nine samples for each date. The results showed a clear increasing time trend of specific conductivity, more evident when considering the last three measurements, which correspond to more than six months of aging. The physico-chemical behavior of EDS strongly supports the significant biological effects observed in a wheat model.

Weak Static and Extremely Low Frequency Magnetic Fields Affect In Vitro Pollen Germination

This study concerns the effects of a weak static magnetic field (MF) at 10 μT oriented downward, combined with a 16-Hz sinusoidal MF (10 μT), on in vitro pollen germination of kiwifruit (Actinidia deliciosa). Extremely low frequency magnetic field (ELF-MF) exposure was carried out by a signal generator unit connected to a copper wire solenoid, inside which samples where placed. Two different kinds of treatment were performed: direct and indirect. In the direct treatment, pollen samples were directly exposed during rehydration, germination, or both. In the indirect treatment, the pollen growth medium was prepared with water aliquots (at standard temperature of 20°C and pH = 6.74) that were exposed before use for 8 or 24 h. The main purpose of our research was to identify a biological marker (in vitro pollen germination in a stressing growth medium without Ca2+) susceptible to the effects of direct or indirect ELF-MF exposure. The working variable was the pollen germination rate, as detected blind after 3 h 30 min by an Axioplan microscope. A directionally consistent recovery of germination percentage was observed both for direct exposure (during germination and both rehydration and germination phases) and water-mediated exposure (with water exposed for 24 h and immediately used). Our results suggest that the ELF-MF treatment might partially remove the inhibitory effect caused by the lack of Ca2+ in the culture medium, inducing a release of internal Ca2+ stored in the secretory vesicles of pollen plasma membrane. Although preliminary, findings seem to indicate the in vitro pollen performance as adequate to study the effects of ELF-MFs on living matter.