Francesco Borghini

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.

Droplet evaporation method as a new potential approach for highlighting the effectiveness of ultra high dilutions

OBJECTIVE This study sought to verify whether the droplet evaporation method (DEM) can be applied to assess the effectiveness of ultra-high dilutions (UHDs). We studied the shape characteristics of the polycrystalline structures formed during droplet evaporation of wheat seed leakages. METHODS The experimental protocol tested both unstressed seeds and seeds stressed with arsenic trioxide 5mM, treated with either ultra-high dilutions of the same stressor substance, or with water as a control. The experimental groups were analyzed by DEM and in vitro growth tests. DEM patterns were evaluated for their local connected fractal dimension (measure of complexity) and fluctuating asymmetry (measure of symmetry exactness). RESULTS Treatment with arsenic at UHD of both stressed and non-stressed seeds increased the local connected fractal dimension levels and bilateral symmetry exactness values in the polycrystalline structures, as compared to the water treatment. The results of in vitro growth tests revealed a stimulating effect of arsenic at UHD vs. control, and a correlation between the changes in growth rate and the crystallographic values of the polycrystalline structures was observed. CONCLUSIONS The results indicate that polycrystalline structures are sensitive to UHDs, and so for the first time provide grounds for the use of DEM as a new tool for testing UHD effectiveness. DEM could find application as a treatment pre-selection tool, or to monitor sample conditions during treatment. Moreover, when applied to biological liquids (such as saliva, blood, blood serum, etc.), DEM might provide information about UHD effectiveness on human and animal health.

Weak Extremely High Frequency Microwaves Affect Pollen-Tube Emergence and Growth in Kiwifruit: Pollen Grain Irradiation and Water-Mediated Effects

OBJECTIVE This study was designed to evaluate the effects of weak-intensity extremely high frequency (EHF) microwaves in a model system-the plant organism pollen grain-lacking the placebo effect, available in large populations, to ensure accurate statistical analysis, and whose sensitivity is closely relevant to animal and human biology. DESIGN This study was blinded using an in vitro pollen germination technique. SUBJECTS AND STUDY INTERVENTIONS: Pollen of kiwifruit (Actinidia deliciosa) was either directly irradiated or grown in a medium prepared with irradiated water, using a CromoStim 2000, (PromoPharma, Republic of San Marino) designed for EHF microwave resonance therapy (MRT). It produces weak intensity EHF radiations (40-78 GHz), either continuous wave (cw) or modulated, at a 10 Hz-frequency, with infrared (IR) carried to 635-950 nm, and with an impedance (IPD) of 10(-21) W/Hz cm(2) and a power supply from 0 to 20 mW. OUTCOME MEASUREMENTS Pollen-tube emergence was expressed as a percent of grains producing a tube and tube elongation was measured at 4 hours of incubation by a turbidimetric assay (A(500)) of cultures, expressed as the net absorbance increase over time 0. RESULTS At days 2 and 4 during aging, both percent of germination and tube growth significantly and consistently improved over controls in kiwifruit pollen grains irradiated for 30 minutes at day 0 at 10 Hz frequency with the CromoStim 2000. Highly significant effects, either stimulant or inhibitory, were also observed on kiwifruit pollen (stressed or not) growing in a medium prepared with water previously irradiated either cw or modulated. Irradiated water affected pollen germination immediately and even after several days following EHF treatment. CONCLUSIONS Either direct or indirect EHF irradiation performed by the CromoStim 2000 is effective on pollen growth processes. In both cases, water seemed to play a primary role. According to the quantum electrodynamical coherence theory, our work could also have implications for homeopathy, suggesting a key to explain the efficacy of high dilutions and succussion procedures.

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.

Extremely Low Doses of Arsenic Affect in vitro Pollen Germination

Introduction: This study deals with the effects of extremely low doses or high dilutions of pharmacological compounds on in vitro pollen germination of kiwifruit (Actinidia deliciosa). Material and Methods: The biological efficacy of As2O3 at the 5th and 45th decimal dilution/succussion level (As 5x and As 45x) was tested on pollen previously stressed with As2O3 150 or 200 μM. The outcome variable was the pollen germination rate, as detected blind after 3 h 30 min by an Axioplan microscope. Results: A directionally consistent recovery of germination percentage was observed in both As 5x and As 45x after stressing with As2O3 150 μM. When pollen was stressed with As2O3 200 μM only as 45x induced a general and significant germination increase. Conclusions: Our results suggest that both treatments might partially remove the inhibitory effect caused by the stressor. Similar effects were observed on the same model using weak extremely low frequency magnetic fields (ELF-MFs) mediated through water. Although preliminary, the findings seem to indicate the in vitro pollen performance as adequate to study the effects of physicochemical subthreshold stimuli (extremely low doses or high dilutions of pharmacological compounds, weak ELF-MFs), mediated through water.