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

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

The history of the Memory of Water.

'Homeopathic dilutions' and 'Memory of Water' are two expressions capable of turning a peaceful and intelligent person into a violently irrational one,' as Michel Schiff points out in the introduction of his book 'The Memory of Water'. The idea of the memory of water arose in the laboratory of Jacques Benveniste in the late 1980s and 20 years later the debate is still ongoing even though an increasing number of scientists report they have confirmed the basic results. This paper, first provides a brief historical overview of the context of the high dilution experiments then moves on to digital biology. One working hypothesis was that molecules can communicate with each other, exchanging information without being in physical contact and that at least some biological functions can be mimicked by certain energetic modes characteristics of a given molecule. These considerations informed exploratory research which led to the speculation that biological signaling might be transmissible by electromagnetic means. Around 1991, the transfer of specific molecular signals to sensitive biological systems was achieved using an amplifier and electromagnetic coils. In 1995, a more sophisticated procedure was established to record, digitize and replay these signals using a multimedia computer. From a physical and chemical perspective, these experiments pose a riddle, since it is not clear what mechanism can sustain such 'water memory' of the exposure to molecular signals. From a biological perspective, the puzzle is what nature of imprinted effect (water structure) can impact biological function. Also, the far-reaching implications of these observations require numerous and repeated experimental tests to rule out overlooked artifacts. Perhaps more important is to have the experiments repeated by other groups and with other models to explore the generality of the effect. In conclusion, we will present some of this emerging independent experimental work.

Effects of pulsed low frequency electromagnetic fields on water using photoluminescence spectroscopy: role of bubble/water interface.

The effects of a pulsed low frequency electromagnetic field were investigated on photoluminescence of well characterized water and prepared under controlled conditions (container, atmospheric, electromagnetic, and acoustic environments). When reference water samples were excited at 260 nm, two wide emission bands centered at 345 nm (3.6 eV) and 425 nm (2.9 eV) were observed. By contrast under 310 nm excitation, only one band appeared at 425 nm. Interestingly, electromagnetic treatment (EMT) induced, at both excitation wavelengths, a decrease (around 70%) in the 425 nm band relative photoluminescence intensity. However, no difference between reference and treated sample was observed in the 345 nm band. Other experiments, performed on outgassed samples (reference and treated), show that the emission bands (position, shape, intensity) under excitation at 260 nm and 310 nm were similar and close to the corresponding bands of the treated nonoutgassed samples. Similar effects were observed on photoluminescence excitation of water samples. Two excitation bands monitored at 425 nm were observed at 272 nm and 330 nm. After EMT and/or outgassing, a decrease (>60%) was observed in the intensity of these two bands. Altogether, these results indicate that electromagnetic treatment and/or outgassing decrease in a similar fashion the photoluminescence intensity in water samples. They also suggest that this effect is most likely indirectly attributed to the presence of gas bubbles in water. The possible role of hydrated ionic shell around the bubbles in the observed extraluminescence is discussed.

Effects of pulsed low-frequency electromagnetic fields on water characterized by light scattering techniques: role of bubbles.

Well-characterized purified water was exposed for 6 h to pulsed low-frequency weak electromagnetic fields. After various time periods, nondegassed and degassed water samples were analyzed by static light scattering. Just after electromagnetic exposure (day 0), a reduction of over 20% in the maximum light scattering intensity at 488 nm wavelength in both nondegassed and degassed samples was observed. By contrast, on day 12 the difference was observed only in nondegassed water samples. The latter effect was attributed to the different geometries of the containers combined with the basic origin of the whole phenomenon due to gas bubbles present in water. By the use of dynamic light scattering, the bubble mean diameter was estimated to be around 300 nm. Our results suggest that the electromagnetic exposure acts on gas nanobubbles present in water and emphasizes the role of the gas/liquid interface. The possibility that exposure to electromagnetic fields disturbs the ionic double layer that contributes to bubble stabilization in water is discussed.

Nutrient transfer by Runoff from sewage sludge amended soil under simulated rainfall.

Wastewater sludges are used in agriculture as soil amendment and fertilizer, with regard to their organic matter and nutrient content. However, availability of nitrogen and phosphorus from sludge-amended soils and their transfer in runoff may lead to eutrophication of downstream surface water. The aim of this study is to establish and compare the effect of two different sludges on these transfers: an anaerobically digested and thermically stabilised sludge (Seine-Aval treatment plant, sludge no. 1), and a limed sludge (Saint-Quentin treatment plant, sludge no. 2). Experiments were performed on 12 sloping micro-plots (1 m x 1 m) submitted to sludge spreading and controlled rainfall simulation. Runoff water was sampled and analysed for concentrations in nitrogen species and phosphorus. Results show that spreading of sludge no. 1 increased both ammonium nitrogen (mean of 1.1 mg L(-1) N-NH4 vs. 0.2 mg L(-1) N-NH4 for control micro-plots) and particulate phosphorus concentrations (mean of 2 mg L(-1) P vs. 1.1 mg L(-1) P for control micro-plots) in runoff water. On the other hand, sludge no. 2 did not induce any significant effect on nutrient concentrations in runoff. These results are related to chemical composition and physical treatment of sludges. This study underlines the existence of a short-term risk of nutrient mobilisation by runoff after sludge spreading on soil, and the need to check precisely the impact of this practice on water quality.

Combinatory chemical and biological approaches to investigate metal elements in agricultural runoff water.

As part of a project studying the interactions between farming practices, soil erosion processes, and fate of agricultural pollutants into runoff waters, we conducted a pilot study to investigate the relationship between metal contents and metallothionein-2A (MT-2A) as a bioindicator of metal exposure. Runoff water samples were collected between May and November 1999 at the point of outlet of an elementary watershed located in the Paris basin. Selected metals (Al, As, Cd, Cr, Cu, Pb, Hg, Ni, and Zn) were analyzed using conventional techniques. In parallel, human T cells were exposed to water samples for 6 and 18 h and then cell viability and MT-2A gene expression were measured. Results show that among the 10 water samples tested, Al and Zn predominate (highest values = 4.9 and 2.6 microM, respectively), while other metals were below the microM level. Five out of 10 samples induced MT-2A gene expression (30-80% increase at 18 h) as compared with the control. When comparing MT-2A induction profile with metals contents, no obvious correlation was found, suggesting that additional components or parameters are involved. Finally, there was an apparent inverse relationship between Ca concentration and MT-2A gene induction. Although still preliminary, in the absence of longer monitoring, this study shows that MT-2A gene expression is a useful tool to complement chemical analysis in assessing metal elements in water. These combinatory approaches will be pursued and integrated in an ongoing watershed field research project.

Wheat gliadin promotes the interleukin-4-induced IgE production by normal human peripheral mononuclear cells through a redox-dependent mechanism.

Increased levels of serum IgE have been described in gliadin-intolerant patients; however, biological mechanisms implicated in this immunoglobulin production remained unknown. In this study, we demonstrated that in vitro crude gliadins and gliadin lysates (Glilys) promoted the IL-4-induced IgE production by human peripheral blood mononuclear cells (PBMC), indicating that the biological process related to gliadin intolerance and/or allergy may lead to IgE production in vivo. It was found that crude gliadin and Glilys potentiated, after 13 days of culture in a dose-dependent manner, IL-4-induced IgE production and, to a lesser extent, the IgG production, while they did not affect IgA or IgM productions. This promoting effect of gliadin and Glilys on the IL-4-induced activation of normal human PBMC was also observed on the early release (2 days) of the soluble fraction of CD23, suggesting its possible involvement in IgE potentiation. The promoting effect of crude gliadin and Glilys appeared to be indirect because they did not modify purified B-lymphocytes IgE production after IL-4 and anti-CD40 monoclonal antibody stimulation. In addition, as revealed by luminol-dependent chemiluminescence, we demonstrated that crude gliadin and Glilys promoted a substantial production of free radicals by normal human PBMC, treated or not with IL-4. This redox imbalance associated with an increased IgE production led us to evaluate the effect of pharmacological antioxidants (N-acetyl-cysteine (NAC) and Cu/Zn-superoxide dismutase (SOD1)) on IgE production by human PBMC. The NAC and the intracellularly delivered SOD1 were found to suppress the IL-4+/-crude gliadin or Glilys-induced IgE production by normal human PBMC. Taken together, these data indicated that gliadin specifically enhanced IL-4-induced IgE production by normal human PBMC, probably by the regulation of redox pathways, and that this 'pro-allergenic' effect could be counteracted by natural antioxidants: thiols and/or vectorized SOD1.