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'''Memory of water''' is a phrase used by [[Homeopathy|homeopaths]] to explain how aqueous ("watery") solutions used as remedies might effectuate the results that they claim to see in their patients. Because homeopathic  remedies deliberately use extremely high dilutions it is unlikely that the solutions contain even a single molecule of substance other than pure water. This has led homeopaths to speculate that a possible explanation for the observed responses is "memory of water". Many widely differing explanations have been proposed how water can supposedly remember the biologically active molecules that it had once been in contact with and that have now been removed altogether from the solution by repeated dilution.
'''Memory of water''' is a phrase used by [[Homeopathy|homeopaths]] to explain how the aqueous (water) solutions they use as remedies might produce the results that they claim to see in their patients. Homeopathic remedies deliberately use extremely high dilutions, so it is unlikely that a therapeutic dose contains even a single molecule of the substance being diluted. This has led homeopaths to speculate that a possible explanation for the observed responses is "memory of water"; the water somehow "remembers" the biologically active molecules that it has once been in contact with, and that "memory" produces therapeutic effects.


The first laboratory observation—and still the most famous—that a homeopathic treatment (succussion and dilution) might have an effect on water as well as on ethanol and propanol was embodied in research by [[Jacques Benveniste]] and his colleagues, published in the prestigious English journal ''Nature'' in June 1988.<ref name=Benveniste>E. Davenas, F. Beauvais, J. Arnara, M. Oberbaum, B. Robinzon, A. Miadonna, A. Tedeschi, B. Pomeranz, P. Fortner, P. Belon, J. Sainte-Laudy, B. Poitevin and J. Benveniste, ''Human basophil degranulation triggered by very dilute antiserum against IgE'',  Nature, Vol. '''333''',  pp. 816-818, 30th June, 1988.[http://www.digibio.com/cgi-bin/node.pl?lg=us&nd=n4_1 Free text on DigiBio site].  [http://www.nature.com/doifinder/10.1038/333816a0  Non-free text on Nature site]</ref> Benveniste purportedly discovered that diluted water might retain some qualities of various materials that had once been dissolved in it. The French newspaper ''Le Monde'' is widely credited with popularizing the phrase that same day in a front-page article that touted as well its ramifications for the overthrow of the foundations of physics.<ref>The ''Le Monde'' article actually called it the ''"La mémoire de la matière"'' (the memory of matter) and not ''"La mémoire de l'eau"'' (the memory of water), that later became famous. The initial ''Le Monde'' article did not contain the "memory of water" phrase at all, but used ''le souvenir de molécules biologiquement actives'' (recollection [by water] of biologically active molecules).</ref> The original research could not be replicated under the scrutiny of a committee constituted by ''Nature''<ref name=Maddox>{{cite journal
Chemists and physicists generally see this notion as nonsense. The consensus of scientists working in the field is that liquid water exists as a continuously rearranging [[hydrogen bond|hydrogen-bonded]] network with motions on the picosecond (10<sup>&minus;12</sup> s) time scale.<ref>F. N. Keutsch, J. D. Cruzan, and R. J. Saykally, Chemical Reviews, Vol.'''103''', pp. 2533-2577 (2003)</ref>. A picture of a quickly rearranging network is very difficult to reconcile with liquid water structures that are sustained for more than a few picoseconds. Accordingly there is no room for a water "memory"  in the current scientific view on the liquid.
 
== The Benveniste study ==
 
In 1988, a French immunologist, [[Jacques Benveniste]], and a group of colleagues published a paper <ref name=Benveniste>E. Davenas, F. Beauvais, J. Arnara, M. Oberbaum, B. Robinzon, A. Miadonna, A. Tedeschi, B. Pomeranz, P. Fortner, P. Belon, J. Sainte-Laudy, B. Poitevin and J. Benveniste, ''Human basophil degranulation triggered by very dilute antiserum against IgE'',  Nature, Vol. '''333''',  pp. 816-818, 30th June, 1988.[http://www.digibio.com/cgi-bin/node.pl?lg=us&nd=n4_1 Free text on DigiBio site].  [http://www.nature.com/doifinder/10.1038/333816a0  Non-free text on Nature site]</ref> in the prestigious English journal ''Nature''. Their data indicated that diluted water, ethanol or propanol might retain some qualities of various materials that had once been dissolved in it. In particular, they claimed to have measured effects on human immune response.
 
Human [[basophil]]s are a rare [[granulocyte]] cell type accounting for 0.1–1% of white blood cells; these cells contain large numbers of "granules" which store inflammatory mediators, including in particular [[histamine]]. These cells can be cultured readily and studied ''in vitro''. In these cells, exposure to anti-human-IgE [[antibodies]] triggers a "degranulation" process in which the granules fuse with the plasma membrane to release their contents, including histamine, into the extracellular fluid. At high concentrations (>10<sup>−6</sup> M) histamine binds to H<sub>2</sub> receptors on the surface of the basophils, and regulates the basophil degranulation by feedback inhibition. Basophil activation can be measured in several different ways. First, degranulated cells can be stained and then counted; this is a subjective measurement and is prone to variable outcomes depending on the observer. Second, histamine release into the culture medium can be measured using fluorimetric assays. Third, the fusion of cytoplasmatic granules leads to the expression of the marker [[CD63]] on the  surface of the basophils; the percentage of basophils that express CD63 can be determined with [[flow-cytometry]], and correlates well with histamine release.
 
Benveniste and his colleagues found evidence that very high dilutions of anti-immunoglobulin E had an effect on the degranulation of human basophils. At the dilutions used, the solutions should have contained only molecules of water, and no molecules of (anti-IgE) at all. Benveniste concluded that the configuration of molecules in water was biologically active. <ref name=Benveniste />
 
The French newspaper ''Le Monde'' covered this, referring to ''"la mémoire de la matière"'' (the memory of matter) and ''le souvenir de molécules biologiquement actives'' (recollection [by water] of biologically active molecules). In English, however, the phrase that became widespread was "memory of water". Le Monde considered the paper important, making it a front page story, and correctly pointing out that if this work were correct, it would overthrow many of the foundations of physics.
 
''Nature'' published the article with two unprecedented conditions: first, that the results must first be confirmed by other laboratories; second, that a team selected by ''Nature'' be allowed to investigate his laboratory following publication. Benveniste accepted these conditions; the results were replicated in Milan, Italy; in Toronto, Canada; in Tel-Aviv, Israel and in Marseille, France, and the article was accompanied by an editorial titled "When to believe the unbelievable." After publication, the follow-up investigation was conducted by a team including the editor of ''Nature'', Dr John Maddox, American scientific fraud investigator and chemist Walter Stewart, and "professional [[pseudoscience]] debunker" [[James Randi]]. With the cooperation of Benveniste's  team, under double-blind conditions, they failed to replicate the results. Benveniste refused to withdraw his claims, and the team published in the July 1988 a detailed critique of Benveniste’s study. <ref name=Maddox>{{cite journal
   | last =Maddox
   | last =Maddox
   | first =John
   | first =John
Line 13: Line 25:
   | date =28 July 1988
   | date =28 July 1988
   | url =http://br.geocities.com/criticandokardec/benveniste02.pdf
   | url =http://br.geocities.com/criticandokardec/benveniste02.pdf
   | doi =10.1038/334287a0 |format=PDF}}</ref> and its conclusions remain extremely controversial. In the two decades since the first article, most molecular physicists and physical chemists have not accepted any  explanations of the "memory of water" as plausible. In spite of their near-universal skepticism, however, Benveniste never retracted his initial claims, going so far, in fact, in 1997, as to say that the memory could even be transmitted across a digital telephone link.<ref name = digibio>Benveniste J ''et al.'' (1997) [http://www.digibio.com/cgi-bin/node.pl?lg=us&nd=n4_3: "Transatlantic Transfer of Digitized Antigen Signal by Telephone Link ''J Allergy Clin Immunol - Program and abstracts of papers presented during scientific sessions AAAAI/AAI.CIS Joint Meeting February 21-26, 1997</ref>
   | doi =10.1038/334287a0 |format=PDF}}</ref> They claimed that the experiments were badly controlled statistically, that measurements that conflicted with the claim had been excluded, that there was insufficient avoidance of contamination, and that there were questions of undisclosed conflict of interest, as the salaries of two coauthors of the published article were paid for under a contract with the French company ''Boiron et Cie''.<ref name=Maddox/>.
 
Subsequent attempts by other labs to reproduce Benveniste's results have failed to reproduce the effects. <ref>S. J. Hirst, N. A. Hayes, J. Burridge, F. L. Pearce, J. C. Foreman, ''Human basophil degranulation is not triggered by very dilute antiserum against human IgE'', Nature vol. '''366''', pp. 525&ndash;527 (1993) [http://dx.doi.org/doi:10.1038/366525a0 doi]
</ref> <ref> Guggisberg AG, Baumgartner SM, Tschopp CM, and  Heusser P (2005) Replication study concerning the effects of homeopathic dilutions of histamine on human basophil degranulation in vitro. ''Complement Ther Med'' 13:91-100.</ref>
 
Benveniste has never retracted his claims. In the same issue of ''Nature'' that carried the critique, Benveniste vigorously attacked the ''Nature'' team’s "mockery of scientific inquiry." <ref>J. Benveniste, ''Dr Jacques Benveniste replies'', News and views, ''Nature'', vol. '''334''' p. 291 (1988) [http://dx.doi.org/doi:10.1038/334291a0 doi]  </ref>. He has maintained his position in later publications as well.
 
== Homeopathic coverage ==


==The ''Nature'' article==
The notion of "memory of water" is taken quite seriously among [[homeopathy|homeopaths]]. For them, it provides an explanation of why some of their remedies work, and raises some very interesting questions &mdash; how does one explain the phenomenon, and what will the explanations tell us about more general issues of the structure of liquids, or indeed of matter? It is clear that we will need some new physics to explain it.


In their ''Nature'' article,  Benveniste ''et al.'' reported<ref name=Benveniste />
To a more orthodox scientist, it is also clear that explaining the phenomenon would require some new physics. What is not clear is that there is anything that needs explaining; the only evidence is the flawed Benveniste work, and that is not nearly enough to contemplate fundamental changes in physics.
that particular [[solution]]s of biologically active compounds subjected to sequential physical processing (shaking) and progressive dilutions appeared to have some biological effects that were different from the "control" effects of the water used as a solvent, even though the solution was diluted so much that the chance that a single molecule of the biologically active solute was left in it was completely negligible. In their original paper Benveniste ''et al.'' state that their results remain unexplained, but suggest that it is somehow related to the molecular organization of water.
Later Benveniste ''et al.''<ref name = digibio/> hypothesized that water somehow "remembers"  the active compounds (together with their biological properties) that it contained before dilution.  The work resulted in considerable controversy, as most other laboratories stated they were unable to reproduce the reported effects, while, on the other hand, an international collaboration led by Professor Madeleine Ennis of Queen's University of Belfast reported confirmation.<ref name = "Belon">P. Belon, J. Cumps, M. Ennis , P. F. Mannaioni, M. Roberfroid, J. Sainte-Laudy and F. A. C. Wiegant (2004) ''Histamine dilutions modulate basophil activation.''  Inflammation Research  '''53:''' 181–188. PMID 15105967. [http://dx.doi.org/10.1007/s00011-003-1242-0 doi]</ref>
Following Benveniste and coworkers, Ennis ''et al.'' studied the effects of homeopathically treated solutions on human [[basophil]]s.  In their paper Ennis and coworkers state emphatically and repeatedly that they cannot explain their findings.
<!--
<ref>Josephson, Brian. Pathological disbelief. [www.lenr-canr.org/acrobat/JosephsonBpathologic.pdf] Josephson is a Physics Nobel laureate who has vigorously taken up the case for many widely denigrated theories, arguing that [http://www.tcm.phy.cam.ac.uk/~bdj10/ "if scientists as a whole denounce an idea this should not necessarily be taken as proof that the said idea is absurd: rather, one should examine carefully the alleged grounds for such opinions and judge how well these stand up to detailed scrutiny."]</ref>
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An overview of the issues surrounding the memory of water and its relationship to homeopathic medicine was the subject of a special issue of the leading journal on homeopathy.<ref name="Homeopathy2007">{{citation
An overview of the issues surrounding the memory of water and its relationship to homeopathic medicine was the subject of a special issue of the leading journal on homeopathy.<ref name="Homeopathy2007">{{citation
Line 29: Line 42:
  | date = 2007
  | date = 2007
  | title = The Memory of Water ''Homeopathy.'' 96:141-230}}  
  | title = The Memory of Water ''Homeopathy.'' 96:141-230}}  
::Copies of the articles in this special issue are freely available on a private website, along with discussion. [http://www.badscience.net/?p=490 Homeopathy Journal Club] hosted by Bad Science, a blog by Ben Goldacre</ref> The articles in this issue propose widely varying mechanisms for water memory, such as: electromagnetic exchange of information between molecules, breaking of temporal symmetry, thermoluminescence, entanglement described by a new quantum theory, formation of hydrogen peroxide, clathrate formation, etc. without any mechanism singularly standing out as the definitive explanation. Some of the proposed mechanisms require revolutionary new physical principles overthrowing much of 20th century physics. Remarkably, all explanations concentrate on water and its alleged special properties, the fact that&mdash;according to Benveniste ''et al.''&mdash;ethanol and propanol also have  memory is completely ignored.
::Copies of the articles in this special issue are freely available on a private website, along with discussion. [http://www.badscience.net/?p=490 Homeopathy Journal Club] hosted by Bad Science, a blog by Ben Goldacre</ref> The articles in this issue propose widely varying mechanisms for water memory, such as: electromagnetic exchange of information between molecules, breaking of temporal symmetry, thermoluminescence, entanglement described by a new quantum theory, formation of hydrogen peroxide, clathrate formation, etc. without any mechanism singularly standing out as the definitive explanation. Some of the proposed mechanisms require revolutionary new physical principles overthrowing much of 20th century physics. Remarkably, all explanations concentrate on water and its alleged special properties, the fact that&mdash;according to Benveniste ''et al.''&mdash;ethanol and propanol also have  memory is completely ignored.
 
The consensus of scientists working in the field is that liquid water exists as a continuously rearranging [[hydrogen bond|hydrogen-bonded]] network with motions on the picosecond (10<sup>&minus;12</sup> s) scale.<ref>F. N. Keutsch, J. D. Cruzan, and R. J. Saykally, Chemical Reviews, Vol.'''103''', pp. 2533-2577 (2003)</ref>. A picture of a quickly rearranging network is very difficult to reconcile with liquid water structures that are sustained for more than a few picoseconds. Accordingly there is no room for a water "memory"  in the modern scientific view on the liquid. If work other than biological effects on human basophils would become available that would support the notion of water memory, and if this work would stand scientific scrutiny, then much of the existing experimental and theoretical data on liquid water would have to be reinterpreted or even rejected. Before this happens, most water researchers do not find it useful to speculate in what way liquid water could store long-lived information.
<!--
==The components of liquid water==
Water is not simply a collection of molecules of H<sub>2</sub>O, it contains several molecular species including ''ortho'' and ''para'' water molecules, and water molecules with different isotopic compositions such as HDO and H<sub>2</sub><sup>18</sup>O. These water molecules as part of weakly-bound but partially-covalently linked molecular clusters containing one, two, three or four hydrogen bonds, and hydrogen ion and hydroxide ion species. In addition, there are ''always'' adventitious solutes in liquid water. Even double-distilled and deionized water always contains significant and variable trace amounts of contaminating ions, and different samples will differ in the contaminants that they contain.
 
==Putative explanations==
There is some support for the notion that water can have properties that depend on how it has previously been processed (that is, water has, in some sense, a kind of "memory"). In particular, water, as a result of repeated vigorous shaking, might include Redox molecules produced from water, dissolved atmospheric gases and airborne contaminants, silicates (i.e., tiny glass "chips"),
nanobubbles and their material surfaces, dissolved ions, including from the glassware. It may also be contaminated by material that adheres to the surfaces of glassware, for example by bacterial material. There might also be some effects of successive shaking on water structure that causes "clustering" of water molecules.
 
These mechanisms are not mechanisms of memory in any cognitive or electronic computing  sense; the term memory here is used as a metaphor, implying only that the past history has a discernible influence on the present properties.
-->
 
==The Benveniste studies==
Human [[basophil]]s are a rare [[granulocyte]] cell type accounting for 0.1–1% of white blood cells; these cells contain large numbers of "granules" which store inflammatory mediators, including in particular [[histamine]]. These cells can be cultured readily and studied ''in vitro''. In these cells, exposure to anti-human-IgE [[antibodies]] triggers a "degranulation" process in which the granules fuse with the plasma membrane to release their contents, including histamine, into the extracellular fluid. At high concentrations (>10<sup>−6</sup> M) histamine binds to H<sub>2</sub> receptors on the surface of the basophils, and regulates the basophil degranulation by feedback inhibition.
 
Basophil activation can be measured in several different ways. First, degranulated cells can be stained and then counted; this is a subjective measurement and is prone to variable outcomes depending on the observer. Second, histamine release into the culture medium can be measured using fluorimetric assays. Third, the fusion of cytoplasmatic granules leads to the expression of the marker [[CD63]] on the  surface of the basophils; the percentage of basophils that express CD63 can be determined with [[flow-cytometry]], and correlates well with histamine release.
 
As mentioned above, the discussion about water memory started when in 1988 Jacques Benveniste (1935-2004), a distinguished French immunologist, published a controversial paper in ''Nature'' reporting on the action of very high dilutions of anti-immunoglobulin E on the degranulation of human [[basophil]]s.<ref name=Benveniste /> At the high dilutions used, the solutions should have contained only molecules of water, and no molecules of (anti-IgE) at all. Benveniste concluded that the configuration of molecules in water was biologically active. 
 
''Nature'' published the article with two unprecedented conditions: first, that the results must first be confirmed by other laboratories; second, that a team selected by ''Nature'' be allowed to investigate his laboratory following publication. Benveniste accepted these conditions; the results were replicated in Milan, Italy; in Toronto, Canada; in Tel-Aviv, Israel and in Marseille, France, and the article was accompanied by an editorial titled "When to believe the unbelievable." After publication, the follow-up investigation was conducted by a team including the editor of ''Nature'', Dr John Maddox, American scientific fraud investigator and chemist Walter Stewart, and "professional [[pseudoscience]] debunker" [[James Randi]]. With the cooperation of Benveniste's  team, under double-blind conditions, they failed to replicate the results. Benveniste refused to withdraw his claims, and the team published in the July 1988 a detailed critique of Benveniste’s study. They claimed that the experiments were badly controlled statistically, that measurements that conflicted with the claim had been excluded, that there was insufficient avoidance of contamination, and that there were questions of undisclosed conflict of interest, as the salaries of two coauthors of the published article were paid for under a contract with a French company, ''Boiron et Cie'', which manufactured homeopathic remedies.<ref name=Maddox/>
 
In the same issue of ''Nature'' (and subsequently) Benveniste vigorously attacked the ''Nature'' team’s "mockery of scientific inquiry." <ref>J. Benveniste,  ''Dr Jacques Benveniste replies'', News and views, ''Nature'', vol. '''334''' p. 291 (1988) [http://dx.doi.org/doi:10.1038/334291a0 doi]  </ref> Subsequent attempts by other labs to reproduce Benveniste's results have failed to reproduce the effects <ref>S. J. Hirst, N. A. Hayes, J. Burridge, F. L. Pearce, J. C. Foreman, ''Human basophil degranulation is not triggered by very dilute antiserum against human IgE'', Nature vol. '''366''', pp. 525&ndash;527 (1993) [http://dx.doi.org/doi:10.1038/366525a0 doi]
</ref>. However other studies have looked at the effects of very low concentrations of [[histamine]] on degranulation induced by anti-[[Immunoglobulin#immunoglobulin|immunoglobulin E (IgE)]] antibodies, and again reported effects at very low concentrations.<ref name = "Belon" /> As degranulation itself produces relatively high concentrations of histamine in the medium, one would only expect an effect with very high concentrations of added histamine&mdash;and indeed the most recent study reported significant effects only at 10<sup>&minus;2</sup> M histamine. These experiments generally involved dilutions of histamine to concentrations of as low as 10<sup>&minus;38</sup> M, and the dilutions were performed conventionally not according to the protocols used in homeopathy.
 
So how is it possible that adding vanishingly low concentrations of histamine to a preparation that is already secreting high concentrations might have any effect? What could possibly explain the extraordinary results reported by Benveniste and others? One difficulty with the basophil preparation is that, in these cells, degranulation can be triggered by many different stimuli, including slight mechanical disturbances and environmental variations in temperature, and is sensitive to small differences in incubation time, making adequate controls very difficult. For example, in the experiments of Guggisberg ''et al.''<ref> Guggisberg AG ''et al.''(2005) Replication study concerning the effects of homeopathic dilutions of histamine on human basophil degranulation in vitro. ''Complement Ther Med'' 13:91-100.</ref> the authors found no significant effects of low dilutions of histamine, but did find significant effects for row numbers of the microtiter plates&mdash;i.e., there was a significant effect simply of the order in which the samples were assayed. They concluded that seemingly trivial differences in the experimental set up can lead to significant differences of the results.
 
Benveniste never retracted his claims.  On the contrary, later he founded the field of "Digital Biology",<ref>[http://www.digibio.com/cgi-bin/node.pl?nd=n1 Overview of DigiBio] Website retrieved May 7, 2009 </ref> which is based on the assumption that molecules emit [[electromagnetic radiation]] in the frequency range 20 Hz to 20 kHz,<ref>[http://www.digibio.com/cgi-bin/node.pl?nd=n3 What is Digital Biology?]  Website retrieved May 7, 2009 </ref> the same range as sound waves audible by humans.<ref>To avoid misunderstanding: [[electromagnetic wave|electromagnetic (EM) waves]] have no relationship to sound waves. Sound waves are propagated by material particles and hence cannot propagate in vacuum, while EM waves can. Furthermore, neither theoretical nor experimental [[molecular spectroscopy]] can explain the existence of molecular EM waves in this region of extremely low frequencies. Moreover, unless it is shown what feeds them, Benveniste's EM waves seem to contradict the principle of conservation of energy</ref> By means of an amplifier, electromagnetic coils and a PC sound card Benveniste claimed that he was able to digitize and store the molecular signals. The digital information (possibly after sending it over the internet) could be replayed to a biological system making it believe that it is in the presence of its "favorite molecule".  As Benveniste's description of his experiments is too vague to even begin thinking about trying to reproduce them, and as his theory is not only primitive and underdeveloped, but also in complete contradiction to the well-established principles of molecular spectroscopy, it is fair to call Digital Biology a [[pseudoscience]].
<!--
On the contrary, in 1997, he declared that the memory could be transmitted across a digital telephone link, suggesting that the memory involved electromagnetic signals.<ref name = digibio/>
 
==Water in living organisms==
Water is essential for living organisms at every level; at the molecular level in living cells, it is essential for functional macromolecular folding, stabilization and activity, transport, membrane formation and protein insertion into membranes; it the intracellular matrix in which biological molecules interact. Understanding exactly how water diffuses when confined in proximity to complex macromolecules inside a cell is therefore an important challenge. The diffusion coefficient of water in biological tissues has been measured using [[NMR spectroscopy|nuclear magnetic resonance]], and these have shown that, within a cell, water diffuses much more slowly than pure water in aqueous media. This is partly explained by tortuosity effects, macromolecular crowding and confinement effects, but some (but not all<ref>Jasnin M ''et al.'' (2008) Down to atomic-scale intracellular water dynamics EMBO reports [http://www.nature.com/embor/journal/v9/n6/full/embor200850.html 9:543–7.("Our data show that the water between macromolecules in the ''in vivo'' intracellular environment has properties that are essentially the same as those of pure water..." )</ref>) have suggested that the interaction with macromolecules might cause "clustering" of water molecules -that it might change the structure of the intracellular water.
 
<blockquote>The cell can be seen, from a somewhat extreme structuralist point of view, as''' organized water'''. There is an incipient order in liquid water, which is given long-range coherence and permanence by the protein framework. In the words of A. Szent-Gyorgyi, '''“Life is water dancing to the tune of solids”.'''</blockquote>
 
==Water structure as a basis for homeopathy==
<blockquote>"Based on this evidence we would be ready to accept that homoeopathy can be efficacious, if only the mechanism of action were more plausible". Kleijnen J ''et al.''(1991) Clinical trials of homeopathy. ''British Medical Journal'' 302:316–23</blockquote>
 
[[Homeopathy]] involves the use of 'remedies' that typically involve "ultradilution" of [[drug]]s; dilution well beyond the point at which ''any'' of the original molecules are still present, combined with vigorous shaking at each stage of dilution. [[Samuel Hahnemann]], the 18th century founder of homeopathy, recognised that the vehicle or solvent (water or alcohol) must be considered as the medicine, rather than the molecule<ref name="pmid14619985">{{cite journal |author=Khuda-Bukhsh AR |title=Towards understanding molecular mechanisms of action of homeopathic drugs: an overview |journal=Mol Cell Biochem |volume=253 |pages=339–45 |year=2003 |pmid=14619985 |doi= |url=http://www.science.smith.edu/departments/Biochem/Chm_357/Articles/homeopathy_molecular%20mechanisms.pdf}}</ref>  Research on the plausibility of homeopathy is thus an attempt to characterize how the behaviour of the molecules of a solvent might differ depending, on the solute that was diluted in it and on the kinetic energy imposed on it (by "shaking"). Liquid water is generally assumed to be a network of H<sub>2</sub>O molecules forming short-lived (on the order of 10<sup>&minus;12</sup> s) [[hydrogen bonds]]. Some scientists have questioned whether the very short life of these bonds determines an equally short life to the structures found in water, at the larger scale of 200 or more H<sub>2</sub>O molecules. At an even larger scale, it can be easily observed that a wave keeps existing despite of the constant doing and undoing of hydrogen bonds, and that ice sculptures are also made of H<sub>2</sub>O molecules constantly bonding and separating. In the same way, water clusters of a hundred or more molecules might have a longer life than the individual bonds composing it.<ref>See the related sections in Martin Chaplin's [http://www.lsbu.ac.uk/water/ Water Structure and Science] resource for animations.</ref>
 
Many 'anomalies' <ref>See Chaplin's [http://www.lsbu.ac.uk/water/anmlies.html web resource</ref> of water reflect a heterogeneity in its structure: liquid water is a combination of different ''[[phase]]s'' (a term in materials science to designate 3D arrangements of molecules or patterns) that are ''not'' short-lived, although, at the smaller, molecular scale, the incessant agitation might evoke the impression that no higher order can exist.


Two physicochemical phenomena have been given particular attention by proponents of homeopathy: [[clathrates]] and [[solitons]]. ''Clathrates'' are complexes of water molecules around low-molecular-weight molecules (e.g., [[methane]]) or atoms (e.g., [[xenon]]) that can cause the growth of other clathrates devoid of central molecules. The presence of clathrates affects the results of mass spectrometry. Thus this is a mechanism whereby low concentration contaminants can influence the apparent properties of water.<ref>{{citation
== Digital biology ==
| journal = Nature
| year = 2007
| volume = 449
| pages = 1033-6
| title =  Clathrate nanostructures for mass spectrometry.
| author =  Northen TR ''et al.''
| url = http://www.ncbi.nlm.nih.gov/pubmed/17960240}}</ref>.


Vigorous shaking of water in glass bottles can cause small amounts of silica (silicate) fragments to fall into it <ref>Demangeat J-L ''et al.'' (2004) Low-Field NMR water proton longitudinal relaxation in ultrahighly diluted aqueous solutions of silica-lactose prepared in glass material for pharmaceutical use. ''Applied Magnetic Resonance'' 26:465–81.</ref> and saturation of water with components of air. Homeopathic drug manufacturers use a double-distilled water in making their medicine, and whatever medicinal substance is placed in the water might interact with the silicate fragments.  
Benveniste never retracted his claims. On the contrary, later he turned to working on what he called "Digital Biology", which is based on the assumption that molecules emit [[electromagnetic radiation]] in the frequency range 20 Hz to 20 KHz, the same range as sound waves audible by humans. He argued <ref>{{citation
| author = Jacques Beneveniste
| title = Understanding Digital Biology
| date = 1998
| url = http://www.homeopathyhome.com/reference/articles/benveniste.shtml
}}</ref>
that electromagnetic vibrations are "signals exchanged among molecules" that are used by living things to convey information. By means of an amplifier, electromagnetic coils and a PC sound card Benveniste claimed that he was able to digitize and store the molecular signals. The digital information (possibly after sending it over the Internet) could be replayed to a biological system making it believe that it is in the presence of its "favorite molecule".


It has been suggested that micro-bubbles and nano-bubbles, caused by vigorous shaking, might "burst" to produce microenvironments of higher temperature and pressure. <ref>Elia V ''et al.'' (2004) Permanent physio-chemical properties of extremely diluted aqueous solutions of homeopathic medicines ''Homeopathy'' 93:144–50.</ref>  Some scientists have estimated that the vigorous shaking involved with making homeopathic remedies changes the pressure in the water, akin to water being at 10,000 feet in altitude.<ref>Roy R''et al.'' (2005) The Structure of liquid water: Novel insights from materials research; potential relevance to homeopathy, ''Materials Research Innovations'' [http://www.rustumroy.com/Roy_Structure%20of%20Water.pdf 9:4].</ref>
To most scientists, these notions are obviously utterly implausible. In the first place, the wavelength of electromagnetic waves at these frequencies is several kilometers, far too long to interact with microscopic molecules. The usual  theories of physics give no reason to expect molecular EM waves in this region of extremely low frequencies and no-one else has observed them. Benveniste's description of his experiments is too vague to even begin thinking about trying to reproduce them. His theory is in complete contradiction to the well-established principles of molecular spectroscopy, and is nowhere near either well enough developed or sufficiently supported by experiment to be taken seriously as a challenge to established theories.
-->


==References==
==References==
{{reflist|2}}
{{reflist|2}}

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Memory of water is a phrase used by homeopaths to explain how the aqueous (water) solutions they use as remedies might produce the results that they claim to see in their patients. Homeopathic remedies deliberately use extremely high dilutions, so it is unlikely that a therapeutic dose contains even a single molecule of the substance being diluted. This has led homeopaths to speculate that a possible explanation for the observed responses is "memory of water"; the water somehow "remembers" the biologically active molecules that it has once been in contact with, and that "memory" produces therapeutic effects.

Chemists and physicists generally see this notion as nonsense. The consensus of scientists working in the field is that liquid water exists as a continuously rearranging hydrogen-bonded network with motions on the picosecond (10−12 s) time scale.[1]. A picture of a quickly rearranging network is very difficult to reconcile with liquid water structures that are sustained for more than a few picoseconds. Accordingly there is no room for a water "memory" in the current scientific view on the liquid.

The Benveniste study

In 1988, a French immunologist, Jacques Benveniste, and a group of colleagues published a paper [2] in the prestigious English journal Nature. Their data indicated that diluted water, ethanol or propanol might retain some qualities of various materials that had once been dissolved in it. In particular, they claimed to have measured effects on human immune response.

Human basophils are a rare granulocyte cell type accounting for 0.1–1% of white blood cells; these cells contain large numbers of "granules" which store inflammatory mediators, including in particular histamine. These cells can be cultured readily and studied in vitro. In these cells, exposure to anti-human-IgE antibodies triggers a "degranulation" process in which the granules fuse with the plasma membrane to release their contents, including histamine, into the extracellular fluid. At high concentrations (>10−6 M) histamine binds to H2 receptors on the surface of the basophils, and regulates the basophil degranulation by feedback inhibition. Basophil activation can be measured in several different ways. First, degranulated cells can be stained and then counted; this is a subjective measurement and is prone to variable outcomes depending on the observer. Second, histamine release into the culture medium can be measured using fluorimetric assays. Third, the fusion of cytoplasmatic granules leads to the expression of the marker CD63 on the surface of the basophils; the percentage of basophils that express CD63 can be determined with flow-cytometry, and correlates well with histamine release.

Benveniste and his colleagues found evidence that very high dilutions of anti-immunoglobulin E had an effect on the degranulation of human basophils. At the dilutions used, the solutions should have contained only molecules of water, and no molecules of (anti-IgE) at all. Benveniste concluded that the configuration of molecules in water was biologically active. [2]

The French newspaper Le Monde covered this, referring to "la mémoire de la matière" (the memory of matter) and le souvenir de molécules biologiquement actives (recollection [by water] of biologically active molecules). In English, however, the phrase that became widespread was "memory of water". Le Monde considered the paper important, making it a front page story, and correctly pointing out that if this work were correct, it would overthrow many of the foundations of physics.

Nature published the article with two unprecedented conditions: first, that the results must first be confirmed by other laboratories; second, that a team selected by Nature be allowed to investigate his laboratory following publication. Benveniste accepted these conditions; the results were replicated in Milan, Italy; in Toronto, Canada; in Tel-Aviv, Israel and in Marseille, France, and the article was accompanied by an editorial titled "When to believe the unbelievable." After publication, the follow-up investigation was conducted by a team including the editor of Nature, Dr John Maddox, American scientific fraud investigator and chemist Walter Stewart, and "professional pseudoscience debunker" James Randi. With the cooperation of Benveniste's team, under double-blind conditions, they failed to replicate the results. Benveniste refused to withdraw his claims, and the team published in the July 1988 a detailed critique of Benveniste’s study. [3] They claimed that the experiments were badly controlled statistically, that measurements that conflicted with the claim had been excluded, that there was insufficient avoidance of contamination, and that there were questions of undisclosed conflict of interest, as the salaries of two coauthors of the published article were paid for under a contract with the French company Boiron et Cie.[3].

Subsequent attempts by other labs to reproduce Benveniste's results have failed to reproduce the effects. [4] [5]

Benveniste has never retracted his claims. In the same issue of Nature that carried the critique, Benveniste vigorously attacked the Nature team’s "mockery of scientific inquiry." [6]. He has maintained his position in later publications as well.

Homeopathic coverage

The notion of "memory of water" is taken quite seriously among homeopaths. For them, it provides an explanation of why some of their remedies work, and raises some very interesting questions — how does one explain the phenomenon, and what will the explanations tell us about more general issues of the structure of liquids, or indeed of matter? It is clear that we will need some new physics to explain it.

To a more orthodox scientist, it is also clear that explaining the phenomenon would require some new physics. What is not clear is that there is anything that needs explaining; the only evidence is the flawed Benveniste work, and that is not nearly enough to contemplate fundamental changes in physics.

An overview of the issues surrounding the memory of water and its relationship to homeopathic medicine was the subject of a special issue of the leading journal on homeopathy.[7] The articles in this issue propose widely varying mechanisms for water memory, such as: electromagnetic exchange of information between molecules, breaking of temporal symmetry, thermoluminescence, entanglement described by a new quantum theory, formation of hydrogen peroxide, clathrate formation, etc. without any mechanism singularly standing out as the definitive explanation. Some of the proposed mechanisms require revolutionary new physical principles overthrowing much of 20th century physics. Remarkably, all explanations concentrate on water and its alleged special properties, the fact that—according to Benveniste et al.—ethanol and propanol also have memory is completely ignored.

Digital biology

Benveniste never retracted his claims. On the contrary, later he turned to working on what he called "Digital Biology", which is based on the assumption that molecules emit electromagnetic radiation in the frequency range 20 Hz to 20 KHz, the same range as sound waves audible by humans. He argued [8] that electromagnetic vibrations are "signals exchanged among molecules" that are used by living things to convey information. By means of an amplifier, electromagnetic coils and a PC sound card Benveniste claimed that he was able to digitize and store the molecular signals. The digital information (possibly after sending it over the Internet) could be replayed to a biological system making it believe that it is in the presence of its "favorite molecule".

To most scientists, these notions are obviously utterly implausible. In the first place, the wavelength of electromagnetic waves at these frequencies is several kilometers, far too long to interact with microscopic molecules. The usual theories of physics give no reason to expect molecular EM waves in this region of extremely low frequencies and no-one else has observed them. Benveniste's description of his experiments is too vague to even begin thinking about trying to reproduce them. His theory is in complete contradiction to the well-established principles of molecular spectroscopy, and is nowhere near either well enough developed or sufficiently supported by experiment to be taken seriously as a challenge to established theories.

References

  1. F. N. Keutsch, J. D. Cruzan, and R. J. Saykally, Chemical Reviews, Vol.103, pp. 2533-2577 (2003)
  2. 2.0 2.1 E. Davenas, F. Beauvais, J. Arnara, M. Oberbaum, B. Robinzon, A. Miadonna, A. Tedeschi, B. Pomeranz, P. Fortner, P. Belon, J. Sainte-Laudy, B. Poitevin and J. Benveniste, Human basophil degranulation triggered by very dilute antiserum against IgE, Nature, Vol. 333, pp. 816-818, 30th June, 1988.Free text on DigiBio site. Non-free text on Nature site
  3. 3.0 3.1 Maddox, John; James Randi and Walter W. Stewart (28 July 1988). "‘High-dilution’ experiments a delusion" (PDF). Nature 334: 287–290. DOI:10.1038/334287a0. Research Blogging.
  4. S. J. Hirst, N. A. Hayes, J. Burridge, F. L. Pearce, J. C. Foreman, Human basophil degranulation is not triggered by very dilute antiserum against human IgE, Nature vol. 366, pp. 525–527 (1993) doi
  5. Guggisberg AG, Baumgartner SM, Tschopp CM, and Heusser P (2005) Replication study concerning the effects of homeopathic dilutions of histamine on human basophil degranulation in vitro. Complement Ther Med 13:91-100.
  6. J. Benveniste, Dr Jacques Benveniste replies, News and views, Nature, vol. 334 p. 291 (1988) doi
  7. Martin Chaplin, ed. (2007), The Memory of Water Homeopathy. 96:141-230
    Copies of the articles in this special issue are freely available on a private website, along with discussion. Homeopathy Journal Club hosted by Bad Science, a blog by Ben Goldacre
  8. Jacques Beneveniste (1998), Understanding Digital Biology