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Interdiscip Sci Comput Life Sci (2009) 1: 81–90
DOI: 10.1007/s12539-009-0036-7




  Electromagnetic Signals Are Produced by Aqueous Nanostructures
              Derived from Bacterial DNA Sequences

                             Luc MONTAGNIER1,2∗ , Jamal A¨
                                                         ISSA1 , St´phane FERRIS1 ,
                                                                   e
                                  Jean-Luc MONTAGNIER , Claude LAVALLEE1
                                                       1                 ´
                 1
                     (Nanectis Biotechnologies, S.A. 98 rue Albert Calmette, F78350 Jouy en Josas, France)
                      2
                        (Vironix LLC, L. Montagnier 40 Central Park South, New York, NY 10019, USA)



     Recevied 3 January 2009 / Revised 5 January 2009 / Accepted 6 January 2009


     Abstract: A novel property of DNA is described: the capacity of some bacterial DNA sequences to induce
     electromagnetic waves at high aqueous dilutions. It appears to be a resonance phenomenon triggered by the
     ambient electromagnetic background of very low frequency waves. The genomic DNA of most pathogenic bacteria
     contains sequences which are able to generate such signals. This opens the way to the development of highly
     sensitive detection system for chronic bacterial infections in human and animal diseases.
     Key words: DNA, electromagnetic signals, bacteria.




   Pathogenic microorganisms in this day of age are not          priate dilutions in water. The emission of such waves is
only submitted to high selective pressure by the im-             likely to represent a resonance phenomenon depending
mune defenses of their hosts but also have to survive un-        on excitation by the ambient electromagnetic noise. It
der highly active antiviral or antibiotic treatments. Not        is associated with the presence in the aqueous dilutions
surprisingly, they have evolved in finding many ways to           of polymeric nanostructures of defined size. The super-
escape these hostile conditions, such as mutations of re-        natant of uninfected eukaryotic cells used as controls
sistance, hypervariability of surface antigens, protective       did not exhibit this property.
biofilms, latency inside cells and tissues.                          In this paper we provide a first characterization of the
   We initially observed (Montagnier and Lavallee, per-          electromagnetic signals (EMS) and of their underlying
sonal communication) that some filtration procedures              nanostructures produced by some purified bacteria.
aimed at sterilizing biological fluids can yield under               In addition to M. pirum, a more classical bacterium,
some defined conditions the infectious microorganism              E. Col i, was utilized for the purpose of the analysis.
which was present before the filtration step. Thus, fil-           The nanostructures produced by HIV will be the sub-
tration of a culture supernatant of human lymphocytes            ject of another paper.
infected with Mycoplasma pirum, a microorganism of                  M. pirum is a peer-shaped small bacterial cell,
about 300 nM in size, through filters of 100 nM or                ressembling M. pneumoniae, which can be grown in syn-
20 nM porosities, yielded apparently sterile fluid. The           thetic enriched medium (SP4) (Tully et al., 1977) but
latter however was able to regenerate the original my-           also mutiplies at the surface of human T lymphocytes.
coplasma when incubated with a mycoplasma negative                  The strain (Ber) used in our experiments was isolated
culture of human lymphocytes within 2 to 3 weeks.                from a T lymphocyte culture derived from the blood
   Similarly, a 20 nM filtration did not retain a minor in-       of an apparently healthy subject (Grau et al., 1993).
fective fraction of HIV, the causal agent of AIDS, whose         The strong mycoplasma adherence to lymphocytes is
viral particles have a diameter averaging 100-120 nM.            mediated by a specific adhesin, whose gene had been
   In the course of investigating the nature of such filter-      previously cloned and sequenced by the authors (Tham
ing infectious forms, we found another property of the           et al., 1994).
filtrates, which may or may not be related to the former:            We used as primary source of the mycoplasma, super-
their capacity to produce some electromagnetic waves             natants of infected human T lymphocyte cultures or of
of low frequency in a reproducible manner after appro-           cultures of the CEM tumor T cell line. All cell cultures
                                                                 were first tested for the lack of M. pirum contamination
   *Corresponding author.                                        by polymerase chain reaction (PCR) and nested PCR,
   E-mail: nadiacpt@yahoo.fr                                     before starting the experiments. Titers of 106 -107 infec-
82                                                                        Interdiscip Sci Comput Life Sci (2009) 1: 81–90

tious Units/ml of M. pirum were readily achieved after          Blaster Card itself connected to a laptop computer,
5-6 days of incubation following deliberate infection of        preferentially powered by its 12 volt battery. Each
both types of cultures.                                         emission is recorded twice for 6 seconds, amplified 500
   Filtration of the clarified supernatant was first per-         times and processed with different softwares for vizual-
formed on 0.45 µM (450 nM) Millipore filters to remove           ization of the signals on the computer’s screen (Fig. 1).
debris, and subsequently on 0.1 µM (100 nM) Milli-                 The main harmonics of the complex signals were an-
pore filters or on 0.02 µM (20 nM) Whatman filters,               alyzed by utilizing several softwares of Fourier transfor-
to remove mycoplasma cells. Indeed, the two 100 nM              mation.
and 20 nM filtrates were confirmed sterile when aliquots             In each experiment, the internal noise generated by
were incubated for several weeks in SP4 medium. Re-             the different pieces of the reading system was first
peated search for traces of mycoplasma DNA by PCR               recorded (coil alone, coil with a tube filled with water).
and nested PCR using specific primers for the adhesin            Fourier analysis shows (Fig. 2(c, d)) that the noise was
gene or for the 16S ribosomal gene was consistently neg-        predominantly composed of very low frequencies, prob-
ative.                                                          ably generated at least in part by the 50/60 Hz ambi-
   However when the filtrates were incubated for two             ent electric current. The use of the 12 V battery for the
weeks (100 nM filtrate) or three weeks (20 nM filtrate)           computer power supply did reduce, but not abolish this
with a culture of human activated T lymphocytes, the            noise, which was found to be necessary for the induc-
mycoplasma was recovered in the medium with all its             tion of the resonance signals from the specific nanos-
original characteristics as previously observed.                tructures.
   The same filtrates were analyzed just after filtra-               When dilutions of the M. pirum filtrate were recorded
tion for production of electromagnetic waves of low fre-        for wave emission, the first obvious phenomenon ob-
quency. For this purpose we used a device previously            served was an increase of the overall amplitude of
designed by Benveniste and Coll (1996; 2003) for the            the signals at certain dilutions over the background
detection of signals produced by isolated molecules en-         noise (Fig. 2(a)) and also an increase in frequencies
dowed with biological activity. The principle of this           (Fig. 2(b)). This change was abolished if the tube to be
technology is shown in Fig. 1.                                  analyzed was placed inside a box sheltered with sheets
                                                                of copper and mumetal (David, 1998).
                                                                   Fourier analysis of the M. pirum signals showed a
                                                                shift towards higher frequencies close to 1000 Hz and
                                                                multiples of it. Profiles were identical for all the dilu-
                                                                tions showing an increase in amplitude (Fig. 2(c) and
                                                                2(d)).
                                                                   The first low dilutions were usually negative, showing
                                                                the background noise only. Positive signals were usu-
                                                                ally obtained at dilutions ranging from 10−5 to 10−8 or
                                                                10−12 . Higher dilutions were again negative (Fig. 3).
                                                                   The positive dilutions varied according to the type of
                                                                filtration, the 20 nM filtrate being generally positive at
Fig. 1   Device for the capture and analysis of electromag-     dilutions higher than those of the 100 nM filtrate.
         netic signals (EMS): (1) Coil: a bobbin of copper         The original unfiltered suspension was negative at all
         wire, impedance 300 Ohms; (2) Plastic stoppered        dilutions, a phenomenon observed for all the microor-
         tube containing 1 mL of the solution to be analyzed;   ganisms studied.
         (3) Amplifier; (4) Computer with softwares.             Size and density of the structures producing the
                                                                signals in the aqueous dilutions:
  Briefly, the 100 nM or 20 nM filtrates are serially di-            An aliquot of the 20 nM filtrate was layered on the
luted 1 in 10 (0,1 +0,9 in sterile water (medical grade).       top of a 5-20% (w/v) sucrose gradient in water and
The first 2 dilutions (1/10 and 1/100) are done in               centrifuged for 2 hours at 35,000 rpm in a swinging
serum-free RPMI medium, in order to avoid eventual              bucket rotor. These conditions had previously been
protein precipitation in deionized water.                       used to obtain the density equilibrium of the intact my-
  Each dilution is done in 1.5 mL Eppendorf plastic             coplasma cells wich formed a sharp bound at 1,21 den-
tubes, which are then tightly stoppered and strongly            sity. Fractions were collected from the bottom of the
agitated on a Vortex apparatus for 15 seconds . This            tubes, pooled 2 by 2 and assayed for signal emission.
step has been found critical for the generation of signals.        Fig. 4 shows that the signal emitting structures were
  After all dilutions have been made (generally 15-20           distributed in a large range of densities from 1.15 to
decimal dilutions), the stoppered tubes are read one by         1.25 and also had a high sedimentation coefficient.
one on an electromagnetic coil, connected to a Sound
Interdiscip Sci Comput Life Sci (2009) 1: 81–90                                                                               83




Fig. 2   Detection of EMS from a suspension of Mycoplasma pirum: Left: background noise (from an unfiltered suspension or
         a negative low dilution). Right: positive signal (from a high dilution D-7 (10-7)). (a) actual recording (2 seconds from
         a 6 second recording) after WaveLab (Steinberg) treatment; (b) detailed analysis of the signal (scale in millisecondes);
         (c) Matlab 3D Fourier transform analyzis (abcissa: 0-20 kHz, ordinate: relative intensity, 3D dimension: recording at
         different times); Frequencies are visualized in different colors; (d) Sigview Fourier transform: note the new harmonics
         in the range of 1 000-3 000 Hz.




Fig. 3   A typical recording of signals from aqueous dilutions of M. Pirum (Matlab software): note the positive signals from
         D-7 to D-12 dilutions.
84                                                                                                   Interdiscip Sci Comput Life Sci (2009) 1: 81–90



                                                                                           10,000 rpm for 15 minutes, the supernatant was fil-
     1.30
                                                                                           tered on 450 nM filter and the resulting filtrate was




                                                                          E.M.S. naneons
                                                                                           filtered again on a 100 nM filter. The final filtrate was
                             1.25                  SN CEM/M.pirum                          found sterile, when plated on nutrient agar medium and
                                                   Filtered 0.02μ                          was analyzed for electromagnetic wave emission, as de-
                                                                                           scribed above for M. pirum. Signal producing dilutions
                                                                                           usually range from 10−8 to 10−12 , with profiles upon
                                        1.20
                                                                                           Fourier transformation, similar to those of M. Pirum
     1.20
                                                                                           (Fig. 5). In one experiment, some very high dilutions
                                                                                           were found positive, ranging from 10−9 to 10−18 . An
                                                                                           aliquot of the unfiltered supernatant did not show any
                                                     1.15
                                                                                           signals above background up to the 10−38 dilution, in-
                                                                              107          dicating again the critical importance of the filtration
                                                                              106          step for the generation of specific signals.
                                                                                              The only difference with M. pirum was that no sig-
     1.10                                                                     105
                                                                                           nal appeared after filtration on 20 nM filters, suggesting
                                                                              104          that the structures associated with the signals were re-
                                                                                           tained by these filters and, therefore, had a size greater
                                                                                           than 20 nM and lower than 100 nM.
                                                                                              We then asked why the lower dilutions, which
      Density




                                                                                           logically should contain a larger number of signal-
                                                                                           producing structures, were “silent”. When we added
                 1   2   3     4    5    6     7    8   9   10 11 12 13                    0.1 mL of a negative low dilution (e.g. 10−3 ) to 0.4
                                                                                           mL or 0.9 mL of a positive dilution (10−8 ), the latter
Fig. 4          Sucrose density centrifugation (35 000 rpm, 2 Hr)
                                                                                           became negative. This indicate that the ”silent” low
                of a 0.02µ filtrate of Mycoplasma pirum suspension.
                The collected fractions were pooled 2 by 2 and di-                         dilutions are self-inhibitory, probably by interference of
                luted up to D-15 and tested for EMS. The bars in-                          the multiple sources emitting in the same wave length
                dicate the fractions positive for EMS.                                     or slightly out of phase, like a radio jamming. Alterna-
                                                                                           tively, the abundance of nanostructures can form a gel
                                                                                           in water and therefore are prevented to vibrate.
  We then turned to a more classical bacterium, E.                                         -Evidence for homologous “cross talk” between
Coli, using the laboratory strain K1.                                                      dilutions
  A culture of E. Coli in agitated (oxygenated) con-                                          We then wonder whether or not it was possible to
ditions, yielded 109 bacterial units/mL, measured by                                       generate new signal-emitting structures from tube to
spectrometry. The suspension was then centrifuged at                                       tube by using wave transfer. The following experiment
Interdiscip Sci Comput Life Sci (2009) 1: 81–90                                                                    85




Fig. 5   EMS from E. Coli 0.1 µ filtrate. EMS positive from dilution D-8 to D-11: (a) Actual recording; (b) millisecond
         analysis; (c) Fourier transform analysis Matlab; (d) Fourier transform analysis SigView. NF: not filtered.
86                                                                       Interdiscip Sci Comput Life Sci (2009) 1: 81–90

which was repeated several times showed that indeed               Finally, two others problems were investigated in the
this was the case.                                             E. Coli system: the first was the role of the initial
  A donor tube of a low “silent” dilution of E. Coli           number of bacterial cells in the induction of the fil-
(10−3 ) was placed side by side close to a receiver tube of    terable signal-producing structures. For this a station-
the positive “loud” highest dilution of the same prepa-        ary culture of E. Coli was counted and adjusted to 109
ration (10−9 ). Both tubes were placed in a mumetal            cells/mL and serial dilutions from 100 to 100 were done
box for 24 hours at room temperature, so that the tubes        down to 1 cell/mL. Each dilution was filtered at 100
were not exposed to external electromagnetic noise, and        nM and then analyzed for signal emission. Surprisingly,
only exposed to the signals generated by the structures        the range of positive dilutions were not strictly depen-
present in the tubes themselves.                               dent on the initial concentration of E. Coli cells, being
  The tubes were then read again by the signal detect-         roughly the same from 109 cells down to 10 cells, sug-
ing device: the donor tube was still silent, however the       gesting that the same final number of nanostructures
receiver tube became also silent.                              was reached at all concentrations. Thus, paradoxically,
  Moreover, when further dilutions were made from the          10 cells are giving the same signals than 109 cells.
receiver tube (10−10 , 10−11 , 10−12 ), these dilutions had       We were also concerned by the possible personal in-
became positive (Fig. 6). These results suggest that the       fluence of the operator in the reading.
receiver tube was made silent by formation of an excess           To address this point, two healthy operators were
of new nanostructures, which could emit signals upon           asked to measure independantly the same dilutions of
further dilution.                                              E. Coli, each one unknowing the results of the other.
                                                               The results of their readings were identical.
                                                                  In addition, the results were independant of the order
                                                               in which the samples were read, whether in descending
                                                               dilutions from to the lowest to the highest or in ascend-
                                                               ing the dilutions from the highest to the lowest.
                                                                  Finally an other laboratory worker placed the diluted
                                                               samples in a random order, the labels being unknown
                                                               from the person reading the samples. The same range
                                                               of positive dilutions was detected again provided each
                                                               tube was well separated from the other, to avoid their
                                                               “cross talk”.
                                                                  We also found that the results were also indepen-
                                                               dant of the location of the reading site: starting from
Fig. 6   Cross-talk between dilutions (from an E. Coli 0.1 µ   the same unfiltered preparation of E. Coli, positive di-
         filtrate), see explanation in the text.                lutions of the filtrates were found to be the same in
                                                               two different locations in France (Paris center and sub-
   This effect was suppressed by interposing a sheat of         urb), one in Canada (Montreal), and one in Cameroun
mumetal between the two tubes during the 24 hours              (Yaound´).
                                                                        e
contact period, pointing to a role of low frequency waves         As shown in the figures, the background noise was
in the phenomenon.                                             variable, according to the location and time of record-
   Emission of similar electromagnetic signals was             ing. It was generally higher in large cities than in iso-
also observed with some other bacterial species such           lated aeras. However, positive signals always clearly
as: Streptococcus B, Staphylococcus aureus, Pseu-              differenciate over the background by higher frequency
domonas aeroginosa, Proteus mirabilis, Bacillus sub-           peaks.
tilis, Salmonella, Clostridium perfringens, all in the         Nature of the aqueous nanostructures:
same range of dilutions observed for E. Coli, and only            Treatments by RNAseA (Promega, 1 µg/ml, 37 ℃ 1
after filtration at 100 nM (and not at 20 nM).                  h), Dnase I (Invitrogen, 10 U/µg DNA, 37 ℃, 18 h),
   Importantly, the transfer effect between two tubes,          Lysozyme (Fisher, 1 mg/mL, 37 ℃ 10 min), Proteinase
one silent, one loud, was only observed if both contained      K (Promega, 0.12 mg/mL, in 1% sodium dodecyl sul-
dilutions of the same bacterial species. In other words,       phate, 56 ℃ 1 h) did not suppress the EMS produc-
a Staphylococcus donor tube could only “talk” with             ing activity of the “loud” dilutions nor did activate the
a receiver tube containing a Staphylococcus dilution,          “silent” dilutions.
and not with a tube of Streptococcus or E. Coli, and              However, heating at 70 ℃ for 30 min suppressed irre-
reciprocally.                                                  versiblly the activity, as well as did freezing for 1 hour
   These results indicate that the transfer effect is medi-     at −20 ℃ or −60 ℃. DMSO (10%), and formamide
ated by species-specific signals, the frequencies of which      (10%) had no effect.
remain to be analyzed.                                            Treatment with lithium cations, known to affect the
Interdiscip Sci Comput Life Sci (2009) 1: 81–90                                                                           87

hydrogen bonding of water molecules, was able to re-             let obtained by ethanol precipitation was resuspended
duce the intensity of the signals, while the range of the        in Tris 10-2 M, pH 7,6 and an aliquot was diluted 1/100
positive dilutions remained unchanged.                           in water. The dilution (10−2 ) was filtered first through
Nature of the bacterial molecules at the origin                  a 450 nM filter and the resulting filtrate was then fil-
of the nanostructures:                                           tered again on a 100 nM filter. The filtrate was further
                                                                 diluted in serial decimal dilutions in water as previously
   In preliminary experiments, we had observed that
                                                                 described.
a pretreatment of a suspension of E. Coli by 1%
                                                                    As for the intact microorganisms, the filtration step
formaldehyde did not alter its capacity to induce the
                                                                 was found to be essential for detection of the EMS in
electromagnetic signals, while killing the bacteria. This
                                                                 the DNA dilutions. In its absence, no signals could be
treatment alters the surface proteins of the bacte-
                                                                 detected at any dilutions.
rial cells without attacking their genetic material, i.e.
                                                                    In contrast to the microorganism suspension, where
double- helical DNA. This suggested that the source of
                                                                 the filtration was supposed to retain the intact cells,
the signals may be the DNA itself.
                                                                 the filtration at 100 nM did not retain the DNA, which
   Indeed, DNA extracted from the bacterial suspension           was still present in the filtrate, as measured by optical
by the classical phenol: chloroform technique was able           density. However, filtration with a 20 nM Whatman
upon filtration and appropriate dilutions in water to             filter retained the nanostructures emitting the EMS,
emit EMS similar to those produced by intact bacteria            suggesting that they have the same range of sizes than
under the same conditions. DNAse treatment of the                those originating from intact bacteria.
extracted DNA solution abolishes its capacity to emit               In the case of DNA, the role of the 100 nM filtration
signals, at the condition that the nanostructures pre-           is probably to dissociate the network of nanostructures
viously induced by the DNA are destroyed. A typical              organized in a gel-like liquid crystal at high concen-
experiment is described as follows:                              trations in water, allowing their dispersion in further
   E. Coli DNA was treated by Proteinase K in the                dilutions. As shown in Fig. 7, the dilutions positive for
presence of SDS (sodium dodecyl sulfate) and further             EMS were in the same range that those observed for
deproteinized by phenol-chloroform mixture. The pel-             the intact bacteria, generally between 10−7 to 10−13 .




Fig. 7   DNAse effect on EMS production. The DNAse treated E. Coli DNA solution and the untreated DNA are diluted
         from D-2 to D-15. Analyzis of the EMS as described in Fig. 5. D-2 dilution (negative for EMS) is shown as control.
         D-9 is positive for EMS (from a range of positive dilutions D-8 to D-11). Note the signal disappearance in the DNAse
         treated DNA.
88                                                                        Interdiscip Sci Comput Life Sci (2009) 1: 81–90

At the high dilution of 10−13 , calculations indicate that      pletely devoid of EMS emission at any dilution (Fig. 7).
there is no DNA molecule of MW larger than 105 in                  Treatment of the DNA solution by a restriction en-
the solution, making it unlikely that the EMS are pro-          zyme acting at many sites of E. Coli DNA (EcoRV) did
duced directely by the DNA itself, but rather by the            not suppress the production of EMS, suggesting that
self-sustained nanostructures induced by the DNA.               this emission is linked to rather short sequences or is
   Generally, all the bacterial species shown to be posi-       associated with rare sequences.
tive for EMS yielded also DNA preparations positive for         Nature of the DNA sequences at the origin of
EMS. Further demonstration that the EMS produced                the EMS:
by bacteria come from their DNA was shown by their                 A non exhaustive survey of the bacterial species and
disappearance after DNAse treatement.                           of their DNA able to display EMS suggests that most
   This inactivation was however only complete when             of bacteria pathogenic for humans are in this category.
the nanostructures induced in the DNA solution which               By contrast, probiotic “good” bacteria as Lactobacil-
are themselves resistant to DNAse were previously fully         lus and their DNA are negative for EMS emission.
destroyed.                                                         In the case of E. Coli, we found that some strains
   This destruction was obtained either by freezing the         used to carry plasmids for gene cloning were also nega-
DNA solution at −20 ℃ for 1 hour or heating it at               tive (Fig. 8).
90 ℃ for 30 minutes.                                               This suggested that only some sequences of DNA are
   After slow cooling to allow the heated DNA to re-            at the origin of the EMS.
anneal, DNAse 1 at a final concentration of 10 U/µg                 As pathogenicity is often associated with the capac-
of DNA was added and the mixture was incubated at               ity of the microorganism to bind eukaryotic cells, par-
37 ℃ for 18 hours in the presence of 5 mM of MgCl2 .            ticulary mucosal cells, we focussed our analysis again
An aliquot of the untreated DNA solution was kept as            to M. pirum DNA, where a single gene (adhesin: 126-
a positive control.                                             kDa protein) is responsible for the adhesion of the my-
   The DNAse-treated preparation was found com-                 coplasma to human cells.




Fig. 8   EMS produced by the 1.5 kb fragment of the adhesin DNA of M. Pirum. The plasmid DNA containing the 1.5 Kb
         fragment was used to transform an E. Coli vector, XL1blue. The whole DNA was extracted and diluted for EMS
         analysis. Left: control background noise of a negative dilution (D-2). Right: positive signal at D-10 (range from
         D-9 to D-12). Bottom: Note the lack of EMS produced by the DNA extracted from the strain transformed by the
         plasmid alone.
Interdiscip Sci Comput Life Sci (2009) 1: 81–90                                                                       89

   This gene had previously been cloned and sequenced       of EMS requires prior 20 nM filtration suggesting that
in our laboratory (Tham et al., 1994). The cloned DNA       the nanostructures produced are smaller that those pro-
existed as two fragments in two plasmids, corresponding     duced by bacterial DNA.
respectively to the N terminal (1.5 Kbp) and the C             In patients infected with HIV, EMS can be detected
terminal (5 Kbp) of the protein.                            mostly in patients treated by antiretroviral therapy and
   The two plasmids (pBluescript SK, Stratagene) con-       having a very low viral load in their plasma. Such
taining the DNA fragments were amplified in a E. Coli        nanostructures persisting in the plasma may contribute
strain, XL1blue.                                            to the viral reservoir which escapes the antiviral treat-
   The DNA of the E. Coli strain (with or without the       ment, assuming that they carry genetic information of
plasmid) alone did not yield EMS at any dilutions.          the virus.
   By contrast when the strain was transformed with ei-        The physical nature of the nanostructures which sup-
ther plasmids carrying an adhesin gene fragment, EMS        port the EMS resonance remains to be determined.
were produced (Fig. 8).                                        It is known from the very early X-ray diffraction stud-
   The two adhesin DNA fragments were then cut by           ies of DNA, that water molecules are tightly associated
specific restriction enzymes (N Terminal: 1.5 kbp/SpeI-      with the double helix, and any beginner in molecular
EcoRI) (C Terminal: 5 kbp/HindIII-XbaI) and isolated        biology knows that DNA in water solution forms gels
by electrophoresis in 0.8% agarose gel. Each DNA frag-      associating a larger number of water molecules.
ment was able to induce EMS (not shown).                       Moreover, a number of physical studies have reported
   We also purified a large fraction of the adhesin DNA      that water molecules can form long polymers of dipoles
from the whole mycoplasma genomic DNA using spe-            associated by hydrogen bonds (Ruan et al., 2004; Wer-
cific primers and amplification by PCR.                       net et al., 2004).
   Again this fragment induced EMS, thus indicating
                                                               However these associations appear to be very short-
that no contaminant DNA coming from the plasmid
                                                            lived (Cowan et al., 2005). Could they live longer, being
carried by E. Coli was involved (not shown).
                                                            self-maintained by the electromagnetic radiations they
                                                            are emitting as previously postulated by Del Guidice,
Discussion
                                                            Preparata and Vitielo (1988)?
   We have discovered a novel property of DNA, that            We have studied the decay with time of the capac-
is the capacity of some sequences to emit electromag-       ity of dilutions for emitting EMS, after they have been
netic waves in resonance after excitation by the ambient    removed (in mumetal boxes) from exposure to the exci-
electromagnetic background.                                 tation by the background. This capacity lasts at least
   Owing to the low sensitivity and specificity of our       several hours, some time up to 48 hours, indicating the
signal capture and analysis, the frequencies emitted are    relative stability of the nanostructures.
all alike, regardless of the bacterial species involved.       Are the latter sufficiently specific of DNA sequences
   However, the experiments of transfer of information      to be able to carry some genetic information?
through plastic tubes suggest that, by refining the             If so, what could be their role in pathogenicity, par-
analysis and eliminating the variability of the excit-      ticularly in the genesis of chronic diseases?
ing signals, we might detect specific differences between        Further studies involving close collaboration between
species, and even between sequences. Indeed, this prop-     physicists and biologists are obviously needed to resolve
erty may be a general one shared by all double-helical      these problems.
DNAs, including human DNA.
   But in our conditions of detection, it seems to be
associated with only certain bacterial sequences.              Acknowledgments We thank Dr A. Blanchard for
   It remains to be seen whether they are restricted to     gift of Mycoplasma pirum DNA and Drs D. Guillonnet,
some genes involved in diseases.                            R. Olivier, L. Thibodeau and J. Varon for helpful dis-
   Experiments to be reported elsewhere indeed indi-        cussion.
cate that this detection applies also at the scale of the
human body: we have detected the same EMS in the            References
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patients suffering of Alzheimer, Parkinson disease, mul-      [1] Benveniste, J., Jurgens, P., A¨  ıssa, J. 1996. Digital
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Montagnier Electromadnetic Signals

  • 1. Interdiscip Sci Comput Life Sci (2009) 1: 81–90 DOI: 10.1007/s12539-009-0036-7 Electromagnetic Signals Are Produced by Aqueous Nanostructures Derived from Bacterial DNA Sequences Luc MONTAGNIER1,2∗ , Jamal A¨ ISSA1 , St´phane FERRIS1 , e Jean-Luc MONTAGNIER , Claude LAVALLEE1 1 ´ 1 (Nanectis Biotechnologies, S.A. 98 rue Albert Calmette, F78350 Jouy en Josas, France) 2 (Vironix LLC, L. Montagnier 40 Central Park South, New York, NY 10019, USA) Recevied 3 January 2009 / Revised 5 January 2009 / Accepted 6 January 2009 Abstract: A novel property of DNA is described: the capacity of some bacterial DNA sequences to induce electromagnetic waves at high aqueous dilutions. It appears to be a resonance phenomenon triggered by the ambient electromagnetic background of very low frequency waves. The genomic DNA of most pathogenic bacteria contains sequences which are able to generate such signals. This opens the way to the development of highly sensitive detection system for chronic bacterial infections in human and animal diseases. Key words: DNA, electromagnetic signals, bacteria. Pathogenic microorganisms in this day of age are not priate dilutions in water. The emission of such waves is only submitted to high selective pressure by the im- likely to represent a resonance phenomenon depending mune defenses of their hosts but also have to survive un- on excitation by the ambient electromagnetic noise. It der highly active antiviral or antibiotic treatments. Not is associated with the presence in the aqueous dilutions surprisingly, they have evolved in finding many ways to of polymeric nanostructures of defined size. The super- escape these hostile conditions, such as mutations of re- natant of uninfected eukaryotic cells used as controls sistance, hypervariability of surface antigens, protective did not exhibit this property. biofilms, latency inside cells and tissues. In this paper we provide a first characterization of the We initially observed (Montagnier and Lavallee, per- electromagnetic signals (EMS) and of their underlying sonal communication) that some filtration procedures nanostructures produced by some purified bacteria. aimed at sterilizing biological fluids can yield under In addition to M. pirum, a more classical bacterium, some defined conditions the infectious microorganism E. Col i, was utilized for the purpose of the analysis. which was present before the filtration step. Thus, fil- The nanostructures produced by HIV will be the sub- tration of a culture supernatant of human lymphocytes ject of another paper. infected with Mycoplasma pirum, a microorganism of M. pirum is a peer-shaped small bacterial cell, about 300 nM in size, through filters of 100 nM or ressembling M. pneumoniae, which can be grown in syn- 20 nM porosities, yielded apparently sterile fluid. The thetic enriched medium (SP4) (Tully et al., 1977) but latter however was able to regenerate the original my- also mutiplies at the surface of human T lymphocytes. coplasma when incubated with a mycoplasma negative The strain (Ber) used in our experiments was isolated culture of human lymphocytes within 2 to 3 weeks. from a T lymphocyte culture derived from the blood Similarly, a 20 nM filtration did not retain a minor in- of an apparently healthy subject (Grau et al., 1993). fective fraction of HIV, the causal agent of AIDS, whose The strong mycoplasma adherence to lymphocytes is viral particles have a diameter averaging 100-120 nM. mediated by a specific adhesin, whose gene had been In the course of investigating the nature of such filter- previously cloned and sequenced by the authors (Tham ing infectious forms, we found another property of the et al., 1994). filtrates, which may or may not be related to the former: We used as primary source of the mycoplasma, super- their capacity to produce some electromagnetic waves natants of infected human T lymphocyte cultures or of of low frequency in a reproducible manner after appro- cultures of the CEM tumor T cell line. All cell cultures were first tested for the lack of M. pirum contamination *Corresponding author. by polymerase chain reaction (PCR) and nested PCR, E-mail: nadiacpt@yahoo.fr before starting the experiments. Titers of 106 -107 infec-
  • 2. 82 Interdiscip Sci Comput Life Sci (2009) 1: 81–90 tious Units/ml of M. pirum were readily achieved after Blaster Card itself connected to a laptop computer, 5-6 days of incubation following deliberate infection of preferentially powered by its 12 volt battery. Each both types of cultures. emission is recorded twice for 6 seconds, amplified 500 Filtration of the clarified supernatant was first per- times and processed with different softwares for vizual- formed on 0.45 µM (450 nM) Millipore filters to remove ization of the signals on the computer’s screen (Fig. 1). debris, and subsequently on 0.1 µM (100 nM) Milli- The main harmonics of the complex signals were an- pore filters or on 0.02 µM (20 nM) Whatman filters, alyzed by utilizing several softwares of Fourier transfor- to remove mycoplasma cells. Indeed, the two 100 nM mation. and 20 nM filtrates were confirmed sterile when aliquots In each experiment, the internal noise generated by were incubated for several weeks in SP4 medium. Re- the different pieces of the reading system was first peated search for traces of mycoplasma DNA by PCR recorded (coil alone, coil with a tube filled with water). and nested PCR using specific primers for the adhesin Fourier analysis shows (Fig. 2(c, d)) that the noise was gene or for the 16S ribosomal gene was consistently neg- predominantly composed of very low frequencies, prob- ative. ably generated at least in part by the 50/60 Hz ambi- However when the filtrates were incubated for two ent electric current. The use of the 12 V battery for the weeks (100 nM filtrate) or three weeks (20 nM filtrate) computer power supply did reduce, but not abolish this with a culture of human activated T lymphocytes, the noise, which was found to be necessary for the induc- mycoplasma was recovered in the medium with all its tion of the resonance signals from the specific nanos- original characteristics as previously observed. tructures. The same filtrates were analyzed just after filtra- When dilutions of the M. pirum filtrate were recorded tion for production of electromagnetic waves of low fre- for wave emission, the first obvious phenomenon ob- quency. For this purpose we used a device previously served was an increase of the overall amplitude of designed by Benveniste and Coll (1996; 2003) for the the signals at certain dilutions over the background detection of signals produced by isolated molecules en- noise (Fig. 2(a)) and also an increase in frequencies dowed with biological activity. The principle of this (Fig. 2(b)). This change was abolished if the tube to be technology is shown in Fig. 1. analyzed was placed inside a box sheltered with sheets of copper and mumetal (David, 1998). Fourier analysis of the M. pirum signals showed a shift towards higher frequencies close to 1000 Hz and multiples of it. Profiles were identical for all the dilu- tions showing an increase in amplitude (Fig. 2(c) and 2(d)). The first low dilutions were usually negative, showing the background noise only. Positive signals were usu- ally obtained at dilutions ranging from 10−5 to 10−8 or 10−12 . Higher dilutions were again negative (Fig. 3). The positive dilutions varied according to the type of filtration, the 20 nM filtrate being generally positive at Fig. 1 Device for the capture and analysis of electromag- dilutions higher than those of the 100 nM filtrate. netic signals (EMS): (1) Coil: a bobbin of copper The original unfiltered suspension was negative at all wire, impedance 300 Ohms; (2) Plastic stoppered dilutions, a phenomenon observed for all the microor- tube containing 1 mL of the solution to be analyzed; ganisms studied. (3) Amplifier; (4) Computer with softwares. Size and density of the structures producing the signals in the aqueous dilutions: Briefly, the 100 nM or 20 nM filtrates are serially di- An aliquot of the 20 nM filtrate was layered on the luted 1 in 10 (0,1 +0,9 in sterile water (medical grade). top of a 5-20% (w/v) sucrose gradient in water and The first 2 dilutions (1/10 and 1/100) are done in centrifuged for 2 hours at 35,000 rpm in a swinging serum-free RPMI medium, in order to avoid eventual bucket rotor. These conditions had previously been protein precipitation in deionized water. used to obtain the density equilibrium of the intact my- Each dilution is done in 1.5 mL Eppendorf plastic coplasma cells wich formed a sharp bound at 1,21 den- tubes, which are then tightly stoppered and strongly sity. Fractions were collected from the bottom of the agitated on a Vortex apparatus for 15 seconds . This tubes, pooled 2 by 2 and assayed for signal emission. step has been found critical for the generation of signals. Fig. 4 shows that the signal emitting structures were After all dilutions have been made (generally 15-20 distributed in a large range of densities from 1.15 to decimal dilutions), the stoppered tubes are read one by 1.25 and also had a high sedimentation coefficient. one on an electromagnetic coil, connected to a Sound
  • 3. Interdiscip Sci Comput Life Sci (2009) 1: 81–90 83 Fig. 2 Detection of EMS from a suspension of Mycoplasma pirum: Left: background noise (from an unfiltered suspension or a negative low dilution). Right: positive signal (from a high dilution D-7 (10-7)). (a) actual recording (2 seconds from a 6 second recording) after WaveLab (Steinberg) treatment; (b) detailed analysis of the signal (scale in millisecondes); (c) Matlab 3D Fourier transform analyzis (abcissa: 0-20 kHz, ordinate: relative intensity, 3D dimension: recording at different times); Frequencies are visualized in different colors; (d) Sigview Fourier transform: note the new harmonics in the range of 1 000-3 000 Hz. Fig. 3 A typical recording of signals from aqueous dilutions of M. Pirum (Matlab software): note the positive signals from D-7 to D-12 dilutions.
  • 4. 84 Interdiscip Sci Comput Life Sci (2009) 1: 81–90 10,000 rpm for 15 minutes, the supernatant was fil- 1.30 tered on 450 nM filter and the resulting filtrate was E.M.S. naneons filtered again on a 100 nM filter. The final filtrate was 1.25 SN CEM/M.pirum found sterile, when plated on nutrient agar medium and Filtered 0.02μ was analyzed for electromagnetic wave emission, as de- scribed above for M. pirum. Signal producing dilutions usually range from 10−8 to 10−12 , with profiles upon 1.20 Fourier transformation, similar to those of M. Pirum 1.20 (Fig. 5). In one experiment, some very high dilutions were found positive, ranging from 10−9 to 10−18 . An aliquot of the unfiltered supernatant did not show any 1.15 signals above background up to the 10−38 dilution, in- 107 dicating again the critical importance of the filtration 106 step for the generation of specific signals. The only difference with M. pirum was that no sig- 1.10 105 nal appeared after filtration on 20 nM filters, suggesting 104 that the structures associated with the signals were re- tained by these filters and, therefore, had a size greater than 20 nM and lower than 100 nM. We then asked why the lower dilutions, which Density logically should contain a larger number of signal- producing structures, were “silent”. When we added 1 2 3 4 5 6 7 8 9 10 11 12 13 0.1 mL of a negative low dilution (e.g. 10−3 ) to 0.4 mL or 0.9 mL of a positive dilution (10−8 ), the latter Fig. 4 Sucrose density centrifugation (35 000 rpm, 2 Hr) became negative. This indicate that the ”silent” low of a 0.02µ filtrate of Mycoplasma pirum suspension. The collected fractions were pooled 2 by 2 and di- dilutions are self-inhibitory, probably by interference of luted up to D-15 and tested for EMS. The bars in- the multiple sources emitting in the same wave length dicate the fractions positive for EMS. or slightly out of phase, like a radio jamming. Alterna- tively, the abundance of nanostructures can form a gel in water and therefore are prevented to vibrate. We then turned to a more classical bacterium, E. -Evidence for homologous “cross talk” between Coli, using the laboratory strain K1. dilutions A culture of E. Coli in agitated (oxygenated) con- We then wonder whether or not it was possible to ditions, yielded 109 bacterial units/mL, measured by generate new signal-emitting structures from tube to spectrometry. The suspension was then centrifuged at tube by using wave transfer. The following experiment
  • 5. Interdiscip Sci Comput Life Sci (2009) 1: 81–90 85 Fig. 5 EMS from E. Coli 0.1 µ filtrate. EMS positive from dilution D-8 to D-11: (a) Actual recording; (b) millisecond analysis; (c) Fourier transform analysis Matlab; (d) Fourier transform analysis SigView. NF: not filtered.
  • 6. 86 Interdiscip Sci Comput Life Sci (2009) 1: 81–90 which was repeated several times showed that indeed Finally, two others problems were investigated in the this was the case. E. Coli system: the first was the role of the initial A donor tube of a low “silent” dilution of E. Coli number of bacterial cells in the induction of the fil- (10−3 ) was placed side by side close to a receiver tube of terable signal-producing structures. For this a station- the positive “loud” highest dilution of the same prepa- ary culture of E. Coli was counted and adjusted to 109 ration (10−9 ). Both tubes were placed in a mumetal cells/mL and serial dilutions from 100 to 100 were done box for 24 hours at room temperature, so that the tubes down to 1 cell/mL. Each dilution was filtered at 100 were not exposed to external electromagnetic noise, and nM and then analyzed for signal emission. Surprisingly, only exposed to the signals generated by the structures the range of positive dilutions were not strictly depen- present in the tubes themselves. dent on the initial concentration of E. Coli cells, being The tubes were then read again by the signal detect- roughly the same from 109 cells down to 10 cells, sug- ing device: the donor tube was still silent, however the gesting that the same final number of nanostructures receiver tube became also silent. was reached at all concentrations. Thus, paradoxically, Moreover, when further dilutions were made from the 10 cells are giving the same signals than 109 cells. receiver tube (10−10 , 10−11 , 10−12 ), these dilutions had We were also concerned by the possible personal in- became positive (Fig. 6). These results suggest that the fluence of the operator in the reading. receiver tube was made silent by formation of an excess To address this point, two healthy operators were of new nanostructures, which could emit signals upon asked to measure independantly the same dilutions of further dilution. E. Coli, each one unknowing the results of the other. The results of their readings were identical. In addition, the results were independant of the order in which the samples were read, whether in descending dilutions from to the lowest to the highest or in ascend- ing the dilutions from the highest to the lowest. Finally an other laboratory worker placed the diluted samples in a random order, the labels being unknown from the person reading the samples. The same range of positive dilutions was detected again provided each tube was well separated from the other, to avoid their “cross talk”. We also found that the results were also indepen- dant of the location of the reading site: starting from Fig. 6 Cross-talk between dilutions (from an E. Coli 0.1 µ the same unfiltered preparation of E. Coli, positive di- filtrate), see explanation in the text. lutions of the filtrates were found to be the same in two different locations in France (Paris center and sub- This effect was suppressed by interposing a sheat of urb), one in Canada (Montreal), and one in Cameroun mumetal between the two tubes during the 24 hours (Yaound´). e contact period, pointing to a role of low frequency waves As shown in the figures, the background noise was in the phenomenon. variable, according to the location and time of record- Emission of similar electromagnetic signals was ing. It was generally higher in large cities than in iso- also observed with some other bacterial species such lated aeras. However, positive signals always clearly as: Streptococcus B, Staphylococcus aureus, Pseu- differenciate over the background by higher frequency domonas aeroginosa, Proteus mirabilis, Bacillus sub- peaks. tilis, Salmonella, Clostridium perfringens, all in the Nature of the aqueous nanostructures: same range of dilutions observed for E. Coli, and only Treatments by RNAseA (Promega, 1 µg/ml, 37 ℃ 1 after filtration at 100 nM (and not at 20 nM). h), Dnase I (Invitrogen, 10 U/µg DNA, 37 ℃, 18 h), Importantly, the transfer effect between two tubes, Lysozyme (Fisher, 1 mg/mL, 37 ℃ 10 min), Proteinase one silent, one loud, was only observed if both contained K (Promega, 0.12 mg/mL, in 1% sodium dodecyl sul- dilutions of the same bacterial species. In other words, phate, 56 ℃ 1 h) did not suppress the EMS produc- a Staphylococcus donor tube could only “talk” with ing activity of the “loud” dilutions nor did activate the a receiver tube containing a Staphylococcus dilution, “silent” dilutions. and not with a tube of Streptococcus or E. Coli, and However, heating at 70 ℃ for 30 min suppressed irre- reciprocally. versiblly the activity, as well as did freezing for 1 hour These results indicate that the transfer effect is medi- at −20 ℃ or −60 ℃. DMSO (10%), and formamide ated by species-specific signals, the frequencies of which (10%) had no effect. remain to be analyzed. Treatment with lithium cations, known to affect the
  • 7. Interdiscip Sci Comput Life Sci (2009) 1: 81–90 87 hydrogen bonding of water molecules, was able to re- let obtained by ethanol precipitation was resuspended duce the intensity of the signals, while the range of the in Tris 10-2 M, pH 7,6 and an aliquot was diluted 1/100 positive dilutions remained unchanged. in water. The dilution (10−2 ) was filtered first through Nature of the bacterial molecules at the origin a 450 nM filter and the resulting filtrate was then fil- of the nanostructures: tered again on a 100 nM filter. The filtrate was further diluted in serial decimal dilutions in water as previously In preliminary experiments, we had observed that described. a pretreatment of a suspension of E. Coli by 1% As for the intact microorganisms, the filtration step formaldehyde did not alter its capacity to induce the was found to be essential for detection of the EMS in electromagnetic signals, while killing the bacteria. This the DNA dilutions. In its absence, no signals could be treatment alters the surface proteins of the bacte- detected at any dilutions. rial cells without attacking their genetic material, i.e. In contrast to the microorganism suspension, where double- helical DNA. This suggested that the source of the filtration was supposed to retain the intact cells, the signals may be the DNA itself. the filtration at 100 nM did not retain the DNA, which Indeed, DNA extracted from the bacterial suspension was still present in the filtrate, as measured by optical by the classical phenol: chloroform technique was able density. However, filtration with a 20 nM Whatman upon filtration and appropriate dilutions in water to filter retained the nanostructures emitting the EMS, emit EMS similar to those produced by intact bacteria suggesting that they have the same range of sizes than under the same conditions. DNAse treatment of the those originating from intact bacteria. extracted DNA solution abolishes its capacity to emit In the case of DNA, the role of the 100 nM filtration signals, at the condition that the nanostructures pre- is probably to dissociate the network of nanostructures viously induced by the DNA are destroyed. A typical organized in a gel-like liquid crystal at high concen- experiment is described as follows: trations in water, allowing their dispersion in further E. Coli DNA was treated by Proteinase K in the dilutions. As shown in Fig. 7, the dilutions positive for presence of SDS (sodium dodecyl sulfate) and further EMS were in the same range that those observed for deproteinized by phenol-chloroform mixture. The pel- the intact bacteria, generally between 10−7 to 10−13 . Fig. 7 DNAse effect on EMS production. The DNAse treated E. Coli DNA solution and the untreated DNA are diluted from D-2 to D-15. Analyzis of the EMS as described in Fig. 5. D-2 dilution (negative for EMS) is shown as control. D-9 is positive for EMS (from a range of positive dilutions D-8 to D-11). Note the signal disappearance in the DNAse treated DNA.
  • 8. 88 Interdiscip Sci Comput Life Sci (2009) 1: 81–90 At the high dilution of 10−13 , calculations indicate that pletely devoid of EMS emission at any dilution (Fig. 7). there is no DNA molecule of MW larger than 105 in Treatment of the DNA solution by a restriction en- the solution, making it unlikely that the EMS are pro- zyme acting at many sites of E. Coli DNA (EcoRV) did duced directely by the DNA itself, but rather by the not suppress the production of EMS, suggesting that self-sustained nanostructures induced by the DNA. this emission is linked to rather short sequences or is Generally, all the bacterial species shown to be posi- associated with rare sequences. tive for EMS yielded also DNA preparations positive for Nature of the DNA sequences at the origin of EMS. Further demonstration that the EMS produced the EMS: by bacteria come from their DNA was shown by their A non exhaustive survey of the bacterial species and disappearance after DNAse treatement. of their DNA able to display EMS suggests that most This inactivation was however only complete when of bacteria pathogenic for humans are in this category. the nanostructures induced in the DNA solution which By contrast, probiotic “good” bacteria as Lactobacil- are themselves resistant to DNAse were previously fully lus and their DNA are negative for EMS emission. destroyed. In the case of E. Coli, we found that some strains This destruction was obtained either by freezing the used to carry plasmids for gene cloning were also nega- DNA solution at −20 ℃ for 1 hour or heating it at tive (Fig. 8). 90 ℃ for 30 minutes. This suggested that only some sequences of DNA are After slow cooling to allow the heated DNA to re- at the origin of the EMS. anneal, DNAse 1 at a final concentration of 10 U/µg As pathogenicity is often associated with the capac- of DNA was added and the mixture was incubated at ity of the microorganism to bind eukaryotic cells, par- 37 ℃ for 18 hours in the presence of 5 mM of MgCl2 . ticulary mucosal cells, we focussed our analysis again An aliquot of the untreated DNA solution was kept as to M. pirum DNA, where a single gene (adhesin: 126- a positive control. kDa protein) is responsible for the adhesion of the my- The DNAse-treated preparation was found com- coplasma to human cells. Fig. 8 EMS produced by the 1.5 kb fragment of the adhesin DNA of M. Pirum. The plasmid DNA containing the 1.5 Kb fragment was used to transform an E. Coli vector, XL1blue. The whole DNA was extracted and diluted for EMS analysis. Left: control background noise of a negative dilution (D-2). Right: positive signal at D-10 (range from D-9 to D-12). Bottom: Note the lack of EMS produced by the DNA extracted from the strain transformed by the plasmid alone.
  • 9. Interdiscip Sci Comput Life Sci (2009) 1: 81–90 89 This gene had previously been cloned and sequenced of EMS requires prior 20 nM filtration suggesting that in our laboratory (Tham et al., 1994). The cloned DNA the nanostructures produced are smaller that those pro- existed as two fragments in two plasmids, corresponding duced by bacterial DNA. respectively to the N terminal (1.5 Kbp) and the C In patients infected with HIV, EMS can be detected terminal (5 Kbp) of the protein. mostly in patients treated by antiretroviral therapy and The two plasmids (pBluescript SK, Stratagene) con- having a very low viral load in their plasma. Such taining the DNA fragments were amplified in a E. Coli nanostructures persisting in the plasma may contribute strain, XL1blue. to the viral reservoir which escapes the antiviral treat- The DNA of the E. Coli strain (with or without the ment, assuming that they carry genetic information of plasmid) alone did not yield EMS at any dilutions. the virus. By contrast when the strain was transformed with ei- The physical nature of the nanostructures which sup- ther plasmids carrying an adhesin gene fragment, EMS port the EMS resonance remains to be determined. were produced (Fig. 8). It is known from the very early X-ray diffraction stud- The two adhesin DNA fragments were then cut by ies of DNA, that water molecules are tightly associated specific restriction enzymes (N Terminal: 1.5 kbp/SpeI- with the double helix, and any beginner in molecular EcoRI) (C Terminal: 5 kbp/HindIII-XbaI) and isolated biology knows that DNA in water solution forms gels by electrophoresis in 0.8% agarose gel. Each DNA frag- associating a larger number of water molecules. ment was able to induce EMS (not shown). Moreover, a number of physical studies have reported We also purified a large fraction of the adhesin DNA that water molecules can form long polymers of dipoles from the whole mycoplasma genomic DNA using spe- associated by hydrogen bonds (Ruan et al., 2004; Wer- cific primers and amplification by PCR. net et al., 2004). Again this fragment induced EMS, thus indicating However these associations appear to be very short- that no contaminant DNA coming from the plasmid lived (Cowan et al., 2005). Could they live longer, being carried by E. Coli was involved (not shown). self-maintained by the electromagnetic radiations they are emitting as previously postulated by Del Guidice, Discussion Preparata and Vitielo (1988)? We have discovered a novel property of DNA, that We have studied the decay with time of the capac- is the capacity of some sequences to emit electromag- ity of dilutions for emitting EMS, after they have been netic waves in resonance after excitation by the ambient removed (in mumetal boxes) from exposure to the exci- electromagnetic background. tation by the background. This capacity lasts at least Owing to the low sensitivity and specificity of our several hours, some time up to 48 hours, indicating the signal capture and analysis, the frequencies emitted are relative stability of the nanostructures. all alike, regardless of the bacterial species involved. Are the latter sufficiently specific of DNA sequences However, the experiments of transfer of information to be able to carry some genetic information? through plastic tubes suggest that, by refining the If so, what could be their role in pathogenicity, par- analysis and eliminating the variability of the excit- ticularly in the genesis of chronic diseases? ing signals, we might detect specific differences between Further studies involving close collaboration between species, and even between sequences. Indeed, this prop- physicists and biologists are obviously needed to resolve erty may be a general one shared by all double-helical these problems. DNAs, including human DNA. But in our conditions of detection, it seems to be associated with only certain bacterial sequences. Acknowledgments We thank Dr A. Blanchard for It remains to be seen whether they are restricted to gift of Mycoplasma pirum DNA and Drs D. Guillonnet, some genes involved in diseases. R. Olivier, L. Thibodeau and J. Varon for helpful dis- Experiments to be reported elsewhere indeed indi- cussion. cate that this detection applies also at the scale of the human body: we have detected the same EMS in the References plasma and in the DNA extracted from the plasma of patients suffering of Alzheimer, Parkinson disease, mul- [1] Benveniste, J., Jurgens, P., A¨ ıssa, J. 1996. Digital tiple Sclerosis and Rheumatoid Arthritis. This would recording/ transmission of the cholinergic signal. Faseb suggest that bacterial infections are present in these Journal 10, A1479. diseases. [2] Benveniste, J., Guillonnet, D. 2003. Method, system Morever, EMS can be detected also from RNA and device for producing signals from a substance bi- viruses, such as HIV, influenza virus A, Hepatitis C ological and/or chemical activity. US Patent N◦ 6 541, Virus. In these cases, optimal filtration for detection 978 B1.
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