Patent Application
20130224788
This invention has for object to reveal latent infections in humans and animals, by showing inhibition, through the examinee, of electromagnetic signals generated by a microorganism.
From the works by Dr. Jacques BENVENISTE and from patent application WO 00/17637, it has been known how to record and digitalize, after analog-to-digital conversion using a computer sound board, an electrical signal characteristic of a molecule possessing a biological activity.
Also known in prior art (WO 09417406) is a process and a device used to transmit biological activity from a first matter, so-called carrier, to a second matter, so-called target, the latter exempted of any traces from said carrier and physically separate from it, and the target not presenting initially the aforementioned biological activity. The method consists in (i) exposing the matter carrying the biological activity of interest to an electrical or electromagnetic signal sensor, (ii) amplifying said electromagnetic or electrical signals characteristic of the emitted biological activity feature, then (iii) exposing the target matter to an emitter of electrical or electromagnetic signals, said emitter being connected to aforesaid sensor through a transmission and amplification circuit, in order to transmit the signal characteristic of biological activity to said target.
In a previous French patent application 05/12686 filed on Dec. 14, 2005, not yet issued to this day, the inventor of this invention was describing a process for characterizing biochemical elements presenting a biological activity, microorganisms in this case, by analyzing low frequency electromagnetic signals, said process bringing improvements to prior art techniques. Said process also relates to biological analysis consisting in recording the electromagnetic or electrical “signatures” corresponding to known biochemical elements, and to compare such pre-recorded “signatures” to that of a biochemical element to be characterized. Said process implicates filtration and dilution steps in order to eliminate microorganisms and cells present within the original sample, the highest dilutions generating the most electrical or electromagnetic signals whereas the least diluted samples don't provide, most of the time, any electrical or electromagnetic signals. The inventor also showed that microorganisms of different nature, such as bacteria and viruses, produce “nanostructures” that persist in aqueous solutions, and that these very “nanostructures” are emitting electromagnetic signals. Said “nanostructures” behaves like polymers of a size less than 0.02 .mu.m for viruses, and less than 0.1/.mu.m for classic size bacteria, and present a density ranging from 1.12 and 1.30 g/ml.
The process described in this application is based on the astonishing observation that in absence of physical contact, the mere vicinity of a closed tube containing a sample of a bacterial or viral filtrate, little diluted and negative with regard to electrical or electromagnetic emitting signals, inhibits the signals produced by a more diluted sample of the same filtrate, initially positive with regard to electrical or electromagnetic signal emission. In this application, such inhibition will be indistinctly called “inhibitory effect” or “negativing effect”. In the same way, in this application, to “inhibit” and “negativate” will be used indistinctly and have a similar meaning. This observation led the inventor to search for the same inhibitory phenomenon from an infected human being. It has been observed, in a patient suffering from an auto-immune microvascularitis of infectious origin, that the diluted samples of his plasma had an inhibitory effect on dilute filtrates of E. coli emitting electromagnetic signals (hereafter EMS), suggesting that the patient was suffering from a chronic infection by this or a related germ. It was also shown that the patient suffering from microvascularitis, as mentioned in the previous example, himself inhibits the EMS emitted by his filtered and diluted plasma, and also inhibits the EMS emitted by a filtered and diluted sample of E. coli culture present in a closed tube. In this case, a 5 minutes contact of a positive dilution in the patient's hand, or 10 minutes at a distance of up to 50 cm, are sufficient to observe said inhibitory effect.
Said inhibitory power thus involves both the emitting structures from one own plasma, and those of a specific bacterial germ, which could thus be used as a universal identification system.
The invention may therefore enable to determine a bacterial or viral origin in illnesses where such germs have not been identified.
A first object of the invention concerns a method for preparing reagents to be used in a test for detecting a microorganism and notably an infection in humans or animals. According to its most general acception, the method includes the following steps:
Analyzing the electrical signals detected using a solenoid and recording digitally aforesaid electrical signal, after analog/digital conversion of aforesaid signal; f) Selecting diluted samples from which the characteristic electrical signals were obtained in (e), by characteristic signals one means signals whose amplitude is at least 1.5 times greater than background noise emitted by water, and/or presenting a frequency displacement towards higher values; g) Introducing the diluted samples selected in step (0 in protective enclosures, which protect said dilutions from very low frequency external electromagnetic fields; h) Distributing one of aforesaid diluted samples from step (g), volume by volume, in two tubes, T1 and T2, with T1 remaining in a protective enclosure protecting said diluted samples from external electromagnetic field interferences, said tube T1 acting as a reference solution, while tube T2, also placed in a protective enclosure, is subsequently being subjected to the presence or contact of a sample suspected of containing said selected specific microorganism.
By “a sample to be tested for presence or absence of aforesaid selected specific microorganism” one means: (i) a human or animal individual suspected to be infected by aforesaid selected specific microorganism, or (ii) a biological specimen or a biological or artificial fluid suspected of containing said selected specific microorganism, or (iii) a food component, a cosmetic, or a pharmaceutical composition susceptible to contain said selected specific microorganism.
By biological fluids, one means any human or animal fluid, e.g. blood, urine, various secretions. By artificial fluid, one means any reconstituted fluid for growing microorganisms, e.g. various culture media for bacteria, yeasts, and molds, and culture media for cells infected by a virus.
Another object of the invention concerns a system for detecting a microorganism within a sample. This system includes:
During detection, tube T2 will be subjected to the presence or contact of sample X to be tested for presence or absence of a selected specific microorganism.
Another object of the invention concerns a method for detecting a microorganism within a sample, characterized in that said method consists of the following steps:
By “a sample X”, one means (i) a human individual or animal suspected of being infected by aforesaid selected specific microorganism, or (ii) a biological specimen, or a biological or artificial fluid, suspected to contain said selected specific microorganism, or (iii) a food component, cosmetic, or pharmaceutical composition susceptible to contain said selected specific microorganism.
The methods according to the invention enable (i) to prepare reagents intended for a test to detect microorganisms implicated in chronic illnesses, and/or intended to detect systemic latent infections under circumstances where a quick and non invasive response is required, as it is in the case of e.g. avian flu virus detection, (ii) the identification of an infection in humans or animals.
Once the responsible microorganism identified, it is possible to confirm the presence of that germ using supersensitive PCR with specific oligonucleotidic promoters from such microorganism.
The invention shall be better understood by reading the following description, presenting in a non restrictive way examples of process embodiment according to the invention.
The figures in annex correspond to non restrictive examples of embodiment.
A Lightly Dilute Bacterial Culture, not Emitting Electromagnetic Signals, “Negates” the Electromagnetic Signals Emitted by a Strong Dilution from the Same Culture
An Escherichia coli (E. coli) bacteria culture in LB (Luria broth) medium is centrifuged at 8000 rpm for 15 minutes in order to eliminate the cells. The bacterial supernatant is then filtered on a 0.45 .mu.m porosity PEVD Millipore filter, and the filtrate is then again filtered on a 0.1 .mu.m porosity Millipore filter.
From the resulting E. coli culture filtrate, which is completely sterile, one prepares a series of samples by diluting the filtrate from 10 to 10 into water down to 10.sup.-15 for injectable preparation. The successive dilutions are strongly agitated with a vortex for 15 seconds between each dilution.
The diluted samples are distributed in 1.5 ml Eppendorf conic plastic tubes. The fluid volume is in general of 1 milliliter.
Each dilute sample is tested for emission of low frequency electromagnetic signals.
The procedure for detecting EMS includes a step aimed at transforming the electromagnetic field from various diluted samples into one signal, namely an electrical signal, using a solenoid for capturing said electromagnetic field.
The transformation of the electromagnetic field coming from the diluted sample analyzed into an electrical signal is done as follows:
Signal detection is carried out using the equipment schematically represented in FIG. 1. The equipment consists of a solenoid reading cell (1) sensitive from 0 to 20000 hertz, placed on a table made of insulating material. Said solenoid used in step (ii) includes a winding comprising a soft iron core. Said winding has an impedance of 300 ohms, an inside diameter of 6 mm, an outside diameter of 16 mm, and a length of 6 mm. The magnetic soft iron core is placed in contact with the external walls of the tube containing the dilution to be analyzed.
The diluted samples to be read are distributed in 1.5 ml Eppendorf (trade mark) conic plastic tubes (2). The fluid volume is in general of 1 milliliter.
Characteristic electrical signal acquisition is performed for a preset duration, i.e. ranging from 1 to 60s. In this example, each sample is read twice successively for 6 seconds.
The electrical signals delivered by the solenoid are amplified and converted into analog-digital signals using a signal acquisition board (sound card) (4) including a computer-built-in analog-to-digital converter (3). Said analog-to-digital converter has twice the sampling rate of the maximal frequency that one wants to digitalize, e.g. 44 kHz.
The digital file corresponding to said converted electrical signal is saved on a mass storage, e.g. as a WAV format audio file.
For processing the characteristic electrical signal, one uses e.g. Matlabs and SigViews (trademarks) software. The recorded digital file may possibly undergo digital processing, i.e. digital amplification for calibrating the signal level, filtering for eliminating undesired frequencies, calculating spectral power distribution (SPD), then such spectral power is truncated, e.g. only keeping frequency bands from 140 Hz to 20 kHz (Matlab), or is transformed in frequency components by Fourier transform (SigView).
The diluted samples presenting characteristic electrical signals are samples diluted to 10.sup.-8, 10.sup.-9, 10.sup.-10. The 10.sup.-2 to l0.sup.-6 dilutions are negative (FIG. 2).
A closed tube containing a 10.sup.-3 dilution aliquot of E. coli is placed side by side with a closed tube containing a 10.sup.-8 diluted sample aliquot of E. coli, in an enclosure surrounded by a Mumetal.®. magnetic shield, and left 24 hours at room temperature. In parallel, a control series is realized. This control series consists of one tube containing a 10.sup.-3 diluted sample aliquot of E. coli, and of another containing a 10.sup.-8 diluted sample aliquot of E. coli that is processed in the same way, but in separate Mumetal.®. enclosures distant from one another. The placement in a Mumetal.®. enclosure eliminates very low active frequencies (5 to 100 Hertz) but not higher frequencies that could come from ambient electromagnetic noise.
After 24 hours, the tubes containing the diluted samples are again analyzed as describes above, revealing that the tube containing a 10.sup.-8 diluted sample aliquot and coupled to the tube containing a 10.sup.-3 diluted sample aliquot, no longer emits any electromagnetic signals, or much weaker ones. On the other hand, the control series tubes remained identical; the tube containing a 10.sup.-8 diluted sample aliquot protected from contact with the tube containing a 10.sup.-3 diluted sample aliquot remained positive for electromagnetic signal emission.
An important particularity of the invention is that the observed negating effect is specific, i.e. the lightly diluted, non-emitting sample and the greatly diluted electromagnetic signal-emitting sample must come from the same microorganism species.
Thus, the diluted E. coli-emitting samples are only “negated” by a weakly diluted non-emitting E. coli sample, but not by a lightly diluted non-emitting Streptococcus or Staphylococcus sample. Similarly, a diluted emitting Staphylococcus sample is only “negated” by a lightly diluted non-emitting sample of Staphylococcus and not by a lightly diluted non-emitting sample of Streptococcus or E. coli.
Quick and Non-Invasive Method for Detecting Infections in Humans and Animals
A blood sample, collected with anticoagulant, preferably heparin, from a patient suffering from a neurological pathology consecutive to a bacterial infection, and an Escherichia coli (E. coli) bacteria K1 culture in suspension in LB (Luria broth) medium are centrifuged in order to eliminate the cells. The bacterial supernatant and/or the plasma collected are then diluted to 10.sup.-2 in RPMI medium. The solutions are filtered on 0.45.mu. Millipore PEVD filter, then the filtrate is again filtered on 0.02 .mu.m Whatman or 0.1 .mu.m Millipore filter.
From the plasma filtrates of infected individual and from the E. coli K1 culture, one prepares a series of diluted samples corresponding to increasing dilution levels, up to 10.sup.-15, in 10 to 10 dilutions in water for injectable preparation under laminar flow hood. The successive dilutions are strongly agitated with a vortex for 15 seconds between each dilution.
The diluted samples are then distributed in 1.5 ml conic Eppendorf plastic tubes. The fluid volume is in general of 1 milliliter.
The selection of the diluted samples emitting characteristic signals, signals whose amplitude is at least 1.5 times greater than the background noise signals and/or are of a frequency higher than background noise, is realized identically to what is described above in example 1, chapter 2. The method described as well as the material are identical to what is described above. Thus, the method includes a step for transforming the electromagnetic field from different dilutions into a signal, namely an electrical signal, by means of a solenoid capturing said electromagnetic field.
The transformation of the electromagnetic field from the analyzed dilution into an electrical signal is done by: (i) Submitting the diluted sample being checked to an electrical, magnetic and/or electromagnetic exciting field; (ii) Analyzing the electrical signals detected using a solenoid, and digitally recording said electrical signal after analog/digital conversion of aforesaid signal; (iii) Selecting the diluted samples presenting characteristic electrical signals, by ‘characteristic’ one means signals whose amplitude is at least 1.5 times greater than background noise signals emitted by water, and/or presenting a frequency displacement towards higher values, and placing them in protective enclosures for protecting said diluted samples against external electromagnetic field interferences.
The diluted samples selected at the previous step (item (iii)), from the plasma filtrate of the infected individual, from E. coli culture filtrate, i.e. the dilutions of filtered sample presenting a characteristic electrical signal, are distributed in Eppendorfs plastic tubes, at a rate of 1 ml per tube, and stored at +4.degree. C. The diluted EMS emitting samples distributed in aliquots are protected from external influences by being placed in an enclosure protected from electromagnetic fields. Preferably, the enclosure is surrounded with a magnetic shield made of Mumetal.®., isolating the enclosure from very low frequency parasitic fields coming from the surroundings.
One of the diluted EMS emitting samples from the plasma filtrate of the infected individual, from E. coli culture filtrate, is distributed volume to volume in two tubes, T1 and T2, with T1 remaining in a protective enclosure protecting said diluted samples from external electromagnetic field interferences, that tube will act as reference solution; tube T2 will be subsequently subjected to the patient and is also placed in a protective enclosure.
Said protective enclosure being preferably surrounded with a Mumetal.®. shield.
The signals corresponding to tube T1 and those corresponding to tube T2, as well as the signals corresponding to water containing tube T3 (background noises) are compared.
The following figures represent the results obtained in the case where the active dilution comes from the examined infected individual plasma:
The analysis by 3 dimensions histogram, respectively for background noise (
Fourier analysis of the positive frequencies generated by tube 1 (
In conclusion, these analyses enable to deduct that the individual examined has a capacity for inhibiting electromagnetic signals emitted by a dilution of his/her own plasma.
Analogous results were obtained with the reference solution, derived from K1 E. coli.
Therefore, this inhibitory capacity concerns not only his/her own plasma but also E. coli emitting structures, suggesting that the individual is infected by an agent producing nanostructures close to those of E. coli.
| Number | Date | Country | Kind |
|---|---|---|---|
| 0605599 | Jun 2006 | FR | national |
This application is a Continuation of U.S. application Ser. No. 12/305,417, filed Nov. 30, 2009 that is a U.S. National Stage entry of International Application No. PCT/FR2007/001042, filed on Jun. 22, 2007, which claims the priority of French Application No. 0605599, filed on Jun. 22, 2006, in France. This application incorporates by reference the full disclosures of all of the above-mentioned applications.
| Number | Date | Country | |
|---|---|---|---|
| Parent | 12305417 | Nov 2009 | US |
| Child | 13785718 | US |
