AU679181B2 - Method for the epigenetic regulation of protein biosynthesisby scale resonance - Google Patents

Abstract

The present invention relates to a method for the epigenetic regulation of protein biosynthesis which consists in using, on the biosynthesis of proteins, scale resonance regulating action of sound transpositions of temporal sequences of quantum vibrations associated with their elongation; said action may be a stimulation or an inhibition of said biosynthesis depending on whether the modulation of the frequences of vibrations used is in phase with or of opposite phase to said elongation. The result is further stabilized by the action of coloured transpositions of groups of quantum vibrations arising from the spacial conformation of proteins from said elongation. The applications, particularly in the agri-foodstuff and health industries, include for implementation purposes a method of defining the metabolic role of proteins from their aminoacid sequences.

Description

OPI DATE 30/12/93 A0 t JP DATE 10/03/94 APPLN. ID 3304/93 IJlljj111 lIII111111111111li ii PCT NUMBER PCT/FR93/00524 I11111 Ili iiIII11111lii1111111111111111111111111111111 AU9343304 DEMIANIDh IN ItRNA[IONALEi PUIILIEE EN VERTU DU TRAITE DE~ COOPERATION EN MATIERC DE BREVETS (PCT) (si) classification Internaionale des brovets 5 (11) Nunifto de publication Internationale: WO 93/24645 C071% J102, A6K 4100 A (43) Date do publication Internationale: 9 dicembre 1993 (09.12.93) C12N 13/00 (21) Num~ro do in demande Internationale: PCT/FR93/00524 Publike (22) Date de d~p6t International: 2juin 1993 (02.06.93) rcapotdrehceinrainl.

Donn~es relatives i In priorit6: 92/06765 4 juin 1992 (04,06.92) FR (71X72) D~posant ctinventeur: STERNHEIMER, Joel JFRI 9 l f FR]; 46, rue de la Montagne Sainte-Genevi~ve, F.75005 7 (74) Niandataire: ORES, Bernard; Cabinet Ores, 6, avenue de Messine, P.75008 Paris (FR).

(81) Etats disgn6s: AU, CA, C, JP, KR, RU, US, brevet europ~cn (AT, BE, CH, DE, DK, ES, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), brevet QAPI (BF, BJ, CF. CG, Cl, CM, GA, GN, ML, MR, NE, SN, TD, TG).

(54)TItle: METHOD FOR THE EPIGENETIC REGULATION OF PROTEIN BIOSYNTHESIS BY SCALE RESONANCE (54)Titre: PROCEDE DE REGULATION EPIGENETIQUE DE LA BIOSYNTHESE DES PROTEINES PAR RESO- NANCE D'ECHELLE (57) Abstract The present invention relates to a method for the epigenetic regulation of protein biosynthesis which consists in using, on the biosynthiesis of proteins, scale resonance regulating action of sound transpositions of temporal sequences of quantum vibrations associated with their elongation; said action may be a stimulation or an inhibition of said biosynthesis depending on whether the modulation of the frequences of vibrations used is in phase with or of opposite phase to said elongation. The result is further stabilized by the action of coloured transpositions of groups of quantum vibrations arising from the spacial conformation of proteins from said elongation. The applications, particularly in the agri-foodstuff and health industries, include for implementation purposes a method of defining the metabolic role of proteins from their aminoacid sequences.

(57) Abr~g6 La pr~sente invention est relative A un proc6d6 de r~gulation 6pig~n~tique de In biosynth~se des prot~ines, qui consiste A utiliser l'action r~gulatrice, par resonance d'6chelle, sur la biosynth~se des prot~ines, des transpositions sonores de s6quences temporelles de vibrations quantiques associ~es A leur 6longation; cette action pouvant 6tre une stimulation ou une inhibition de cette biosynth~sc, suivant que Ia modulation des fr~quences des vibrations utilis~es est en phase ou en opposition de phase avec cette longation; le r~sultat obtenu 6tant en outre stabilis6 par Ilaction des transpositions color~es do groupemnents de vibrations quantiques d~coulant de Ia. conformation spatiale des prot6ines issues de cette 6longation. Les applications, notamment dans les domoines de 1'agro-alimentaire et do la sant6, incluent pour leur raise en muvre un proc~d6 permettant do d~limiter los r6les m~taboliques des prot~ines d partir de leur s~quence en acides amin~s.

SUMMARY AND FIELD OF INVENTION The present invention relates to a method for epigenetic regulation of in situ protein biosynthesis, and to the applications of this method, especially in the fields of agronomy and health. It consists in using the regulating action, on the biosynthesis of proteins, by scale resonance, of transpositions into sound of temporal sequences of quantum vibrations associated with their elongation. This action may be either an increase of the rate of this synthesis with at the same time a regulation of its rhythm, or 1 o a reduction of this rate, depending upon whether the modulation of the vibration frequencies used is in phase with, or in phase opposition to said elongation (this being true for the quantum vibrations as well as for their transposition into sound). The result hitherto obtained is further stabilized by the action, again through scale resonance, of colored light transpositions 1 5 of grouped quantum vibrations arising from the spatial conformation of proteins issued from this elongation.

This method applies in a specific way to every protein of known structure. Its use is however all the more appropriate when the synthesis of this protein is even more dependent upon epigenetic factors, that is to 2 0 say external to the DNA of the system to which it belongs, and especially in the present case, upon acoustic and electromagnetic factors. In addition it requires for its practical implementation, the determination of metabolic agonisms and antagonisms of these proteins due to scale resonance phenomena naturally associated with their biosynthesis. The 2 5 characterization of these proteins in their associated metabolic subsets (thus delimiting their metabolic role using their aminoacid sequence) is another feature of the present invention.

The identification of proteins designed to be regulated as part of a given application includes other criteria such as a correspondence I I 1 between acoustic and electromagnetic phenomena of which effects can be observed on living beings, and the transposed proteic sequences whose criteria are also an application of the present invention.

BACKGROUND OF INVENTION I. The demonstration of the musical properties of elementary particles Sternheimer, C. R. Acad. Sc. Paris 297, 829, 1983), as well as underlining the necessity of a coherent theory for the latter, has especially allowed to suggest an important role for the scale at which the phenomena 1 0 happen, scale taken as an autonomous dimension with respect to spacotime. Later developments (initiated in J. Sternheimer, Colloque International "Louis de Broglie, Physicien et Penseur", Ancienne Ecole Polytechnique, Paris, November 5-6, 1987) have led to conclude to the physical existence of quantum waves associated to particles and 1 5 propagating themselves, not only in space-time, but also in that scale dimension, thus linking together successive levels of the organization of matter. These waves, for which we have been able to write and solve the propagation equations, hereby allow an action of one scale onto the other, between phenomena that are similar enough to constitute, in a 2 0 mathematically well-defined sense, harmonics of a common fundamental tone Sternheimer, Ondes d'echelle [scaling waves], I. Partie Physique, 1992, to be published; II. Partie Biologique, the summary of which follows).

The theoretical reasons for their existence, as well as the conformity to experiment of different consequences of their properties, make scaling 2 5 waves appear as a universal phenomenon whose function is at first to ensure the coherence between the different scales of a quantum system, and that especially takes shape and can be described in the process of protein biosynthesis. The peptidic chain elongation effectively results from 7 the sequential addition of aminoacids that have been brought onto the 3 ribosome by specific transfer RNAs (tRNAs). When an aminoacid, initially in a free state, comes to affix itself to its tRNA, it is at this instant already stabilized enough with respect to thermal agitation while keeping a relative autonomy because it is linked to the tRNA by only one degree of freedom for its de Broglie wavelength to reach the order of magnitude of its size; this gives it wave properties, and the interference between the scaling wave then associated to it and those similarly produced by the other aminoacids, results in a synchronization, after a very short period of time (which can be evaluated to be about 10 -12.5 second), of the proper 1 0 frequencies associated with these aminoacids according to one and same musical scale, which therefore more precisely depends upon the transfer RNA population. Nevertheless, to within the approximation of the tempered scale, this scale is universal, especially due to the very peculiar distribution of aminoacid masses, which is already very close to it. (In a 1 5 similar way, the DNA nucleotides are also tuned to a same musical scale, as can easily be seen from their masses).

But the phenomenon which we refer to appears in an even more explicit way when, the aminoacid being carried by its tRNA, the latter in turn fixes itself onto the ribosome. It is in fact at this moment, that is to say 2 0 up to the transfer which fixes it to the peptidic chain, that the stabilization with respect to thermal agitation becomes such that the wavelength of the aminoacid outgrows its size by a full order of magnitude. The scaling wave which is then emitted interferes, at the scale of the protein in formation, with similar waves previously emitted by the other aminoacids. This draws 2 5 constraints of a musical type for the temporal succession of the proper frequencies associated to these waves, so that the scaling waves may (if we generalize the previous situation) continue their itinerary, and thus insure coherence and communication between different levels of the organism for example, the only succession of these waves has for ~'g consequence to minimize the dissonance (harmonic distance) and the frequency gaps (represented by melodic distance) between successive aminoacids; even more so, because each scaling wave appears as a superposition of waves linking two given levels (and therefore that of each aminoacid to that of the protein) in twice, thrice...the time taken by the fastest, this implies the existence of periods of minimization of harmonic distances notably, showing punctuations in the temporal succession of frequencies; which other levels will complete with correlations all the more rich and marked that they themselves are more numerous to influence the 1 0 protein synthesis. As a consequence, this provides a remarkable prediction: proteins must possess, in the very succession of the proper quantum frequencies associated to the sequence of their aminoacids, musical properties all the more clear and elaborate that their biosynthesis is more sensitive to epigenetic factors in general; and conversely, it must 1 5 be possible to act epigenetically, in a specific way for each protein onto that biosynthesis.

The observation of protein sequences described in literature, (cf. M.

O. Dayhoff, Atlas of proteir sequence and structure, volume 5 and supplements, N.B.R.F. (Washington) 1972-78; the updating of which is 2 0 accessible through CITI 2, 45 rue des Saints-Peres, Paris) allows to confirm that things are as such; not only all proteins do possess musical properties in the sequence of their aminoacids, but these properties are all the more developed that the proteins are, in a general way, more epigenetically sensitive. In addition, the acoustic transposition of the 2 5 series of proper frequencies corresponding to the production of scaling waves in phase with the elongation of a given protein, has a stimulating action onto the biosynthesis of this protein in vivo, and in a correlative way it has, on the contrary, an inhibiting action for scaling waves in phase opposition. These actions which reproduce at our scale the similar I actions that already take place at a quantum scale between proteins during their synthesis (and therefore play an important role in their metabolism: the proteins that are musically similar are thus, in a systematic way, metabolically agonist) seem effectively common to all living beings who are sensitive to sound vibrations, and we have had many occasions to observe them.

In the case of animals having a nervous system, it seems possible to give (at least for vertebrates, where the "microphonic potentials" that faithfully reproduce the shape of an applied wave have effectively been 1 0 observed) the following description of these phenomena: the sound wave is transformed into electromagnetic impulses of same frequency, right from the starting point of the auditory nerve. These impulses, by virtue of the scale invariance of scaling wave equations applied to the photon (which generalize Maxwell's equations), then have a direct action, by scale 1 5 resonance, on their quantum transpositions; because the squared quantum amplitudes are proportional to the number of proteins that are simultaneously synthesized, the resonance phenomenon results, in the case of scaling waves in phase, in an increase of the rate of synthesis, as well as a regulation of its rhythm; and in the case of scaling waves in 2 0 phase opposition, in a reduction of this rate. [As we can notice, it is because the mnicrophonic potentials take place, in the auditory nerve, before the start of the actual nerve impulse (cf. P. Buser and M. Imbert, Audition, Hermann tditeur, Paris, 1987), that the mechanism referred to here, does not require at this point the cerebral analysis of these impulses].

2 5 Among plants, the (mechanical) sensitivity to sounds is well visible specially through interferometry and the scaling waves behave theoretically in a similar way, The solution to the scaling wave equation, which effectively shows the existence of scaling waves having a range close to Avogadro number I I 6 (as it is the case for the above-mentioned transpositions), in addition leads to anticipate similar properties for the scaling waves drawn from the spatial distribution of aminoacids (whose de Broglie wavelength is then comparable to their size) inside the protein after it has been synthesized, this time with a range approximating the square root of that number: the observation of their tertiary structu-res confirms the existence of harmonies within vibratory frequencies of aminoacids spatially nearby inside proteins (and especially at their surface, as it can be expected from their wavelength), and at the same time we have effectively been able to 1 0 observe, through the colored transpositiors of these frequencies, an appreciable stabilization of the effects obtained with the use of the musical transpositios.

The present invention is drawn from these observations.

DESCRIPTION OF INVENTION II. In order to decode proteins, we proceed as follows: 1. We determine the frequency series in the following way: to each aminoacid corresponds a keynote, the exact frequency of which is 2 0 obtained from the proper frequencies of aminoacids in their free state (proportional to their masses), by minimizing the global harmonic distance Z ij Pi Pj logsup (pi, qj calculated for all possible pairs of notes, (pi qj) being the harmonic intervals globally the closest to the corresponding proper frequency ratios, taking into account their respective proportions 2 5 Pi' Pj in the environing population of transfer RNAs; while respecting the condition 5 f< A f/2 where 5 f is the displacement of the initial frequency towards its synchronized value, and A f is the interval between the two successive synchronized frequencies of the obtained scale, which encompass this initial frequency; then (according to the method described

I

_I in the French patent number 8302122 by the same inventor), transposition into the field of audible frequencies.

Within the approximation of the tempered scale, one thus obtains a universal code for the stimulation of protein synthesis: Gly low A; Ala C; Ser E; Pro, Val, Thr, Cys F; Leu, Ile, Asn, Asp G; Gin, Lys, Glu, Met A; His B flat; Phe, as well as SeC B; Arg, Tyr sharp C; Trp sharp D and another one for its inhibition, which is deduced from the preceding one by symmetrization of the logarithms of the frequencies around their central 1 0 value: Trp C; Arg, Tyr D; Phe, SeC E flat; His E; Gin, Lys, Glu, Met F; Leu, Ile, Asn, Asp G; Pro, Val, Thr,Cys A; Ser B flat; Ala sharp D; Gly sharp F the application of which globally results in scaling waves respectively in 1 5 phase with and in phase opposition to those taking place during the synthesis process. (By "universal code", we hereby mean that this code is identical for all proteins to within the approximation of the tempered scale; the low A, for a central frequency located 76 octaves below the centre of gravity of the initial frequencies of leucine, isoleucine, and asparagine, is at 2 0 220 Hz. The definition of harmonic distance given above recovers and extends the definition suggested by Y. Hellegouarch, C. R. Math. Rep.

Acad. Sci. Canada, Volume 4, Page 227, 1982). More finely, the exact values depend on the proportions of the groups of the above-mentioned aminoacids among the transfer RNA population surrounding the protein 2 5 biosynthesis, and they may be calculated in every such situation.

2. We determine the period (or the periods) appearing in the molecule.

The very existence of these periods results directly, as said in part I, from that of scaling waves. An indication for at least some of them is ILl" usually given by the presence of obvious cadences (such as GG, F-S that is to say F closely followed by S as well as the cadence ending the signal peptide when it is present, for stimulation; series of R or Y, for inhibition; exceptionally, relative pauses induced by harmonic variations which would otherwise be too straight; and in all cases, cadences expressing the return to the tonic note) producing punctuations in the musical development. We then determine more precisely the similar passages, either by the direct repetition of notes (when it is the case, the period is given by a simple calculation of autocorrelations of notes; or more 1 0 finely by minimizing the frequency differences between notes by the number that minimizes the average on the protein of melodic distances between notes located an integer number of intervals apart), either of melodic movements (the period is then given by a calculation of autocorrelations of signatures or frequency variation signs from one 1 5 note to the next; or more finely, by a calculation of autocorrelations of the melodic distances from one note to the other, the distances being counted with their sign, i. e. multiplied by the corresponding signatures; or even more finely, by the number which minimizes the average on the protein of step by step melodic distances variations, to within an integer number of 2 0 intervals apart; the repetition of the melodic contours being precised by a calculation of autocorrelations of pairs, or even better, of triplets of signatures), either again by the logic of the harmonic movement, that reproduces the notes or the melodic movement to the nearest simple harmonic transposition (octave, fourth or fifth in general; the period is then 2 5 given by the number that minimizes the average on the protein of harmonic distances between notes located an integer number of intervals apart).

Sometimes also, when an "alignment" of similar sequences especially among different species is available, the period appears in the additions or in the deletions of certain of these sequences. The result must give a L r melodically and harmonically coherent progression. In order to do that, we take into account the fact that the last notes of each period or member of phrase usually the second half, and more particularly the last note as well as those situated on the strong beat (the characteristic of which will be described in section 4) are the most important for this progression. The final result is then the most significant (that is to say that we balance these dii'lrent elements according to their relative importance in the protein, and especially the harmonic and melodic distance by the square of the ratio of their normalized standard deviations) respecting the whole of 1 0 these criteria; there is usually one that is distinctly more significant than the others, as it so happens when we try to determine by calculus the spatial foldings of molecules; cases similar to allosteria nevertheless exist, and have a biological meaning (stimulation or inhibition by such molecule or such other one during the metabolism), but influence more frequently the 1 5 position of the measure than the period (metabolic function different according to the context, for instance, CG rich or AT rich; the measure bars depending upon the composition of the DNA, as the "Christmas trees" that can be seen during certain syntheses cf. B. Alberts and al., Molecular biology of the cell, 2nd edition, Garland Publ. Co 1989, page 539 2 0 clearly display).

3. If necessary, we rectify such or such period especially so that the melodic passages that are related (that is to say which repeat or follow one another) can be found in the same place inside the measure: we deduce therefrom the individual lengths of notes. (This operation of adjusting the 2 5 phrasing to the measure is comparable to the well known phenomenon of lengthening the vowels of a sung text).

In practice, operations described in sections 2 and 3 can be performed most easily with a keyboard such as a Casio TM equipped with a "one key play" device, or with a computer programmed especially r I for that purpose, in which one has previously stored in memory the sequence of notes as obtained in section 1, and where one can then directly play the sequence of notes, which allows control and adjustment of these operations. Then they nevertheless require some precautions.

Prudence implies, among other things, to also decode the same molecule or a musically similar molecule, in the direction of inhibition (or in any case in the direction opposite from the initial one), taking into account the fact that molecules very often have a preferential decoding direction; it is in particular frequent for pairs of molecules that sensibly exert the same 1 0 function, to find one being more musical in inhibition and the other one in stimulation (that is especially the case in the metabolism of irmmunity and auto-immunity); in this case, the presence and the distribution of cadences (that differ in stimulation and in inhibition, cf section 1) normally allows to recognize them at first, and consequently to preserve oneself.

1 5 4. We check the rhythmic style through the distribution of th bases of DNA: first, if needed, through their autocorrelations (when the molecule is musical enough, the period of these autocorrelations corresponds to that of the protein; then they determine in princio the measure bars, the ranks of base triplets or more precisely of 2 0 bases in third position in these triplets for which the peaks of autocorrelation are the highest, corresponding to the most accentuated notes); then by referring to codon usage, in comparison with known molecules (already decoded, or more regular and thus raising less difficulties) having the same supposed rhythmic style: the style of 2 5 musical rhythm (which by constraining the accentuation of notes, hereby influences the choice of bases in third position) determining (at least approximately) in a univoque way the codon usage, molecules of the same style must therefore have (very sensibly) the same codon usage.

If necessary, we correlatively correct the decoding of some passages.

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We then try to determine the tone quality. It is in principle different for every molecule, and in any case for every distribution of notes. In theory, it mainly depends upon the molecule itself but it also depends upon all the levels of the organism which retroa t on the harmonic structure of aminoacid vibrations. A first approach is given by adjusting the distribution of molecule keynotes to the theoretical graph of that distribution (as can be deduced from the scaling wave equation, as well as it corresponds to what can be observed in average, on the whole of proteins). This leads to deduce (as in the French patent number 8302122) which harmonics 1 0 are amplified and which are softened in the wanted tone; we then select the closest tone quality in a palette of given ones (like a voice memory for sampler, or as one can already find included in many expanders and musical softwares). Tis implies to distinguish more precisely between three situations: 1 5 distribution of notes constant along the molecule (we then have a relatively fixed harmonic structure); straight distribution changes (we then have different successive tones of instrument, for instance cytochrome C with several organ registers); progressive distribution change (the distribution then reproduces the time 2 0 evolution of the harmonic structure of one keynote, for example myosin where this evolution clearly indicates a timbre of trumpet), Apart from this, determining the tempo gives no real problem to the technician because it normally follows from the rhythmic style determined above; it is generally all the faster that there are important redundincies 2 5 in the proteic sequence, as it is the case for fibrous proteins.

6. We then determine the colors by applying the code, which is also universal at first approximation, and is deduced from vibration frequencies of individual aminoacids through the formula (drawn from scaling wave theory): ~III i v vo Argch (e (fifo) Logch where Io represent the proper quantum frequencies associated with aminoacids as previously, and v, Vo those of colors, the index o showing central values; this gives the following code relating to the stabilization of proteins synthesized in situ (the code related to tie stabilization of their inhibition is deduced as in section 1 by symmetnzation of the logarithms of frequencies with respect to the central lemon yellow): Gly dark red; Ala bright red; Ser orange; Pro, Val, Thr, Cys ochre; Leu, Ile, Asn, Asp lemon yellow; Gin, Glu, Lys, Met green: 1 0 His emerald; Phe blue; Arg, Tyr indigo; Trp purple, these frequencies then being moved towards red or purple according to the global repartition of the molecule frequencies in a way similar to the description for tone quality as above. The spatial position of colors then being the same as those of the aminoacids in the tridimensional spatial 1 5 representation of the molecules.

I Examples.

One will find below some examples of musical and colored decodings of protein sequences. (In these examples as well as in the figures, to make 2 0 things easier, we use the one-letter notation for aminoacids: Gly G; Ala A; Ser S; Pro, Val, Thr, Cys P, V, T, C respectively; Leu, Ile, Asn, Asp L, I, N, D; Gin, Glu, Lys, Met Q, E, K, M; His H; Phe F; Arg, Tyr R, Y; Trp W).

1) Example of a protein regular from beginning to end. On the evolutive alignments of a protein that has been particularly well studied, cytochrome C, we can observe a constant deletion of eight aminoacids (sometimes seven) among animal proteins when compared to plants. Observing the autocorrelations of notes and melodic contours confirms this first indication of the value of the musical period; if, in fact, we count the occurrences of the same note, as well as of the same direction of pitch variation occurring three times in a row (the same triplet of signatures), which are distant from an integer number k of notes, we obtain the following result: Values of k 1 2 3 4 5 6 7 8 9 10 11 12 Note autocorrelations 19 15 15 20 19 1517 21 14 17 18 13 Melodic contour autocorr. 1 7 4 6 5 10 8 13 5 4 4 4 Total 20 22 19 26 24 2525 34 19 21 22 17 the peak at k 8 being worth about 2.5 standard deviations (as compared 1 0 to its expectation value 22.3 4.7 determined from the repartition of notes of the molecule); the significance of this peak is even more so reinforced when using melodic distances as described in II, 2 (it outgrows distinctly 3 standard deviations if we include the autocorrelations of melodic intervals, by taking as a definition of the melodic distance between two 1 5 notes, the absolute value of the difference of the ordinal ranks of their tempered frequencies in the scale obtained in II 1, arranged in ascending order this definition being derived from the usual nomenclalure: second, third, etc., for the notes of a musical mode; the secondary peak at k 7 then becomes slightly significant, corresponding, as we shall see later, to the relative stretching of the seventh note which tends to precede the return to the tonic; whereas the one at k 4 is reinforced when we use harmonic distances as described in II 2, because it corresponds, as we shall see later, to spatial foldings of the molecule). The observation of the cadences also confirms this value, as well as that of the internal similarities 2 5 (thus, the last five notes of the first, second and third group of eight produce together an exact harmonic superposition, in other words a canon for three voices). More precisely, these last two investigations show first of all a greater relative importance of the seventh (F-S cadence on the second period) and eighth notes (back to the A minor tonic) for each period, the r 14 latter once more prevailing over the former (the perfect S-Q cadence on the sixteenth note prevails over the preceding F-S cadence with the recovering of the initial tonality); the division (as economic as possible while taking into account the preceding constraints) of the period results into six semiquavers, one quaver, one crotchet (which means relative lengths 1-1-1-1-1-1-2-4, with a 6:8 rhythm; cf. figure One will notice the coherence of the melodic progression (wherefrom the observed regularity mainly proceeds) as well as the richness of the harmonic progression, the A minor tonality being accompanied with modulations in E 1 0 minor (second bar), G minor (eighth bar), and F major (third and ninth bar), notably.

If we then examine the distribution of DNA bases, we can see that the first and seventh notes of each period distinctly foster, respectively, adenine and thymine in third position, whereas the third and eighth notes 1 5 foster in the same way cytosine and guanine. As well as confirming the above division for the period and the relative lengthsof notes (that is to say the fact that the seventh and eighth notes have length that are respectively twice and four times the first), this shows furthermore that in an AT-rich environment, strong beats are on the first and seventh notes, and therefore 2 0 the measure bars are on the first, whereas in a CG-rich environment, the musical sequence starts on an anacrouse (strong beat on the third and eighth notes, measure bar on the third). We can conclude that the protein must have distinct metabolic roles, depending on its environment. Actually, the range of its metabolic action is first demonstrated by the degree of its 2 5 musical evolution (in comparison with the sequence of Euglena gracilis, for example, where one can observe, in the three first measures, an improvement of 56% of the melodic [regularity] level and of 16% of the harmonic [regularity] level, defined from the minimization of the respectively melodic and harmonic distances between successive notes); id the search of musical similarities with other proteins then shows on one hand the possibility to superpose cytochrome C onto endozepine, with a musical reading frame compatible with the measure bar on the first note, and which is effectively a (slightly) AT-rich molecule: which allows one to precise an anti-depressive role for the cytochrome (and its music), through the eventual desinhibition of neurotransmission; as well as, on the other hand, a musical enchainment (then beginning on an anacrouse) with cytochrome-oxidase, which is effectively (slightly) CG-rich, and which ends the respiratory chain, other metabolic role of cytochrome C located 1 0 just before cytochrome oxidase in this chain.

As for tone quality, because tonality is here in A (minor), the quasi absence of the fourth and the relative weakness of the fifth (E) compared to the distinct dominance of the tonic note and to the abundance of the octave (low A medium A) will privilege harmonics 1 and 2, to the 1 5 prejudice of the followings, indicating an organ timbre, with in fact slightly different registers according to the passages. Eventually, colors effectively group themselves into colored stains onto the mature protein (cf. figure 2) with, as in the case for music, remarkable harmonic responses. (Notice that the color determination is useful to confirm the musical decoding, 2 0 insofar as some autocorrelations of notes are translated not into the musical period but in the spatial folding of the molecule: then we must eventually subtract them if we want to determine by this mean the musical periods; it is the case here where a secondary peak of these autocorrelations k 4, due to the a-helix of the beginning which can be 2 5 seen on figure 2 corresponds to these foldings. Conversely, the musical decoding may then give indications about the spatial structure of a protein).

2) Example of control of the decoding of a protein showing rhythmical variations. As we have just seen, the decoding of a ~II~ I protein may be controlled at different levels, including the decoding of molecules known to be metabolically agonist, and including as well the coherence of the conclusions that may be drawn, on a metabolic level, from the musical similarities that can be observed. We can thereby rebuild step by step large sections of the metabolism at the molecular scale. As we shall see, this also facilitates the decoding: in the previous example, the "rhythmic formula" of cytochrome C may be transcribed as follows: 6/8 GDVEKGK:K:::IIFIMKCS:Q:::CHTVEKG:G:::, etc.

where the underline the strong beats, and the indicate the place of measure bars, whereas the indicate the lengthening of notes.

In subunit II of cytochrome oxidase, which is musically chained to cytochrome C, the beginning is, on the contrary, clearly a four-time 1 5 formula as is simply shown by the internal similarities (so the notes 7 to 22, which remind in their contours the manner of Bach, split into grps of four notes, each one being superposable to the next). At theea measure, one finds another measure which is not only superposable, for all its strong beats, onto the first measure of cytochrome C, but 2 0 is in fact, even practically identical to the third measure of the same cytochrome. This implies a lengthening of the eighth measure (as the cadence seen at the end of this measure already indicates in itself), in a sixtime measure (figure 1): 14/8 MTHQSHAYIHMVKPSPW PLTGALSAILLMTSGLAI S+ 4 4 MWFHFHSM ITLLMLGLLI TNTLTMYQII6/8 WWRDVTR: 4 ESTYQGH:H:: :ITPPVQKG:::: I S+ I i This change in rhythm (from 4/8 to 6/8) is also well visible in base autocorrelations of the DNA where, at this point, the prominent peak goes from the fourth to the sixth base triplet (and despite the fact that the ternary rhythm of the bases, which usually prevails in base autocorrelations of the coding parts of DNA, i s slightly less distinct here).

(In figure 1 we have the sequence start on an anacrouse, thus emphasizing, as indicated above, the strong beat on the third note, in view of the enchainment with the CG-rich rhythmic variant of cytochrome C).

3) Example of reconstitution of a metabolic chain including 1 0 stimulations and inhibitions. Here is another example of reconstitution, step by step, of a metabolic chain. The decoding of histone 4 is particularly easy: the periodicity of 7 is clearly visible on the sequence at the outset of the molecule, the repetition of G within a two aminoacid interval indicates a binary rhythm, and the GG cadences that end the two first periods 1 5 specify right away a four-time rhythm: ISGRGKGG: KGLGKGG: S+ this pattern goes on until the end of the sequence, with the only exception of the last measure which is syncopated in order to recover, through an 2 0 internal similarity, the rhythm of the first two measures (figure The global repartition of the notes shows a harmonic structure cc-responding to the tone of a flute, and the "skip of notes" repeated from the beginning, which suggests a sound with an attack, even allows to specify a timbre similar to that of Pan's pipes.

2 5 Histone 4 is one of the most conserved proteins among the animal and plant kingdoms. This does not mean that its metabolic action, obviously essential, doesn't sometimes need to be tempered; thus the theme of its first two measures is found again, in inhibition and transposed to the fourth, in the conserved part of the beginning of chalcone synthase, which I I is the pigmentation enzyme of many flowering plants (figure This may be compared to the supposed role of chromatin, which histone 4 is part of, in the process of magnesium fixation: during spring, plants need a lot of magnesium (for photosynthesis) and its fixation needs to be stimulated (including by the bird songs evoked by the theme of this molecule); chalcone synthase is then inhibited; whereas during the fall, the weaker stimulation of histone desinhibits chalcone synthase and allows the replacement of the green of the leaves by brighter colors of that season, the diversity of which, so much praised by the poets, becomes thus more 1 0 understandable through their epigenetic component.

Actually, when listening to the musical transposition of histone 4, auditors reported several times "an urge to eat chocolate" which contains magnesium (some even found that "it produces the same effect as that of granulated magnesium, except that this effect is immediate in this case").

1 5 This, we can point out, presents some inconvenience for people having a slightly too high rate of cholesterol. And actually, the musical decoding of chalcone isomerase, metabolically agonist of chalcone synthase, but which "works better" musically in stimulation, includes a series of themes and variations whose succession reproduces, in flowering plants, themes of the 2 0 full metabolic chain regulating cholesterol in man: listening to that "second degree" antagonist of histone 4 then allows (following a method which usually applies to this type of situation) to possibly correct the secondary effect mentioned. In addition, the frequency of the ascending fourths in chalcone isomerase tends to approximate that observed in the 2 5 alcali light chain of mammalian myosin, which stimulates muscular contraction (while magnesium, as we know, acts as a muscular decontractant). Listening to it therefore also encourages physical exercise which is another well-known way to help cholesterol regulation. In fact, this last example underlines the importance of a quasi-general -II I II I I phenomenon, that is, the epigenetic co-operation of different factors in the stimulation of protein synthesis, which accounts for the aspect meaningful in itself of the musical sequencej: in this way, for example, listening to myosin will generally suggest a military march.

4) Example of the biochemical analysis of an epigenetic cooperation involving harmonic superpositions. When it is possible, the biochemical analysis of these epigenetic cooperations is a valuable help for decoding. So, another well-known way to stimulate epigenetically the muscular decontraction is heat, whose healing action for rheumatism, for 1 0 example, is well known. The action of heat is conveyed by a group of proteins called heat shock, generally synthesized together. This suggests that they should show harmonic superpositions: and in fact, the hsp 27, which appears to be the most musical, es superpose itself onto the beginning of hsp 70, the most abundant, which sort of plays here the role of 1 5 a bass line. These two molecules are again superposable together with the beginning of troponin C, which regulates calcium in muscular contraction, and for which we are thus led to anticipate a role all the more important, as an anti-rheumatic, that its musical level is high (figure Nevertheless, it is advisable to underline that many other molecules, also of a high musical 2 0 level and epigenetically sensitive, may be implicated in this type of ilment, from the stimulation of prolactin and beta-lipotropin (precursor of betaendorphin) to the inhibition of estrogen receptor, including the inhibition of IgE and interleukin 1 beta.

These few examples clearly show how large sections of the 2 5 metabolism can be reconstituted step by step, with many ways to check or control the coherence of the results obtained, and thereby to precise the musical decoding of the concerned proteins.

I I IV Applications.

For implementation purposes, we will use the transcriptions in the form of either musical scores, or of recordings of the obtained musical sequences, as well as colored spatial representations of proteins, together or separately, on all type of support: record, compact disk, floppy, audio or video cassette, paper, fabric or others, specially as far as the colored images are concerned.

The recordings of musical sequences may be realized from musical scores as described in II (for which we gave some examples in by 1 0 using one of the methods evaluated in B. H. Repp, J. Acoust. Soc. Am. 88, p, 6 2 2 (1990); the most precise of these methods having been used in the examples hereby given.

1) In the fields of agronomy and textile industries, the possibility to stimulate certain specific protein synthesis, concerning for example bovine 1 5 lactation, fermenting of baker's yeast, the sweet taste of some fruits, animal or plant fibres (keratine of sheep's wool, fibroin of silkworm, etc.), as well as the proteins specific to certain medicinal plants; and in the field of environment, for example, the assimilation of industrial effluents through plants, by stimulating the biosynthesis of the corresponding proteins. We 2 0 have hereby been able to observe on a cow who regularly, during days and at the time of milking, listened to recordings of musical transcriptions of the aminoacid sequences of bovine prolactin, lactoglobulin, and lactalbumin a reduction, by a ratio of 3, of the relative quantity of whey, resulting in a milk highly enriched in proteins, and consequently, in a 2 5 particularly savoury cheese. In the same way, among tomatoes that were given in their time of growth, a "cocktail" of musical transpositions of different proteins including: specific virus inhibitors, various extensines, then a flowering enzyme (LAT 52), an antibacterial protein from which we also expected, because of its musical similarity with thaumatin, an improvement of sugar percentage (P 23), and at last, inhibitors of fruit softening enzymes (pectinesterase and polygalacturonase) we observed a distinct increase in size and number of fruits (summing up to a ratio of about 3,5) as well as a sensitive increase of the sweet taste in a significant proportion of the fruits that had particularly received P 23. These noteworthy results nevertheless go along with a certain amount of precautions: there hereby exist some counterindications to an excess of stimulation, especially of prolactin, which must be cautiously taken into col.ideration by breeders that carry out these 1 0 methods, as well as for the animals themselves who may be fragilized.

Thus, in the well-known experiences carried out in Israel on cows with Mozart music bovine prolactin has in fact, apart from a "musical level" particularly high which can here define in a mathematically simple way (from the melodic and harmonic levels, cf. I paragraph some musical 1 5 turns that can be qualified as "typically Mozartian" the rate of mammites could seem worrying: in such a case (which we have also been able to observe), one ought to complete the hearing of prolactin with that of alpha-1 antitrypsin, whose musicality is also very elaborate and whose metabolism is complementary on this point. Similarly for tomatoes 2 0 receiving outside stimulations, one must be cautious not to interrupt the cycle too suddenly.

As they are, these results still give an indication of the order of magnitude of results we can obtain in such conditions, and clearly show the interest of the invention.

2) In the therapeutic and preventive fields, many ailments are characterized by a specific metabolic weakness, and can therefore be efficiently prevented or treated with the help of the present invention.

Because the minimal length of a musically active sequence is of the order of that of a signal peptide from several aminoacids to a few tens this

I

action may be very fast and appear already after a few seconds or a few minutes. Nevertheless, the complete integration of the produced effect, which is metabolically complex, can take slightly more time, or even require. in case of a strong cultural conditioning, a certain initial training (apparently corresponding to "re-learning" to listen through the conveying scaling waves proceeding from the cochlear microphonic current): but usually, this is accomplished rather rapidly for the obvious benefit of the persons concerned.

For a responsible use of the described method, it is therefore 1 0 important to really well know the metabolic role of the molecules involved.

And it is of course one of the interests which obviously outpasses the sole therapeutic frame mentioned here of the musical decoding of proteins (associated to the corresponding colors) to allow, by systematically spotting the similarities and counter-similarities of melodies 1 5 (and colors) from the protein sequences that are known and available in data banks, to select proteins that are metabolically agonist and antagonist of a given protein, for which the degree of musical elaboration also gives an indication of the importance of its metabolic role. The described method therefore allows to precise particular indications for some proteic 2 0 sequences (as we gave some examples above in Ill).

Let us recall in connection to this, that we often observe in animal or plant proteins, especially among the most musical ones, successive melodic fragments of human metabolic chains, and that in consequence, the transpositions which are active on man are not limited to human 2 5 molecules, as shown by the examples we gave also in III 3. On the contrary, the metabolism of those species seems in some way more "specialized" for the production of certain molecules, and it is indeed the generally most musical proteins that will be the most important for the applications. Of course, these correspondences between different species I r facilitate further more the delimitation of the metabolic role, and the decoding of proteic sequences.

It is opporrunate to point out that the musicality of a molecule implies in itself that its epigenetic stimulation is (generally), in principle, preferable for a therapeutic use, (because of the range of its metabolic interactions), to its direct absorption: the "most musical" molecules are generally those for which either the production by genetic engineering, or the therapeutic use which derives from it, will meet some problems, such as of transportation to the site of action, or of stability, or more specifically of 1 0 secondary effects related to doses that should be much more imlortant that what they are in the body to obtain comparable effects, because then, the scaling waves naturally associated to their production are not present any more. This, of course, is particularly true for the inhibition of proteins, when the natural inhibitor is, for example much heavier or simply 1 5 when the production needs to be reduced at a given time (cf. 11 3) or in a systematic way.

Eventually, concerning the use of transcriptions of proteic sequences, th', ery quickness of their action may allow, by differential comparison, especially bipolar, of their positive and negative effects, to precise which- 2 0 one is the most appropriate in a given situation. [It is to be noted that the possibility thus conferred to anyone to acknowledge by himself and very rapidly the action of these transcriptions, and the recognition of the self which comes out as a result, do not constitute their least interest]. This identification may itself be facilitated by the comparison with transcriptions 2 5 of known proteic sequences, of acoustic or electromagnetic phenomena exhibiting distinct series of frequencies, and for which some effects may have been observed in a similar situation.

I

24 As will be appreciated from the above, the invention is in no way limited to those methods of putting it into effect, of construction and of application which have been described above in detail; on the contrary, it covers all versions which may be conceived of by workers skilled in the art, without exceeding either the framework or the scope of the present invention.

Claims (9)

1. Method for determining musical sequences corresponding to the amino acid sequences of proteins, through the decoding and transposition into sound of temporal series of quantum vibrations associated with elongation of the proteins to epigenetically regulate the biosynthesis of these proteins by scale resonance, comprising a succession of the following stages: a) To each amino acid of a given protein a keynote is associated, the frequency of which results from the application of a code obtained from the proper frequencies of the amino acids in their free state, proportional to their masses, by minimising the global harmonic distance between the frequencies of these amino 15 acids for all possible pairs of amino acids taking into account their proportion in the population of transfer RNAs while respecting the condition that the displacement of the initial frequency towards its synchronised value, be 2 inferior to half the interval between the two synchronised S 20 frequencies which surround that initial frequency, then a transposition of the frequencies thus obtained in the auditive range, the code thus obtained being relative to the biosynthetic stimulation of that protein, the code relative to its inhibition being obtained by symmetrisation of the logarithms of heretofore obtained frequencies with respect to their central value considered as the origin; b) determining the musical periods by observing the similar sequences; c) determining the lengths of the notes by rectifying collectively then individually the periods determined in b) by adjusting the phrasing to the measure; d) determining the tone quality through the retroaction of all the amino acids of the global protein on the harmonic structure of each protein by adjusting the repartition of the notes of the protein to the average s*&OroApedI43CU3O.STERNHEVAER 202 I 26 repartition of these notes for the whole of proteins, determining from what was deduced which harmonics must be raised and which must be lowered in order to obtain the tone quality corresponding to the given protein, the musical sequences able to regulate the biosynthesis of that protein being thus obtained at the end of this fourth stage.

2. The method according to claim 1, wherein a code is obtained during stage a) which code is specific to the stimulation of protein biosynthesis, and universal to within the approximation of the tempered scale, which consists in the association to the different aminoacids, of the following keynotes: Gly low A; Ala C; Ser E; Pro, Val, Thr, Cys F; 15 Leu, Ile, Asn, Asp G; Gln, Lys, Glu, Met A; His B flat; Phe, as well as SeC B; Arg, Tyr Sharp C; Trp Sharp D.

3. The method according to claim 1, wherein a code is obtained during stage a) which code is specific to 20 inhibition of protein synthesis and universal to within the •.approximation of the tempered scale, obtained by taking the notes of the tempered scale which are symmetrical to those of the code according to claim 2, with respect to the central G. 25 4. The method according to claim 1, characterised by the fact that each sound transposition of quantum vibrations associated to the biosynthesis of a given protein is completed by the colour transposition of quantum vibrations associated to the mature protein after it is spatially folded back over itself, according to a code specific to the stabilisation of that protein or to the inhibition of its biosynthesis obtained through a musical sequence realised according to claim 1, which code is deduced from the code obtained during the stage a) of claim 1, by application of the formula V VO Argch(ef"o Logch l where f, fo are the musical frequencies and V, V 0 the stafthrorlkaptrsed433o.93.STERNIEt4MER 2.2 ~'I C 27 frequencies of colours, with the index o showing the central values. The method according to claim 4, wherein a code is obtained specific to the stabilisation of proteins stimulated by the musical sequences obtained according to claim 1 by using the code according to claim 2, which consists in the association to the different amino acids of the following colours: Gly dark red; Ala bright red; Ser orange; Pro, Val, Thr, Cys ochre; Leu, Ile, Asn, Asp lemon yellow; Gln, Glu, Lys, Met green; His emerald; Phe blue; Arg, Tyr indigo; Trp purple.

6. The method according to claim 4, wherein the code obtained is specific to the stabilisation of the inhibition 15 of proteins obtained by using musical sequences realised according to claim 1 or claim 3, obtained by symmetrisation of the logarithms of the wavelengths of the code according to claim 5 with respect to the logarithm of the wavelength of central lemon yellow considered as the origin. 20 7. Transcriptions in the form of music scores, colour representations, and/or recording on all appropriate support, of the musical sequences when obtained by working out the method according to claim 1 and/or of the transposed colours obtained by working out the method 25 according to claim 4.

8. Application of the method according to any one of claims 1 to 6 in an agronomic, environmental, textile, therapeutic or preventive fields, by stimulation and/or inhibition of the appropriate plant protein synthesis comprising administering musical sequences to plants.

9. The method of claim 8 wherein the musical sequences are obtained through the method according to claim 1, then stabilisation through the colour transpositions obtained by working out the method according to claim 4. staflhrorikeepspeci43304.93.STERNHEIMER 20.2 1 I c~- I O 28 Application of the transcriptions and especially of the recordings of the musical sequences and/or of colour transpositions according to the method of claim 7 in a therapeutic, and preventive aim, as well as agronomic, environmental and textile.

11. A method for the characterisation of protein sequences fit to be regulated by using the method according to claim 7 for the applications or method according to any one of claims 8 to 10, characterised in that one delimits the metabolic role of the protein by decoding with the method according to any one of claims 1 to 3, hereby showing the mr'sical similarities and anti-similarities that they present with other proteins, the harmonic superpositions with other protein melodies, or a combination of these factors, from which the agonisms and antagonisms they have with these other proteins can be deduced.

12. A method according to claim 11, in which the characterisation according to claim 11, for a given 20 application or method according to any one of the claims 8 to 10, of the protein sequences fit to be regulated in view of this application or method, is refined by bipolar differential comparisons with the positive or negative effects obtained by using transcriptions according to claim 7.

13. A method according to claim 11, in which the characterisation according to claim 11, for a given application or method according to any one of the claims 8 to 10, of the protein sequences fit to be regulated in view of this application or method, is refined by identification, through musical similarity or anti- similarity according to claim 11, of the proteins involved during positive or negative effects due or associated to acoustic or electromagnetic phenomena exhibiting distinct series of frequencies. statlhronskeopopUid4330.93.S1ERNHEIMER 20.2 29 DATED THIS 20TH DAY OF FEBRUARY 1997 JTOEL STEENHEIMER By Its Patent Attorneys: GRIFFITH HACK Fellows Institute of Patent Attorneys of Australia 0 OV .sees stafuinorlkeospd43304.93.STERNHEMER 20.2 .I PATENT APPLICATION In the name of Joel STERNHEIMER Invention: Joel STERNHEIMER METHOD FOR EPIGENETIC REGULATION OF PROTEIN 1 0 BIOSYNTHESIS BY SCALE RESONANCE ABSTRACT OF THE DISCLOSURE The present invention relates to a method for epigenetic regulation of protein biosynthesis which consists in using the regulating action, on the biosynthesis of proteins, by scale resonance, of transpositions into sound of temporal sequences of quantum vibrations associated with their 2 0 elongation. This action may be a stimulation or an inhibition of this biosynthesis, according to whether the modulation of the vibration frequencies used is in phase or in phase opposition to said elongation; the result hitherto obtained is further stabilized by the action of colored transpositions of grouped quantum vibrations arising from the spatial 2 5 conformation of proteins issued from said elongation. Applications, especially in the fields of agronomy and health, include for implementation purposes, a method allowing to delimit the metabolic roles of proteins using their aminoacid sequence.