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Original Text Copyright © 1996 by Astro, Ltd. English Translation Copyright © 1996 by åÄàä ç‡ÛÍ‡ /Interperiodica Publishing (Russia).
LASER METHODS IN CHEMISTRY, BIOLOGY, AND MEDICINE
Is It Possible to Create a Laser Based on Information Biomacromolecules?
A. A. Berezin*, P. P. Garyaev*, V. S. Gorelik**, S. A. Reshetnyak**, and V. A. Shcheglov**
*Department of Theoretical Problems, Russian Academy of Sciences, Moscow, Russia **Lebedev Physical Institute, Russian Academy of Sciences, Leninskii pr. 53, Moscow, Russia
e-mail: optdep@sci.ﬁan.msk.su Received August 2, 1996
Abstract—A method of enhancement of luminescence from information biomacromolecules by the introduction of an activator (donor) into the samples under study is proposed. The experimental and theoretical studies of two pairs of mixtures (DNA + dimedrol and nucleohiston + dimedrol) demonstrate that a superﬂuorescence regime can be implemented in these two-component mixtures under two-photon excitation. Several intrinsic metabolites of biosystems (nucleotides, vitamins, alkaloids, etc.) can function in vivo as dimedrol-like activators. We formulate a hypothesis that the laser pumping of genetic structures is implemented in vivo due to the energy of the corresponding ATP systems.
The truth always wins, but sometimes plays a draw.
Several decades have past since the Nobel Prize winners Academicians A.M. Prokhorov and N.G. Basov (Russia) and Ch. Townes (USA) put forward and implemented the idea of quantum oscillators. At present, one can hardly name a branch of science and technology where quantum oscillators are not applied (from biology and medicine to laser thermonuclear fusion). The successors made a great contribution to the development of this concept. This paper is a priority one. We pose the question as to whether it is possible to create in vitro a laser based on information biomacromolecules, ﬁrst of all, DNA, RNA, and chromosomes. It is difﬁcult to expect the construction of high-power lasers based on these structures. The question is posed in a different manner: we would like to know what new data about DNA, RNA, and chromosomes can be obtained with the help of such a laser through the study of the parameters of its radiation. In our opinion, these investigations will provide fundamentally new results. For example, one can obtain the data on the nonlinear dynamics of DNA, RNA, and chromosomes, including the soliton-type dynamics, rovibrational oscillations, modulation of the dispersion of optical rotation and circular dichroism, energy transfer, and some others layers of information unavailable earlier (with the use of such methodology). Dynamic modiﬁcations of the laser beam of this type can be of the semantic–genetic–biosymbolic type and, therefore, can possess a tremendous biological activity. Earlier, we suggested the initial ideas on this subject [1–3]. Speciﬁcally, we discussed the idea of the construction of a laser system using the Frölich modes [1, 2]. Yu.N. Zhivlyuk (Russia) expressed an interesting idea about the possibility of construction of a biolasers based on cellular structures (private communication).
This idea is based on the possibility of application of phase transitions at the secondary, tertiary, and higher structural levels of biomacromolecules. The difﬁculty of the demonstration of the correctness of all these ideas is associated with the fact that the majority of genetic structures containing aromatic and heterocycle rings are “transparent” within the characteristic spectral range of 350–400 nm. Another difﬁculty is that high-power pumping would inevitably destroy “fragile” biostructures, which has been demonstrated experimentally by the research group headed by S.G. Rautiman, the Corresponding Member of the Russian Academy of Sciences. To implement some of the discussed concepts, we carried out an in vitro investigation of the spectra of two-photon-excited luminescence (TPEL) of gel–liquid-crystal samples of nucleohiston, which is a sum fraction of chromosomes where histon proteins and DNA dominate (standard high-polymer preparations of Sigma). To substantially increase the intensity of TPEL of genetic structures, we suggested a method of activation of luminescence by the introduction of activators (donors) of TPEL into the samples under study. The absorption spectra of organic molecules used as activators are close to those of DNA and nucleohiston. These molecules are characterized by a high intensity of emission spectra that lie within the areas of absorption of DNA and nucleohiston. We used crystalline dimedrol as an activator. Its structure contains a pair of benzene rings, which provides an intense asymmetric band of TPEL in a broad spectral range of 280–350 nm (the dashed line in Fig. 1). We used a copper-vapor laser for photon pumping of the samples under study. This laser operated in a standard pulse–periodic regime at the repetition rate of
1212 I, rel. units 100 ×10 ×1 50 1 2 290 310 330 350 ×10
BEREZIN et al.
370 390 λ, nm
detection system consisted of a ﬁlter for the selection of laser lines (510.8 and 578.2 nm), a ﬁlter for the selection of luminescence in the UV and violet ranges (with suppression of laser radiation), a monochromator (MDR-2) for spectral scanning within a broad range (from UV to visible), a two-coordinate plotter for spectra recording, and a unit for the control of the reference signal and the determination of the efﬁciency of the detected signal. We used a strobe pulse with a duration of 25–30 ns synchronized with the excitation pulse to suppress thermal ﬂuctuations. The secondary emission pulse was detected by means of an FEU-130 photomultiplier. Figure 1 presents the spectra of a gel–liquid-crystal preparation of DNA mixed with dimedrol (DNA–DL) and of nucleohiston with dimedrol (NH–DL). It is seen that the amplitude of the TPEL spectrum of DNA–DL is only an order of magnitude lower than that of the TPEL spectrum of pure dimedrol. It provides a substantial increase in the TPEL intensity of the DNA–DL mixture in comparison with a pure sample of DNA in the form of solid gel . This spectrum also reveals some other speciﬁc features of the mixtures under study. The quantum yield of TPEL for the NH–DL mixture was lower than that for the DNA–DL mixture. Another typical feature is the increase and quenching of TPEL in time. For NH–DL, the TPEL intensity increased in time (Fig. 2). Curve 1 corresponds to the beginning of the experiment. Curves 2 and 3 (with the increase of luminescence) were detected 30 and 50 min after the beginning of the experiment, respectively. An opposite effect is observed in the case of DNA–DL (Fig. 3). We should note the presence of a vibronic structure in the spectra of TPEL in the form of separate overlapping bands in the range of 310–370 nm, which is especially clearly pronounced for DNA–DL (Fig. 3). Such a structure is close to that observed earlier for the spectra of TPEL of nucleoside triphosphates . A fast quasi-resonant energy transfer from excited molecules of dimedrol to genostructures under study can be the reason for abrupt increase in the quantum yield of TPEL of nucleohiston and DNA in the presence of an activating donor. A ﬁne multiband structure of DNA spectra observed in this case correlates with the type of vibronic bands for a series of aromatic and heterocyclic compounds, including pure nucleoside triphosphates and DNA . This splitting of spectra may be due to transitions of electrons in biomacromolecules from the electronic term S1 to excited vibrational levels of the ground state S0. In this case, a population inversion at the transitions S1–S0 can be implemented if the population of the S1 term is sufﬁcient. Let us estimate the intensity I0 of laser radiation that is necessary for inversion (superﬂuorescence) under conditions of the experiments performed. The conditions of inversion can be written as N1gν / Nν g1 > 1,
LASER PHYSICS Vol. 6 No. 6 1996
Fig. 1. Normalized spectra of two-photon excited luminescence (TPEL) of (dashed line) dimedrol, (1) DNA–dimedrol mixture and (2) nucleohiston–dimedrol mixture.
I, rel. units 50 ×10 2 1 300 320 340 360 380 λ, nm 3
Fig. 2. Dynamics of the increase of TPEL of the nucleohiston–dimedrol mixture in time: spectra are measured (1) at the beginning of the experiment, (2) 30 min, and (3) 50 min after the beginning of the experiment.
I, rel. units 100 ×10 50
1 2 3
380 λ, nm
Fig. 3. Dynamics of the quenching of TPEL of the DNA– dimedrol mixture in time: spectra are measured (1) at the beginning of the experiment, (2) 30 min, and (3) 50 min after the beginning of the experiment.
10 kHz, mean power of 1–3 W, peak power of 106 W, lasing wavelengths of 510.8 and 578.2 nm (green and yellow lines), and pulse duration of 10–20 ns. The laser beam was focused into a spot 2–3 mm in diameter at the sample. This laser was rather efﬁcient for the excitation of TPEL in the studies of electron–vibrational spectra of proteins, DNA, nucleohiston, and their components (purines, pyrimidines, and amino acids [3, 4]). The
IS IT POSSIBLE TO CREATE A LASER BASED ON INFORMATION BIOMACROMOLECULES?
where N1 is the density of working molecules in the S1 state, Nν is the density of molecules in the S0 state, and g1 and gν are the corresponding statistical weights of the quantum levels. The relevant population density can be estimated as (2) N1gν / Nν g1 = P1 / U1 , where P1 (s–1) is the rate of population of the S1 level and U1 (s–1) is the decay rate of this level due to radiative and (or) nonradiative processes. The quantity P1 is estimated as P1 = W /2hντVeffN0 , (3) where W and τ are the energy and the duration of the laser pulse, respectively; Veff = S / βI0 is the effective volume of the medium where two-photon absorption occurs (S is the cross section of the focused light beam at the sample and β is the effective depth of penetration of radiation into the sample); and N0 is the density of biomacromolecules (DNA or nucleohiston). Taking into account relations (1)–(3), one can represent the condition of population inversion for superﬂuorescence as I0 > 2hνN0 SU1τ / Wβ. 10–19 Using typical values, N0 ≅ (1/3) × 1026 m–3, hν = 5 × J (λ = 400 nm), τ ≅ 10 ns, S ≈ 10–5 m2, U1 ≈ 108 s–1, W = 0.2 J, and β ≈ 5 × (10–9–10–10) m Bt–1, we obtain an estimate I0 ≥ 3(1011–1012) Bt m–2, which is close to the value of the intensity in our experiments. Both experimental studies and the corresponding theoretical estimates allow us to assume rather conﬁdently that luminescence enhancement in genostructures is implemented in vitro under the applied regimes of two-photon excitation with the use of dimedrol as an activator. In other words, the emission of DNA and nucleohiston is of a superﬂuorescent type. It is not improbable that endogenic compounds that directly or indirectly interact with DNA and chromo-
somes (steroid hormones; carbohydrates; nucleoside mono-, di-, and triphosphates; some vitamins, e.g., riboﬂavin; aromatic and heterocyclic amino acids; catechol- and indolealkylamines; some antibiotics; drugs, e.g., endogenic morphines, such as ethanol metabolites and peptide endorphynes), alkaloids, toxins, enzyme cofactors, heme-containing proteins, and other multiple organic compounds containing benzene and heterocyclic components play the role of dimedrol-like compounds and act as activators in biosystems. The conditions when in vivo electronic population inversion may occur in genostructures, similar to the conditions implemented in the regimes of TPEL, are not clear. These conditions can be achieved in biosystems, for example, due to photon–phonon interactions in DNA within the framework of the Dicke theory . However, such interactions are related to purely physical mechanisms. As for the physicobiochemical processes resulting in laser pumping of DNA and chromosomes in vivo, we can predict (P.P. Garyaev) the existence of high-power ATP systems in biosystems that deliver energy for the transition of gene structures into Frölich biocoherent states, similar to the states that were obtained as a particular case in our work. This work was supported by the Russian Foundation for Basic Research, project no. 96-02-18855a. REFERENCES
1. Garyaev, P.P., 1994, Wave Genome (Moscow: Obshchaya Pol’za) (in Russian). 2. Blagodatskikh, V.I., Gariaev, P.P., Maslov, M.U., et al., 1996, Laser Phys. (in press). 3. Garyaev, P.P., Gorelik, V.S., Kozulin, E.A., and Shcheglov, V.A., 1994, Kvantovaya Elektron., 21, 603. 4. Agal’tsov, A.M., Garyaev, P.P., Gorelik, V.S., and Shcheglov, V.A., 1993, Kvantovaya Elektron., 20, 371; Gorelik, V.S. and Kozulin, E.A., 1994, Kvantovaya Elektron., 21, 499. 5. Li, K.-H., 1992, Coherent Radiation from DNA Molecules, Recent Advances in Biophoton Research and Its Applications, Popp, F.A., Li, K.H., and Gu, Q., Eds. (Singapore: World Scientiﬁc), p. 157.
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