J Mol Evol DOI 10.1007/s00239-014-9641-0

REVIEW

Life Origination Hydrate Theory (LOH-Theory) and the Explanation of the Biological Diversification Victor E. Ostrovskii • Elena A. Kadyshevich

Received: 9 March 2014 / Accepted: 14 August 2014  Springer Science+Business Media New York 2014

Abstract The Life Origination Hydrate Theory (LOHTheory) considers the life origination process as a sequence of thermodynamically caused regular and inevitable chemical transformations regulated by universal physical and chemical laws. The LOH-Theory bears on a number of experimental, thermodynamic, observation, and simulation researches. N-bases, riboses, nucleosides, and nucleotides and DNAs and RNAs are formed repeatedly within structural cavities of localizations of underground and underseabed honeycomb CH4-hydrate deposits from CH4 and nitrate and phosphate ions that diffused into the hydrate structures; proto-cells and their agglomerates originated from these DNAs and from the same minerals in the semiliquid soup after liquation of the hydrate structures. Each localization gave rise to a multitude of different DNAs and living organisms. The species diversity is caused by the spatial and temporal repeatability of the processes of living matter origination under similar but not identical conditions, multiplicity of the DNA forms in each living matter origination event, variations in the parameters of the native medium, intraspecific variations, and interspecific variations. The contribution of the last to the species diversity is, likely, significant for prokaryotes and those eukaryotes that are only at low steps of their biological organization; however, in the light of the LOH-Theory, of available longterm paleontological investigations, and of studies of V. E. Ostrovskii (&) Karpov Institute of Physical Chemistry, Vorontsovo Pole str. 10, Moscow 105064, Russia e-mail: [email protected]; [email protected] E. A. Kadyshevich Obukhov Institute of Atmospheric Physics RAS, Pyzhevsky side-str. 3, Moscow 119017, Russia e-mail: [email protected]

reproduction of proliferous organisms, we conclude that, in toto, the contribution of interspecific variations to the species diversity was earlier overestimated by some researchers. The reason of this overestimation is that origination of scores of «spores» of different organisms in any one event and multiple reproductions of such events in time and Earth’s space were not taken into consideration. Keywords Living matter origination LOH-Theory  Origins  Biological diversification explanations  Life origination thermodynamics  Evolution biological  Monochirality of DNA

Introduction: Interconnection Between the Problems of Life Origination and Biological Diversification Allow us to begin our consideration from the life concept definition. This problem has been under spirited discussion since 1935, when Stanley pioneered isolation and crystallization of the tobacco mosaic virus. It is considered by us in (Ostrovskii and Kadyshevich 2007), where the opinions of different authors are presented. The problem consists in the fitting of the boundary line between the living and mineral substances. Today, viruses and even viroids are regarded as biological substances and are studied by biologists. Each of these species contains DNA of a specific composition. Therefore, we regard the formation of nucleic acids from minerals as the onset of the simplest pre-cellular life. If a living system were devoid of nucleic acid, with its protein content preserved, the vital activity would surely cease; if a living system were devoid of its protein, with the nucleic acid preserved, the vital activity of the system supplied with nutrients might normalize with time. Apparently, once nucleic acids had originated and

123

J Mol Evol

propagated and a medium appropriate for their existence and replication had appeared, the appearance of cellular life was merely a matter of time. Thus, in our terminology, DNAs and RNAs are the simplest forms of pre-cellular living matter, and cells and cellular agglomerates are the valid living organisms, while nitrogen bases (N-bases), riboses, nucleosides, and nucleotides are the living matter simplest elements (LMSEs) or constituents of living matter. Just the understanding of the role of nucleic acids and revealing of their chemical composition and structure were the acme of perfection in the twentieth century. Although nucleic acids were discovered by F. Miescher as early as in 1869 (Dahm 2008), their critical importance for the existence of living matter was established as late as in the midtwentieth century. Before this, an opinion existed that protein is the key substance characteristic for living matter. However, chemical studies of protein have given no effective Ariadne’s clews to unravel the naturalistic way that could lead to living matter origination. Through the centuries, the Biblical version prevailed, and, along with it, hypotheses of spontaneous generation, putrefaction of preexisting organic compounds, Oparin’s hypothesis of 1924, and those akin to the last arose and retired from the pages of scientific journals. Authors of the last group of hypotheses took arbitrarily that external energy is necessary for syntheses of living matter from minerals and considered different sources of external energy, such as atmospheric electrical discharges, thermal springs, etc. Meanwhile, thermodynamic calculations (Lide 1996; Ould-Moulaye et al. 2001, 2002; Ostrovskii and Kadyshevich 2006) showed that no external energy is necessary for syntheses of biologically active substances from simple minerals; moreover, significant amounts of energy evolve during such processes. In the mid-twentieth century, hypotheses of cosmic origination of the Earth’s living matter were proposed, but they considered not the way but the place of origination of living matter sources. This list of available hypotheses is not exhaustive. Ch. Darwin (1809–1882), one of the greatest naturalists and geologists of the nineteenth century, refuted putrefaction of pre-existing organic substances and did not exclude spontaneous generation. He was by no means consistent materialist. Darwin, apparently, did not believe in the possibilities of a definite scientific solution of the life origination problem and believed that, originally, only one or a few living forms appeared on the Earth; for example, he wrote in 1868 that ‘‘…facts have as yet received no explanation on the theory of independent Creations; they cannot be grouped together under one point of view, but each has to be considered as an ultimate fact. As the first origin of life on this earth, as well as the continued life of each individual, is at present quite beyond the scope of science, I do not wish to lay much stress on the

123

greater simplicity of the view of a few forms, or of only one form, having been originally created, instead of innumerable miraculous creations having been necessary at innumerable periods….’’ (Darwin 2010, v. 1, p. 12) Thus, Darwin’s Evolutionary Theory was not associated with any concrete mechanism of living matter origination and Darwin knew nothing about the role of nucleic acid molecules, whose chemical compositions determine the species characteristics of organisms capable of developments on the basis of multiple replications of these nucleic acids. By the way, we know at present that origination of nucleic acid molecules of a definite composition should precede the origination of any species of living organisms and that each nucleic acid molecule has a number of features inherent in all nucleic acids and a number of specific features inherent in each individual species. If different DNA molecules and different cellular organisms could originate in any one localization and if there were a lot of such localizations, the requirements to the subsequent DNA modifications in the course of the evolution could be multiply decreased as compared with the case considered by Darwin. Darwin died more than 130 years ago. For these years, multiple paleontological studies and experiments with proliferous organisms were performed, principally new approaches to the problem of living matter origination and development were proposed, and new hypotheses of living matter origination and development were published. The results of these studies and, in particular, new interpretation that is given to the mechanisms of origination of DNAs and proto-cells by the Life Origination Hydrate Theory (LOH-Theory) (Kadyshevich and Ostrovskii 2007, 2009; Ostrovskii and Kadyshevich 2006, 2007, 2011, 2012a, b) can modify the role of evolution in the variety of Earth’s flora and fauna and can harmonize the real historical role of evolution with the results of the paleontological studies and experiments with proliferous organisms. According to this new interpretation, living matter had appeared at the Earth repeatedly and in different underseabed and underground localizations; therewith, a lot of different living cells with different DNAs had originated in each of these localizations. We will show below that the LOH-Theory is capable of clearing up the living matter origination and its earlier development history and of revealing the biological diversification additional sources and that the conclusions following from this theory are in the movement of the conclusions following from a number of the paleontological and laboratory findings. The LOH-Theory is developed on the basis of a general approach, which was also applied by us to clarify the mechanisms of such natural global processes and phenomena as the cellular mitosis and DNA replication (Mitosis and Replication Hydrate Theory, MRH-Theory)

J Mol Evol

(Kadyshevich and Ostrovskii 2007; Ostrovskii and Kadyshevich 2011), solar system formation (Physically Formed Objects–Chemically Formed Objects Hypothesis of Solar System Formation, PFO–CFO hypothesis) (Kadyshevich 2009; Kadyshevich and Ostrovskii 2007, 2010, 2011), stellar transformations (Kadyshevich and Ostrovskii 2013; Ostrovskii and Kadyshevich 2013), and Earth’s natural gas formation (Ostrovskii 2010). Briefly speaking, this approach is as follows. We state that all natural global processes and phenomena result from thermodynamically conditioned, natural, and inevitable chemical transformations governed by universal physical and chemical laws. If a phenomenon looks as a random one, this testifies that our information about it is too scant and it is sufficient to increase the time and space scales to be sure of its consistency with natural regularities and, in particular, of a decrease in the free energy of the system as a result of this phenomenon. Just as a result of the thermodynamically caused directedness of natural phenomena and processes, researchers are principally capable of mental doubling back the way the nature went and, thus, of revealing the main milestones in Nature’s movement. A naturalist must search for a ‘‘hook’’ in the environment in order to catch on it and, having the thermodynamic laws as the guiding thread, to guess the logics used by Nature in its temporal transformations. At the present time, in contrast to the nineteenth century and first five decades of the twentieth century, there are no doubts that just DNAs represent the simplest form of the precellular living matter and that, depending on the sequence of the N-bases, a DNA molecule, being located in a suitable medium under suitable conditions, is capable of producing an absolutely predictable plant or animal living entity. Therefore, apparently, it should be obvious that the problem of decoding of the natural mechanism of living matter origination cannot be solved without decoding of the mechanism of chemical formation of DNAs from minerals. Meanwhile, this is only one of the sub-problems of the general living matter origination problem. The second sub-problem is decoding of the development of the mechanism of extended self-reproduction of the DNA molecules. The LOH-Theory and MRH-Theory together are aimed at solution of these two sub-problems. Meanwhile, this review is dedicated to consideration of the degree of compatibility between the LOHTheory and some available experimental data, on the one hand, and the notions of the biological diversification, on the other hand. To make this, we consider necessary to demonstrate the experimental, philosophical, and observational sources of the LOH-Theory and to present some materials that count in its favor. In more detail, our theory and related subjects are considered in (Ostrovskii and Kadyshevich 2012b) and in the publications that are referred there. Notice the following. Today any other form of life, but that existing around us, is unknown and, therefore, when

writing about origination of life, we keep in mind the process that could proceed no matter where over the Universe under the corresponding conditions and, of course, under the occurrence of all necessary mineral substances.

Chemical Sources of Living Matter: Each Natural Phenomenon or Entity is Produced from Minimal Variety of Simpler Ones Figure 1 presents some information on the carbon-containing components of DNAs and RNAs. This information will be useful for the subsequent consideration. Figure 2a presents a fragment of the DNA mono-strand structure, where Ad, G, Cy, and Th are N-bases; adenine and guanine are purines; and cytosine and thymine are pyrimidines. Figure 2c presents the DNA double helix scheme. In it, each spiral ribbon symbolizes the DNA chain similar to that presented in Fig. 2a and each horizontal segment symbolizes the hydrogen purinepyrimidine bond. Figure 2b presents the chemical compositions and structures of the hydrogen-bound purinespyrimidine ‘‘liaisons,’’ which connect the DNA mono-strands in double helixes (the circles shown around the N-bases will be explained below). Each DNA represents a polymer molecule and consists of radicals of a sugar [desoxy-D-ribose (DDR)] joined to one another through phosphate bridges and connected with N-base radicals as shown in Fig. 2a. RNA molecules are similar to DNAs in their structure, but contain uracyl (U) instead of Th and sugar D-ribose (DR) instead of DDR. The DNA and RNA molecules, which are characteristic for some organisms, can also contain other N-bases, for example, xanthine (X) and hypoxanthine (HX). It is important that all DNA and RNA molecules include only five chemical elements, namely H, C, N, O, and P. Therewith, all N-bases and riboses that enter into the compositions of DNA and RNA molecules are formed by four chemical elements: H, C, N, and O. Just this feature allows the conclusion about the chemical nature of the minerals which are the sources of nucleic acids. To reveal these minerals, we used the Newton’s principle ‘‘Nature is simple and does not luxuriate in excesses’’ and approached to the subject with the following question: ‘‘Are there two minerals capable of forming, as a result of their chemical interaction, the N-bases and riboses which enter into the composition of all DNA and RNA molecules?’’ As sources of carbon, we assumed methane, ethane, or propane, because these hydrocarbons are the simplest and most abundant carbon-containing minerals and because they contain hydrogen, which also is one of the building materials for DNA and RNA molecules. As sources of nitrogen and oxygen, we assumed a niter, namely, NaNO3 or KNO3, by reasons considered below. The fact is that the

123

J Mol Evol Fig. 1 N-bases and riboses that are the precursors of the nucleic acids

123

J Mol Evol

Fig. 2 a Fragment of a DNA molecule; b scaled schematic representation of pairing between N-bases of two DNA molecules in the double helix structure; the specified circles mark the corresponding atoms, unspecified circles correspond to H-atoms, circles of diameter 0.69 nm correspond to the free diameter of the large cavity in hydrate structure II; c schematic representation of a double helix formed by

two DNA molecules; d scaleless scheme of pairing between polyacrylamide (acrylamide) molecules; e scaled schematic representation of a phosphate group inside a small cavity of hydrate structure II (a circle of diameter 0.48 nm corresponds to the free diameter of the small cavity)

process of formation of LMSEs consists of redox reactions. Therefore, the nitrogen valence state in the oxide is of prime importance for determining the feasibility of the process under consideration. We stated that, indeed, the reaction of CH4, C2H6, or C3H8 with niter is capable of giving all phosphatefree LMSEs that enter into the DNA and RNA compositions (Ostrovskii and Kadyshevich 2006, 2007, 2012b). From ethylene and niter, the full set of N-bases and riboses, which is necessary for formation of DNAs and RNAs, cannot be synthesized; for this aim, the third component, namely N2, is necessary (Ostrovskii and Kadyshevich 2006). Thus, we

stated that the reaction between a nitrate and CH4, C2H6, or C3H8 could be the most laconic way of formation of all necessary phosphate-free LMSEs. This conclusion is confirmed by the example of the reactions written below, where CH4 is used as the source of carbon. Before considering these reactions, we will give some preliminary notes. It is known that the molar ratios Ad/Th and G/Cy in DNA molecules and the molar ratios Ad/U and G/Cy in RNA molecules are equal to unity. Meanwhile, in different species, the relative numbers of Ad and G are not the same (for example, in the human sperm, the Ad, G, Th, and Cy

123

J Mol Evol

percentages are about 31, 19, 31, and 19). We consider below a hypothetical averaged situation, when the molar ratio Ad/G is equal to unity, and solve the question of whether such a situation can arise on the basis of reactions between CH4 and KNO3 in a medium that contains no other chemically active elements. In other words, we consider the following two pairs of reactions: reactions (1) and (2) between KNO3 and CH4, which lead to formation of equimolecular quantities of Cy, G, Ad, and Th and to simultaneous formation of DDR in a molar ratio of 1/1 to the sum of these N-bases (this is the material for the subsequent DNA formation) and reactions (3) and (4) between KNO3 and CH4, which lead to formation of equimolecular quantities of Cy, G, Ad, and U and to simultaneous formation of DR in a molar ratio of 1/1 to the sum of these N-bases (this is the material for the subsequent RNA formation). These reactions could be also written for any other molar ratios Ad/G and Th/Cy in DNA molecules and Ad/G and U/Cy in RNA molecules; however, just these forms of Eqs. (1)–(4) are the most convenient for their further consideration. Reactions (1) and (3) are written for the conditions when KNO3 is in excess, and reactions (2) and (4) are written for the conditions when CH4 is in excess. In this instance, we analyze the feasibility of the stoichiometric balance for such reactions, i.e., the principal possibility of their going and consider neither their thermodynamics nor their kinetics, i.e., neither their direction nor their rate. 28:2 KNO3 þ 39 CH4 ! C5 H6 N2 O2 ðThÞ þ C4 H5 N3 O ðCyÞ þ C5 H5 N5 O ðGÞ þ C5 H5 N5 ðAdÞ þ 4 C5 H10 O4 ðDDRÞ þ 28:2 KOH þ 33:4 H2 O þ 6:6 N2 þ 1:5 O2 ;

ð1Þ

27 KNO3 þ 39 CH4 ! C5 H6 N2 O2 ðThÞ þ C4 H5 N3 O ðCyÞ þ C5 H5 N5 O ðGÞ þ C5 H5 N5 ðAdÞ þ 4 C5 H10 O4 ðDDRÞ þ 27 KOH þ 34 H2 O þ 6 N2 ; ð2Þ 29:4 KNO3 þ 38 CH4 ! C4 H4 N2 O2 ðUÞ þ C4 H5 N3 O ðCyÞ þ C5 H5 N5 O ðGÞ þ C5 H5 N5 ðAdÞ þ 4 C5 H10 O5 ðDRÞ þ 29:4 KOH þ 31:8 H2 O þ 7:2 N2 þ 1:5 O2 ; ð3Þ 28:2 KNO3 þ 38 CH4 ! C4 H4 N2 O2 ðUÞ þ C4 H5 N3 O ðCyÞ þ C5 H5 N5 O ðGÞ þ C5 H5 N5 ðAdÞ þ 4 C5 H10 O5 ðDRÞ þ 28:2 KOH þ 32:4 H2 O þ 6:6 N2 ð4Þ It is seen that, indeed, reactions of methane with niter (i.e., ?5-valence nitrogen in combination with -4-valence carbon) could be the sources of nucleic acids under the conditions that they are thermodynamically allowed and

123

proceed rather slowly in such a way that the thermodynamic equilibrium has time to establish over the system at each reaction step, i.e., if their rates are determined by the movement of the thermodynamic front. It is important that NH3 is incapable of supplying redox reactions, which can lead to formation of all LMSEs; moreover, NH3, as will be shown below, is, under definite conditions, a product of the KNO3 and CH4 interaction. Of course, the interaction in the system (KNO3 ? CH4) could lead to a maximum decrease in the free energy per one KNO3 mol, if the reaction proceeded up to CO2 formation by the equation KNO3 þ CH4 ! CO2 þ NH3 þ KOH:

ð5Þ

The causes of inhibition of this reaction and of preferable formation of nucleosides through reactions (1)–(4) are considered in detail in (Ostrovskii and Kadyshevich 2012a, b). In short, they are as follows: (1) low temperature and, as a consequence, the thermodynamic front, i.e., slow steadystate C (-4) oxidation by N (?5); (2) DNA kinetic stability (the DNAs of the frozen mammoths are available even today); (3) elimination of N2 (produced at the step of DNA formation) from the reaction zone, where it could be principally capable of providing the full DNA oxidation up to CO2. These features summed with the occurrence of excessive CH4-hydrate provide the stability of nucleosides.

The LOH-Theory as the Unique Thermodynamically Grounded Theory of Living Matter Formation A detailed thermodynamic consideration of this problem was given earlier (Ostrovskii and Kadyshevich 2006, 2007, 2012a, b; Kadyshevich and Ostrovskii 2009). Below, we give general results obtained in these works. For a reaction i, we calculated DiG = Di(DfHj) T DiSj, where DiG, Di(DfHj), and DiSj are the standard Gibbs free energy, enthalpy, and entropy changes in this reaction. For reactions (3) and (4), the following results were obtained (kJ/mol): D3G = D3(DfHj) - T D3Sj = -9,156 ? 298.15 • 3.471 = -8,121 and D4G = D4 (DfHj) - T D4Sj = -9,410 ? 298.15 • 3.787 = -8,281. It is seen that the enthalpy and Gibbs free energy changes are negative for both reactions and so great in their magnitudes that variations in the ambient conditions within wide limits could not make them positive. Remind that these thermodynamic calculations are performed for ([Ad]/ [G]) = 1. It was shown by us (Ostrovskii and Kadyshevich 2012b) that [Ad]/[G] variation in RNA (and DNA) from 0.0625 to 16.0 leads to change in the DiG value by 109 kJ/ mol, i.e., by 1.3 % only. Thus, practically under any conditions, reactions (3) and (4) should proceed from left to right. On the basis of thermodynamic calculations, we also

J Mol Evol

showed (Ostrovskii and Kadyshevich 2012a, b; Kadyshevich and Ostrovskii 2009) that the products of methane oxidation by niter should contain Th, Ad, G, Cy, U, DR, and DDR all together; therewith the partial concentrations of these products depend on the ambient conditions and methane/niter ratio. This means that methane and niter could react predominantly by reactions (1), (2), (3), or (4) in different epochs. We should denote an intriguing feature of these reactions. We see that all they produce N2, i.e., the gas, which is the main component of the present Earth’s atmosphere. When niter is in excess, N2 and O2 are produced together and, therewith, the volumetric N2/O2 ratio is equal to 0.815 and 0.828 for reactions (1) and (3), respectively. In the present atmosphere, the volumetric ratio (N2/O2) = 0.788. These values are closely related, and this fact makes one think; however, we abstain at present from its discussing because this goes beyond the scope of the problem under our consideration. On the basis of thermodynamics, a number of other problems, which relate to the living matter origination, were solved by us. One of them is the following. Why do only five N-bases usually enter into the DNA and RNA compositions, and why are the other ones random? To understand the principle that could underlie this ‘‘lusus naturae’’ (caprice of Nature), we consider the reaction between G and water with formation of X. Let the reactions leading to formation of N-bases and riboses proceed in a closed system and an equilibrium be established after a time. It is clear that equilibrium in the reaction system suggests equilibrium between all its components, in agreement with the detailed equilibrium principle; this principle allows us to elucidate whether X can exist in the system containing G and H2O. C5 H5 N5 O ðGÞ þ H2 O $ C5 H4 N4 O2 ðXÞ þ NH3

ð6Þ

Calculations show that, under standard conditions, D6G = 7.32 kJ/mol. This estimate means that equilibrium (6) is shifted to the left and X formation is thermodynamically disadvantageous. Apparently, the analogous cause hampers entering of other N-bases but Ad, G, Th, Cy, and U into DNA and RNA molecules. For equilibrium (6), the absolute change in the free energy is small; therefore, nucleic acids may contain X under certain conditions differing from the standard ones. Indeed, X sometimes enters into the compositions of natural nucleic acids. Ammonia might be also obtained from CH4 and niter in the absence of xanthine. In a closed system, i.e., under conditions when N2 is not thrown off from the reaction zone and the equilibrium is not shifted to its formation in the course of the reaction between niter and methane, this reaction goes through the state of the system that can be

described as follows (Ostrovskii and Kadyshevich 2007, 2012b). 23:25 KNO3 þ 38 CH4 ! C4 H5 N3 O ðCyÞ þ C5 H5 N5 O ðGÞ þ C5 H5 N5 ðAdÞ þ C4 H4 N2 O2 ðUÞ þ 4 C5 H10 O5 ðDRÞ þ 23:25 KOH þ 22:5 H2 O þ 8:25 NH3 : ð7Þ For this reaction, D7G = -6,146 kJ/mol, i.e., the full set of the RNA precursors can be produced from CH4 and KNO3 with liberation of NH3. A similar result can be obtained for the LMSE set that is necessary for the DNA synthesis. Reactions (6) and (7) show that NH3 can be a side product in the process of living matter formation. The thermodynamic consideration allows the following important conclusions: (i) the chemical interaction between niter and methane, ethane, or propane can provide formation of the entire set of N-bases and riboses necessary for DNA and RNA origination; (ii) the relative yields of different members of this set depend on the conditions and can vary in wide ranges; (iii) just the thermodynamics is instrumental in the selection of N-bases to be further incorporated in nucleic acids; (iv) formation of N-bases, DR, and DDR from hydrocarbons and niters is associated with liberation of O2, or N2, or NH3 or with simultaneous liberation of O2, N2, and NH3 and with enrichment of the Archaean atmosphere with these gases in proportions dependent on the underground reaction conditions (complete reduction of KNO3 by CH4 can also lead to CO2 liberation); (v) a portion of produced NH3 can be used for formation of amino-acids; and (vi) reactions between niters and hydrocarbons could proceed in different epochs and in different CH4-hydrate localizations over the globe. The thermodynamic feasibility of reactions between ribose, N-bases, and phosphoric acid under the standard conditions follows from (Ould-Moulaye et al. 2002). In the course of polycondensation processes, water is liberated, and there are no doubts in their thermodynamic feasibility. A ‘‘crude product’’ obtained from CH4 and niter can wait arbitrarily long until phosphate ions, which are necessary for DNA (and RNA) formation, diffuse to it. Diffusion of phosphates from any remote source led to formation of DNA- and RNA-like molecules and proto-cells as will be shown below.

Something About the Natural DNA and RNA Sources According to the LOH-Theory, DNA and RNA molecules were synthesized by Nature within the phase of the methane-hydrate deposits from methane and nitrate and

123

J Mol Evol

Fig. 3 Intra-structural cavities of hydrate structures I, II, and H: the vertexes are the O-atoms of waters, and the length of each edge corresponds to the sum of the lengths of the O–H valence bond in any water and HO hydrogen bond between this and an adjacent water;

above each cavity, its free diameter, the number of its facets (the superior figures) restricted with the definite number of the edges (the lower-case figures), and the indexes of the hydrate structures into which this cavity is included are given

phosphate ions. Consider some information on the natural abundance of these substances and on their chemical characteristics. Methane hydrate is one of the natural gas hydrates. The underground deposits of CH4 and other hydrocarbons resulted from the reaction between H2 and CO2; CO2 could be produced from carbonates as a result of their thermal decomposition induced by the gravitational compression of the young-Earth crust (Ostrovskii 2010). Hydrogen could be desorbed from the solid aggregates of which the young Earth was composed; these aggregates had adsorbed H2 from the near-presolar star space before they were captured by the Earth’s gravitational force in the period of the Earth origination as a planet body (Ostrovskii and Kadyshevich 2012b). Thus, the living matter sources are H2, carbonates, nitrates, and phosphates, which had resulted from transformation of the presolar-star substances. The CH4 deposits exist predominantly in the form of gas hydrates. Gas hydrates are solid or semi-liquid, mineral clathrate substances characterized by the honeycomb cubic (structure I, a = 1.20 nm), face-centered cubic (structure II, a = 1.73 nm), or hexagonal (structure H, a = 1.23 nm and c = 1.02 nm) lattice composed of large and small cavities, where the waters (hosts) are the vertices of the cavities and other atoms, molecules, or atomic groups (guests) are housed within the cavities (Atwood et al. 1996; Carroll 2003; Chaplin 2013). The first structures of such a kind were discovered early in the nineteenth century. Those substances contained molecules of gases as the guest particles, and, therefore, they were termed gas hydrates. At present, several hundred gas hydrates are known. As guests, particles of one type or two different types can be housed within the large cavities and, in addition, particles of a third type can be housed within the small cavities. Gas hydrates that contain guest particles of two or more different chemical natures are termed mixed gas hydrates. The

structure type depends on the size of the guest particles or on the sizes of the guest particles if particles of two or three types are housed within the cavities. Gas-hydrate structures can exist only under the condition that some guest particles are housed within no less than 75–80 % cavities of, at least, any one type; otherwise, the loose structure collapses and transforms to the usual dense ice. In the hydrate structure II, the ideal water-to-guest ratio is equal to 17 and the critical one is equal to 20721. In gas hydrates, the guest– H2O interactions are provided by the Van-der-Waals forces. Figure 3 shows scaled schemes of the gas-hydrate cavities inherent in structures I, II, and H. Structures I, II, and H contain 512 and 51262, 512 and 64, and 512, 435663, and 51268 cavities, respectively (above each scheme, the lowercase figures mean the numbers of the edges for a facet and the superior figures mean the numbers of the facets that terminate the corresponding cavity). In these schemes, each vertex responds to the O atom of any water and each edge responds to the sum of the O–H valence bond of any water and the HO hydrogen bond of this water with any adjacent one. Figure 3 gives also the sizes of particles capable of being housed within the cavities of any type. Structures I and II have cavities of two types; structure H has cavities of three types. Many clathrate compounds of structure II with 17 ‘‘host’’ waters per one ‘‘guest’’ molecule are well known, e.g., C4H4O17H2O (Stackelberg and Meuthen 1958), (CH2)4 O17H2O (Pinder 1965), CH3Cl17H2O (Stackelberg and Muller 1954), C3H617H2O (Clarke et al. 1964), and socalled mixed hydrates, such as C3H82H2S17H2O (Platteeuw and Van-der-Waals 1959), (CH2)4O2H2S17H2O (Pinder 1964), C3H82CH417H2O (Van-der-Waals and Platteeuw 1959), etc. According to (Byk and Fomina 1970), each unit crystal cell of structure II has a size of 1.74 nm and consists of 136 waters, which form 16 small and 8 large

123

J Mol Evol

cavities with free diameters of 0.48 and 0.69 nm, respectively; the small and large cavities represent somewhatdepressed penta-dodecahedrons and almost spherical hexadecahedrons, respectively. Just these structural parameters were used by us for the calculations presented below. Hydrate structures have a unique peculiarity. The free energy inherent in different hydrate structures is almost the same, and, therefore, they can metamorphose depending on the conditions and on the nature of the guest particles. Variations in the concentrations of the host (water) and/or guest (substrate) components can change the aggregate (structural) state of the systems. For example, when waters slowly enter into an amorphous dried water–substrate system capable of structuring, a gas-hydrate structure arises and develops with a decrease in the free energy up to hydrate formation over the entire system; however, entering of excessive water leads to the disruption of the hydrate structure. This peculiarity is explained by the fact that the structuring is caused by the weak Van-der-Waals forces and, therefore, the molar free energies of the structured and unstructured states are rather close. It will be shown below that this peculiarity of the gashydrate structures is of principal importance for the living matter origination and development; moreover, it is possible that it is the cause of a number of derangements in the regular functioning of living cells and multicellular aggregates (Kadyshevich and Ostrovskii 2007; Ostrovskii and Kadyshevich 2011, 2012b). At present, there are many underseabed CH4-hydrate deposits (Ginsburg and Soloviev 1998; Barenbaum 2007). In 2004, the CH4 mass in the proven marine CH4-hydrate deposits was estimated as (175) million km3 (Milkov 2004), and this estimate grows continuously. Underground CH4-hydrate deposits are also known. The CH4-hydrate deposits are rather deep under the seabed or under the Earth’s surface in the high-latitude regions, where the surface temperature is low (the seabed temperature at depths of more than 400 m is about 274 K, and the stratum temperature at the Earth increases with depth by 3 K per each 100 m). However, it is quite possible that, over colder periods of the Earth’s history, the underground CH4hydrate deposits were throughout the land. The capability for hydrate formation is a fundamental property of water; it can reveal itself in solid and highly concentrated semi-liquid systems at sufficiently low temperatures and sufficiently high concentrations (or pressures) of particles of the sizes that correspond to the free sizes of the gas-hydrate cavities; the chemical nature of the guest particles is not of principal importance for the question on the possibility of hydrate formation in any substrate–water system. Consider some information on the NO3-- and PO43-ions and on their natural sources.

Fig. 4 Nitro-methane– nitronic-acid equilibrium

It is well known that HNO3 is capable of reacting with alkanes under their pressure with formation of nitroalkanes as the primary products (Konovalov’s reaction) (Konovalov 1893). Under room temperature, Konovalov’s reaction proceeds with CH4 very slowly on the scale of duration of the usual laboratory experiments, but Nature has nowhere to hurry. The slowness of these natural reactions and the constancy of the conditions are necessary for thermodynamic controlling the sequence of the reaction steps, for uniform filling all structural cavities by guest molecules of any one composition, and for grading the transformations of the guest molecules in the direction ‘‘from the simple to the complex’’ over the entire volume of a gas-hydrate localization. Nitro-methane is a very active substance; it is capable of subsequent reacting with CH4 and NO3--ions giving more complicated organic substances just around 273 K, i.e., in the temperature range in which CH4-hydrate deposits occur. Nitro-methane reacts in the form of nitronic acid H2C = N?(OH)(O)- (IUPAK term: azinic acid), which exists at temperatures about 273 K and below. Nitronic acid exists in two forms, according to the following equilibrium (Fig. 4). Nitronic acid anions, H2C = N(O2)2-, are degraded to ketones and can be alkylated on oxygen and used as a dipole in the Huisgen’s reaction of the 1,3-dipolar cycloadditions, which leads to formation of pentamerous rings and can initiate different other chemical reactions (Carey and Sundberg 2004). Thus, two substances, NO3- and CH4, being together inside gas-hydrate matrixes, could produce all N-bases and riboses which are necessary for living matter origination. We showed above that thermodynamics allows such interactions and that the occurrence of the water matrix terminates the subsequent chemical transformations of the LMSEs within the hydrate matrix after full filling of the hydrate cavities by the N-bases and riboses. Localizations of phosphates exist everywhere over the globe, and, if there are doubts relative to mineral origin of some of them, it is commonly accepted that apatite Ca5 Y(PO4)3 (Y = F, Cl, or OH) is of mineral origin (Nekrasov 1965). In addition, phosphate ions are dissolved in different water sources. They could be transported to the Earth during and just after the period of its formation, or phosphor-containing compounds could be oxidized in the period of cooling of the young planet.

123

J Mol Evol

It is not impossible that phosphorus, which, according to (Nekrasov 1965), had slowly reacted as follows (the heat effect is given for the standard conditions): P4 þ 16 H2 O ! 4 H3 PO4 þ 10 H2 þ 1306 kJ:

ð8Þ

Other hypotheses of formation of the phosphorus compounds of biological significance are also available (e.g., de Graaf et al. 1995; Gull et al. 2014). The solubility of phosphates in water at 293 K is about 3 mg/l. In ocean water, the contents (mg/l) of N, P, and C are 15.5, 0.088, and 28.0, respectively (Turekian 1968), and, at a depth of 70 m, PO43- content is about only 0.02 mg/l (Atkins 1926). In our days, an enormous number of living organisms exist on the Earth’s surface, near the seabed, and in underseabed layers; therewith, each cell of these organisms contains DNA molecules and each DNA molecule includes a multitude of phosphate radicals. The total amount of phosphorus in vegetable and animal organisms existing now is very large. This means that, formerly, this entire phosphorus was accumulated in the Earth’s subsurface and under-seabed layers. The amount of mineral phosphorus at the primordial Earth was much larger than that at the actual Earth. Today, in the near-surface layers, the average concentration of phosphorus is significantly lower than that of carbon, nitrogen, or hydrogen. This peculiarity in the structure of the Earth will possibly lead to the following phenomenon. In future, just the ‘‘resources’’ of phosphorus will limit the total mass of living organisms on the Earth (in the case that phosphorus will not diffuse to the surface from deeper layers). But there can be no doubt that, in the days of the earlier life, phosphorus was present at the Earth in amounts sufficient for life origination. The seawater thickness is populated by living organisms very sparsely in comparison with the near-seabed layers just because of low concentration of phosphate ions in water; however, the near-seabed layers, even at depths of many kilometers, are abundantly populated, as living organisms get their phosphate ions from the seabed soil. In our time, the plants get phosphate ions from soil and these ions diffuse to their roots, although the phosphorus concentration in the soil is low and phosphorus is sparingly soluble. Other ions do not bind chemically in plant cells, and, therefore, they do not diffuse there from soil. In the same way, phosphate ions diffused into the hydrate structures filled with nucleosides. Nature does not like emptiness. If a definite element binds chemically in some spatial region, a ‘‘vacuum’’ relative to this element arises in the vicinities of this region and atoms of this element diffuse into this ‘‘vacuum’’ from the environment. Therewith, the entropy increases. Just due to the process of intra-soil diffusion, clumps of plants occur on the land and deep in

123

the water, near the seabed. Plants get phosphorus from the soil, and animals get phosphorus by eating plants, which had accumulated it, and by eating other animals, which accumulated phosphorus by eating plants and animals. Some living organisms that have features of both plants and animals are also known. However, plants appeared primarily and, then, some of them ‘‘learned to live’’ and reproduce progeny using plants as ‘‘half-products.’’ The fact that phosphate minerals are sparingly soluble in water does not conflict with the life origination problem solution proposed by us. According to the LOH-Theory, the life developed not in water, but in the methane hydrate solid phase and, then, in the semi-liquid structured supercytoplasm, where nitrate and phosphate ions diffused. All subsequent processes of DNA replication and transformation also proceed in structured protoplasm rather than in water. A doubt may arise whether there are areas where methane, niter, and apatite deposits are located at rather small distances from each other. We cannot resolve this doubt entirely for the Earth’s crust state that had occurred 4000 Myr ago. However, at present, such regions occur. For example, several CH4-hydrate localizations occur along the shelf of California, Oregon, Peru, etc. (Barenbaum 2007; Ginsburg and Soloviev 1998). Meanwhile, several localizations of sodium and potassium niters are known over the coastal continental heights of Chile, Bolivia, California, etc. (Frye 1982; Ericksen 1983) and localizations of apatite are available over Northern Chile, Brasil, and Mexico (Treloor and Colley 1996). All these deposits are rather extensive and their origin gives rise to a lot of talk. For example, Ericksen (1983) writes about the Chilean sodium niter deposits: ‘‘Because of their obscure origin, the nitrate deposits have been of lively discussions for more than 100 years, but none of the dozens of published reports about them gives an adequate or wholly acceptable explanation for the sources for their sole net constituents and their modes of emplacement.’’ There was an opinion that NaNO3 of Atacama Desert resulted from decomposition of the bird metabolism products (BMPs). Meanwhile, it is known that, in the BMPs, the percentage of phosphoric acid (13 %) is higher than that of nitrogen (9 %). If birds in abundance would had visited this region in former times, this should mean that there had been a sufficient amount of water and that wellsoluble niter rather than poorly soluble phosphate should be predominantly washed out by water from the BMPs; if the birds were ‘‘abstemious’’ and this region was always desert, the niter should be mixed at present with phosphate. Meanwhile, the Atacama Desert ore contains NaNO3 (8–27 %), KNO3 (1.5–2.5 %), Na2SO4 (about 2 %), Mg2 SO4 (about 1.6 %), iodides (up to 0.4 %), etc., but contains no phosphate. In addition, the NaNO3 storage in the

J Mol Evol

Atacama Desert is very large: from 1830 to 1980, about 23.4 million metric tons of NaNO3 was produced, and its today storage is about 200 million metric tons (Ericksen 1983). Apparently, there are no doubts that this niter is of natural origin. The Atacama Desert niter deposit is by no means unique. According to the PFO–CFO Solar System Formation Hypothesis (Kadyshevich 2009; Kadyshevich and Ostrovskii 2011), the near-surface localizations of Earth’s minerals were obtained over the period of the Solar System formation, when the hot cores of the terrestrial planets were buried by the cold remote small celestial objects formed by lighter chemical elements.

Experimental Works that Preceded Formulation of the LOH-Theory The principal idea of the LOH-Theory arose as a result of our extensive studies of water interaction with different polymers and monomers. The most intriguing results were obtained by us when studying polyacrylamide (PAA) moistening and drying. These works were induced by our desire not only for understanding the mechanisms of these processes as such, but also for clarifying the effect of water concentration on the processes of formation and destruction of the purinepyrimidine bonds between the DNA mono-strands in double helixes in the process of DNA replication. The fact of the matter is that clarification of the mechanism of DNA replication on the basis of H2O sorption and desorption in the DNA–H2O system is questionable because of the occurrence of a masking effect from the hydrophilic phosphate and ribose groups which enter into the composition of the DNA molecules. Therefore, we used PAA as the model substance, because it is the polymer that contains functional groups similar to the amido-groups (AGs) of DNA and contains no other functional groups capable of H2O sorbing. The similarity between the DNA– DNA and PAA–PAA intermolecular binding is illustrated by Fig. 2b, d, respectively. To make certain that the functional groups of PAA and DNA should be similar in their chemical properties, we studied the available data on the valence angles and lengths of the valence bonds in the AGs of these two substances and had concluded that they are identical (Ostrovskii and Kadyshevich 2000). We studied water vapor sorption and desorption in the PAA–H2O system by adsorption and microcalorimetric methods (Ostrovskii and Tsurkova 1997, 1998a, b; Ostrovskii et al. 2000, 2001); most of the experiments were performed in undisturbed highly concentrated aged semiliquid aqueous media at about 290 K. The processes of H2O sorption and desorption were also studied in the alanine–water and glycine–water systems (Kadyshevich and

Ostrovskii 2007; Ostrovskii and Kadyshevich 2012b). The techniques and procedures of these experiments are detailed in the works cited above in this paragraph; the original FOSKA microcalorimeter is described in (Ostrovskii 2002). Some results and principal schemes of the portable sorption and desorption vacuum glass apparatuses are presented in Fig. 5. As far as we know, neither the differential heats of H2O sorption nor the differential rates of H2O sorption by PAA were studied up to these experiments and the phenomena discovered by us were unknown earlier. The earlier measurements of the integral heats of PAA dissolution in water led to negative and very small magnitudes (Silberberg et al. 1957; Day and Robb 1981). In our experiments, the following specific features of H2O vapor sorption by polymers with functional AGs were revealed. The differential molar heat of H2O desorption (Fig. 5e) at high degrees of dilution is n-independent (n is the waters-to-AGs ratio in the PAA–H2O phase after water vapor sorption); at n [ 17, the differential heat of water vapor desorption measured in seven successive experiments is equal to the tabulated value of the heat of H2O vaporization (QL, horizontal line in Fig. 5e) from the pure H2O surface. The peculiarities in the differential heats of H2O sorption and in other parameters of water sorption and desorption are observable at 0 \ n \ 17. In this n range, the differential heat of water sorption depends on the n value in a complicated way, and this shows that the specific enthalpy and entropy of the system as well as its structural state vary significantly with the n value. At about n = 17, some increase in the differential heat of H2O sorption was observed (Ostrovskii and Tsurkova 1997, 1998a, b) and, as it was shown in (Ostrovskii and Kadyshevich 2012b), the rate of H2O vapor desorption into a degassed volume is decreased by a factor of several units as compared with the system states characterized by either higher or smaller n values. When H2O vapor contacts with PAA in air of almost 100 % humidity at about 290 K (Fig. 5b), water sorption proceeds very slowly and terminates or, at least, moderates critically after a lapse of 400 days at a stoichiometry of (17718) H2O molecules per one AG group; this result was confirmed in two experiments. The procedure of these experiments is simple; it is illustrated by Fig. 5a. Similar experiments showed that the approximately constant rates of H2O vapor sorption by glycine and alanine measured under the same conditions decreased by a factor of several units at n [ 20 (Kadyshevich and Ostrovskii 2007; Ostrovskii and Kadyshevich 2012b). Extensive discussions of these results led us to the conclusion that functional polymers are capable of forming structured hydrates in highly concentrated undisturbed water systems at rather low temperatures. Waters are capable of spontaneous entering into such systems with a gain in the Gibbs free energy up to approximately 17

123

J Mol Evol

Fig. 5 a Water vapor sorption by polyacrylamide (PAA) from air of 100 % humidity at about 290 K; b degree of PAA wetting versus duration of sorption; c apparatus for sorption experiments in the deaired PAA–water system: (1) calorimetric ampoule, (2) top of the calorimeter, (3) test tube, (4) sample for wetting, (5) ‘‘breaker,’’ (6) sealed water-containing glass sphere, (7) neck, (8) mercury manometer, and (9) tube to vacuum setup; d apparatus for desorption experiments: (5) neck, (4) wetted sample or liquid (for other notation,

see c); e calorimetric molar heats of H2O sorption: sorption by sample 8 (filled circle) at 292 K and desorption from samples 7 (filled square), 9 (filled triangle), 10 (open diamond), and 11 (open circle) at 292, 288, 297, and 291 K, respectively (samples 7, 9, 10, and 11 are aged before the experiments for 14, 9, 6, and 16 days, respectively); QL is the heat of H2O vaporization from the pure water surface at 290 K

waters per one functional group of a polymer; the subsequent forced introduction of waters into the water/substrate systems leads to disruption of the systems and to formation of solutions. Functional organic monomer molecules are also capable of forming such structures; however, the molar free energy gain at formation of such hydrates is smaller than that in the case of functional polymers. In the review works (Ostrovskii and Kadyshevich 2000, 2002a, 2011; Ostrovskii et al. 2000, 2001; Kadyshevich and Ostrovskii 2007), we arrived at the conclusion that the scientific explanation of the living matter origination and development was veiled by Nature in the hydrogen interactions between water and chemical substances and in physicochemical laws that govern these interactions. It was stated that, in the in vivo quasi-equilibrium DNA-doublehelix–H2O systems with n & 17, each N-base is encircled by a water envelop, which consists of about 17 H-bound waters; the subsequent forced introduction of waters into the system leads to continuous neutralization of the pair interactions between the DNA strands as a result of formation of a water continuum. Therewith, the DNA double helixes become steadily unwound and the DNA mono-

strands develop self-identical chemical structures at their one-dimensional surfaces, whereupon, the newly formed double helixes float away from each other. This conclusion is of fundamental importance for understanding the living matter origination mechanism and also the mitosis and DNA replication mechanism.

123

The Life Origination Hydrate Theory: From Minerals to Protocells The scientific studies considered above led us to the following principal conclusions, which initiated formulation of the LOH-Theory and were used as its basis. (1)

The most laconic and thermodynamically allowed process that could lead to formation of different DNA and RNA molecules should include the chemical interaction between three minerals widely distributed on the Earth: methane, niter, and phosphate; these minerals are also sufficient for formation of all principal amino-acids, proteins, and other principal biologically important compounds.

J Mol Evol

(2)

(3)

(4)

(5)

The components of the DNA (RNA) molecules are in size conformity with the cavities of the gashydrate structure II. Methane hydrate is a widely distributed mineral, and, on the Earth, there are regions where CH4hydrates, niter, and phosphates neighbor. In highly concentrated semi-liquid PAA–water and aminoacid–water systems, the host–guest structures, which are similar to those in the solid gas hydrates, form at rather low temperatures and under undisturbed conditions. In the in vivo quasi-equilibrium H2O–DNA double helix systems with n & 17, each N-base is encircled by a water envelop, which consists of about 17 H-bound waters, and the subsequent forced introduction of waters into the system leads to a continuous neutralization of the paired interactions between the DNA strands as a result of formation of a water continuum; therewith, the DNA double helixes become steadily unwound and the DNA mono-strands develop self-identical chemical structures at their one-dimensional surfaces, after which the newly formed double helixes float away from each other.

The LMSEs, DNAs, RNAs, amino-acids, and proto-cells originated repeatedly and, possibly, originate in our days from CH4 (or C2H6, C3H8) and nitrate and phosphate ions under the seabed or under the Earth’s surface within gashydrate structures under stationary ambient conditions. The reactions of LMSEs formation could proceed under CH4 pressure and at a temperature of about 273–293 K, i.e., under the conditions when CH4-hydrate is stable and aqueous solutions are not frozen. As we stated, gas-hydrate structures are typical not only for solids but also for highly concentrated semi-liquid undisturbed substrate–water systems at rather low temperatures. Meanwhile, the movement of thermodynamic fronts proceeds in semi-liquid systems much more smoothly than in solid ones. The maintenance of thermodynamic fronts is of primary importance for production of LMSEs and for subsequent production of DNAs and RNAs. The processes under consideration proceeded in semi-liquid structured systems. It is important to take into account that the hydrate formation processes and, in particular, the CH4-hydrate formation, proceed with a decrease in the Gibbs free energy of the systems. Therefore, the CH4-hydrate crystallization/destruction process goes at a higher temperature than the water freezing/ melting process. The LMSEs formation processes went by the following schemes. Pressed CH4 penetrated into underseabed or underground karst cavities or into sand stratums filled with nitrate and phosphate solutions produced as a result of

dissolving distant deposits of these minerals. When the CH4 pressure increased up to a definite critical level, CH4hydrate structures with included nitrate and phosphate ions arose. Therewith, the above-considered chemical reactions leading to formation of the LMSEs and DNA and RNA molecules proceeded over the entire volume of the systems. It is also possible that the nitrate and phosphate solutions diffused from outside into the volumes filled preliminary with CH4 and that diffusion of these solutions led to structuring of the systems. The situations when two or several flows of CH4 and solutions enriched with PO43and NO3- met together within any underseabed or underground cavity or within any porous stratum and initiated formation of DNAs and RNAs are also possible. It is important that CH4 molecules reacted selectively with NO3--ions, because methane is characterized by extremely low reaction ability and is capable of interacting almost exclusively with nitrate ions under the ambient conditions appropriate for the CH4-hydrate formation process; as for the phosphate ions, they also represent one of rather rare substances that are capable of reacting in the system under consideration. Just the thermodynamics is instrumental in selection of N-bases to be further incorporated in the composition of nucleic acids. As an example, we showed above that, for the reaction between G and H2O with formation of X and NH3, D(G) = ?7.32 kJ/mol. This value means that, in a system, where reactions proceed in the vicinity of 273 K and so slowly that the equilibrium relations between reacting components keep constant in time, G should usually prevail over X but, under some conditions, the relative amounts of the last may be noticeable. This is the cause of the usual significant prevailing of G over X in the DNA molecules (Kadyshevich and Ostrovskii 2009). The thermodynamic front governs the processes inside the hydrate structure. At the beginning, nitrate ions react with CH4; the reactions go in different cavities indiscriminately; but, when any small cavity becomes filled up, the resulted molecule ‘‘jumps’’ into an adjacent large cavity, and the thermodynamically directed reaction continues within this cavity up to its full occupation. In the first period, all molecules grow by the same thermodynamically caused plan; but then they begin to hinder one another and occupy the residual empty space in different manners. The formation of cyclic molecules is most economy relative to the occupied space and to the integrated gain in the free energy; therefore, just cyclic molecules form (as was noted above, reactions of nitronic acid anions with CH4 lead to formation of ring structures). When the subsequent increase in the size of the cycles becomes impossible, the side groups fill the opens in the large cavities. Therewith, all N-atoms are with many uses because of their different

123

J Mol Evol

valences and high gain in the free energy resulted from their entering into the newly formed molecules. There are no prizes for guessing that the composition of N-bases within the large cavities at their full filling cannot be identical. Each N-base joins the excessive hydrocarbon and oxy-hydrocarbon radicals and grows into the adjacent small hydrate cavity with the gradual formation of a nucleoside. These processes are governed by the thermodynamic front and reactivity of the radicals that form under the reaction conditions and are limited by the sizes of the hydrate cavities. In Fig. 2b, e, the diameters of the large and small circles respond to the free volumes of the hydrate structure II large and small cavities, respectively. It is seen that the sizes of N-bases and phosphate ions correlate well with the sizes of the large and small cavities. The large and small cavities as if are created as the molds for the N-bases and phosphate ions, respectively. In order that the H-bond would arise between purine and pyrimidine bases, the distance between them should be strictly fixed. Our two-dimensional consideration allows the conclusion that the hydrate structure with its systematically located large cavities creates the geometric conditions necessary for formation of H-bonds between these N-bases. If two molecules of purine bases were formed inside two neighboring large cavities of the hydrate structure, the distances between the oppositely polarized groups belonging to these two molecules would be smaller than the equilibrium ones and, therefore, the polar groups of these molecules would inevitably ‘‘turn away’’ from each other. Therefore, the two-dimensional consideration gives grounds to assume that the H-bound purine–pyrimidine associates originate at the step of their formation from mineral substances within the gas-hydrate structures. Phosphate ions form bridges between nucleosides, joining them to each other by polycondensation reactions, which lead to formation of DNA- and RNA-like polymer molecules. These molecules are similar, because each nucleoside consists of one ribose-like radical and one purine or pyrimidine radical, but not identical, because nucleosides contain different ribose radicals and different purine and pyrimidine radicals and because the nucleosides enter into nucleic acids in different successions. Apparently, DNA molecules originate in the form of dimers within the hydrate structures. In (Ostrovskii and Kadyshevich 2012a), it was shown that a combination of the effects of hydrate-structure matrix and thermodynamic controlling of the chemical reactions as applied to the system under consideration creates conditions for specific and by no means random locations of N-bases in the resulted DNA-like and RNAlike molecules and that a variety of different and specific nucleic acids originated in any one CH4-hydrate localization. The importance of nonrandom arrangements of N-bases was noted in (Abel and Trevors 2006; Trevors and

123

Abel 2004). At present, we cannot explain why molecules of two types, DNAs and RNAs, formed. We think that this feature is conditioned by delicate geometric peculiarities, which exert themselves in a specific filling of the structural cavities at the last steps of formation of N-bases and riboses within the hydrate structure. Under the influence of water that diffused into the system and/or evolved as a result of the polycondensation reactions, the solid structure transforms to a structured semi-liquid soup in which, on the basis of methane and of nitrate and phosphate ions that diffused into the system, the simplest living organisms began the long history of their development, complexification, perfection, symbiosing, etc., and expansion over the world. By analogy with the cellular cytoplasm, we term this soup super-cytoplasm (Kadyshevich and Ostrovskii 2007; Ostrovskii and Kadyshevich 2011). In the super-cytoplasm, all the substances necessary for the existence and development of the primary DNA- and RNA-like substances could be synthesized on the basis of CH4 and of phosphates and niters that diffuse from the environment. Under appropriate conditions, this leads to an increase in the concentrations of nucleic acids and organophosphorous substances within the super-cytoplasm. Increasing in the concentrations to a certain critical level leads to precipitation of phosphor-containing membranes around DNAs and to origination of proto-cells. Thus, in addition to the super-cytoplasm, intracellular cytoplasm appears. After that, nucleic acids begin to develop and replicate inside the cells and the cells begin to divide similarly to the cells of the present prokaryotes. It has been demonstrated (see, e.g., Orgel 1992, 2000; Cech and Bass 1986; Li and Nicolaou 1994) that not only nucleic acids but even simpler organic substances are capable of self-replicating with consumption of certain chemical elements from the surrounding and excretion of nonutilized molecular residues. The hypothetical mechanism of self-replication of the DNA-like molecules is described in more detail in (Kadyshevich and Ostrovskii 2007; Ostrovskii and Kadyshevich 2011). Briefly, it is as follows. Water molecules sorb actively around nonwatered purine–pyrimidine (–H2 N?-O=) H-bond (Ostrovskii and Kadyshevich 2000, 2002a, 2006; Ostrovskii et al. 2000, 2001). The bonds of such a type are characteristic for pair-wise joining of N-bases belonging to nucleosides, nucleotides, and nucleic acids in highly concentrated aqueous media. Although the intermolecular H-bonds in the super-cytoplasm are watered to different degrees, it may happen in any moment that several molecules, which actively sorb waters, are localized in a minor volume of super-cytoplasm. This event may lead to the occurrence of the shell zone of a decreased water concentration around these several organic molecules. Such a water concentration decrease may be

J Mol Evol

sufficient for precipitation of an organo-phosphate semipermeable membrane-like shell at some distance from the molecules under consideration. If the water concentration over the system is rather low and the rate of formation of new organic molecules is rather high, a multitude of such membrane shells may precipitate simultaneously. They are the first proto-cells. Later on, each cell can obtain water and organic substances only as a result of their diffusion through the cell membrane. Thus, the intracellular cytoplasm appears in addition to the super-cytoplasm. After that, the nucleic acids replicate inside the cells. A multitude of similar but different proto-cells containing similar but different DNA- and RNA-like molecules originate within any one localization. After our thermodynamic calculations, there is no doubt that the processes of living matter origination require no energy; furthermore, they evolve much energy. As biota accumulated in the environments, some living organisms began to ‘‘feed on’’ other organisms that were dissipated in the super-cytoplasm soup. The ‘‘organismskillers’’ destroyed the bodies of the ‘‘organisms-victims’’ up to N-bases, PO43-- and NO3--ions, and riboses and used this material for their own extended reproduction. They were the first animals. Other organisms, i.e., plants, continued to use the mineral food. Possibly, origination of the animals and plants was dependent on the DNA concentration in the hydrate matrixes before their liquation and in the semi-liquid super-cytoplasm soup. This ‘‘food revolution’’ increased the rate of the extended reproduction of the ‘‘animals,’’ because the necessity of the LMSEs syntheses disappeared and the heat evolution decreased as a consequence of the necessity of the endothermic food destruction. After the ‘‘heat revolution,’’ the process of the DNA replication became almost autothermal, because the chemical states of the food and of the newly formed organisms were closely related in their chemical composition and structure. Amino-acids could originate in the super-cytoplasm; the processes of their origination require no additional source substances besides those occurring in the super-cytoplasm and in the gas phase over it. We showed above (reaction (5)) that this soup can contain some amount of ammonia along with other necessary substances. It is known that DNAs, being in the water solution or in the crystal form, show a dextrorotatory activity, i.e., they rotate the plane-polarized light to the right. The DNA optical activity, as such, surprises nobody, because DNA molecules include asymmetric four-substituted carbon atoms and form double helixes and because the planepolarized light rotation phenomenon is characteristic for organic molecules that are asymmetric relative to a plane, axis, or center (most frequently, atom), have helical form, or contain the so-called topological bonds; therewith, the

occurrence of an asymmetric carbon atom in an organic molecule is the most frequent but by no means unique cause of its chirality (central chirality). But the fact of DNA monochirality, i.e., the absence of laevorotatory DNA optical isomers, was always mysterious and initiated a set of papers and a number of very non-trivial assumptions on its source, including even the effects of the fundamental asymmetry of the Universe, the so-called ‘‘weak’’ interactions, and other phenomena of the Universal scale. The point is that the researchers assumed that DNAs are produced by Nature from N-bases, phosphates, and DDRs. Meanwhile, the D- and L-enantiomers of desoxy-ribose are synthesizable in experiments together and, usually, in almost equivalent quantities. Therefore, the mechanism of the natural selection of DDRs from the racemic mixture of desoxy-D- and desoxy-L-riboses was incomprehensible. There is the widely distributed opinion that this riddle is credited with the problem of living matter origination (for example, in (Bonner 2007)). Such an opinion is natural since the cause of such a selection is hidden in the mechanism of living matter appearance and of its initial development and since it is difficult to reveal it, bearing on the DNA features only. Analogous difficulties are typical when solving any reverse problem. The LOH-Theory allows a new natural view on the way of the DNA dextrorotatory problem solution, which, in our opinion, makes possible the introducing of the living matter origination phenomenon into the common system of the present knowledges about nature. According to the theory, no riboses as such form when DNAs are being produced from CH4 and nitrate and phosphate ions inside the hydrate structure. The N-base radicals ‘‘sprout’’ steadily into the neighboring small structural cavities, obeying the thermodynamic front, when they are being synthesized inside the hydrate large cavities from CH4 and nitrate ions. Thus, the nucleosides form. Just the thermodynamic front governs formation of such a chemical substance which is then capable of being a three-leg junction between one N-base radical and two phosphodiester radicals. This junction is similar in the chemical composition to the DDR radical but is not identical to it in the threedimensional structure. Namely, its pentamerous ring is somewhat twisted (Sundaralingam and Jensen 1965; Ostrovskii et al. 2014) as compared to the ribose ring and the geometry of its side HOCH2-group somewhat differs from that of desoxy-ribose. It is seen that, according to the LOHTheory, the DNA origination process does not require the DDR selection from the mixtures of enantiomers. Figure 6 illustrates our consideration. It presents the structure of the four-radical desoxycytidine—3,50 – bis (dimethyl phosphate) complex arranged within the gashydrate structure II on the basis of the PC program designed by A. Dzyabchenko. The X-ray parameters of the

123

J Mol Evol

Fig. 6 Scaled 3D image (Mercury PC program) of the desoxycytidine – 3,50 – bis (dimethyl phosphate) complex within the CH4hydrate structure II; gray, violet, red, white, and yellow sticks are the atomic radii of the intra-molecular atoms or the atomic diameters of the end atoms for the C, N, O, H, and P atoms, respectively; the H2O hydrate matrix is rejected (Color figure online)

free-complex structure are taken from (Sundaralingam and Jensen 1965). In (Ostrovskii et al. 2014; Kadyshevich et al. 2014), it is shown that this complex can be housed within the gas-hydrate structure II cavities. The waters of the hydrate matrix are rejected for the complex to be wellobservable. It is rotated by us together with the hydrate structure by the PC Mercury program to show in detail the central pentamerous ring and its surrounding. The occurrence of methyl radicals does not influence noticeably the nucleoside structure, and, thus, this figure shows the nucleoside structure in the DNA composition. It is seen that C-atoms 1, 3, and 4 are the chiral ones, because each of them is bound with four different chemical groups, and C-atoms 2 and 5 are non-chiral similarly to the corresponding atoms in the free desoxy-ribose composition. It is also seen that the desoxy-ribose-like radical that joins cytosine with the phosphodiester radicals is not identical to the free ribose molecule in its structure, because the pentamerous ring of this radical is significantly non-planar. Other nucleosides are characterized by analogous arrangements within gas-hydrate structure II. This figure is, apparently, sufficient for explaining the chirality of nucleosides and DNA molecules within hydrate matrixes, because each nucleoside has chiral atoms and has neither a plane nor a center of symmetry. However, apparently, the chirality of the ribose-like radicals as such is not the only cause capable of initiating the observable chirality of the DNA–hydrate-matrix system. According to the recent works (Dryzun et al. 2009; Dryzun and Avnir 2012), ‘‘the proportion of non-biological chiral crystals is

123

as high as 23 %; and only about 6 % of these are labeled as chiral,’’ i.e., the natural crystals are chiral much more often than that it was earlier considered. This conclusion was made on the basis of detailed studies of different zeolites and quartzes; their chirality is determined by the imperfectness of the crystals or/and by the occurrence of structural and absorbed asymmetrical foreign particles within the crystal structures. It might seem as if this discovery relates to crystals but not to the biologically active molecules as such. However, DNAs and other functional biological molecules, being in water systems at rather low temperatures, are surrounded by the ordered structured gashydrate coats and can reveal themselves like imperfect crystals. It is possible that the effect similar to that observed in the above-cited works reveals itself in the imperfectly structured gas-hydrate structures containing guest sub-structures in their cavities, i.e., the DNA chirality is not the effect of the DNAs as such but is the combined effect of the DNAs and water matrix. It is important that the above-cited authors have observed experimentally just the mono-chiral natural crystals. It is no wonder in the light of the present understanding of the phenomenon of chirality. Indeed, though the equilibrium mixtures of enantiomers are racemic, i.e., it contains equimolecular quantities of the dextrorotatory and levorotatory enantiomers, a potential barrier exists between enantiomers of opposite signs and its value can vary over a wide range (Kisel’ and Burkov 1980; Krupcik et al. 2003). Therefore, an enantiomer, being formed at a low temperature, may be in excess supply for a long time if the activation energy of its interconversion is sufficiently high and the ambient temperature is rather low. In connection with the consideration of the DNA chirality problem, notice the following data. As is written in ‘‘Something about the Natural DNA and RNA Sources’’ section, we state that the process of DNA origination within CH4-hydrate localizations starts from methane-nitrate formation. It is shown experimentally that methane-nitrate is capable of forming from CH4 and HNO3 at about 273 K and that, in this reaction medium, its chiral nitronic-acid modification (see above Fig. 4 in ‘‘Something about the Natural DNA and RNA Sources’’ section) produces secondary products (nitronic esters, oximes, etc.), which retain the chiral configuration (Kang and Yin 1997). Thus, apparently, chirality reveals itself even at the initial steps of transformation of methane and nitrate ions into nucleosides. Apparently, the above consideration of the DNA chirality problem represents the first realistic solution of the riddle that exists factually for about 160 years from 1848 when chirality of some biologically active substances was discovered by L. Pasteur. The LOH-Theory allows for answering the following questions (Kadyshevich and Ostrovskii 2009, 2010;

J Mol Evol

Ostrovskii and Kadyshevich 2011). (1) In what phase did the LMSEs form? (2) From what substances did the LMSEs form? (3) By what mechanism did the N-bases, riboses, and nucleosides form? (4) Is Nature capable of synthesizing LMSEs from minerals with no external energy? (5) How had methane hydrate originated? (6) How had CH4 and NO3- met together? (7) Why did no substance but NO3- react with CH4-hydrate? (8) How did DNA- and RNA-like molecules form from nucleosides? (9) Is there a relation between DNA and RNA formation, on the one hand, and the atmosphere composition, on the other hand? (10) Why do only five chemical elements usually enter into the DNA and RNA composition? (11) Why are N-bases incorporated into DNA and RNA similar in their composition and structure? (12) Why are N-bases and riboses limited in size? (13) Why are N-bases not identical? (14) Why do only five N-bases usually enter into the DNA and RNA composition and why do other N-bases, such as X, sometimes enter into the DNA and RNA compositions? (15) Could DR, DDR, Th, and U exist simultaneously in a reaction mixture containing CH4 and niter? (16) How had it happened that the sequences of N-bases in DNA and RNA molecules are not random? (17) How did proto-cells originate? (18) Why is the volume density of living organisms over the world ocean column much less than that over the soil, near-surface, and near-seabed layers? (19) What is the cause of the DNA monochirality? Most of these questions are answered above. Some aspects of the problem of living matter origination are first considered in this paper. Some questions that are listed here have a new elucidation. In conclusion of this section of the paper, we give the principal scheme of living matter origination (Fig. 7).

Observations and Simulations Counting in Favor of the LOH-Theory When formulating the LOH-Theory, we showed in the twodimensional coordinate system that the LMSEs sizes correlate well with the sizes of the gas-hydrate structure II large and small cavities. However, two-dimensional consideration gives insufficient knowledge about the possibility of housing DNA components within the three-

dimensional hydrate structure. We noted repeatedly that a group of tasks can be put for computer simulation of the rearrangement of different DNA components within the hydrate structure. At present, several tasks of such a kind are solved by us on the basis of an original crystallographic ‘‘a structure within another structure’’ program developed by A. Dzyabchenko and described in (Dzyabchenko and Kadyshevich 2013). On the basis of three-dimensional computer simulation, it was confirmed that the N-bases can be housed within the large cavities and riboses and phosphate groups can be housed within the small cavities of hydrate structure II (Kadyshevich et al. 2013, 2014; Ostrovskii et al. 2013, 2014). Housing of G (purine) and Cy (pyrimidine) into the adjacent large cavities allows location of these N-bases at such distances of one another that the lengths of the H-bonds between them (nm, 0.287, 0.295, and 0.288) (Kadyshevich et al. 2013; Ostrovskii et al. 2013) almost coincide with those calculated earlier in (White et al. 1978) (nm, 0.293, 0.296, and 0.293) and in (Ycˇas 1969) (nm, 0.284, 0.292, and 0.284) on the basis of X-ray measurements of crystallized DNA double helixes. Housing of G and DDR radicals into the adjacent large and small cavities, respectively, allows their arrangement within these cavities with no structural destruction at a distance of 0.148 nm (Kadyshevich et al. 2013); this distance correlates well with the G–DDR chemical bond length (0.151 nm) calculated from the X-ray study of crystallized DNA double helixes (Takusagawa et al. 1982). Meanwhile, housing of two purine N-bases into the gashydrate adjacent large cavities leads to a significant tension of the structure and to its partial destruction. Apparently, two purines cannot be housed within two adjacent large cavities, while a purine and a pyrimidine can be housed within two adjacent large cavities. This result counts in favor of origination of DNA–DNA double helixes within the gas-hydrate structures. Thus, the geometric compatibility of the CH4-hydrate structure II and components of the DNA and RNA molecules, which was earlier stated on the basis of the two-dimensional consideration, is now confirmed for the three-dimensional system. All simulated pictures described in the previous paragraph are available for free in the corresponding publications. In Fig. 8, we consider, as an example, the G–Cy complex structure. In Fig. 8a, the non-scaled scheme of

Fig. 7 General scheme of living matter origination

123

J Mol Evol

Fig. 8 The H-bound G–Cy complex: a scheme; b, c simulated scaled 3D images within two adjacent large hydrate cavities (Mercury PC program); the N(Cy)O(G), N(Cy)N(G), and O(Cy)N(G) H-bond

lengths are (nm) 0.287, 0.295, and 0.288, respectively. The corresponding X-ray data are (nm) 0.293, 0.296, and 0.293 (White et al. 1978) and 0.284, 0.292, and 0.284 (Ycˇas 1969)

this H-bound complex is given; in Fig. 8b, such an H-bound complex is placed within gas-hydrate structure II; and, in Fig. 8c, the cytosine and guanine molecules are placed within the adjacent large cavities of the same structure and are there turned up to their positions at which the structural cavities and the guest molecules can be observed each separately. The white, green, violet, or red colorations show atomic radii (in Fig. 8b) or atoms (in Fig. 8c) of H, C, N, or O atoms, respectively. The thin lines nearby G and Cy represent the H2O-hydrate matrix. The chemical nature of each atom is also seen in Fig. 8a. The right picture shows that the sizes and configuration of a gas-hydrate structure large cavity is appropriate to house G or Cy in it, i.e., such molecules, being housed within large cavities, fill them completely and, therewith, do not destroy the neighboring hydrate structure. Then, they can be turned within their cavities in such a way that the distance between them allows formation of hydrogen bonds as it is said in the previous paragraph. The PC program of turning is based on minimization of the potential energy of the system through variation of all degrees of freedom of the hydrate structure by the method described in (Dzyabchenko and Kadyshevich 2013). As early as 2002 (Ostrovskii and Kadyshevich 2002b), we prognosticated that the processes of simplest living matter origination on Earth had proceeded in ancient times and, possibly, proceed at present in those underseabed and underground localities where water and simplest mineral substances necessary for syntheses of living entities are available and the temperature and pressure conditions are suited for hydrate formation. Three years later, huge colonies of prokaryotes were discovered in the underseabed soil over the Pacific Ocean open areas and Central America coastline under the water column from 427 to 5,086 m (Schippers et al. 2005). The underseabed soil was drilled to a depth of 400 m. It is

stated that the concentration of living organisms in the soil samples taken at different points of the Ocean is from a few thousand to several million per cubic centimeter and does not depend on the depth of drilling. Everywhere, the submarine bacteria-iferous grounds contain CH4-hydrate. It is noteworthy that the bacteria are found near the American continent with its large niter deposits (see ‘‘Something about the Natural DNA and RNA Sources’’ section). The authors of the paper believe that these bacteria emit methane and serve as a source of methane hydrate deposits. However, any living entity requires the chemical sources for its reproduction; methane, as the unique source of carbon capable of DNA forming in the system under consideration, is necessary for synthesizing new DNAs and for the cellular replication. Chemistry is the stubborn science. It requires the explanation of chemical transformations by chemical equations. If a system contains bacteria and contains no other source of carbon but methane, this means that these bacteria ‘‘eat’’ methane to form DNAs for their ‘‘children’’ according to the chemical Eqs. (1)–(4) and subsequent polycondensation reactions with phosphate ions. Apparently, the discovery of the bacteria under the sea bottom in a close proximity to methane hydrate and niter deposits under the sea bottom confirms our theory. The following trait counts in favor of such a conclusion. Temperatures of up to 299 K measured by the authors of this discovery in the bacteria-iferous ground layers require a special explanation. Normally, the seabed temperature at depths of more than 400 m is about 274 K and increases by 3 K per each 100 m of seabed vertical drilling. Therefore, the temperature 400 m below the sea bottom must be only 286 K. The most probable explanation of the unusually high temperature in the zone of prokaryote activity is the heat production as a result of DNA formation from CH4 and NO3-- and PO43--ions within the CH4-hydrate phase. We showed above that reactions (1)–(4) are highly

123

J Mol Evol

exothermic; it is known that the subsequent polycondensation reactions are also exothermic, though their heat production is not as high as that of the reactions of N-bases and riboses formation. The active bacterial life was also discovered in CH4containing pores as a result of deep drilling down to 6,820 m in Siberia; the authors write that the rate of biomass formation from mineral carbon-containing substances is about 1–10 ton per year at 1 km2 of the Earth’s surface (Oborin and Khmurchik 2008). According to (Schippers et al. 2005), the mass of the today underseabed and underground living matter that is replicating on the basis of methane is of the same order of magnitude as the mass of the overseabed and ground-based living matter. There is one more repeated observation in support of our concept. It was shown that gas samples taken from CH4hydrate deposits for analyses contain much N2 and little O2: 4 and 0.005 %, respectively, according to (Davidson et al. 1986) and 11.4 and 0.2 %, respectively, according to (MacDonald et al. 1994). It is seen that the N2 to O2 ratio is much higher than that in the Earth’s atmosphere; evidently, the samples could not have been contaminated by atmospheric nitrogen during their collection and storage. Potential sources of elemental N2 in the Earth’s crust are few. Therefore, it is quite possible that nitrogen was produced by the reduction of methane or other hydrocarbons by niter in the deposits of hydrocarbon-hydrates as a result of the reactions described in ‘‘Chemical Sources of Living Matter: Each Natural Phenomenon or Entity Is Produced from Minimal Variety of Simpler Ones’’ and ‘‘The LOHTheory as the Unique Thermodynamically Grounded Theory of Living Matter Formation’’ section.

Discussion: The LOH-Theory as the Ground for Consensus Between Anti-Darwinists and NeoDarwinists The LOH-Theory was published at different steps of its development in (Ostrovskii and Kadyshevich 2000, 2002a, b, 2006, 2007, 2009, 2011, 2012a, b; Ostrovskii 2010; Kadyshevich and Ostrovskii 2009, 2010, 2012), as was mentioned above, and was presented in the form of lectures and oral talks to a number of audiences over the world at more than 30 international physical, chemical, thermodynamic, biological, geological, and specialized conferences. It considers the problem of the mechanism of living matter origination and does not consider the problem of the living matter existence on other, but the Earth, celestial bodies. We think that, wherever living matter similar to the Earth’s one would originate, the mechanism of its origination would be similar to that described by this theory.

An analysis of the living matter phenomenon performed by us in the period of formulation of the LOH-Theory, MRH-Theory, and PFO–CFO hypothesis led us to the following fundamental conclusions relative to the life origination process at the Earth: (1) The DNA occurrence and reproduction is the principal feature of living matter; the proteins are side products. (2) DNAs and cellular life had originated from inorganic and simplest organic minerals as inevitable products of the atomistic world. (3) Stable undisturbed conditions favored living matter origination. (3) Living matter originated at the Earth repeatedly in different periods of the Earth’s history when its origination was favored with the ambient conditions. (4) In each appropriate period, living matter originated in a number of localizations. (5) The reacting systems transformed from mineral substances to DNAs and proto-cells so slowly that they passed all possible states step by step in the direction of gradual decrease in the Gibbs free energy; i.e., just the thermodynamic front governed the movement from minerals to living matter. (6) In each localization, a set of different DNAs and different proto-cells which gave start to development of a set of different organisms originated. (7) The differentiation of the living matter into the plant and animal organisms could start at the step of the primary super-cytoplasm soup. (8) The diversity of the available forms of living matter is caused by the spatial and temporal repeatability of the processes of living matter origination under similar but not identical ambient conditions, multiplicity of the DNA forms in each event of living matter origination, variations in the parameters of the native medium, intraspecific variations, and interspecific variations. The LOH-Theory and the well-known evolutionary theory interpret differently the living matter origination and development and the present species diversity. Apparently, the occurrence of differences is natural because the volume of knowledges in biology, paleontology, physics, and chemistry changed significantly for a century and a half from the time of publication by Darwin of his outstanding book ‘‘On the Origin of Species.’’ Although the Darwinian views underwent changes for this period and the neoDarwinism differs from the original Darwin’s views, nevertheless, this modified elucidation of the species diversity, apparently, is also in need of modification. Meanwhile, we hope that the LOH-Theory allows a consensus between the today neo-Darwinists and anti-Darwinists. The notion on evolutionary accumulation of minor changes in the DNA of an organism and revolutionary manifestation of summed minor changes in the appearance of a new species is scarcely justified. At the root, Darwin’s evolutionary theory is based on the assumption that living entities appeared primarily on the Earth in a very small number and in the simplest forms and is aimed at explanation of the present species diversity. Indeed, apparently,

123

J Mol Evol

it would be difficult to propose any other mechanism for the diversification of the species if such an assumption is regarded as of paramount importance. However, this assumption did not become more evident with time. Factually, the general method proposed by Darwin and by his followers for confirmation of the evolutionary theory is based on compilation of the sequences of conjectural biological transformations through systematization of the modern flora and fauna. However, the evidential standard of this method as such is under question. Indeed, when a very wide set of entities characterized by numerous and dissimilar parameters is under analysis, a number of series in which different parameters vary in some order can be constructed. At this, it is difficult to find convincing proofs of whether these series are arbitrary or they reflect some fundamental dependences. The occurrence of numerous gaps in the series proposed by Darwin and by advocates of his theory stimulates doubts about the presence of the fundamental content in these series. According to the evolutionary theory, the fossil record should be rife with examples of transitional forms leading from the less to more evolved species. Quite the contrary, ‘‘…instead of filling the gaps in the fossil record with socalled missing links, most paleontologists found themselves facing a situation in which there were only gaps in the fossil record, with no evidence of transformational intermediates between documented fossil species’’ (Schwartz 1999). This opinion written 15 years ago reflects the today state of the problem. Advocates of the theory of the evolutionary transformation of the world from the mineral desert with several living entities produced about 3.5–3.8 billion years ago by any mysterious way to the dwelling place for about 3.0 million plant and animal species (it is the total amount of the extant and deleted species), including innumerable individuals, presented no satisfactory arguments for their assumption. As before, ‘‘… today, many evolutionists assume that a large number of small mutations can account for macroevolution. This conclusion is not based on experimental evidence, but on the assumption that the evidence for microevolution can be extrapolated to macroevolution. The empirical evidence, however, is clear—neither macromutations nor micromutations can provide a significant source of new genetic information.’’ (Bergman 2007). Apparently, this Bergman’s conclusion is proved by the experiments with drosophila and other rapidly reproducing species. These experiments have been widely performed over the world in many biological laboratories. They were started early in the twentieth century by W. Castle (Carroll 2007) and were aimed just at solving the evolutionary problem. For more than 100 years, the researchers attentively but fruitlessly searched for new species among the

123

numerous generations of the drosophila and other proliferous insects. Drosophila melanogaster are usually used for this aim but other insects are also not forgotten. The ontogenesis period for drosophila is about 10 days at 298 K and about 20 days at 291 K, and one female fly produces about 400 eggs for its short life. Thousands of researchers and their students in many tens of laboratories over the world tried to discover among the fly posterity even if one organism of any new species; many hundreds of thousands and, may be, millions of flies were attentively inspected. But all hopes of the observers were doomed to disappointment. Today, the following ambiguous conclusion can be made in the certainty: if interspecific variations exist, they are extremely rare. However, the advocates of the neo-Darwinian idea of accumulation by organisms of micromutations, which, after their critical number, can lead to a macromutation and to the formation of a new species, do not surrender although they have no real proofs of their correctness. The LOH-Theory rejects the bench mark of the Darwinian logic, namely his assumption on the primary origination of a unique living entity or a small quantity of living entities. Though, it gives a unique possibility for bridging and achievement of a consensus between the neoDarwinism and anti-Darwinism. We believe that the consensus in the biological community relative to the socially important problem of living matter origination would be useful not only for the community but also for the social medium. The fundament for this consensus is as follows. On the one hand, the LOH-Theory, apparently, should satisfy anti-Darwinists, because it, contrary to Darwin’s state, assumes the origination of different DNA and RNA molecules and different proto-cells within any natural ‘‘incubator,’’ simultaneous occurrence of a number of such ‘‘incubators,’’ repeatability of the ‘‘periods of incubation’’ in the Earth’s history, and many-fold decreasing in the ‘‘load’’ on subsequent processes of the species diversification and on the number of the steps in this process. The last point neutralizes the important and just objection of the today anti-Darwinists against Darwin’s theory that the period of living matter existence on the Earth is too short for species diversification from several living entities to about 3.0 million plant and animal species and innumerable number of entities. On the other hand, the LOH-Theory, owing to the decrease in the ‘‘load’’ on the processes of the species diversification and on the number of the steps in this process, makes realistic and deserving subsequent study the assumption of the neoDarwinists about the possibility of diversification through accumulation by organisms of micromutations, which, after their critical number, can lead to a macromutation and, in the sequel, to the formation of a new species.

J Mol Evol

Fig. 9 Generalized paleontological information on the temporal connection between the Earth’s glaciations and periods of massaged flora and fauna distribution (numerals are given in Mya)

Conclusion The LOH-Theory differs principally from all living matter origination theories published earlier. These differences are not only factual but also represent a different world-outlook and philosophy. We consider the life origination process as a sequence of thermodynamically caused regular and inevitable chemical transformations, which are regulated by universal physical and chemical laws. Not proteins and not amino-acids but the DNA molecules are considered as the first carriers of life. LMSEs, DNAs, and RNAs formed within the underground or underseabed honeycomb CH4-hydrate structure from CH4 and nitrate and phosphate ions that diffused into the hydrate structure, and proto-cells and cellular agglomerates originated from the same minerals within the semi-liquid soup that formed after liquation of the hydrate structures. Living matter originated repeatedly in different localizations, and each localization gave rise to a multitude of different DNAs and different living organisms. The DNAs and the cells are principally similar in their constitution, because they are built by Nature on the basis of the same mineral materials and of the same physical and chemical laws. Living matter originated and can originate now everywhere over the Universe where necessary minerals and necessary ambient conditions exist and where these conditions are continuous for long periods of time. We think that Earth’s type living matter based on DNA replication can originate wherever by no mechanism that differs principally from that described by the LOHTheory. The LOH-Theory is based on our discoveries of the gas-hydrate structure formation in highly concentrated semi-liquid water/substrate systems (where substrate is a functional polymer) and strict dimensional correspondence of large and small hydrate cavities with N-bases and with riboses and phosphate-groups, respectively, and on numerous thermodynamic calculations, which confirmed the feasibility of DNAs and cells formation and of reactions between nitrate ions and CH4 under conditions of CH4-hydrate existence (the so-called Konovalov’s reactions), etc. The dimensional correspondences are confirmed by the three-dimensional simulation. The theory includes an important notion of a ‘‘thermodynamic front’’ whose

temporal movement determines the slow (on the human life duration scale) stepwise filling of the gas-hydrate cavities, i.e., formation of purine and pyrimidine nuclei within the large cavities, of riboses within small cavities, and of substituting groups in the purine and pyrimidine nuclei and subsequent formation and lengthening of the DNA- and RNA-like molecules. The most appropriate conditions for origination of the living species governed by the most lengthy and complicated DNAs occurred during the completion phases of the Earth’s history cold periods. This statement relates in the first place to the surface and flying species, because the CH4-hydrate deposits could be located in these periods near the Earth’s surface. Owing to the persistence of the Earth defrosting, these natural thermostats could be at the optimum temperature for a long time, and, thus, the DNA communities had sufficient time to reach their perfection within the hydrate structures and could appear at the Earth’s surface after liquation of hydrate structures. Apparently, the most favorable conditions for origination of new multi-cellular living species arose at the end of the Neoproterozoic Era, in the period between 570 and 525 million years ago. Indeed, paleontological studies show that a majority of the modern multi-cellular animals appeared in the chronicle of the fossils over this period. The species diversity is caused mainly by the spatial and temporal repeatability of the processes of living matter origination under similar but not identical conditions, multiplicity of the DNA forms in each living matter origination event, variations in the parameters of the native medium, intraspecific variations, and, in a less degree, interspecific variations. The contribution of the last to the species diversity is, likely, significant for prokaryotes and those eukaryotes that are only at low steps of their biological organization; however, in the light of the LOHTheory, of available long-term paleontological investigations, and of studies of reproduction of proliferous organisms, we conclude that, in toto, the contribution of interspecific variations to the species diversity was overestimated significantly. The reason of this overestimation is that origination of scores of «spores» of different organisms in any one event and multiple reproductions of such

123

J Mol Evol

events in time and Earth’s space were not taken into consideration. The conclusive scheme (Zak 2014) presented in Fig. 9 generalizes the available paleontological information on the temporal connection between the post-glacial periods in the Earth’s history and the periods of massaged flora and fauna distribution over the Earth or over its separated regions. The explosive living matter expansion proceeded late in cold periods, about (Mya) 3900 (after the faint Sun period), 2100, and 542. Just late in the cold periods, the conditions arose when the CH4-hydrate localizations were rather close to the Earth’s surface and the temperature was sufficiently high for a rather long time to unbrake the DNA formation processes that were ‘‘frozen’’ earlier. In the course of each interglacial Earth’s history period favorable for syntheses of the LMSEs, DNAs, and cells and for development of plant and animal organisms, the diversity and multiplicity of the living species were caused by the following main factors: (1) a multiplicity of the DNA modifications that were produced within each natural ‘‘incubator’’; (2) the occurrence of a great number of ‘‘incubators’’; (3) in the periods of climatic catastrophes, the survival of some organisms and some DNA modifications that existed during previous favorable periods; (4) variability of DNAs under the effect of the natural selection. In our opinion, the LOH-Theory gives ground for the consensus between anti-Darwinism and neo-Darwinism in explanation of the species diversification, because it leaves the door open for the interspecific variations albeit limits their role in the evolution of the species. Acknowledgments We are grateful to our Reviewer for calling our attention to insufficient explanation of the phenomenon of DNA monochirality in the initial manuscript and thus stimulating us to more comprehensive consideration of this topical problem and to Niles Lehman, Editor in Chief, for repeated reading of the manuscript and useful advices. The work is partially supported by the Russian Foundation for Basic Research, project no. 12-05-01082.

References Abel DL, Trevors JT (2006) Self-organization vs. self-ordering events in life-origin models. Phys Life Rev 3:211–228 Atkins WRG (1926) The phosphate content of seawater in relation to the growth of the algae phytoplankton. Part III. J Mar Biol Assoc 14:447–467 Atwood JL et al (eds) (1996) Comprehensive supramolecular chemistry. Pergamon Press, New York Barenbaum AA (2007) On possible relationship between gas-hydrates and submarine groundwater. Water Resour 34:587–592 Bergman J (2007) Creative evolution: an anti-Darwin theory won a Nobel. Acts & Facts 36(7) Bonner WA (2007) Origins of chiral homogeneity in Nature. In: Eliel EL, Wilen SH (eds) Topics in stereochemistry, vol 18. Wiley, Hoboken, pp 1–96

123

Byk SSh, Fomina VI (1970) Gas hydrates. In: Progress in science and technology. Ser Chemistry, Physical Chemistry, 1968, VINITI, Moscow, Russia Carey FA, Sundberg RJ (2004) Organische chemie. Wiley, Weinheim Carroll JJ (2003) Natural gas hydrates: a guide for engineers Amsterdam. Gulf Professional Publ, Amsterdam Carroll SB (2007) The making of the fittest: DNA and the ultimate forensic record of evolution. WW Norton & Co, New York Cech TR, Bass BL (1986) Biological catalysis by RNA. Ann Rev Biochem 55:599–629 Chaplin M (2013) Water Structure and Science; http://www.lsbu.ac. uk/water/clathrat2.html Clarke EC, Ford RW, Glew DN (1964) Propylene gas hydrate stability. Can J Chem 42:2027–2029 Dahm R (2008) Discovering DNA: Friedrich Miescher and the early years of nucleic acid research. Hum Genet 122(6):565–581 Darwin CR (2010) The variation of animals and plants under domestication. Cambridge University Press, Cambridge Davidson DW, Garg SK, Gough SR et al (1986) Laboratory analysis of a naturally occurring gas hydrate from sediment of the Gulf of Mexico. Geochim Cosmochim Acta 50:619–623 Day JC, Robb ID (1981) Thermodynamic parameters of polyacrylamides in water. Polymer 22:1530–1533 de Graaf RM, Visscher J, Schwartz AW (1995) A plausibly prebiotic synthesis of phosphonic acids. Nature 378(6556):474–477 Dryzun C, Avnir D (2012) On the abundance of chiral crystals. Chem Commun 48:5874–5876 Dryzun C, Mastai Y, Shvalb A, Avnir D (2009) Chiral silicate zeolites. J Mater Chem 19:2062–2069 Dzyabchenko AV, Kadyshevich EA (2013) Life Origination Hydrate Hypothesis (LOH-Hypothesis): the theory and procedure of simulation of basic DNA units in hydrate structure. EPSC Abstracts 8:EPSC2013–284 Ericksen GE (1983) The Chilean nitrate deposits. Am Sci 71:366–374 Frye K (ed) (1982) The encyclopedia of mineralogy. Stoutsburg, Pen, USA Ginsburg GD, Soloviev VA (1998) Submarine gas hydrates. Okeanologia, St. Petersburg Gull M, Zhou M, Fernandez FM, Pasek MA (2014) Prebiotic phosphate ester syntheses in a deep eutectic solvent. J Mol Evol 78:109–117 Kadyshevich EA (2009) PFO–CFO hypothesis of planet formation. M&PS 44:A105 Kadyshevich EA, Ostrovskii VE (2007) Hypothetical physicochemical mechanisms of some intracellular processes: the hydrate hypothesis of mitosis and DNA replication. Thermochim Acta 458:148–161 Kadyshevich EA, Ostrovskii VE (2009) Hydrate hypothesis of living matter origination (LOH-hypothesis): thermodynamic grounds of formation of living matter simplest elements from hydrocarbons and niter. J Therm Anal Calorim 95:571–578 Kadyshevich EA, Ostrovskii VE (2010) Hydrate hypothesis of living matter origination: logical and thermodynamic grounds. In: Dmitrievskiy AN, Valyaev BN (eds) Degassing of the Earth: geotectonics, geodynamics, geofluids; oils and gas; hydrocarbons and life. GEOS, Moscow, pp 195–198 Kadyshevich EA, Ostrovskii VE (2011) Development of the PFO– CFO hypothesis of Solar System formation: why do the celestial objects have different isotopic ratios for some chemical elements? In: Bonanno A, de Gouveia dal Pino E, Kosovichev A (eds) Proceedings of the International Astronomical Union (Advances in Plasma Astrophysics). Cambridge University Press, Cambridge, pp. 95–101. http://journals.cambridge.org/ action/displayAbstract?fromPage=online&aid=8295090

J Mol Evol Kadyshevich EA, Ostrovskii VE (2012) Life Origination Hydrate hypothesis (LOH-hypothesis): CH4-hydrate matrix as a necessary condition for life origin, EPSC Abstracts 7:EPSC2012-113 Kadyshevich EA, Ostrovskii VE (2013) PFO–CFO Hypothesis of Solar System Formation: the presolar star as the only source of chemical elements for the Solar System. EPSC Abstracts 8:EPSC2013-38 Kadyshevich EA, Dzyabchenko AV, Ostrovskii VE (2013) Life Origination Hydrate hypothesis (LOH-hypothesis): computer simulation of rearrangement of different DNA components within CH4-hydrate structure II. Proceedings of the European Planetary Science Congress 2013, EPSC2013-285, Copernicus Gesellschaft, Gottingen, Germany; http://presentations.coperni cus.org/EPSC2013-285_presentation.pdf Kadyshevich EA, Dzyabchenko AV, Ostrovskii VE (2014) Life Origination Hydrate Theory (LOH-Theory) and Mitosis and Replication Hydrate Theory (MRH-Theory): three-dimensional PC validation. EPSC Abstracts 9:EPSC2014-53-2 Kang F-A, Yin C-L (1997) Studies on the chemistry of the chiral nitronic acid and nitronic esters. Tetrahedron: Asymmetry 8:3585–3589 Kisel’ VA, Burkov VI (1980) Gyrotropy of crystals. Nauka, Moscow Konovalov MI (1893) Nitrating action of nitric acid on hydrocarbons of saturated character. J Russ Physicochem Soc 25:446–454 Krupcik J, Oswald P, Ma´jek P, Sandra P, Armstrong W (2003) Determination of the interconversion energy barrier of enantiomers by separation methods. J Chromatogr 1000:779–800 Li T, Nicolaou KC (1994) Chemical self-replication of palindromic duplex DNA. Nature 369:218–221 Lide DR (1996) Handbook of chemistry and physics, 76th edn. CRC Press, New York MacDonald IR, Guinasso NL, Brooks JM, et al (1994) Seafloor gashydrates: video documenting oceanographic influences on their formation and dissociation. In: Near Surface Expression of Hydrocarbon Migration. AAPG Hedberg Research Conference Abstracts, Vancouver, B.C., Canada Milkov AV (2004) Global estimates of hydrate-bound gas in marine sediments: how much is really out there? Earth Sci Rev 66:183–197 Nekrasov BV (1965) Foundations of general chemistry, vol 1. Khimiya, Moscow, pp 432–440 Oborin AA, Khmurchik VT (2008) Chemosynthesis at deep fluids as an additional source of organic matter in the lithosphere. In: Dmitrievskiy AN, Valyaev BN (eds) Degassing of the Earth; geodynamics, geofluids, oil, gas, their parageneses. GEOS, Moscow, pp 366–367 Orgel LE (1992) Molecular replication. Nature 358:203–209 Orgel LE (2000) Self-organizing biochemical cycles. Proc Nat Acad Sci 97:12503–12507 Ostrovskii VE (2002) Differential microcalorimeter for isothermal measurements of heat effects in two-phase systems and examples of its application. Rev Sci Instrum 73:1304–1312 Ostrovskii VE (2010) Hypothesis of the natural gas abiotic origin in the context of living matter origination. In: Vladimirov AI, Lapidus AL (eds) Gas chemistry at the present stage of development. Gubkin RSU NG, Moscow, pp 35–69 Ostrovskii VE, Kadyshevich EA (2000) Use of data on the polyacrylamide–water system in analyzing the equilibrium DNA–water structure. Russ J Phys Chem 74:1114–1124 Ostrovskii VE, Kadyshevich EA (2002a) Hydrate model of the equilibrium DNA–H2O system. Int J Nanosci 1:101–121 Ostrovskii VE, Kadyshevich EA (2002b) The DNA–water system: review and a new concept. In: 12th International Symposium on Supramolecular Chemistry, ISSC-XII, Eilat, Israel, Extended Abstracts, P-48

Ostrovskii VE, Kadyshevich EA (2006) Thermodynamics of formation of nitrogen bases and D-ribose from mineral substances in light of the problem of origination of simplest elements of living matter. Thermochim Acta 441:69–78 Ostrovskii VE, Kadyshevich EA (2007) Generalized hypothesis of the origin of the living-matter simplest elements, transformation of the Archean atmosphere, and the formation of methane-hydrate deposits. Phys Uspekhi (Adv Phys Sci) 50:175–196 Ostrovskii VE, Kadyshevich EA (2009) Life origination hydrate hypothesis (LOH-hypothesis). Orig Life Evol Biosph 39:219–220 Ostrovskii VE, Kadyshevich EA (2011) Mitosis and DNA replication and life origination hydrate hypotheses: common physical and chemical grounds. In: Seligmann H (ed) DNA replication— current advances. InTech, Rijeka, pp 75–114 Ostrovskii VE, Kadyshevich EA (2012a) Life Origination Hydrate Hypothesis (LOH-Hypothesis). Life 2:135–164. http://www. mdpi.com/2075-1729/2/1/135 Ostrovskii VE, Kadyshevich EA (2012b) Life Origination Hydrate Hypothesis (LOH-Hypothesis): original approach to solution of the problem. Global J Sci Front Res (A) 12(6):1–36 https:// globaljournals.org/GJSFR_Volume12/1-Life-OriginationHydrate-Hypothesis.pdf Ostrovskii VE, Kadyshevich EA (2013) PFO–CFO Hypothesis of Solar System formation: the notion of the Sun-like stars and their transformations. EPSC Abstracts 8:EPSC2013-145 Ostrovskii VE, Tsurkova BV (1997) Differential heat effects and mechanisms of interaction of polyacrylamide with water. Russ J Phys Chem 71:967–973 Ostrovskii VE, Tsurkova BV (1998a) The system polyacrylamide– water. Differential heat effects, rates and molecular mechanisms of wetting and drying. J Therm Anal 51:369–381 Ostrovskii VE, Tsurkova BV (1998b) The polyacrylamide–water system: application of differential calorimetry to study the mechanisms of dissolution. Thermochim Acta 316:111–122 Ostrovskii VE, Tsurkova BV, Kadyshevich EA, Gostev BV (2000) Differential heat effects, kinetics, and mechanisms for the drying and water vapor wetting of the acrylamide–water system. Russ J Phys Chem 74:191–201 Ostrovskii VE, Tsurkova BV, Kadyshevich EA, Gostev BV (2001) Comparison study of the acrylamide–water and polyacrylamide– water systems: differential heat effects, kinetics, and mechanisms of drying and vapor-phase wetting. J Phys Chem B 105:12680–12687 Ostrovskii VE, Kadyshevich EA, Dzyabchenko AV (2013) Life Origination Hydrate Hypothesis (LOH-Hypothesis): underground laboratories of nature as a place for synthesis of the first living organisms. In: Modern problems of theoretical, experimental and applied mineralogy, Geoprint, Syktyvkar, pp 472–475 Ostrovskii VE, Kadyshevich EA, Dzyabchenko AV (2014) Life origination hydrate theory (LOH-Theory): the DNA monochirality nature. EPSC Abstracts 9:EPSC2014-833 Ould-Moulaye CB, Dussap CG, Gros JB (2001) A consistent set of formation properties of nucleic acid compounds. Purines and pyrimidines in the solid state and in aqueous solution. Thermochim Acta 375:93–107 Ould-Moulaye CB, Dussap CG, Gros JB (2002) A consistent set of formation properties of nucleic acid compounds. Nucleosides, nucleotides and nucleotide-phosphates in aqueous solution. J Therm Anal 387:1–15 Pinder KL (1964) Time dependent rheology of the tetrahydrofuranhydrogen sulphide gas hydrate slurry. Can J Chem Eng 42:132–138 Pinder KL (1965) A kinetic study of the formation of the tetrahydrofuran gas hydrate. Can J Chem Eng 43:271–274

123

J Mol Evol Platteeuw JC, Van-der-Waals JH (1959) Thermodynamic properties of gas hydrates II: phase equilibria in the system H2S-C3H8-H2O at -3 C. Rec Trav Chim 78:126–133 Schippers A, Neretin LN, Kallmeyer J et al (2005) Prokaryotic cells of the deep subseafloor biosphere identified as living bacteria. Nature 433:861–864 Schwartz JH (1999) Sudden origins. Wiley, New York, p 89 Silberberg A, Eliassaf J, Katchalsky A (1957) Temperature-dependence of light scattering and intrinsic viscosity of hydrogen bonding polymers. J Polymer Sci 23:259–284 Stackelberg M, Meuthen B (1958) Solid gas-hydrate VII. Watersoluble ether. Z Electrochem 62:130–131 Stackelberg M, Muller H (1954) Solid gas-hydrate II. Structure and stereochemistry. Z Electrochem 58:25–39 Sundaralingam M, Jensen LH (1965) Stereochemistry of nucleic acid constituents. I. Refinement of the structure of cytidylic acid b. J Mol Biol 13:914–929 Takusagawa F, Dabrow M, Neidle S, Berman HM (1982) The structure of a pseudo intercalated complex between actinomycin and the DNA binding sequence d(GpC). Nature (L) 296:466–469

123

Treloor PJ, Colley H (1996) Variations in F and Cl contents in Apatites from magnetite-apatite ores in Northern Chile and their ore genetic implications. Mineral Mag 60:285–301 Trevors JT, Abel DL (2004) Chance and necessity do not explain the origin of life. Cell Biol Int 28:729–739 Turekian KK (1968) Oceans. Prince-Hall, Englewood Van-der-Waals JH, Platteeuw JC (1959) Clathrate solutions. Adv Chem Phys 2:1–57 White A, Handler P, Smith EL, Hill RL, Lehman IR (1978) Principles of biochemistry, 6th edn. McGraw-Hill, New York Ycˇas M (1969) The biological code. North-Holland Publishing Co., Amsderdam Zak C (2014) http://www.corzakinteractive.com/earth-life-history/32_proter ozoic.htm; source: University of California Museum of Paleontology Tour of Geologic Time  Wikipedia - Geologic Time Scale