I. Ignatov,
O.V. Mosin

It is believed that the big bang explosion 13,7 billion years ago produced the universe that was much denser and hotter, and made almost entirely of one element - hydrogen. Deuterium (D) was formed during the next moments of evolution of the universe as a result of collision of one neutron and proton at temperatures of about one billion degrees. Furthemore, the two formed deuterons stuck together into helium nuclei contained two protons and two neutrons.

Thus, deuterium can be served as a feasable indicator of evolution, as natural amount of deuterium is constant. Up to the present time it has been considered, that during the formation of helium, almost all the deuterim atomd combined to form helium nuclei, leaving a tiny remant to be detected today so that only one in 10 000 deuterons remained to be unpaired. In 2007 the American astronomer G. Linski using FUSE ultra-violet telescope detected the increased contents of deuterium in a galaxy of the Milky Way. Proceeding from the amount of deuterium, natural prevalence of deuterium makes up no more than 0,015 atom% and depends strongly on both the uniformity of substance and the total amount of matter formed in course of early evolution. Now it is obvious, that the natural amount of deuterium is at least twice more, than it was supposed, and makes up approxumately 0,003 %. In particular, on formation of stars it was spent not 1/3, but only 15 % of deuterium and it is distributed non-uniformly. These data testify that for formation of stars it was required much less hydrogen, turned then into helium. This new information can change radically our theoretical representations not only about formation of stars and galaxies, but also about molecular evolutions. A constant source of deuterium are explosions of nova stars and thermonuclear processes going inside the stars. Probably, it could explain the fact why the quantity of deuterium increased slitely during the global changes of climate for worming conditions.

The second item is chemical processes going on with the participation of deuterium as a result of which heavy water (D2O) is formed from gaseous deuterium and oxyden at high temperatures. We proposed that primary water could contain more deuterium in the early stages of evolution of a life. Thus, in primary atmosphere of the Earth which was consisted, basically, from carbonic monoxyde (CO), hydrogen (H2), nitrogen (N2), ammonia (NH3) and methane (CH4), there was not oxygen forming protective ozone layer, capable to protect a surface of the Earth from rigid short-wave radiation of the Sun carrying huge energy. Therefore, radiation freely passed by through an oxygen-free atmosphere and reaching hydrosphere, caused photolysis and radiolysis of water. Energy of radiation, volcanic geothermal processes on a hot surface of the Earth and electric discharges in atmosphere, could lead to the enrichment of hydrosphere by deuterium in form of semiheavy water (HDO) that evaporates more slowly then H2O, and condenses faster.

Gravitational field of the Earth is insufficiently strong for retaining of lighter hydrogen, and our planet is gradually losing hydrogen as a result of its disosiation into interplanetary space. Hydrogen evaporates faster than heavy deuterium which is capable to be collected in the Earth hydrosphere. Therefore, as a result of this natural process of fractionation of isotopes throghout Eath evolution there should be an accumulationof deuterium in surface waters, while in atmosphere and in water vapor the deuterium contents are lower. Thus, on the planet there is going on a universal natural process of isotope separation, playing an essential role in maintenance of life on the planet. That is why during evolution process there should be an accumulation of deuterium in hydrosphere and surface waters.

According to the international standard VSMOW corresponding to Pacific ocean water that is rather stable on isotopic structure, the absolute contents of deuterium in oceanic water compiles D VSMOW/H VSMOW = (155,76±0,05) ×10 6 (155,76 ppm) [7]. For the international standard of natural water of Antarctic Region SLAP, which contains less deuterium, the absolute contents of deuterium compiles D SLAP/H SLAP = 89×10 6 (89 ppm).

In nature hydrogen and deuterium are widespred in the proportion 1:6400. In natural waters the contents of deuterium is distributed non-uniformly: raging from 0,02-0,03 mol.% for river and sea water, up to 0,015 mol.% for water from the Antarctic ice, - the most deuterium depleted natural water with the deuterium contents being in 1,5 times smaller, than in sea water. Thawed snow and glacial waters in mountains and some other regions of the Earth are also contain less heavy water, than usual drinking water. On the average, 1 ton of river water contains approximately 150-300 g of deuterium.

In D2O-H2O mixures there is occured an isotope exchange leading to the formation of semiheavy water” (HDO): Н2O + D2O = 2НDO. Therefore, deuterum at small amounts is present at water almost completely in form of НDO, and at high - in form of D2O. Physical properties of D2O differ from those for usual water [1]. It boils at 101,44 0С, freezes at 3,82 0С, has density at 20 0С 1,105 г/sm3, and the maximum of density is not on 4 0С, as for usual water, but on 11,2 0С (1,106 г/sm3). The above-mentioned factors together with the structure, density and viscosity of D2O in comparison with H2O lead to change of rates and kinetisc of enzime reactions in D2O-H2O mixures [9]. However, there are also such reactions which rates in D2O are higher, than in Н2O. Basically, they are reactions catalizing by ions D3О+ or H3О+ or ODand OH-.

The important fact is that carbon bonds forming by deuterium are stronger and stable, than carbon bonds forming by hydrogen. The big durability of D-O bonds, in comparison with H-O bonds causes distinctions in kinetics of chemical reactions between D2O and H2O. Thus, according to the theory of absolute speeds, break up of H-O bonds can occur faster, than D-O bonds, mobility of ion D3O+ is lower on 28,5 % than Н3Oion, and ОD-ion is lower on 39,8 % than OH-, the constant of ionization of D2O is less than constant of ionization of H2O [12]. It means, that in primary water on the Earth self-organizing deuterated structures could be existed longer in time. There are bases to believe, that during that epoch there was a process of structurization in heavy water environment of organic molecules because structuring properties and stabilizing influence of D2O on chemical bonds due to isotopic effects of deuterium are more expressed, than those for H2O (Mosin, 1996). As it was shown by our studies, the maximum kinetic isotopic effect which can be expected at ordinary temperatures in a chemical reaction leading to rupture of bonds involving hydrogen can be readily calculated, and the maximum ratio kh/kd in macromolecules is in the range of 7 to 9 for C-H versus C-D, N-H versus N-D, and O-H versus O-D bonds [8]. However, maximum ratios are seldom observed for a variety of reasons, but values of kh/kd in the range of 2 to 5 are common (Wiberg K. B., 1955). Deuterium located at positions in a macromolecule other than at the reaction locus can also affect the rate of a reaction. Such an effect is a secondary isotope effect and is usually much smaller than a primary isotope effect.

As have shown by our studies, biological objects sensitively react to change of isotopic composition of water. It is shown, that at placing a cell in D2O, not only H2O removes from a cell due to reaction of isotopic exchange Н2О-D2О, but also there is occured fast isotope (H-D) exchange in hydroxyl (-OH), sulfhydryl (-SH) and amino groups (-NH2) of all organic substances, including proteins, nucleinic acids, carbohydrates and lipids. It is known, that in these conditions only covalent C-H bond is not exposed to an isotopicxchange and, hereof only substances with bonds such as C-D can be synthesized de novo [10].

Depending on what position is occupied by deuterium atom in a molecule there are distinguished primary and secondary isotope effects of deuterium, stipulated by intermolecular interactions. In this aspect the most important for structure of a macromolecule are dynamic short-lived hydrogen (deuterium) bonds. They are formed between the neighbor atoms of hydrogen (deuterium) and heteroatoms of oxygen, carbon, nitrogen, sulfur, and play a leading role in maintenance of spatial structure of macromolecules and in intermolecular interactions.

Other important property is defined by the three-dimentional structure of D2O having the tendency to pull together hydrophobic groups of macromolecules to minimize their disruptive effect on the hydrogen (deuterium)-bonded network in the presence of D2O. This leads to stabilization of the structure of protein and nucleinic acid macromolecules in the ence of D2O.

Furthermore, the substitution of hydrogen atom with deuterium atom also affects the stability and geometry of hydrogen bonds in apparently rather complex way and may, through the changes in the hydrogen bond zero-point vibrational energies, alter the conformational dynamics of hydrogen (deuterium)-bonded structures of DNA and proteins in D2O. It may possibly cause disturbances in the DNA-synthesis, leading to permanent changes in its structure and consequently in the cells genotype. The multiplication which would occur in macromolecules of even a small difference between a proton and a deuteron bond would certainly have the effect upon its structure. The sensitivity of enzyme function to structure and the sensitivity of nucleic acids function (genetic and mitotic) to its structure would lead to a noticeable effect on the metabolic pattern and reproductive behavior of the organism in the the presence of D2O. And next, the changes in dissociation constants of DNA and protein ionizable groups when transfer the macromolecule from H2O to D2O may perturb the charge state of the DNA and protein.

Deuterated cells of various microorganisms adapted to the maximal concentration of D2O in growth media are convenient objects for evolutional and adaptational studies. During the growth of cells on D2O media there are synthesized macromolecules in which atoms of hydrogen in carbon skeletons are almost completely replaced on deuterium. As it has been shown by us, such deuterated macromolecules undergo the structural-adaptive chainges necessary for normal functioning of a cell in the presence of D2O.

We investigated isotope effects of deuterium in cells of various taxonomic groups of microorganisms realizing methylotrophic, hemoheterotrophic, photoorganotrophic and photosynthetic ways of assimilation of carbon substrates (methylotrophic bacteria, halobacteria, blue-green algae) in D2O. It is shown, that the effects observed at the growth of cells on D2O possess complex multifactorial character and are connected to changes of morphological, cytologic and physiological parameters - magnitude of the log - period, time of cellular generation, outputs of biomass, a ratio of synthesized amino acids, proteins, carbohydrates and lopids, and with an evolutionary level of the organization of investigated object as well. The experimental data testify that the cells realizing the special adaptive mechanisms promoting functional reorganization of work of the vital systems in the presence of D2O. Thus, the most sensitive to replacement of Н+ on 2H+ are the apparatus of biosynthesis of macromolecules and a respiratory chain, i.e., those cellular systems using high mobility of protons and high speed of break of hydrogen bonds. Last fact allows to consider adaptation to D2O as adaptation to the nonspecific factor effecting simultaneously functional condition of several numbers of cellular systems: metabolism, ways of assimilation of carbon substrates, biosynthetic processes, transport of substances, structure and functions of macromolecules.

Furthermore, deuterium induces physiological, morphological and cytological alterations in the cell. This leads to the formation in D2O of large atypical cells. They are usually 2-3 times larger in size and have a thicker cellular wall compared to the control cells, which are grown on ordinary water. Besides of that, the structure of DNA in deuterated cells in heavy water may alters; distribution of DNA in them was non-uniform (Mosin, 1996), (Mosin, Ignatov, 2012).

Biological experiments with D2O and its structure enable to analyze conditions at which life has evolved.

It is difficult to admit, that life could arise in chaotic" water. The structure of water testifies that life has evolved in the information water environment. The most favorable for origin of life are alkaline mineral waters, which interacte with calcium carbonate (CaCO3), and then sea waters (Ignatov, 2012). Once appeared in these waters. self-organizing of primary organic forms in the water environment was supported by thermal energy of magma, volcanic activity and solar radiation.

Let’s review the following reactions:

(1) CO2 + 4H2S + O2 = CH2O + 4S + 3H2O

(2) СаСО3+ HOH + СО2 = Ca(HCО3)2

The first equation shows how some chemosynthetic bacteria use energy from the oxidation of hydrogen sulfide (H2S) to sulfur (S).

The second equation is related to one of the most common processes in nature.

In the presence of water and carbon dioxide, calcium carbonate transforms into calcium hydrogencarbonate.

In the presence of hydroxyl OHions, the cellular processes are activated. Kagava demonstrates that an effect of improving the conductivity of the cell membrane is observed. The valid reaction is:

(3) CO2 + ОН- = HCО3-

(4) 2 HCO3- + Ca2+ = CaCO3 + CO2 + H2O

It is assumed that the second reaction has been valid upon the origination of the stromatolites.

When considering the issue of self-organization in nature, there is an exceptionally interesting example found in the karst springs in Zlatna Panega, Teteven district. Algae are surrounded with bubbles 3-5 mm in size. These bubbles are retained long enough – from hours to days. Water itself, which is similar in its spectrum to plants, “seeks” to preserve the self-organizing structures. During the study, the ambient temperature was 5 °C. It is known that if Ca2+ ions are added to a solution of pectin molecules, the solution is gelatinised. The reason is that Ca2+ ions bind to pectin molecules and cellulose microfibrils are formed. There is evidence that this kind of Ca2+ - sutures play a crucial role in the unification of the different components of the cell wall and influence its compactness and strength. Cytoplasm consists of 99% water, ions and other elements that form its basic structure.

In 2003, Arge and McKinnon received the Nobel Prize in chemistry for the discovery of water channels for water penetration into the cells. The aquaporin protein that regulates water exchange plays a role here. By increasing the concentration of calcium ions this process is more active.

cabanero and team carried out experiments, which show the role of Ca2+ in the permeability of the cell wall of plant cells.

In the most ancient organisms in evolution a silicon skeleton is observed. These are the sea sponges, the radiolaria and the diatomeae. Silicon was gradually displaced by the more active element calcium. The organisms reached the silicon-calcium skeleton and the most evolutionarily advanced of them – the calcium skeleton.

Molecular biology, almost like a dogma, adopts the model for the structure of the cell membrane. The main part of the membrane is made up of a double phospholipid layer.

In the Sargasso Sea a phenomenon is observed. Membranes of blue-green algae cells are made of phosphorus-containing lipids. The reason is that there is almost no phosphorus and nitrogen in the Sargasso Sea. The unicellulars have found a smart way to use sulfur instead of phosphorus in their lipid membranes. Apparently, in “self-organization” the structures choose the optimal components for them.

In late 2009 and early 2010 were carried out experiments with control “deionized” water, mineral water, sea water, and mountain water from Bulgaria. Water from karst springs was also studied. The experiments were made with Antonov’s device for spectral analysis of water. Cactus juice was studied too (Ignatov, 2009). The cactus was selected as a model system because the plant contains about 90% water. Also, photosynthesis is carried out by the enlarged stems, which serve for storage of water as well. Mineral water from different springs was examined.

Closest to the spectrum of tap, however, is the spectrum of mineral water, which reacts with calcium carbonate (СаСО3). Karst springs have a similar spectrum. Closest to the spectrum of sap is the spectrum of karst springs water, which interacts with mineral water.

In nature there are places with different water and identical external conditions. 25 km away from Teteven, Bulgaria there is the Zlatna Panega karst spring with a temperature from 10 to 12,5 °C. Three mineral springs with an average temperature of 21 °C pour into the lake formed by the spring. Only a few kilometers away is the Vit river with an average temperature of around 15 °C. There isobvious difference between the plant life in the water of the river and the karst spring. This is visible proof of the optimally good place for the active life of algae under identical external conditions. The difference is only in the structure of water.

Circulating in bowels on cracks, crevises, channels and caves karst waters are enriched with Ca(HCO3)2, actively cooperated with live matter and contain the information on a life in later geological periods.

As an atmosphere of the Earth during its evolution changed from reducive to oxidizing, gradually there was purification of hydrospheres from deuterium. At later stage of development water could be cleared from deuterium by means of rocks. The big role in this process, apparently, was played by karst rocks. Now the contents of deuterium in karst rocks are equaled 30-35 g/ton (Олег, скажи, ето правильно ли написано?), while in sea water its contents compose 150-200 g/ton. It is obvious, that when water contains less of deuterium, evolution may be proceed faster. If on the Earth would not begin natural process of deuterium purification, evolution could be stiffen at very low level.

Close in value peaks in the amplitudes of the spectra of sap and of karst and mineral springs, which interact with calcium carbonate are observed. Peaks are found at -0.1112, -0.1187, -0.1262, -0.1287 и -0.1387 еV. The average energy of the hydrogen bonds between water molecules in the creation of cluster formations is measured. Similar amplitudes in the spectrum between the sap and the mountain and sea water is observed at -0.1362 еV. The spectrum of the control “deionized” water is substantially different from that of tap, mineral and mountain water.

The evidence shown indicates that the emergence of life depends on the properties and structure of water and also on additional conditions (Ignatov, Mosin, 2009). Mineral water, which interacts with calcium carbonate is closest to these conditions and has left a trace in plants with its structure, and entropy. Next in line with regard to quality are sea and mountain water (Ignatov, 2010). In warm and hot mineral waters the peaks in the differential non-equilibrium energy spectrum (DNES) (Antonov, 1993) are more pronounced in comparison to the peaks in the same water with a lower temperature. This signifies that there is more energy for the preservation of a self-organized structure. The spectral range of DNES is in the middle infrared range from 8 to 14 micrometers. There is the earth atmosphere’s window of transparency for the electromagnetic radiation in the close and middle infrared range. In this interval energy is radiated from the Sun towards the Earth, and from the Earth towards surrounding space. Water changes with a cosmic rhythm. The likelihood of origination of life is biggest in warm and hot water with a specific structure (Ignatov, 2009, 2010). In January 2010, American scientist David Ward and colleagues described fossilized stromatolites in the Glacier National Park in the USA. They are studying microbes in Yellowstone National Park in USA, which are building stromatolites in hot water similar like ancient organisms. Rotorua in New Zealand is a similar place. Stromatolites have lived in warm and hot water in zones of volcanic activity. Warm and hot waters can be heated by magma as well. Their age is 3.5 billion years. These are the first organisms with fossil evidence. This is a confirmation and of the concept based on biophysical analyses for origination of life in warm and hot mineral waters and geysers (Ignatov, 2009, 2010) with more deuterium to the present time (Mosin, 2010). In June 2010, an article with this evidence was published at an Euromedica congress in Hanover, Germany. In common parlance, the Russian scientist Mosin calls the analyses “Water for the Origination of Life.” In September 2010, the American scientists Stockbridge, Lewis, Yung Yuan and Wolfenden published an article with the popular title “Is the Origin of Life in Hot Water?”. They consider the probability of faster biochemical reactions in hot water. Mosin thinks that in the beginning of evolution there was much more deuterium in water and this is a significant fact regarding its information abilities about the preservation of life.

In September 2011 a team of Japanese scientists led by Tadashi Sugawara also brought us closer to the secret that life has originated in warm or, more likely, hot water. They have created a proto cells, witch are similar of bubbles. For this purpose, they have made an aqueous solution of organic molecules, DNA and synthetic enzymes. The solution was heated to a temperature close to water’s boiling point – 95 degrees. Then its temperature was lowered to 65 degrees. A formation of a proto cells wits membrane was also observed. These proto cells are multiplying. This is a step for creation of synthetic cell. This laboratory experiment is an excellent confirmation of the possibility that life hasoriginated in hot water (Ignatov, 2010), (Ward, 2010). Upon changing the temperature is changing the middle energy of hydrogen bonds among water molecules, as an information bearer of life. (Ignatov, Antonov, 1998).

Our studies prove that life originated in containing deuterium hot mineral alkaline water.