Hot mineral water with more deuterium for origin of life and living matter. Compositions of the water for origin of lifeI. Ignatov, O.V. Mosin
It is believed that the big bang explosion occurred 3,7 billion years ago produced the universe that was much denser and hotter than today, and made up almost entirely of hydrogen (H). 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. Furthermore, the two formed deuterons stuck together into helium (4He) nuclei containing two protons and two neutrons. Thus, deuterium can serve as a feasible indicator of evolution, as natural amount of deuterium tends to be constant. It has been calculated, that during the formation of helium, almost all deuterium atoms combined to form helium nuclei, leaving a tiny remnant to be detected today, so that only one in 10 000 deuterons remained to be unpaired. Proceeding from the amount of deuterium, natural prevalence of deuterium makes up approximately 0,015 at. % D, and depends strongly on both the uniformity of substance and the total amount of matter formed in the course of early evolution. In 2007 the American astronomer J. Linski using FUSE ultra-violet telescope detected the increased contents of deuterium in the Milky Way galaxy . It became obvious that the natural amount of deuterium in outer Space is at least twice more, than it was expected. In particular, on the formation of stars it was spent not 1/3, but only 15 % D, turned then into 4He, and it is distributed non-uniformly. This new information can change radically our theoretical representations not only about formation of stars and galaxies, but also about molecular evolution. Constant sources of deuterium are explosions of nova stars and thermonuclear processes occurring inside the stars. Probably, it could explain a well known fact why the amounts of deuterium are increased slightly during the global changes of climate in worming conditions. 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 dissociation into interplanetary space. Hydrogen evaporates faster than heavy deuterium which is capable to be collected by the hydrosphere. Therefore, as a result of this natural process of fractionation of isotopes throughout the process of Earth evolution there should be an accumulation of deuterium in hydrosphere and surface waters, while in atmosphere and in water vapor deuterium contents are lower. Thus, on the planet there is going on a natural process of isotope separation, playing an essential role in maintenance of life on the planet.
According to the international standard VSMOW corresponding to Pacific ocean water which is rather stable on isotopic structure, the absolute contents of deuterium (isotopic shifts, δ) compile D VSMOW/H VSMOW = (155,76±0,05) ×10-6 (155,76 ppm) . For the international standard of natural water of Antarctic Region SLAP containing less deuterium, the absolute contents of deuterium compile D SLAP/H SLAP = 89×10-6 (89 ppm). The average ratio of H/D in nature compiles 1:6400. In natural waters the contents of deuterium are distributed non-uniformly: from 0,015 at.% D for water from the Antarctic ice - the most deuterium depleted natural water with deuterium contents in 1,5 times smaller, than in sea water, up to 0,02-0,03 at.% D for river and sea water. Thawed snow and glacial waters in mountains and some other regions of the Earth usually contain on 3-5% less deuterium, than usual drinking water. On the average, 1 ton of river water contains approximately 150-300 g of deuterium. Other natural waters contain varying levels of deuterium from δ = +5.0 D,%, SMOW (Mediterranean Sea) up to δ = -105 D,%, SMOW (Volga River).
We proposed earlier that primary water could contain more deuterium in early stages of evolution of life, and deuterium was distributed non-uniformly in hydrosphere and atmosphere . The reason of this is that the primary reductive atmosphere of the Earth, consisted basically from mixture of gases (CO, H2, N2, NH3, CH4), was lacked O2–O3 layer protecting the Earth surface from rigid short-wave solar radiation carrying huge energy. This simplifies radiation to be freely passed by through O2-free atmosphere and reaching hydrosphere, may be the cause of further radiolysis and photolysis of water. Furthermore, energy of radiation, volcanic geothermal processes on a hot Earth surface and electric discharges in atmosphere, could lead to the enrichment of hydrosphere by deuterium in form of HDO that evaporates more slowly then H2O, and condenses faster. The formation of HDO occurs in D2O-H2O mixtures via isotopic exchange: Н2O + D2O = 2НDO, causing deuterium at small amounts to be present in water almost completely in form of НDO, and at high amounts - in form of D2O. Physical properties of D2O differ from those for H2O: D2O 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). These factors as well as the structure, density and viscosity of D2O in comparison with H2O lead to the changes of kinetic rates of enzymes reactions in D2O-H2O mixtures . However, there are also such reactions which rates in D2O are higher, than in Н2O. Basically, they are reactions catalyzing by ions D3О+ or H3О+ or ODand OH-.
According to the theory of chemical bond, breaking up of H-O bonds can occur faster, than D-O bonds, mobility of ion D3O+ is lower on 28,5 % than Н3Oion, and ОDion is lower on 39,8 % than OH- ion, the constant of ionization of D2O is less than constant of ionization of H2O. It means that in primary water on the Earth self-organizing deuterated living structures could have been existed longer in time. There are bases to believe, that during that epoch there was a process of structuring in 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 . As it was shown by our studies, the maximum kinetic isotopic effect which can be observed at ordinary temperatures in a chemical reaction leading to rupture of bonds involving H and D was calculated, and the maximum ratio kh/kd in macromolecules is in the range of 6 to 8 for C-H versus C-D, N-H versus N-D, and O-H versus O-D bonds . However, maximum ratios are seldom observed for a variety of reasons, and average values of kh/kd in the range of 2 to 5 are more common. Deuterium located at positions in a macromolecule other than at the reaction locus can also affect the rate of a reaction. This effect is a secondary isotopic effect and is usually much smaller than a primary isotopic effect. In this aspect the most important are dynamic short-lived hydrogen (deuterium) bonds formed between the neighbor atoms of hydrogen (deuterium) and O, C, N, Sheteroatom, playing a leading role in maintenance of spatial structure of macromolecules and intermolecular interactions. Furthermore, the substitution of H with D 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 protein in D2O. It may cause disturbances in the DNA-synthesis, leading to permanent changes in DNA structure and consequently in cell 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 the structure. The sensitivity of enzyme function to the structure and the sensitivity of nucleic acid function (genetic and mitotic) would lead to a noticeable effect on the metabolic pathways and reproductive behavior of an organism in the presence of D2O. And next, the changes in dissociation constants of DNA and protein ionizable groups when transferring the macromolecule from H2O to D2O may perturb the charge state of the DNA and protein molecules. Other important property is defined by the three-dimensional structure of D2O molecule having the tendency to pull together hydrophobic groups of macromolecules to minimize their disruptive effect on the hydrogen (deuterium)-bonded network in D2O. This leads to stabilization of the structure of protein and nucleic acid macromolecules in the presence of D2O .
Our studies have demonstrated that biological objects sensitively react to change of isotopic composition of water. It is shown, that at placing a cell in D2O, not only H2O is removed from a cell due to reaction of isotopic exchange Н2О-D2О, but also there is occurred fast isotopic (H-D) exchange in hydroxyl (-OH), sulfhydryl (-SH) and amino groups (-NH2) of all organic substances, including proteins, nucleic acids, carbohydrates and lipids. It is known, that in these conditions only covalent C-H bond is not exposed to isotopic (H-D) exchange and, thereof only substances with bonds such as C-D can be synthesized de novo .
Deuterated cells of various microorganisms adapted to the maximal concentration of D2O in growth media (95-98 vol.% D) are convenient objects for evolutional and adaptation studies as well as structural-functional studies. During the cellular growth on D2O media there are synthesized macromolecules in which hydrogen atoms in carbon skeletons are almost completely replaced on deuterium. As it has been shown earlier, such deuterated macromolecules undergo the structural-adaptive modificational changes necessary for normal functioning of cells in the presence of D2O.
We have investigated isotopic effects of deuterium in prokaryotic and eukaryotic 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 with using 1H-NNR-, IR-, and mass-spectrometry technique. It was demonstrated, that the effects observed at the cellular growth on D2O possess complex multifactorial character to be connected to the changes of morphological, cytologic and physiological parameters - magnitude of the log-period, time of cellular generation, outputs of biomass, a ratio of amino acids, protein, carbohydrates and lipids synthesized in D2O, and with an evolutionary level of organization of investigated object as well. The experimental data testify that cells realize 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 D+ 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 breaking up of hydrogen bonds. Last fact allows 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, and transport function, structure and functions of macromolecules. There is evidence that during adaptation to D2O the ration of synthesized metabolites is changing. 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 grown on H2O. Besides of that, the structure of DNA in deuterated cells in D2O may alters; distribution of DNA in them was non-uniform . The data obtained confirm that adaptation to D2O is a phenotypical phenomenon as the adapted cells return back to normal growth after some log–period after their replacement into H2O. At the same time the effect of convertibility of growth on H2O/D2O does not exclude an opportunity that a certain genotype determines displaying of the same phenotypical attribute in D2O.
Biological experiments with D2O and structural-conformational studies enable to analyze conditions at which life has evolved. It is difficult to admit, that life could arise in "chaotic" non-informative water. The unique structure of water testifies that life has evolved in the informative water environment. The most favorable for origin of life are alkaline mineral waters interacting with CaCO3 and then sea waters . Circulating in bowels on cracks, crevices, channels and caves karst waters are enriched with Ca(HCO3)2, actively cooperating with live matter and contain the information about life in later geological periods. Once appeared in these waters the process of self-organization of primary organic forms in water solutions may be 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. Contemporary chlorophyll contains the elements C, H, O, N, Mg.
These data have proved that life originated in hot mineral alkaline water containing more deuterium. The evidence shown indicates that the emergence of life depends on the properties and structure of water and also on additional conditions. 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 . The data obtained also have shown a possible way of transition from synthesis of small organic molecules due to the energy of ultra-violet solar radiation and thermal activity to more complex organic molecules as protein and nucleic acids. Protein molecules are composed from one or several polypeptide chains, consisting of a big number of various amino acids. Their subsequent condensation into polypeptide chains can take place under certain conditions after their formation. The important factor in reaction of condensation of two molecules of amino acids is allocation of H2O molecule when peptide chain is formed. As reaction of polycondensation of amino acids is accompanied by dehydratation, the H2O removal from reactional mixture speeds up the reaction rates. This testifies that early evolution of life may occur nearby active volcanoes, because at early periods of geological history volcanic activity occurred more actively than during subsequent geological times. However dehydratation accompanies not only amino acid polymerization, but also association of other blocks into larger organic molecules, and also polymerization of nucleotides into nucleic acids. Such association is connected with the reaction of condensation, at which from one block removes proton Н+, and from another – hydroxyl group (OH-) with formation of H2O molecule.
The possibility of existence of condensation-dehydratation reactions under conditions of primary hydrosphere was proven by Calvin in 1965 . From all chemical substances hydrocyanic acid (HCN) and its derivatives – cyanoamid (HNCN2) and dicyanoamid HN(CN)2 possess dehydratation ability and the ability to catalyze the process of linkage of H2O from primary hydrosphere . The presence of HCN in primary hydrosphere was proven by Miller's early experiments. Chemical reactions with HCN and its derivatives are complex with chemical point of view; in the presence of HCN, HNCN2 and HN(CN)2 the condensation of separate blocks of amino acid molecules accompanied by dehydratation, can proceed at normal temperatures in strongly diluted H2O-solutions (Fig. 1, Fig. 2). Furthermore, polycondensation of amino acids in the presence of HCN and its derivatives depends on acidity of water solutions in which they proceed . In acid water solutions (рН 4-6) these reactions do not occur, whereas alkaline conditions (рН 8-9) promote their course. There has not been unequivocal opinion up till now, whether primary ocean had alkaline composition, but it is quiet probable, that such a value of рН was possessed by mineral waters adjoining with basalt, and these reactions could occur at contact of water with basalt rocks.
Fig. 1. Reactions of condensation and dehygratation catalised by HCN and its derivatives, resulting in formation from separate molecules of larger organic molecules. The top three equations: condensation and the subsequent polymerization of amino acids in proteins, carbohydrates in polycarboxydrates and acids into lipids. The bottom equation - condensation of adenibe with ribose and Н3РО3, leading to formation of nucleo base .
Fig 2. Prospective mechanisms of purine formation - adenine, guanine and xantine from water mixture NH3 and HCN (above) and adenine from water mixture NH3-HCN (below) at +95 0С. (total reaction: 5HCN = adenine) .
In model experiments amino acid mixtures were subjected to influence of temperatures from 60 0C up to 170 0С with formation of short protein-like molecules resembling early evolutionary forms of proteins . The best results on polycondensation were achieved with the mixes of amino acids containing aspartic and glutamic acids, which are essential amino acids occurring in all modern living organisms. During synthesis short proteins were formed from them similar to the natural proteins, subsequently designated as thermal proteinoids. They are consisted of large molecules with molecular weight up to 300000, consisting of the same amino acids, as natural proteins. On quantitative structure they usually contain 18 of 22 amino acids usually occurring in protein hydrolysis’s of modern organisms that correspond to the general definition of protein. The synthesized proteinoids are similar to natural proteins on a number of other important properties, for example on linkage by polinucleobases, on suitability for food to bacteria and rats, on ability to cause the reactions similar to those catalyzed by enzymes in living organisms, such as decarboxylation, amination, deamination, and oxidoreduction. Thus, synthesized proteinoids are capable to catalytically decompose glucose  and to have an effect similar to the action of a-melanocyte-stimulating hormone .
For last years it has been made much progress in studying the structure and properties of proteinoids. While treating a hot mixture of proteinoids in water solutions of salts, in the reaction mixture are formed elementary membrane like proteinoid microspheres . The size of proteinoid microspheres is relatively small; their diameter makes up approximately 5-10 µm (Fig. 3). On morphological features proteinoid microspheres remind a cellular membrane which may be as well double.
Fig. 3. Microphotograps of protenoid microspheres in water solution .
In view of the preceding data the origin of life looks as follows. The initial stage of evolution, apparently, was formation in primary hydrosphere at high temperatures from mixtures of amino acids and nitrogenous substances - analogues of protein and nucleic acids. Such synthesis is possible under conditions of reductive atmosphere at presence in water solutions H3PO3 at high temperatures and ultra-violet radiation . The next stage is polycondensation of amino acids into thermal proteinoids at temperatures 65-100 0С at the presence of phosphates and aspatric and glutamic acids in water. Then in a mix of proteinoids in hot water solutions were formed membrane like structures. The ability of proteinoids to perform some functions similar to enzymic functions is expressed that they can in the presence of Zn2+ split down ATP. Additionally, proteinoid microspheres may possess the ability to synthesize RNA coding short protein.
In September 2011 a team of Japanese scientists led by Tadashi Sugawara also brought us closer to the fact that life has originated in warm or, more likely, hot water. They have created a proto cells, wich were similar to the 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 0С, then its temperature was lowered to +65 0С. Under these experimental conditions the formation of proto cells with membrane was observed. These proto cells are multiplying that is a further step for creation of synthetic cell. This laboratory experiment is an excellent confirmation of the possibility that life originated in hot water (Ignatov, 2010; Ward, 2010).
The important role in a process of origin of life evidently was played by the most widespread mineral of the earth's crust - quartz (SiO2). The crystal of quartz possesses of tetrahedral structure from which there can be derived various chained and tape silicate structures bound with each others (Fig. 4). Uniqueness of quartz consists in that its crystals are optically active, i.e. are capable to interact with polarized light. Therefore, on surface of L-and D-enantiomer quartzes crystals was possible selective absorption L-and D-isomers, that can explain stereoselectivity of evolution.
Fig. 4. Crystal structures of quartz (left) and tetrahedral structure of water (right).
The prospective structure of water could be caused by its ancient geological connection with quartz and other calcium-silicate minerals prevailing in the earth’s crust, in contact with which water were placed in. That is why the development of the most ancient forms of life is connected to calcium-silicate minerals. The most ancient proofs of existence of living organisms with layered limy structure are dated back to 3,5 billion years. These most ancient limy (dolomite) formations of Precambrian period - stromatolites were building their skeletons of limestone and SiO2. Stromatolites were formed at the bottom of superficial reservoirs in Arching during the most ancient geological epoch of the Earth - 2,5-3,5 billion years ago. Studying of these formations is very important as stromatolites carry data about life origination and organic structure of the first living organisms - numerous colonies of cyanobacteria, diatom algae and oil assimilating bacteria existing in thicknesses of limestones and dolomite in the muzzles of volcanoes and thermal sources. These planktonic forms of microorganisms lived in the top layers of sea water together with other organisms possessing limy (Foraminifera) and chitinous skeletons the size of which reaches 10 µm. After death these organisms sedimented on a sea-bottom, and the substance of their skeleton was interacted chemically with sea water. CaCO3 of Foraminifera and chitin of other planktonic microorganisms were dissolved in water better than SiO2 of diatoms and radiolaria, forming silica deposits. Siliceous slates with adjournment of these siliceous microorganisms were formed during of Phanerozoic geological epoch in deep ocean hollows, on depths about 1-2 km deep. Temporary blossoming of an organism with SiO2 skeleton could lead to such congestion of silicon in oceanic waters. Then SiO2 could crystallize into limestone in the centers of crystallization, gradually replacing CaCO3 molecules by SiO2. Later on, the organisms possessing limy skeletons - Foraminifera began to absorb Ca2+ from calcareous breeds.
When considering the processes of self-organization in nature, there is an exceptionally interesting phenomenon found in the karst springs in Zlatna Panega, Teteven district (Bulgaria). Algae found there are surrounded with bubbles 3-5 mm in size. These bubbles are retained long enough – from hours to days. It is most likely that water itself, which is similar in its IR-spectrum to the IR-spectrum of plant sap, tends to preserve the self-organizing structures. It is known that if Ca2+cations are added to a solution of pectin molecules, the solution is gelatinised. The reason for this is that Ca2+cations bind to pectin molecules and cellulose microfibrils are formed. There is evidence that these Ca2+-complexes play a crucial role in the unification of the different components of the cell wall and influence its compactness and strength.
In late 2009 and early 2010 there were carried out experiments at the Research Centre for Medical Biophysics in Bulgaria with samples of
deionized water (control), mineral water, sea water, water from karst springs, and mountain water from Bulgaria, analysed with using Antonov’s device for IR-spectral analysis of water. Mineral water samples from different Bulgarian springs were also examined. Cactus juice was selected as a model system because the plant contains about 90% H2O. The data demonstrated that closest to the IR-spectrum of tap water, however, was the IR-spectrum of mineral water reacted with СаСО3. Water samples from Karst springs have a similar IR-spectrum. Closest to the IR-spectrum of sap is the IR-spectrum of karst spring water. Peaks measured by IR-spectroscopy were detected at -0.1112, -0.1187, -0.1262, -0.1287 and -0.1387 еV. Similar amplitudes in the IR-spectrum between the sap and the mountain and sea water were observed at -0.1362 еV. The IR-spectrum of the control
deionized water is substantially different from that of tap, mineral and mountain water. Further on, the average energy of the H...O-bonds between H2O molecules in the formation of water cluster associates (H2O)n was measured at – 0.1067±0.0011 eV. Upon changing the temperature it is changing the middle energy of H...O-bonds among H2O-associates, that makes it possible information transferin.
The results indicate that the emergence of life depends on the properties and structure of water and on additional conditions as well as temperature and pH. Mineral water, which interacts with CaCO3 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 [10, 18]. In warm and hot mineral waters the IR-peaks in the differential non-equilibrium energy IR-spectrum (DNES) (Antonov, 1993) are more pronounced in comparison to the IR-peaks received in the same water with a lower temperature. This may signify that there is more energy for the preservation of self-organized structures in time. The spectral range of DNES was in the middle infrared range from 8 to 14 micrometers. It is thought that 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 may change its structure and physical-chemical properties with cosmic rhythms. The likelihood of origination of life is biggest in warm and hot water with a specific structure . In January 2010, American scientist David Ward and his colleagues described fossilized stromatolites in the Glacier National Park in the USA. They studied microorganisms in Yellowstone National Park in USA, which formed stromatolites in hot water by similar like ancient organisms. Stromatolites were formed in warm and hot water in zones of volcanic activity, which can be heated by magma. This fact is a confirmation of the concept based on biophysical analyses for origination of life in warm and hot mineral water and geysers  with more deuterium to the present time. In June 2010, an article with this evidence was published at Euromedica congress in Hanover, Germany. In common parlance, the Russian scientist O. V. 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? in which they considered the probability of faster biochemical reactions in hot water. O.V. Mosin thinks that in the beginning of evolution there was much more deuterium in water, and this is a significant fact regarding thermal stability of deuterated macromolecules in the preservation of life under thermal conditions.
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