Reproducible Peak Clusters on Differential Mouse Mortality Curves and Their Relation to the Gompertz Model.

It is shown that differentiation of mouse mortality curves (number of animals that died at a certain age plotted versus their lifespan) results in the appearance of eight clearly distinguished clusters of peaks corresponding to increased mortality rates. Smoothing of the original mortality curves and subsequent transformation of the differential mortality curves according to the Gompertz model makes the peaks and the corresponding clusters less pronounced and drives the logarithm of the force mortality curve toward a straight line. The positions of the clusters on the lifespan axis (expressed in days) were calculated as weighted means by dividing the sum of the products of multiplication of the peak heights and their position on the lifespan axis by the sum of the peak heights within a cluster. To prove that the peaks and their clusters are not random, we have demonstrated that the positions of the clusters on the lifespan axis do not depend on the extent of mortality curve smoothing or the group of mice analyzed.

New Data on Programmed Risks of Death in Normal Mice and Mutants with Growth Delay.

Study of the lifespans of normal (non mutant) mice and growth delay mutants has shown that mortality rates for both kinds of animals exhibit reproducible fluctuations. In the case of the mutant mice, the positions of peaks on the differential mortality curves (mortality rate plotted against lifespan) coincided in different-sex groups of animals and in same-sex subgroups of animals. Differential mortality curves of the mutant mice also had a peak at 1 month of age that was absent from the differential mortality curves of the normal mice. In the case of normal animals, positions of most peaks were the same in the studied independent subgroups of males, and to a lesser extent - independent subgroups of females, which might be explained by a shift in mortality peak positions due to the reproductive activity of females. Similar positions of mortality rate peaks in the differential mortality curves for animals from independent groups and subgroups indicate the existence of increased risks of death at specific ages. The observed pattern could be due to the programming in the genome of both the periods of increased risk of death and the intermitting intervals of stable development.

Age fluctuations in mortality of mice with mutation causing growth retardation.

Lifespan of mice over a number of consecutive generations of descendants of a male with a mutation causing growth retardation was studied. The mutant and normally developing (normal) mice were obtained by crossbreeding of mutant males with normal females from the same brood. The mutant females were infertile. Mortality of the mutant and normal mice was shown to fluctuate depending on age. The curve of dependence of lifespan on their serial number in a series of lifespan increase (mortality rank curve) had the form of evident steps for the mutant mice, while in normal mice this feature was less pronounced. These steps indicate that in the course of development of mice stages with low mortality are alternately replaced by stages with increased mortality. One month after birth, the first stage of stable development of mutant males and females is replaced by a stage with abnormally high mortality, which coincides with the period of their maximal backlog in weight compared to the normal animals. Within two months, surviving mutants catch up in weight with normally developing mice and externally become indistinguishable from them. The steps are reproduced on mortality rank curves in mutant and normal mice, both in groups of mice of different sexes and in parallel same-sex groups. The observed phenomenon is interpreted within the hypothesis of a genetic aging program in mice that provides periodic changes when stages of great viability are followed by stages of increased sensitivity to the external risk factors causing death. Less-expressed steps on mortality rank curves of normal females were shown to be enhanced by the removal from the sample of parous females and animals with tumors. Results of the study indicate the possibility of detecting in humans of ontogenesis-programmed stages of high and low sensitivity to external influences and the prospect of the development of effective measures to prevent risks of premature death.

[Graduated change of life expectancy in mice in ontogenesis].

Life expectancy of descendants of a normal female mouse and a male with an inherited growth inhibition mutation discovered in a laboratory population was investigated. The hereditability of the characteristic allows us to consider it a result of mutation. It was shown that, in mice, the curve of dependence of life expectancy on their serial number in a row of increase in life expectancy (curve of rank distribution) has step-like shape for mutant males and females, as well as for males with normal development. The first grade of mice death on the curve of rank distribution was observed at one month after their birth and was characteristic only of males and females with a mutation during the period of maximum lag in weight as compared with their normal relatives. The surviving mutants catch up to the normally developing individuals within two months and externally become indistinguishable from them. The subsequent grades of death in mutants and normal males coincide on the time axis. The steps are absent on the rank curves of life expectancy of normally developing females. The time intervals between the steps are reproduced in parallel groups of mice and, hence, are not casual deviations from theoretical curves and are of a regular nature. The discovered phenomenon is interpreted within the scope of a hypothesis about the realization of the genetic program of ontogenesis, which provides periodic change of vitality stages with stages of sensitivity to external risk factors, which increase the probability of death, by mice. Absence of such stages in the group of normally developing females can be explained by shifts in development, which are produced by the irregular performance of reproductive functions.

[Influence of carbon dioxide on the impregnation of the nervous tissue by silver].

It has been shown that 4% carbon dioxide (CO2) in the air above reaction mixture inhibits the initiation of the formation of silver nanoparticles from complexes with biogenic amines (noradrenaline and serotonin). At the same concentration of CO2 in the air above solution of AgNO3, which is used for staining nerve tissues by the method of Golgi, neurons are preferentially stained, whereas at a concentration of 0.06%, vessels and poor neurons are stained. It is suggested that the entry of free silver ions to neurons is due to the inhibition of sites of initiation of silver nanoparticles in vessels at high CO2 concentrations, while the lack of inhibition leads to silver precipitation in vessels at low CO2 concentrations. It can be assumed that, for stable silver impregnation, the concentration of CO2 must be controlled.

[The different effects of carbon dioxide on the toxicity of silver ions for prokaryotic and eukaryotic microorganisms].

The effect of carbon dioxide on survivability of bacteria Escherichia coli and the germination ability ofconidia of the fungus Neurospora crassa in the presence of silver nitrate was studied. It was shown that carbon dioxide increased the toxic effect of silver ions on prokaryotic cells of E. coli but did not change the survivability of spores of the eukaryote N. crassa.

[Carbon dioxide of air inhibits the formation of silver nanoparticles initiated by proteins in polyacrylamide gel and in solution].

It was shown that the detection of proteins in polyacrylamide gel by silver is inhibited by contact with air of the ammonia complex with silver ions used at the first stage of detection. It was proved by experiments on the reduction of silver by ethanolamine from a complex with ethanolamine and by formaldehyde from a complex with ammonia that the formation of silver nanoparticles initiated by proteins is inhibited by air carbon dioxide. The participation of carbon dioxide in this process is discussed. It was found that even the breathing of an experimenter can induce variations in carbon dioxide concentration sufficient to adversely affect the reproducibility of the silver staining techniques. It was concluded that, for stable staining of proteins by silver in polyacrylamide gel, it is necessary to maintain a low concentration of carbon dioxide in air over the detection solutions.

[Isolation of cell membranes from Saccharomyces cerevisiae for evaluation of the protein composition]. / Poluchenie kletochnykh obolochek Saccharomyces cerevisiae s tsel'iu opredeleniia ikh belkovogo sostava.

We describe a simple method for the isolation of membrane fractions from Saccharomyces cerevisiae yeasts, containing the complex of plasma membranes and cell walls. The method is based on cell disruption on an INBI flow disintegrator. This device spares subcellular structures, which simplifies the isolation of cell membranes. The membrane fraction obtained by this method was suitable for studies of protein composition of these structures by means of two-dimensional electrophoresis.

Structure-chemical approach to organization of information on metabolic charts.

A nontraditional approach to construction of metabolic charts is proposed. It is based on the discovery of symmetry in the structure of the network of metabolic reactions. So if compounds and reactions are located on the metabolic chart according to their chemical features, the chart structure will acquire a periodic pattern. The charts thus created have a natural two-dimensional coordinate system of the metabolic network. Points on the X-axis correspond to number of carbon atoms in the carbon skeleton of compounds in columns and points on the Y-axis correspond to number of -COOH groups in compounds filing in series of rows on the charts. As a result this coordinate system sections the field of the charts into rectangular blocks each of which containing compounds with the same numbers of carbon atoms and the same numbers of -COOH groups. The latter substantially improves the charts and makes them a more valid source of metabolic data possessing heuristic properties. The periodicity of the metabolic network structure enables us easily to remember information about biochemical reactions and their products. The charts can also be used as a universal key for any biological database information that is systematically connected with the metabolic information. The charts can be important for medicine and pharmacology because they can help to understand the metabolic processes involved in decomposition of a particular drug or to find the chain of reactions blocked or initiated by administering this drug into a living organism.

[The theory of phyllotaxis. II. Crystallographic interpretation of the interrelation between lower and superior phyllotaxis forms]. / Teoriia fillotaksisa. II. Kristallograficheskaia interpretatsiia vzaimosviazi mezhdu nizshimi i vysshimi formami fillotaksisa.

Cross-opposite phyllotaxis forms are defined as superior with respect to the alternate ones and verticillate phyllotaxis forms as superior with respect to the opposite ones. Different phyllotaxis forms can be interpreted as a result of stretching of crystal-like structures of the embryo formed by dense packing of rudiments. Based on hypothetical concepts of the properties of plant rudiments and embryos, possible mechanisms of the formation of superior phyllotaxis forms from the lower ones have been analyzed. It was shown that the superior phyllotaxis forms can be considered as the results of additive summation of the lower forms. The theoretical conclusions are confirmed by the examples of polymorphic phyllotaxis in conspecific plants and by the facts of accidental splitting of superior phyllotaxis forms into the corresponding lower forms in nature and in experiment. The mechanisms underlying the formation of multiple forms of helical phyllotaxis have been proposed. The concept of a new type of mixed hexagonal-tetragonal phyllotaxis has been formulated and the mechanism of its formation has been considered. The forms of corn grain packaging in the corncob and leaf arrangement on the strawberry tomato stem are given as examples of true hexagonal-tetragonal phyllotaxis in nature.

[Theory of phyllotaxis. I. A geometric model for helical forms of consecutive phyllotaxis]. / Teoriia fillotaksisa.I. Geometricheskaia model' obrazovaniia spiral'nykh form ocherednogo fillotaksisa.

We have developed a geometric model for helical forms of consecutive phyllotaxis on the basis of an axiomatic approach. It follows from the model that rudiment growth and the movement of the cylindrical rudiment surface in the absence of a displacement in the direction along the rudiment axis leads to a repeating transition of tetragonal packaging of the rudiment into hexagonal packaging and vice versa. Under these conditions, sequences of rudiments produce left-handed and right-handed helices, the number of which at the circumference of the cylinder corresponds to adjacent numbers of the Fibonacci series. We demonstrate that the left-handed and right-handed isomers of helical forms of the consecutive phyllotaxis appear as a result of the transition of an unstable symmetric structure of the embryo at early developmental stages into stable left-handed or right-handed structures.

Structural symmetry of the metabolic reaction network. I. Carboxylic acid metabolism.

A new approach to systematization of the information on metabolism, based on the symmetry observed in the pattern of the metabolic reaction network, is discussed. A theoretical substantiation of symmetry existence is given. A symmetrical metabolic scheme is developed for metabolism of carboxylic acids. The symmetry of the metabolic reaction network can be used for systematization of biochemical, physiological, medical and other data associated with metabolism as well as for prediction of the new information.

[Structural theory of phyllotaxis. II. Relationship between lower and higher forms of phyllotaxis]. / Strukturnaia teoriia fillotaksisa. II. Vzaimosviaz' mezhdu nizshimi i vysshimi formami fillotaksisa.

Opposite phyllotaxis forms are defined as superior ones in relation to alternate phyllotaxis forms, and verticillate phyllotaxis forms are defined as superior ones in relation to opposite phyllotaxis forms. On the basis of hypothetical notions about the properties of plant bumps and embryos, the probable mechanisms of creation of superior phyllotaxis forms from the lower ones are analyzed. It is shown that superior phyllotaxis forms can be considered to result from the combination of lower ones and that the superior forms can be split into the corresponding lower ones under artificial or natural influences.

Effect of light on generation of colored silver colloids in protein solutions.

The exposure to light (20 mW/cm2, an incandescence lamp) of weakly alkaline protein solutions which contained silver nitrate and formaldehyde initiated reduction of silver ions with the subsequent generation of colored silver colloids. At light intensities lower than 0.2 mW/cm2 the generation of colored silver colloids was delayed. The rate of silver reduction depended on the protein type and on the light spectral structure. In particular, solutions which contained prealbumin, lysozyme, gamma-globulin, and transferrin were more photosensitive than solutions which contained albumin, pepsin, and beta-amylase. The formation of [Ag(NH3)2]+ complex after an addition of ammonium ions into the solutions preferentially suppressed silver reduction in the dark and under exposure to red light, thus resulting in a significant difference in the time of appearance of colored silver colloids when the solutions were exposed to violet or red light. These findings are promising for the elaboration of selective silver development of proteins in polyacrylamide gels.

[Regularity in the pattern of the metabolic reaction network]. / Reguliarnost' v strukture seti reaktsii metabolizma.

A basically new approach to the presentation of information on metabolism based on the regularity in the pattern of metabolic reaction network is discussed. The existence of such regularity is theoretically substantiated. The metabolic charts of carbohydrates, carboxylic acids and nitrogen-containing compounds with a regular structure are given. The possibilities of exploiting the regularity of the metabolic reaction network in the systematization of information relevant to the metabolism and prognostication of new information are considered.

[Kinetics of DNA-dependent RNA synthesis: couple synthesis of di-, tri- and tetranucleotides in the presence of a limited set of substrates]. / Kinetika DNK-zavisimogo sinteza RNK: sopriazhennyi sintez di-, tri- i tetranukleotidov v prisutstvii minimal'nogo nabora substratov.

The qualitative and quantitative characteristics of the short oligonucleotides synthesis by Escherichia coli RNA polymerase on A1 promoter of the bacteriophage T7 in the presence if incomplete set of nucleoside triphosphates were studied. The binding of the fourth substrate with enzyme-template complex was shown to occur after binding of the third substrate only. The curves of di-, tri- and tetranucleotide synthesis as the function of CTP and GTP concentration were constructed. The empiric formulas for the rates of the coupled synthesis if tri- and tetranucleotides were derived from these curves. A kinetic scheme describing the experimental data was proposed.

[Kinetics of DNA-dependent RNA synthesis: coupled synthesis of di- and trinucleotides in the presence of a minimum complement of substrate]. / Kinetika DNK-zavisimogo sinteza RNK: sopriazhennyi sintez di- i i trinukleotidov v prisutstvii minimal'nogo nabora substratov.

The qualitative and quantitative characteristics of the synthesis of the short oligonucleotides by Escherichia coli RNA polymerase on A1 promoter of the bacteriophage T7 deletion mutant delta D111 DNA in the presence of the incomplete set of nucleoside triphosphates were studied. It was shown, that in conformity with the structure of A1 promoter the oligonucleotides pppApU, pppApUpC were synthesized in the presence of ATP, UTP, CTP; the oligonucleotides pApU, pApUpC-in the presence of AMP, UTP, CTP and oligonucleotides pApU, pApUpC, pApUpCpG-in the presence of AMP, UTP, CTP, GTP. The curves of di- and trinucleotide syntheses as the functions of the substrate concentrations were obtained. The analytical formulas for the rates of the coupled synthesis were derived from these curves. A kinetic scheme that is in conformity with the experimental data was proposed. This scheme includes the stage of the reversible, random and release of di- and trinucleotides from the enzyme-template complex.

[Kinetics of DNA-dependent synthesis of RNA: correlation between abortive and productive initiations]. / Kinetika DNK-zavisimogo sinteza RNK: sootnoshenie reaktsii abortivnoi i produktivnoi initsiatsii.

A general equation of abortive initiation kinetics is inferred at its coupling to a productive initiation. It describes the abortive initiation dependence upon time and concentrations of the initiating nucleotide and the following nucleosidetriphosphate (NTP). The equation analysis revealed the abortive initiation dependence upon NTP's concentration in a characteristic manner possessing a maximum at the time it comes to saturation. The kinetic constants were estimated experimentally from the plot when the extent of pApU synthesis was plotted first against time in presence of a complete set of NTP's and second, against AMP and UTP concentration in the absence of the other three NTPs. The estimated values of the constants were introduced into the equation to analyze the dependence of pApU synthesis upon NTPs concentration ([ATP]=0.2 [UTP]=[GTP]=[CTP]=[NTP]). The theoretically calculated curve was compared to the experimentally obtained. Good coincidence of these curves supports the correctness of the general equation. The general equation of abortive initiation gives possibility to infer an equation of transcription involving the initiation and the elongation studies.