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Some properties of the cell-free DNA (cfDNA) of peripheral blood plasma were assessed in 153 employees of atomic industry enterprises. The contents of ribosomal repeat (rDNA) and its concentration in plasma increased in cfDNA of the group of persons in comparison with non-irradiated individuals. The contents of satellite III in cfDNA of donors and of irradiated persons do not differ and less than in DNA nucleus. The correlation between cumulative dose of radiation, contents of rDNA in cfDNA and the frequency of lymphocytes bearing mutations at T-cell receptor (TCR) locus was obtained. The definition of three indications in irradiated persons: the contents of ribosomal genes in cfDNA, TCR-mutant cell frequency and concentration of ribosomal genes in blood plasma--may be useful for revealing individuals in organism of which an intensive cell apoptosis takes place and there is an increased probability of carcinogenesis and of progress of disease of immune system.

At the initial stages of an adaptive response the transposition of the homologous chromosome loci from the peripheral parts of the nucleus and their approach happens. It is necessary for the repair of DNA double strand breaks in the process of the homologous recombination. Was shown that the chromosome loci transposition and accompanied by the nucleolus activities took place first in the irradiated (X-rays, 10 cGy) G0-lymphocytes, and then in the intact (bystander) cells incubated in the growth medium of irradiated lymphocytes. If there is a bystander effect the quantity of irradiated cells may be three order less than the bystander cells that affirms the great capacity of stress-signalization system. Moreover, the DNA fragments (the factors of stress signaling) were obtained from the growth medium supernatant of the irradiated and of the intact lymphocytes. In other independent experiments they were inoculated into the growth medium of recipient cells. Was demonstrated that there is loci transposition of homologous chromosomes loci and of nucleus activity after introducing the DNA fragments of irradiated cells. After introducing the DNA fragments of non-irradiated cells the both effects were not observed. In the work the characteristics of the obtained factors and the possible ways of stress signaling between the irradiated and the bystander lymphocytes were discussed.

During the process of the realization of the bystander effect the trans of the Signal from irradiated cells to the intact cell (bystander cells) happens. So both type of cells (irradiated and intact cells) have the same damages and reactions. There are new data about bystander effect as the transduction mechanism of the adaptive response and we have investigated this phenomenon. There are an incubation of the intact (bystander cells) and the exposed (X-radiation of 10 cGy) human lymphocytes and we analyze the location of the centromeric loci of the first chromosome. We observed hat for the first time that after X-ray exposition of the adaptive doses the transposition of the chromosome loci from the peripheral to the central parts of the nucleus in intact (bystander cell) G0-lymphocytes which were incubated in the growth environment cells with irradiated cells removal. We support that the starting states of the adaptive response is the loci extrication of the matrix, the transposition and the approach homologous chromosomes. This process is necessary for the DNA double strand breaks reparation (in the case of injured dose X-radiation) with the participation of the homologous recombination.

Below were demonstrated the differences in the cell reaction of the chromosome loci transference induced by adaptive doses X-radiation in the cells nucleus in norm cells and in cells with repair process defect of the oncological patients and patients with hereditary disease. It was supposed that the transference of the homologous chromosome loci is necessary for the realization of the correct DNA's double strand repair. The chromosome loci transference in normal cells could be induced by different factors such as X-radiation, RNA-polymerize II repression by alpha-amanitin and possible by other factors. In cells with BSCA1 and BRCA2 genes mutations the chromosome loci transference could not be induced by the X-radiation, but it could be induced by the RNA-polymerize II repression by alpha-amanitin. The defect of the chromosome loci transference condition on genetics or the another determinatives and this defect could be the one of the important reasons of the genome instability. There was suggested the new conception of the mechanisms of the cell differentiation and cell chronological senescence. The vector of the cell differentiation mechanisms is the progressive chromatin condensation determined by the physical mechanisms, i.e., transformation from less probable (the stem cells) to more probable cell system state (the differentiated cells) and as a result of it changing of the genes transcription. The continued chromatin condensation (most pronounced in the old cells) decreases the probability of the realization of the DNA repair as the reason of the chromosome loci transference limitation. In this work are presented experimental and theoretical information that proves our conception.

A study was made of the effect of the oxidizing agent potassium chromate (K2CrO4, PC) on cultured dermal fibroblasts of a healthy donor and three patients with rheumatoid arthritis (RA). Characteristics of the rRNA gene (RG) complex-RG copy number, active RG (ARG) dosage, and 18S rRNA content--were determined for each cell line. In cells of the healthy donor, oxidative stress caused by low doses of PC (2-4 microM, 1-4 h) induced an early response, including a 50-80% increase in total RNA and rRNA. An appreciable activation of the nucleolus was observed cytochemically, by silver staining and morphometry. The early response grew considerably lower with the increasing passage number and/or PC concentration. Exposure to 6-12 microM PC for 24 h led to a progressive cell death (late response). The existence and intensity of the early response correlated positively with the cell survival during further culturing. Cells of the RA patients displayed almost no early response even at early passages: total RNA did not increase, and rRNA increased by no more than 10%. Cell disruption (apoptosis) during further culturing was more intense than in the line originating from the healthy donor. The apoptosis intensity characterized by the increase in the content of DNA fragments in the culture medium and in the caspase 3 activity, was inversely proportional to the ARG dosage in the genome. The results provide the first quantitative characterization of the early and late responses of cells to PC-induced oxidative stress and suggest a role of the ARG dosage in cell survival in stress.

As we demonstrated earlier, the adapting X-ray doses (3 and 10 cGy) induced movement of chromosome centromeric loci in G0-lymphocyte nuclei. In the present study we investigated the influence of X-rays with 3 and 10 cGy doses on the content of total, 18S and 45S rRNA in human G0-lymphocytes because it is known that the transcription products participate in nucleus organization. It was shown that 3 h after irradiation the content of both total and 18S RNA was significantly increased. The 3 cGy dose induced higher level of the rRNA than 10 cGy dose did in cells of some individuals. At the same time, the 45S RNA content was not changed significantly. This result may suggest that process of rRNA transcription and primary transcript (45S rRNA) processing have been completed during 3 h after irradiation. The data about an activation of rRNA synthesis were confirmed by cytological observation. Under 3 and 10 cGy doses both nuclei diameter and area of the Ag-stained granules were increased, depending on dose. These data also may be connected with an initiation of rRNA transcription because of correlation of Ag-painting with nucleolus activity. Thus, adapting X-ray doses induce displacement of chromosome loci in lymphocyte nuclei and activation of rRNA transcription. Further investigations are required for understanding of these phenomena interconnection.

The 3H-thymidine incorporation in human lymphocytes of healthy donors induced by UVC radiation under doses 0.0008-60 J/m2 was investigated. It was shown that the incorporation of 3H-thymidine increases under doses in interval 0.1-20 and is constant under doses higher than 20 J/m2. Under doses in interval 0.006-0.03 J/m2 near a half of all samples had the level of incorporation increased in comparison with control samples. We connect the presence, absence or variability of this effect with individual peculiarities of cells and with different activity of cell subpopulations that are different on morphological and physiological characteristics. The hypothesis about the role of this factor in the influence of low doses of pathogenic agents (UVC and X-radiation, chemical compounds) on human lymphocytes is discussed.

In the present work it is shown that the phenomenon of interphase chromosome centromeric region displacement, earlier revealed by the authors, is not realized in G0-lymphocytes with heterozygous BRCA1/2 gene mutations. The role of these genes in DNA double strand break (DSB) reparation is known. It is concluded that chromosome locus displacement is necessary for DSB repair, at least in the process of homologous recombination. In accordance with our data, some feature (pericentromeric cluster disintegration and displacement, the nucleus size increasing) characteristic for S- and G0-lymphocytes are observed in normal G0-lymphocytes treated with 3 and 10 cGy. However, the size of nucleus in G0-lymphocytes is restored through 6 hours after irradiation in opposite to the process in dividing cells. It was proposed that some typical for resting cell functions of G0-lymphocytes after inducing by adaptive doze of radiation are stopped as similarly as after stimulation of cells. Interestingly, that the process of the induced chromosome loci displacement is correlated with the decreasing of DNA reparation possibilities under UV-irradiation. The induced apoptosis level also decreases when chromosome loci are displaced. The possible mechanisms of the revealed phenomenon are discussed. This research supported by RFBR grant (No. 01-04-49180).

It is concluded that a dose range from background dose to several cGy may be separated into two parts: a) first--the interval of small doses limited from above by D* which is determined from (1 g (D*/Doc)) < -0.51 g (n/2)), where Dse is an average dose of a single event, n--quantity of irradiated cells; in this interval only one track intersects a sensitive volume; b) second--the interval of low doses, in which in average one track intersects the volume and which is ranged from top D* to bottom Dse. Because events in this region qualitatively are similar to background events, cells in the dose range b) may be adapted to the influence of radiation. The first stage of the adaptive response of cells is associated with chromosome loci (centromere) movement in a cell nucleus and as we suggest the latter is the fundamental mechanism for repairing DSB DNA and switching of gene transcription. Because the movement of chromosome loci both in the resting cells under the adapting doses and in the normal dividing cells is much the same (but the latter lose their function characteristic for differentiated resting cells), it could be assumed that the resting cells under the adapting doses also lose their functional parameters. Under chronic exposure to low doses this functional changes can be principal for discussion on the influence of low doses on health.

The rDNA transcribed region (TR) was tested for accessibility to RsaI recognizing 15 TR sites, DNase I, and photoinducible arylazide (N-(4-azido-2-hydroxybenzoyl)-N,N'-diaminoheptane acetate) in isolated nuclei and, with arylazide, in intact cells. Arylazide entered cells well and did not appreciably affect the chromatin structure. Its photolysis products efficiently modified DNA in accessible sites. Single-strand breaks made by DNase I were not transformed in double-stranded in rDNA TR, suggesting the necessity of denaturing electrophoresis for such an analysis. About 70% of all rDNA copies proved poorly inaccessible to endonucleases and arylazide, the accessibility being higher in their 18S and 5.8S rRNA gene regions than in the regions of the external transcribed spacers (ETSs) and the 28S rRNA gene. Proteinase K disrupted this structure, and the corresponding copies were extracted from nuclei. This explained why in situ hybridization occasionally fails to reveal rDNA in the nucleolar fibrillar center (FC) on electron microscopic preparations. In other rDNA copies, TR (excluding 5'-ETS) was accessible to nucleases and arylazide. These copies were not extracted from nuclei treated with proteinase K. Some of their RsaI sites were protected by tightly bound proteins. Seven such regions were identified in TR. Possible association of the molecular structure, nucleolar location, and functional state of rDNA is discussed.

The mutation process has many stages. The information presented in this article suggest that a cell exposed to low LET radiation in the low-dose range (up to 1 cGy) must almost completely repair all spontaneous and radiation-induced DNA lesions. But reparation of DNA double-stranded breaks (DSB), which are the basis of genome instability has peculiarity. We have shown that the mechanisms of action of low doses (which initiate natural antimutagenic reactions of resting cells--an adaptive response) are associated with chromosome loci (centromere) movement in a cell nucleus. We suggest that the movement of chromosome loci in cell nucleus is the fundamental mechanism for repair of DSB and switching of the transcription of gene (it is known that in case of lymphoid cells Ikaros-complexes repressor is colocolizated with centromere loci); in particular, of nucleolar transcription activities because the latter is dependent on centromere arrangement. Because the movement of chromosome loci in both the mitotic cycle and under adapting dose on resting cells is much the same it could be assumed that in latter case the cells also lose their functional characteristic for differentiated resting cells. Under chronic exposure to low doses the functional changes can be the cause of organic changes if adapting dose affects the sufficient part of the cells. The role of cells of evolutional or ontogenetic reserve in mutation process is considered.

It was shown by blot-hybridization with corresponding DNA probes after electrophoretic separation of control and experimental samples of human genome DNA that accumulation of single-strand breaks in the chains of double-strand fragment of transcribing range of ribosomal gene (TRrDNA) does not result in double-strand breaks. That differs from the other studied DNA sequences (cluster of histon genes, Alu-repetition, telomeric repetition and satellite III). Single-strand breaks and double-strand breaks were induced by endonucleases and by gamma-radiation. In spite of higher chemical modification of TRrDNA by arylazide and dimethylsulfate (because of high content of GC-pairs), under the following fragmentation TRrDNA was found to be more resistant to double-strand breaks than other studied DNA sequences. At the same time in the range of non-transcribing spacer (NTS) of ribosomal gene, the section with higher sensitivity to double-strand breaks was found. Higher resistance of TRrDNA to double breaks makes it possible to identify these fragments in cell material from different tissue after death or in DNA samples after prolonged storage. Resistance of TRrDNA to formation of double-strand breaks can be used for its detection in biological fluids after cell death, including the death initiated by ionizing radiation.

Increasing of 3H-thymidine incorporation in lymphocytes of human peripheral blood which depends non-linearly on X-ray dose (3 cGy max) and interleukin-2 (IL-2) concentration (17.5 Units/ml) is shown. However addition of IL-2 (17.5 U/ml) into the medium of cells after irradiation (3 cGy) decreases almost to the control the effects induced by independently shown actions. Lymphocytes subpopulation responsible for the described phenomena are isolated during the fractionation of lymphocytes in the density gradient and pH (V-fraction BSA). Cell fraction less than 1-2% from the isolated lymphocytes is characterized by increasing of spontaneous corporation of 3H-thymidine, large sizes (d > 8 mkm), decreasing repair after UV-irradiation. It is believed that low dose irradiation and IL-2 activate this cell subpopulation of "last reaction", and higher doses of these factors and this both actions stopping 3H-thymidine incorporation initiate apoptosis. The relation of this sell subpopulation and before proposed ontogenetical reserve cells is discussed.

Novel biophysical and biological aspects of eukaryotic cell response to low doses of low-LET radiation are considered in the present paper. The model of universal nuclear target in eukaryotic cells is proposed. The target is considered as a spherical volume homogeneously filled of DNA with adjacent water layer. Thickness of water layer equals to diffusion length of hydroxil radicals hitting DNA. Two subregions are proposed of the cellular membrane with different ability to respond to low doses of ionizing radiation. An original mechanism of repair of DNA double strand breaks (dsb) is described. This mechanism suggests directional conformational movement of separate locuses of homological chromosomes through narrow tube ("throat") between specialised nuclear regions. The fluxes of nuclear liquid induce a force which is responsible for highly elastic chromosome deformation and chromosome movement resulting in proximity of homological chromosomes in the throat. Close proximity is required for DNA dsb repair. All of DNA dsbs have to be repaired in low doses range owing to high genomic stability of eukaryotic cells. However, it has been hypothesized an existence of a small subpopulation of cells with unstable genome named as cells of evolutionary or/and ontogenetical reserve (ER or/and OR). Genomic instability is induced in these cells by epigenetic program. This program results in DNA damage mainly in chromosomal "hot spots". As a result new genetic variants are originated. The ability of cell population in vivo to eliminate or not eliminate Er/OR cells determinates individual sensitivity of organism and hence health consequences to low dose radiation exposure.

DNA double strand breaks in chromatin induced by high LET ionizing radiation are analysed on the basis of biophysical modelling and computer simulation. Theoretical and experimental data are presented arguing in favour of nonrandom clustered distribution of DNA double strand breaks in chromatin.

Human lymphocytes were X-irradiated at doses 0-10(-2) Gy and allowed to repair for some time. Nuclei were prepared and digested with restriction endonuclease Rsa I to selectively release 28S RNA gene fragment fraction, containing the DNA-binding protein, Hpa II digestion of nuclei was used to investigate the methylation of the 28S RNA gene fragment. There was a differential enrichment of 28S RNA gene binding protein for different X-ray doses with maximum enrichment for dose 2-3 x 10(-2) Gy with following diminish to 10 x 10(-2). The enrichment of less methylated fractions of 28S RNA gene was observed during X-irradiation. This might be explained by a different X-ray-induced changes of methylated and unmethylated rDNA binding with nuclear proteins. The possible mechanism for this phenomena are discussed.

It was shown that the maximum adaptive response to damaging irradiation dose would be achieved after preirradiation when for latter the product of the dose by the dose rate (i. e. D*D/t) is equal to constant value. The radiobiological constant is the reciprocal of this product, i. e. k = t/D2 = tau/delta 2, where delta is an average dose received by the cell for single pass of ionizing particle through the cell sensitive volume with a given density. The triggering signal for the adaptive response program initiated by preirradiation is a certain reaction, which is completed for a time interval tau between two consistent acts of absorption. The radiobiological constant is determined by this time interval tau and by a dose delta. In human lymphocytes in the case of signal initiated by X-rays the value of tau is within the range of 1--5 s.

The model predicting quantitatively a dose response for genetic lesions in low dose range is presented. The model is based on the concept of programmed initiation of the genetic damage in the subpopulation of specific cells of "evolutionary reserve" (CER). It is demonstrated that complex shape of dose-response curve results from superposition of processes in different subpopulations within exposed cell population, mainly in CER. The model allows us to explain the minimum observed in the dose-response curve at doses corresponding to one (on the average) event of energy deposition per CER with spontaneous aberrations. The equation for dose-response function for dicentrics describes satisfactory experimental observations. Advantage of proposed equation is that it does not deal with free parameters to fit the data. They are determined independently by using known physical and biological characteristics. The data described allow us to suggest that sensitive body for CER is a cell as a whole. Probably, programmed initiation of genetic lesions in CER requires both two hits at cell membrane and radiation effect on cell nucleus.

Effects of X-ray low doses (0.5-25 cGy) with following UV light (254 nm, 20 J/m2) on human peripheral blood lymphocytes were studied. Reparation response registered by unscheduled DNA synthesis (UDS) activity was demonstrated to be the most intensive after action of X-rays in dose ranges 2-3 cGy and 15-20 cGy, and least intensive after 10 cGy. In those cells where UV light was followed by X-rays, dose ranges 2-3 cGy and 15-20 cGy cause essential decrease of UDS as compared with UV-light action only. The most intensive UDS was in those UV-irradiated lymphocytes which where previously exposed to 10 cGy of X-rays. At the same time 10 cGy cause minimal reparation response without UV light. Possible mechanisms of discovered phenomena are discussed. In particular, reaction on 2-3 cGy might be a reorganization of genome for adaptive response or an evolution reserve cells response. From 10 cGy the reason of reaction might be a reparation, induced by radiation.

The dependence of UV-induced unscheduled DNA synthesis (UDS) in the unstimulated lymphocytes of human peripheral blood on the ionic strength (mu) of the culture medium has been shown. In the level of mu lower or higher than the physiological (mu ph) one, UDS significantly decreases. The effect of modification of mu due to the changes of ionic strength is absent in the lymphocytes of the classic form of xeroderma pigmentosum. The phenomenon may become useful in the development of a new test for revealing cells with a genetically or physiologically changed system of UV-induced DNA repair. The mechanisms of investigated phenomenon, particularly their dependence on the structure of chromatin, as well as the influence of ionic strength on binding of the repair enzymes with DNA are discussed.

A concept is proposed where the events induced by low doses of ionizing radiation with low linear energy transfer have different script than those induced by high doses. Its essence consist in the existence of a small fraction (subpopulation) in clonal heterogeneous population whose cells are determined for a specific programmed response induced by a number of action that are potentially able (at high doses) to decrease the survival of the given population. The function of the postulated cell is a deliberate sacrifice necessary for formation of evolutionary reserve. A mechanism of this response is based on an autogeneration of wide-scale genome rearrangements in cells and is not connected with direct influence of genetically hazardous agents on DNA. This leads to arising of a number of genetic variants (evolutionary reserve cells). Selection among them enhancing survival of the population may in the same time lead to harmful consequences for the organism (if there are malfunctions of the immune system). Medical-biological consequences of the effect of low doses are associated with the appearance of such clones and biologically active products secreted from the exposed cells. Mechanisms responsible for the above phenomena are discussed.

The dependence of UV-induced unscheduled DNA synthesis (UDS) in non-stimulated lymphocytes of human peripheral blood on the ionic strength (mu) of the culture medium has been shown. With the level of mu lower or higher than physiological (mu ph) the UDS significantly decreases. The effect of modification of mu due to changes in ionic strength is absent in the lymphocytes of patients with the classic form of xeroderma pigmentosum. This phenomenon may become useful for development of a new test revealing cells with genetically or physiologically changed system of UV-induced DNA repair. Mechanisms of investigated phenomenon, particularly their dependence on the chromatin structure, as well as the influence of ionic strength on binding the repair enzymes with DNA are discussed.

Low-level radiation either from external, or from incorporated sources is shown to cause a non-monotonous change in some intranuclear parameters of human peripheral blood lymphocytes. For instance, radiation induces changes in the parameters that characterize the location of perecentromeric regions within the interphase nucleus and a non-monotonous increase in nuclear sizes. The exposure to doses exceeding 5 cGy impairs the relationship between the nuclear sizes and location therein of the perecentromeric regions of interphase chromosomes which exist in the control and after exposure to 2.5 cGy. Differences between the dose ranges of 1.5-3.5 and 17-25 cGy are manifested by the kinetics of restoration of the pattern of distribution of lymphocyte nuclear sizes.

A probe containing full-size DNA copy of influenza A/USSR/90/70 virus protein gene M labeled with biotin on 32P was used for influenza A virus RNA detection by dot hybridization method. For labeling with biotin, a new method of its administration by chemical modification of nucleic acid was employed. In homologous DNA:DNA hybridization the sensitivity of determinations was less than 1 pg in the biotin-treatment of the probe and 1.25 pg in its radioactive labeling. Hybridization of DNA probe with cytoplasmic RNA isolated from influenza A virus-infected (strains A/USSR/90/77 and A/Texas/77) MDCK cells revealed RNA in the dot corresponding to 4.5-5.5 1g ID50 of virus present in 2 x 10(4) cells. The probe did not bind with negative controls in any dot in all the tests. The results of the study indicate that DNA probes labeled with biotin and 32P and used in dot hybridization for influenza A virus RNA detection in infected cells show the similar sensitivity and specificity.

The paper treats of the correlations between the position geometry of the extended successions of the residue of basic amino acids and those nonbearing any positive charge along the polypeptide chains of the histone H2A, H2B, H3, H4 molecules forming nucleosomes protein frame. The number of histone packing variants in the tetramere conforming to the conditions of the revealed complementarity reached the value of 10(4), and in respect to histone octamere--10(8). A hypothesis was put forward on structural heterogeneity of nucleosomes (of functional significance) the choice of whose protein frame packing variant could be realized by the mechanisms of allosteric regulation of the primary structure of DNA molecule section serving as the nucleosome element.

It has been shown for the first time that heparin, one of the natural polyanions, is capable of dissociating the histones H1, H2A and H2B from chromatin in a medium of "physiological" ionic strength: 0.15 M NaCl + 0.7 mM Na-phosphate buffer, pH 7.0. Under the same conditions heparin produces chromatin fractionation, leading to the appearance of slow-sedimenting DNA, augments viscosity and reduces turbidity of chromatin suspension. This evidences the decompactization of deoxyribonucleoprotein fibrils.

It was shown with the model supramolecular chromatin systems that the activity of the factor of chromatin condensation changes in the process of liver regeneration. No activity of the factor of chromatin condensation was found 20 hrs after operation. Within 32-33 hrs (in the period of increased mitotic frequency) its activity approaches that prior to the operation. The nature of the activity of the factor of chromatin condensation is discussed with respect to the functional state of chromatin.

Electrophoretic analysis of histones bound to DNA and remaining free in the mixtures of DNA with the total histone of chromatin in a medium of physiological ionic strength has shown that even the minimal weight excess of the total histone with reference to DNA (1.1:1) leads to the formation of nucleohistone impoverished in HI fraction because of histone competition for DNA. Within the histone/DNA ratio equal to 1.4, H3, H4, H2A and H2B are bound to DNA without competition, i.e. at a ratio in which they are added to DNA. Provided the histone/DNA ratio is higher in the mixture, there form nucleohistones enriched with H3 and H4 fractions. The role of histone competition for DNA in eu- and heterochromatin assembly is discussed.

To study the energy absorption in cell nuclei the analysis was made of the level of registration of beta-particles, emitted by the incorporated tritium, using a liquid scintillation counter. It was shown that the treatment, modifying the chromatin structure, changed the number of beta-particles registered. The pattern of the spectral changes indicated that the effect observed was not the result of the quenching processes. The effect was absent when the source of a higher energy (14C) was used. The authors submit the conceptions concerning the mechanisms of the phenomenon observed which are confirmed by the theoretical studies.

By using fluorescent microscopy and acridine orange staining it was shown in the studies on short-term culture of human cells that the melting patterns of chromatin DNA of intact lymphocytes of healthy individuals represented the curves with 6 maxima (F530) at the temperature ranges of 45 degrees C, 65 degrees C, 85 degrees C, and 92 degrees C (P less than 0.01). The melting patterns of lymphocytes from patients with Down's disease represented curves with 4 maxima at the temperature ranges of 65 degrees C, 85 degrees C, 88 degrees C, 92 degrees C (P less than 0.01). No decline in the fluorescence intensity at the temperature intervals of 78 degrees C-85 degrees C was apparently due to a greater degree of condensation of definite regions of the trisomal cell chromatin complex. Possible mechanisms accounting for structural readjustments of the interphasic human lymphocyte chromatin occurring under thermal effects are discussed.

Protein with a molecular mass of 53000 daltons undetectable in healthy persons was identified by electrophoresis in peripheral blood leukocytes of patients with Down's syndrome. The protein was completely extracted with 0.4 N HCl from leukocyte homogenates and was found to be identical, as regards electrophoretic mobility, to protein detected in two patients with chronic myeloblastic leukemia. The causes of discrepancy between theoretically expected and electrophoresis-revealed differences in protein composition of normal and trisomal cells.

Relationships have been studied between call thymus chromatin isolated in a medium of low ionic strength (0.7 mM Na-phosphate buffer, pH 7) and total histone which was added in different amounts (up to a 40-fold excess) with respect to chromatin DNA. The maximum protein/DNA ratio in chromatin-histone complexes being formed was 3.27. It has been demonstrated for the first time that with an elevation in the amount of supplementary total histone there took place a release of natural histones of chromatin, first of HI and then of H2A and H2B. It is concluded that supplementary histone interacts with DNA, modifying its interplay and chromatin protein even though the dissociation is not fixed yet.

A thermomechanical method was applied to the study of the interaction of the model desoxyribonucleoprotein systems (DNP-systems) of chromatin with the blood sera of healthy and sick persons. It was demonstrated that the blood sera of healthy persons caused decondensation of the DNP-systems. The intensity of this action varied when the sera with the composition altered as a result of pathology were used: the sera of the patients suffering from systemic lupus erythematosus increased the condensation; in comparison with the sera of healthy persons, the sera of schizophrenic patients intensified the DNP-system decondensation. The sera action was not connected with the activity of the serum enzymes. The role played by the serum components in regulation of the structural-functional chromatin properties is discussed.

The interaction between acridine orange (AO) and diluted and concentrated solutions of DNA, DNP systems and chromatin suspension at the physiologic ionic strength was investigated. The effect of AO on DNP systems was also investigated. It was shown that highest possible number of AO molecules bound to DNA made up 70% of the total number of nucleotides. The model of AO binding to DNA is proposed and used for calculation of constants of stronger and weaker AO-binding capacities equal to 6-10(6) M-1 and 1,7-10(5) M-1, respectively. The AO-DNA binding constants in DNP-complex are five as low. The primary number of binding sites in chromatin suspension made up 10% of the corresponding sites in DNA and increased as AO was adsorbed. AO induced the supercontraction of oriented DNP systems at the physiologic ionic strength and the appearance of the low-temperature melting hump.

The influence of blood sera from patients with Down's syndrome and healthy ones, and different serum fractions on the structure of the deoxyribonucleoprotein systems (DNP-system) has been studied. It was demonstrated that non-fractionated sera of the patients produce a condensation effect on the DNP-system in contradistinction to the sera from healthy people. The analysis of the action of single serum fractions showed that different condensation effect results from the activity of high molecular nondialysable thermosensitive components whose action disappears at gel-filtration of serum proteins. A possibility of the humoral control over chromatin structural organization in vivo is discussed in terms of the evidence on the similarity of serum proteins and chromosomal nonhistone proteins.