Characteristics of DNA replication in the long-term culture of human cells at the stationary phase

Peculiarities of DNA replication in cultured human diploid fibroblasts in logarithmic and stationary phases were studied using DNA autoradiography. The rate of DNA replication fall from 30-36 mu/hour at active proliferative phase to 18-20 mu/hour at late stationary phase. This phenomenon is characteristic of stationary cultures after stimulation to proliferate by changing medium as well as by culturing without stimulation. Possible mechanisms of DNA replication rate alteration in senescent human cells are discussed.     

Unusual case of 18p- syndrome: diagnosis using a cloned DNA fragment

The patient with atypical clinic picture of 18p- syndrome is described. The in situ hybridization technique was used to localize chromosome 18-specific cloned sequence to metaphase chromosomes of the proband. The predominant hybridization was found in pericentromeric regions of homologous chromosome 18. The amount of pericentromeric DNA measured by in situ hybridization was different in homologous chromosomes and the number of radioactive grains was statistically greater in the normal chromosome 18 than in the chromosome 18p-. The cause of asymmetrical hybridization of probes to homologous chromosomes 18 is discussed. The results obtained indicate that this probe may be useful in clinical cytogenetics for identification of chromosome 19 in metaphase and interphase cells, determination of breakpoints or studies of pericentromeric DNA polymorphisms.     

A cytogenetic study of the human chorion for the purpose of the prenatal diagnosis of hereditary diseases

Cytogenetical investigation of 50 diagnostic chorionic villus samples from women with a high risk of giving birth to babies with chromosomal and genic pathology, and of 128 chorionic samples obtained from medical abortions, both on the 8-12th weeks of gestation was performed by means of original direct chromosomal analysis. Chromosomal anomalies were found in 6 cases of diagnostic chorion biopsies (12%) and in 4 cases (3%) of medical abortions. The former group included 5 embryos with autosomal trisomy (4--Ts21 and 1--Ts13) and one embryo with monosomy 18. The latter group contained 2 embryos with X-chromosome monosomy and 2 other with chromosomal mosaicism. A significant prevalence of the female sex was found in the diagnostic group (sex ratio 0.56), but not in the medical abortion one (sex ratio 1.0). Analysis of routine chromosomal preparations and those after in situ hybridization with X-chromosome alfoid-probe YAP 1-10 revealed polyploidy in average in 0.8-1% chorion cells. The feasible causes of sex ratio distortion in embryos of diagnostic group and factors responsible for the rate of polyploidy are discussed. High reliability of originally elaborated direct "shaking-blotting" method of chromosomal preparations from chorionic villus samples is stressed.     

A case of deletion of the short arm of the chromosome 21 (21p-) (christchurch chromosome) discovered prenatally:clinical and cytogenetic data

Results of cytogenetic research of placental villi and amniotic fluid cells culture of the 22-weeks-old fetus with multiple congenital malformations (MCM) are presented. The absence of the short arm in one of the homologue of the chromosome 21 was revealed. Cytogenetic analysis of the fetus father's blood lymphocytes determined the similar chromosome. Further research of the father's karyotype made by FISH-method using specific DNA samples had discovered the absence of subtelomeric parts in the short arm of the chromosome 21 that might be considered as a deletion. It was suggested that the effect of position and interaction of genes could play a key role in appearing of MCM in the fetus in the case when the 21p-chromosome was transferred to it from the healthy parents.     

Disruptions in the blood system upon exposure to low-dose ionizing radiation depending on duration of emotional stress

In experiments on rats, disturbances in the development of blood system adaptive reactions on emotional stress were found. Under the combined effect of preliminary (before stress) exposure to 0.8 Gy of gamma-radiation and the emotional stress of various duration (2, 4 and 8 days), a compensatory capability of the blood system decreased. The degree of the disturbances directly depended on the duration of the emotional stress.     

Study of alpha-satellite DNA in cosmid libraries, specific for chromosomes 13, 21, and 22, using fluorescence in situ hybridization

Fluorescent in situ hybridization (FISH) was employed in mapping the alpha-satellite DNA that was revealed in the cosmid libraries specific for human chromosomes 13, 21, and 22. In total, 131 clones were revealed. They contained various elements of centromeric alphoid DNA sequences of acrocentric chromosomes, including those located close to SINEs, LINEs, and classical satellite sequences. The heterochromatin of acrocentric chromosomes was shown to contain two different groups of alphoid sequences: (1) those immediately adjacent to the centromeric regions (alpha 13-1, alpha 21-1, and alpha 22-1 loci) and (2) those located in the short arm of acrocentric chromosomes (alpha 13-2, alpha 21-2, and alpha 22-2 loci). Alphoid DNA sequences from the alpha 13-2, alpha 21-2, and alpha 22-2 loci are apparently not involved in the formation of centromeres and are absent from mitotically stable marker chromosomes with a deleted short arm. Robertsonian translocations t(13q; 21q) and t(14q; 22q), and chromosome 21p-. The heterochromatic regions of chromosomes 13, 21, and 22 were also shown to contain relatively chromosome-specific repetitive sequences of various alphoid DNA families, whose numerous copies occur in other chromosomes. Pools of centromeric alphoid cosmids can be of use in further studies of the structural and functional properties of heterochromatic DNA and the identification of centromeric sequences. Moreover, these clones can be employed in high-resolution mapping and in sequencing the heterochromatic regions of the human genome. The detailed FISH analysis of numerous alphoid cosmid clones allowed the identification of several new, highly specific DNA probes of molecular cytogenetic studies--in particular, the interphase and metaphase analyses of chromosomes 2, 9, 11, 14, 15, 16, 18, 20, 21-13, 22-14, and X.     

Dynamic components of interrelation among the parameters of heart preload, venous return and central venous pressure

In acute experiments on cats, in applying of original methodical approach--control of systemic circulation by the aid of computerized negative feedback loop changing the volume of circulating blood (method of biological feedback)--first were experimentally measured and analyzed the dynamic characteristics of relationship between central venous pressure and venous return of blood to the right heart. The following positions are offered and validated in the work. (1) It is shown that the passive component (mechanical compliance) is more important than active one (active myogenic component) in the small circle of circulation being compared to large one. (2) Venous return plays the leading role in forming the shifts of central venous pressure directly during developing of the transition processes of systemic circulation caused by the norepinephrine injection and the linear type of this link is proved directly during the development of the cardiovascular shift. (3) The dynamic characteristics of relationship between venous return and central venous pressure during the geodynamical reaction caused by the shifts of intravascular blood volume are experimentally measured and mathematically analyzed. It is revealed that dynamic summands of this link may overbalance the static ones known before in influence on the total shifts in developing of the systemic reaction of circulation and this influence increases when the velocity of changes in studied parameters of circulation becomes more.