[Analysis of gene expression pattern in peripheral blood leukocytes during experimental heat wave]. / Analiz izmeneniia ékspressii genov v leikotsitakh krovi cheloveka vo vremia ispytaniia, modeliruiushchego usloviia teplovoi volny.

The conditions of Moscow 2010 summer heat wave were simulated in an accommodation module. Six healthy men aged from 22 to 46 years stayed in the module for 30 days. Measurements of gene expression in peripheral blood leukocytes before, during and 3 day after simulated heat wave were performed using qRT-PCR. We observed a shift in the expression level of certain genes after heat exposure for a long time, and rapid return to the initial level, when volunteers leaved the accommodation module. Eight genes were chosen to form the "heat expression signature". EGR2, EGR3 were upregulated in all six volunteers, EGR1, SIRT1, CYP51A1, MAPK9, BAG5, MNDA were upregulated in 5 volunteers.

[Phospholipase A2, group IIa, as an earlier unknown immunohistological marker of cardiac myxoma].

Phospholipase A2, group IIA, gene expression has been analyzed in primary heart tumors. High expression has been demonstrated through several ways: reverse-transcriptase chain polymerase chain, Northern blotting hybridization at the RNA level and immunoblotting, immunohistochemical assay at the protein level. Human cardiac myxoma exhibits highly positive phospholipase A2, group IIA, immunophenotype (100% positive cases). The immunophenotype is unique among human primary cardiac tumors. Phospholipase A2, group IIA, can be proposed as a tissue marker for pathological examination after heart tumor resection.

Gene expression analysis to identify mRNA markers of cardiac myxoma.

cDNA expression arrays were used to identify mRNA expression markers for cardiac myxoma. The RNA profile analysis suggests that cardiac myxoma should be considered as a stand-alone tissue rather than a pathological modification of particular normal tissue. The analysis reveals a set of genes which are highly and steadily expressed in cardiac myxomas and can serve as an mRNA expression markers of the tumour. Marker status of selected genes was confirmed by reverse transcriptase polymerase chain reaction analysis. Genes MIA (melanoma inhibitory activity) and PLA2G2A (phospholipase A2, group IIA) show the highest specificity as cardiac myxoma markers, since they have more than 10-fold higher RNA level in cardiac myxomas than in any one of 15 normal tissues tested. Among markers of myxoma at least three are participants of phospholipid metabolism: ANXA3, PLA2G2A, and phospholipid transfer protein. Tissue inhibitor of metalloproteinase 1 and secretory leucocyte protease inhibitor are inhibitors of proteases degrading extracellular matrix proteins and participating in cell proliferation regulation. MIA, SPP1, fibromodulin are modulators or participants of the interaction between extracellular matrix proteins and their cell surface receptors. SOX9 is a transcription factor required for chondrocyte differentiation. Calretenin (CALB2) is an intracellular calcium-binding protein with poorly understood function.

[PRKAR1A gene mutations in two patients with myxoma syndrome (Carney complex)]. / Mutatsii v gene reguliatornoi sub"edinitsy 1-alpha tsiklo-AMF-zavisimoi proteinkinazy A (PRKAR1A) u dvukh bol'nykh s miksomnym sindromom (kompleksom Carney).

Carney complex is an autosomic dominant disorder initially described as the association of cardiac myxomas, spotty skin pigmentation and endocrine overactivity and considered as a multiple neoplasia and lentiginosis syndrome. Mutations in the tumor suppressor gene PRKAR1A, coding for the type 1-alpha regulatory subunit of cAMP-depended protein kinase A have been previously identified in about half of the Carney complex kindreds. In this paper we report identification of the molecular defect in PRKARIA gene in two Carney complex patients. A new mutation (403delAC) located in a 3rd exon of PRKARIA gene has been observed in one case, and a previously described mutation in exon 7 (847delTC) in the second case.

[Cloning and analysis of the nucleotide sequence of the segment in the Mycoplasma gallisepticum genome containing the gene for the ATP-binding subunit of DNA topoisomerase type II (topIIB)]. / Klonirovanie i analiz posledovatel'nosti nukleotidov uchastka genoma Mycoplasma gallisepticum, soderzhashchego gen ATP-sviazyvaiushchei sub''edinitsy DNK-topoizomerazy tipa II (topIIB).

M. gallisepticum genome fragment carrying complete coding sequence for ATP-binding subunit of topoisomerase II (topIIB), partial coding sequence for N-terminal part of A-subunit of topoisomerase II and region upstream of topIIB gene (open reading frame encoding 99 amino acids) was sequenced. The nucleotide sequence of topIIB has significant homology with previously reported gyrase genes and parE gene of E. coli. No protein sequence significantly similar to the open reading frame upstream from the gene topIIB was found in GeneBank data.

[Identification of a Mycoplasma strain using fingerprinting of restriction enzyme hydrolysates of chromosomal DNA]. / Identifikatsiia shtamma Mycoplasma s pomoshch'iu fingerprintirovaniia restriktaznykh gidrolizatov khromosomnoi DNK.

The heterogeneity among Mycoplasma gallisepticum strains was characterized by comparing genomic DNA from different strains. Two criteria were used for the comparison. 1. Analysis of SmaI and BglI restriction patterns. A method using a dialysis sack for preparation of mycoplasma DNA suitable for pulsed-field gel electrophoresis is described. 2. Comparison of "fingerprinting" patterns obtained as a result of Southern transfer of HindIII-digested DNA followed by hybridization with cloned repetitive M. gallisepticum sequences. It was shown that both types of patterns analyzed are strain-specific; moreover, we were able to identify as a A5969 the strain of M. gallisepticum which was wrongly referred to as S6 in our previous works (Mol. Biol. 1991. V. 25. P. 1643-1649; FEBS Lett. 1991. V. 291. P. 71-74).

[Mutual location of the rRNA operon and tuf gene in the Mycoplasma gallisepticum strain S6 genome]. / Vzaimnoe raspolozhenie operona rRNK i gena tuf v genome Mycoplasma gallisepticum, shtamm S6.

The genomic library of Mycoplasma gallisepticum was constructed and two clones, selected by hybridization on E. coli 16S rRNA, were analyzed. The restriction map of the clones indicate that both clones belong to the same region of the M. gallisepticum genome. The results of Southern hybridization with either E. coli 16S rRNA, or E. coli 23S rRNA, or oligonucleotide synthesized as a part of M. gallisepticum 5S rRNA, led to the conclusion that the unspliced rRNA operon was cloned. The order of genes in the operon is common for eubacteria: 16S-23S-5S. The tuf gene of M. gallisepticum was mapped inside the cloned region 5 kb upstream of 16S gene by hydridization on E. coli tufA gene and oligonucleotide synthesized on the basis of M. gallisepticum tuf gene sequence. The direction of transcription of the gene and the expected direction of transcription of the rRNA operon coincide.

[Specific protection of DNA by distamycin A, netropsin and bis-netropsins against the action of DNAse I]. / Spetsificheskaia zashchita DNK distamitsinom A, netropsinom i bis-netropsinami ot deistviia DNKazy I.

Interaction of netropsin, distamycin A and a number of bis-netropsins with DNA fragments of definite nucleotide sequence was studied by footprinting technique. The nuclease protection experiments were made at fixed DNA concentration and varying ligand concentrations. The affinity of ligand for a DNA site was estimated from measurements of ligand concentration that causes 50% protection of the DNA site. Distribution pattern of the protected and unprotected regions along the DNA fragment was compared with the theoretically expected arrangement of the ligand along the same DNA. The comparison led us to the following conclusions: 1. Footprinting experiments show that at high levels of binding the arrangement of netropsin molecules along the DNA corresponds closely to the distribution pattern expected from theoretical calculations based on the known geometry of netropsin--DNA complex. However, the observed differences in the affinity of netropsin for various DNA sequences is markedly greater than that expected from theoretical calculations. 2. Netropsin exhibits a greater selectivity of binding than that expected for a ligand with three specific reaction centers associated with the antibiotic amide groups. It binds preferentially to DNA regions containing four or more successive AT pairs. Among 13 putative binding sites for netropsin with four or more successive AT pairs there are 11 strong binding sites and two weaker sites which are occupied at 2 D/P less than or equal to 1/9 and 2 D/P = 1/4, respectively. 3. The extent of specificity manifested by distamycin A is comparable to that shown by netropsin although the molecule of distamycin A contains four rather than three amide groups. At high levels of binding distamycin A occupies the same binding sites on DNA as netropsin does. 4. The binding specificity of bis-netropsins is greater than that of netropsin. Bis-netropsins can bind to DNA in such a way that the two netropsin-like fragments are implicated in specific interaction with DNA base pairs. However, the apparent affinity of bis-netropsins estimated from footprinting experiments is comparable with that of netropsin for the same DNA region. 5. At high levels of binding bis-netropsins and distamycin A (but not netropsin) can occupy any potential site on DNA irrespectively of the DNA sequence. 6. Complex formation with netropsin increases sensitivity to DNase I at certain DNA sites along with the protection effect observed at neighboring sites.