Expression of micrornas in molecular genetic breast cancer subtypes.

BACKGROUND: It is shown that each type of human malignancies has a unique set of expressed miRNAs, and tumor-specific miRNAs in biological tissues of a patient are stable. The aim of this study was to determine the differences in the expression of miRNAs in tumor tissue of invasive breast carcinoma compared to normal tissue, as well as to analyze the variable expression of miRNAs in molecular genetic subtypes of breast cancer. METHODS: We determined differences in mRNA expression in 35 biopsies of tumor tissue of various molecular genetic subtypes of breast cancer and 35 biopsies of adjacent conventionally normal breast tissue by RT-PCR in real time. We assessed the expression levels of miRNA-21, 221, 222, 155, 205, 20a , 125b and 200a. RESULTS: A significant increase in the level of expression of the oncogenic miRNA-20a (p=0.000141) and miRNA-221 (p=0.037777) in the triple negative cancer in comparison with the luminal A and luminal B/HER2/neu-negative breast cancer subtypes was established. Assessment of significance of the results was conducted using ROC analysis. For miRNA-221 AUC value was 0.772, for miRNA-20a AUC value was 0.949. The obtained results suggest the possibility of using the levels of miRNA-21, 155, 205, 125b expression in tumor tissue to assess a malignant potential of a breast carcinoma. The levels of expression of oncogenic miRNA-221 and miRNA-20a are increased in TNBC compared with luminal A and luminal B/HER2/neu-negative breast cancer subtypes, supporting the characteristic of TNBC as the most aggressive subtype of breast cancer. MiRNA-20a is a marker of TNBC compared with luminal subtypes of breast cancer. MICRO ABSTRACT: To identify markers for breast cancer with triple-negative phenotype, we evaluated expression levels of siRNA-21, 221, 222, 155, 205, 20a, 125b, 200a and 146b in the tumor tissue of 35 patients by RT-PCR. AUC value equal to 0.949 in the ROC-analysis allows us to recommend the miRNA-20a as a marker of triple negative breast cancer to differentiate it from the luminal subtypes.

Straightforward measurement of anisotropic thermal properties of a Bi2Se3 single crystal.

We demonstrate here a simple measurement protocol which allows the thermal properties of anisotropic crystalline materials to be determined. This protocol is validated by the measurement of Bi2Se3, a layered material consisting of covalently bonded sheets with weak van der Waals bonds between each layer, which has highly anisotropic thermal properties. Thermoreflectance microscopy measurements were carried out on a single-crystal Bi2Se3 sample, firstly on the bare sample and then after capping with a 100 nm thick gold layer. Whereas on the bare sample lateral heat diffusion is dominated by the in-plane thermal diffusivity, on the metal-capped substrate heat diffusion perpendicular to the sample surface dominates. Using a simple theoretical model, we show how this double measurement protocol allows the anisotropic thermal conductivity coefficients of bulk Bi2Se3 to be evaluated.

Two-dimensional metal-chalcogenide films in tunable optical microcavities.

Integration of quasi-two-dimensional (2D) films of metal-chalcogenides in optical microcavities permits new photonic applications of these materials. Here we present tunable microcavities with monolayer MoS2 or few monolayer GaSe films. We observe significant modification of spectral and temporal properties of photoluminescence (PL): PL is emitted in spectrally narrow and wavelength-tunable cavity modes with quality factors up to 7400; a 10-fold PL lifetime shortening is achieved, a consequence of Purcell enhancement of the spontaneous emission rate.

Expression of microRNAs, CYP1A1 and CYP2B1 in the livers and ovaries of female rats treated with DDT and PAHs.

AIMS: In this study, we determined the expression level of miRNAs and the induction of CYP1A1 and CYP2B1 in the livers and ovaries of female Wistar rats treated with DDT, benzo[a]pyrene (BP), and 3-methylcholanthrene (MC). This study compared CYP1A/2B induction and miRNA expression levels to cast light on a possible role of miRNA in the tissue-specific induction of CYPs. MAIN METHODS: The induction of CYP1A1/2B1 enzymes was detected by ethoxy-, pentoxyresorufin O-dealkylation and Western blot analysis. The CYP1A1/2B1 gene expression was determined by RT-real time PCR. Relative levels of expression for selected in silico miR species were determined by real time PCR with small nuclear U6 RNA employed as a reference gene. KEY FINDINGS: After bioinformatic analysis, miR-21, 221, 222, and 429 were chosen as potential post-transcriptional regulators of rat CYP1A and CYP2B. It was shown that miR-21, 221, 222, and 429 expression levels decreased in the liver of DDT-, BP-, and MC-treated rats, whereas increases were observed in CYP1A1 and CYP2B1 mRNA expression levels and protein content, and EROD and PROD activities. Conversely, a tendency for elevated levels of miRNAs in the ovaries of inducer-treated female rats was observed. In the ovaries, a high level of CYP1A1 and CYP2B1 mRNA expression was observed, although protein content and enzyme activity were not visible. SIGNIFICANCE: These data suggest a potential involvement of miRNA in the post-transcriptional regulation of CYP1A and CYP2B in the livers and ovaries of chemically induced rats.

[Microrna, evolution and cancer].

MicroRNAs are known as a posttranscriptional negative regulators of gene expression by binding to the 3'UTP of target mRNAs in cytoplasm. More than 1600 microRNAs expressed in human cells, are involved in the regulation of embryogenesis, differentiation, cell cycle, apoptosis, senescence, thus determining cell fate. Up to 60 % of protein coding genes are under their control. Various sets of microRNAs found in different human tissues under normal and pathological conditions, including cancer, suggest that miRNAs are involved in most cellular pathways. To date, there is no doubt that regulatory potential of the genome is largely determined by miRNAs. In our study, we performed a comparative phylogenetic analysis of the origin and evolution of the total set of 1048 miRNAs in the human genome and investigated the role of certain miRNAs in carcinogenesis of thyroid and mammary glands, as potential diagnostic and prognostic biomarkers of malignancy. Analysis of phylogenetic distribution of miRNAs in the human genome has shown four peaks of appearance of new miRNA genes in the evolution from Methazoa to H. sapiens. The highest amount of new miRNA genes appeared after divergence of H. s. from common ancestor with P. t. Expansion of transposable elements in genome was accompanied by the origin of new miRNA genes on the basis of their sequences. More than 14 % from 1600 miRNAs of human genome originated from mobile elements and still remain. Profiles of expression of 5 miRNAs, pertaining to oncomicroRNAs - miR-21, -221, -222, -155 and -205 - allow distinguishing ductal invasive carcinoma of mammary gland and thyroid papillary carcinoma. The data obtained suggest different ways and roles of participation of the same miRNAs in carcinogenesis of thyroid and mammary glands. So, these miRNAs and profiles of their expression might be used in the diagnosis and prognosis of cancer.

Cell cycle arrest in the thymus and spleen in male mice under conditions of chronic social defeat stress: effects of diazepam.

The effects of chronic social defeat stress on the percentage of cells in different phases of the cell cycle and in apoptosis in the thymus and spleen of male mice were studied by the method of flow cytofluorometry. In stressed males, thymus weight decreased, the percent of proliferating thymocytes was significantly lower, and the percentage of G0-G1 cells was higher than in intact males. Stress substantially reduced the percentage of splenocytes in the G0-G1 phase and apoptotic cells, but the percentage of S and G2-M cells and proliferation index significantly increased. Chronic administration of anxiolytic diazepam prevented the majority of the changes in the percentage of cells in different phases of the cell cycle, but apoptosis in the thymus increased under these conditions. Possible association between cell cycle disorders, impairment of cell immunity, and chronic anxiety developing under conditions of long-term social defeat stress is considered.

[Comparative organization and the origin of noncoding regulatory RNA genes from X-chromosome inactivation center of human and mouse].

After the radiation of primates and rodents, the evolution of X-chromosome inactivation centers in human and mouse (XIC/Xic) followed two different directions. Human XIC followed the pathway towards transposon accumulation (the repeat proportion in the center constitutes 72%), especially LINEs, which prevail in the center. On the contrary, mouse Xic eliminated long repeats and accumulated species-specific SIN Es (the repeat proportion in the center constitutes 35%). The mechanism underlying inactivation of one of the X chromosomes in female mammals appeared on the basis of trasnsposons. The key gene of the inactivation process, XIST/Xist, similarly to other long noncoding RNA genes, like TSIX/Tsix, JPX/Jpx, and FTX/Ftx, was formed with the involvement of different transposon sequences. Furthermore, two clusters ofmicroRNA genes from inactivation center originated from L2 [1]. In mouse, one of such clusters has been preserved in the form of microRNA pseudogenes. Thus, long ncRNA genes and microRNAs appeared during the period of transposable elements expansion in this locus, 140 to 105 Myr ago, after the radiation of marsupials and placental mammal lineages.

[Exon-intron structure of the Xist gene in elephant, armadillo, and the ancestor of placental mammals].

The Xist gene belongs to the class of long noncoding regulatory RNA genes which play a key role in the process of inactivation of one of the X chromosomes in females of placental mammals. Based on interspecific comparative sequence analysis performed using a set ofbioinformatic programs and approaches, the exon-intron gene structure was first described in two species, elephant and armadillo, belonging to the most primitive placental mammal groups, Afrotheria and Xenarthra. Using multiple sequence alignment of the species representing all main groups of placental mammals (12 species), consensus sequence of the ancestral gene was reconstructed. In the gene structure four evolutionary conserved regions with the identity level of 90% and the sizes of more than 100 bp were identified. Substantial contribution of transposable elements to the gene origin, as well as mosaic evolution of certain elements of the Xist locus was demonstrated. It is likely that the ancestral gene consisted often exons and was formed before the radiation of placental mammals, in the period from 140 to 105 Myr ago.

Magnetic resonant mode in the single-layer high-temperature superconductor Tl2Ba2CuO(6+delta).

An unusual spin excitation mode observed by neutron scattering has inspired numerous theoretical studies of the interplay between charged quasiparticles and collective spin excitations in the copper oxide high-temperature superconductors. The mode has, thus far, only been observed in materials with crystal structures consisting of copper oxide bilayers, and it is absent in the single-layer compound La(2-x)Sr(x)CuO(4+delta). Neutron-scattering data now show that the mode is present in Tl(2)Ba(2)CuO(6+delta), a single-layer compound with a superconducting transition temperature of approximately 90 kelvin, demonstrating that it is a generic feature of the copper oxide superconductors, independent of the layer sequence. This restricts the theoretical models for the origin of the resonant mode and its role in the mechanism of high-temperature superconductivity.

Characterization of the genomic Xist locus in rodents reveals conservation of overall gene structure and tandem repeats but rapid evolution of unique sequence.

The Xist locus plays a central role in the regulation of X chromosome inactivation in mammals, although its exact mode of action remains to be elucidated. Evolutionary studies are important in identifying conserved genomic regions and defining their possible function. Here we report cloning, sequence analysis, and detailed characterization of the Xist gene from four closely related species of common vole (field mouse), Microtus arvalis. Our analysis reveals that there is overall conservation of Xist gene structure both between different vole species and relative to mouse and human Xist/XIST. Within transcribed sequence, there is significant conservation over five short regions of unique sequence and also over Xist-specific tandem repeats. The majority of unique sequences, however, are evolving at an unexpectedly high rate. This is also evident from analysis of flanking sequences, which reveals a very high rate of rearrangement and invasion of dispersed repeats. We discuss these results in the context of Xist gene function and evolution.

[The characteristic properties of Vibrio cholerae eltor isolated from environmental objects on the territory of the former USSR during the 7th cholera pandemic]. / Kharakteristika nekotorykh svoistv Vibrio cholerae eltor, vydelennykh iz ob''ektov okruzhaiushchei sredy na territorii byvshego SSSR vo vremia sed'moi pandemii kholery.

The properties of 22,382 V. eltor strains isolated from environmental objects on the territory of different climatic and geographical zones during the period of 1970-1988 were studied. The study was made on the morphology of their colonies, the agglutinability of the strains by cholera O serum, type-specific serum and RO serum, their capacity for being lyzed by V. eltor bacteriophage, their hemolytic activity and virulence. Differences in the occurrence of strains with any of the above-mentioned properties, depending on the object from which they were isolate, the climatic and geographical zone and the intensity of the epidemiological situation with regard to cholera.

MEC: a transposable element from Chironomus thummi (diptera).

Two genomic clones, pC1.2 and p20D (containing inserts of 2.0 and 1.6 kb, respectively) were isolated from the A2b region to polytene chromosome IV of Chironomus thummi thummi salivary gland cells. Upon in situ hybridization to polytene chromosomes of C. thummi thummi and C. thummi piger, p20D DNA hybridized mainly over the A2b region of chromosome IV, whereas pC1.2 DNA hybridized to at least 90 sites distributed over all the chromosomes. A partial nucleotide sequence analysis showed that these clones were very similar and allowed the detection of a 596 bp insert in the pC1.2 clone. This insert possesses all of the essential features of a Class II transposable element and was called MEC. It carries a nearly perfect 107 bp terminal inverted repeat containing one mismatch and is flanked by a 5 bp direct repeat. The 372 bp central region contains a short open reading frame with a coding capacity of 58 amino acids.

A tissue-specific puff (Balbiani ring a) in Chironomus thummi may contain a gene encoding a 67-kDa protein which exhibits non-tissue-specific expression.

A 2.3-kb genomic clone has been isolated from the region where the tissue-specific puff, Balbiani ring a (BRa), is found on chromosome IV of the special lobe of Chironomus thummi salivary gland cells. The clone was characterized by nucleotide sequence analysis. Two clusters of direct tandem repeats were identified, as well as large and small open reading frames (ORFs). The large ORF was fused to an Escherichia coli lacZ gene. Antibodies against the beta-galactosidase/ORF fusion protein reacted selectively on Western blots with a 67-kDa protein. Western-blot analysis and immunoelectron microscopy showed that this protein was distributed in the cells of all larval tissues examined. We concluded that BRa, a tissue-specific puff, whose activity correlates with the synthesis of 160-kDa secretory protein [Kolesnikov et al., Chromosoma 83 (1981) 661-677], may also contain a gene which is not expressed in a tissue-specific manner.

A new transposable element in Chironomus thummi.

A 1.7 kb long transposable element called TECth1 was found in the 3' flanking region of a Chironomus thummi Balbiani ring gene. As shown by sequence comparison with a second copy, TECth1 is characterized by a perfect terminal inverted repeat of 17 bp flanked by a duplicated target site of 8 bp, four internal imperfect inverted repeats of 17 to 26 bp and terminal regions of about 0.25 kb with a high number of short direct repeats of the consensus sequence ACTTT or permutated and mutated forms such as TTTAC or ACTAT. The terminal inverted repeats and the 8 bp target site duplication are reminiscent of Drosophila P and hobo elements but no long open reading frame starting with ATG is present, suggesting that the two TECth1 copies studied represent deletion derivatives of a longer element coding for its own transposase. In situ hybridization revealed about 75 labelled sites distributed over all chromosomes with the Balbiani ring locus most strongly labelled. Fifty percent of the sites are specific for a given individual, and these variable sites are often heterozygous for the element.

[The composition of a population of cloned cultures of revertant Vibrio cholerae L forms]. / Izuchenie sostava populiatsii klonovykh kul'tur revertantov L-form kholernykh vibrionov.

The process of L-transformation and L-transformed state duration have been studied for their effect on variability of main characters of revertant cultures of choleric vibrions L-forms at the population level with the use of cloned cultures of the choleric vibrions. The study was conducted on two strains of the choleric vibrion of the eltor biovar in different periods of storage in the L-transformed state (1, 3, 6 months). It has been revealed that characters of the species and biovar remained stable despite the influence of L-transforming agents. The characters of clone cultures characterizing virulence (sensitivity to KhDF phages, hemolytic activity, toxin production and virulence for sucking rabbits proved to be subjected to variability to the greatest extent with simultaneous preservation of the toxin-production gene. A resistant change of the serovar (from Inaba to Ogava) is observed only in one revertant-subculture of the virulent strain.

Cell-specific synthesis and glycosylation of secretory proteins in larval salivary glands of Chironomus thummi.

Synthesis and glycosylation of larval salivary gland secretory proteins of Chironomus thummi were analyzed with respect to cell specific differences in the Balbiani ring (BR) pattern and glycoprotein composition of secretion formerly detected by histochemical staining procedures. In the secretion of a special cell type in salivary glands, which is characterized by the appearance of an additional BR, and additional polypeptide with a relative molecular weight (Mr) of 160 kD was found differing in its antigenic properties and tryptic fingerprint pattern from main cell secretion proteins. This so-called ssp-160 component is preferentially synthesized and glycosylated in the special cells. In the same cells, both the synthesis and glycosylation of all other major secretory proteins was found to be diminished or even repressed. In contrast to the conspicuous cell-specific differences at the level of protein synthesis. RNA analyses show the prominent synthesis of 75S RNA in both main and special cells and gave no clear indication of the synthesis of a smaller RNA fraction as expected from the size of ssp-160 component.--These and further data on synthesis and properties of secretory proteins as well as expression of BR DNA are discussed with regard to the assumption that at least some of the eight major secretory polypeptides are coded for by BR DNA. The BR gene(s) might have originated by manifold duplications and modifications of short repetitive prototype DNA sequences, which are coordinatively expressed.

[Functional activity of the salivary glands of Drosophila melanogaster in the course of metamorphosis: a comparative electrophoretic analysis of the proteins of the salivary glands, fat body and hemolymph]. / Funktsionl'naia aktivnost' sliumnykh zhelez Drosophila melanogaster v khode metamorfoza: sravnitel'nyi élektroforeticheskii analiz belkov sliunnykh zhelez, shirovogo tela i gemolimfy.

The proteins of the salivary gland secretion at the moment of puparium formation and the protein composition of the salivary glands, fat-body and hemolymph during the III larval instar and 12 hours prepupal period were investigated by means of electrophoresis in polyacrilamide gel, in the presence of urea. Both the qualitative and quantitative differences in the protein composition of the salivary glands were found which appeared to be due to the change of functions of the salivary glands in the process of development. 69 protein fractions were isolated in the salivary gland secretion, 40 of them being PAS-positive. A correlation was noted between the appearance of the first PAS-positive fractions at the stage of 95-100 hrs (from the moment of egg laying) and the cytological demonstration of PAS-positive secretion granules. New glycoproteins appeared in the salivary glands in prepupae (2 hrs after the puparium formation) and, later, in the hemolymph. A complicated pattern of the secretion formation in the salivary glands of the III instar larvae and possible change of its composition during metamorphosis are discussed.