Detection of Y-chromosome marker in plasma of pregnant women using real time PCR: diagnostic accuracy depending on gestation age.

The evaluation of the clinical significance of the test for the detection of the Y-chromosome marker in the plasma of a pregnant woman at different stages of pregnancy by real-time PCR was carried out. The blood samples of 4616 women at 4 to 32 gestation weeks were studied. Identification of the Y-chromosome marker was carried out based on the amplification of a region of the TSPY gene. The Y-chromosome marker was unambiguously identified in 2131 samples, which accounted for 46.2% of the total number of analyzed samples. In 233 samples (5%), the Y-chromosome marker was detected with reduced reliability, and in 15 samples (0.3%), an unambiguous conclusion about the presence or absence of Y-specific DNA in plasma could not be made during the initial study. The diagnostic accuracy of the Y-chromosome marker determination in the plasma of a pregnant woman at the 4-6th gestation week was 95.5%, and from the 7th week and at later stages of pregnancy it reached 97.3-98.2%. Testing from the 7th gestation week may be recommended for reliable prenatal sex determination of the fetus by real-time PCR analysis of extracellular circulating fetal DNA.

A simplified and sensitive immunoprecipitation mass spectrometry protocol for the analysis of amyloid-beta peptides in brain tissue.

In the field of Alzheimer's disease, there is an urgent need for novel analytical tools to identify disease-specific biomarkers and to evaluate therapeutics. Preclinical trials commonly employ amyloid beta (Aß) peptide signatures as a read-out. In this paper, we report a simplified and detailed protocol for robust immunoprecipitation of Aß in brain tissue prior to mass spectrometric detection exemplified by a study using transgenic mice. The established method employed murine monoclonal and rabbit polyclonal antibodies and was capable of yielding well-reproducible peaks of high intensity with low background signal intensities corresponding to various Aß forms.

Persistent histone modifications at the BDNF and Cdk-5 promoters following extinction of nicotine-seeking in rats.

Drugs of addiction lead to a wide range of epigenetic changes at the promoter regions of genes directly implicated in learning and memory processes. We have previously shown that the histone deactylase inhibitor, sodium butyrate (NaB), accelerates the extinction of nicotine-seeking and provides resistance to relapse. Here, we explore the potential molecular mechanisms underlying this effect. Rats received intravenous nicotine or saline self-administration, followed by 6 days of extinction training, with each extinction session followed immediately by treatment with NaB or vehicle. On the last day of extinction, rats were killed and the medial ventral prefrontal cortex retained for chromatin immunoprecipitation and quantitative polymerase chain reaction (qPCR). A history of nicotine exposure significantly decreased H3K14 acetylation at the brain-derived neurotrophic factor (BDNF) exon IV promoter, and this effect was abolished with NaB treatment. In contrast, nicotine self-administration alone, resulted in a significant decrease in histone methylation at the H3K27me3 and H3K9me2 marks in the promoter regions of BDNF exon IV and cyclin-dependent kinase 5 (Cdk-5). Quantitative PCR-identified changes in several genes associated with NaB treatment that were independent of nicotine exposure; however, an interaction of nicotine history and NaB treatment was detected only in the expression of BDNF IV and BDNF IX. Together these results suggest that nicotine self-administration leads to a number of epigenetic changes at both the BDNF and Cdk-5 promoters, and that these changes may contribute to the enhanced extinction of nicotine-seeking by NaB.

Specific features of root aerenchyma formation in Sium latifolium L. (Apiaceae).

Types of aerenchyma formation in adventive roots of wild plant S. latifolium L. are described for the first time. Specific cell divisions and patterns of cell growth in the inner and outer root cortex at the initial stages of aerenchyma formation are highlighted in detail. Destructive processes in cells occurred under aerenchyma formation are considered in the view of known stages of programmed cell death in plants.

[Medical and hygienic aspects of instrumental supervision system over nuclear materials and radioactive substances transport on Russian Federation territory].

Hygienic evaluation of radiation situation in operation of mobile and stationery elements within a project of national system for instrumental supervision over nuclear materials and radioactive substances transport, created with a Global initiative against nuclear terrorism. Levels of exposure to ionizing radiation of the screening complexes appeared to match requirements on radiation safety for service personnel and general population.

[Interrelation of microtubules and microfilaments in the distal elongation zone of Arabidopsis thaliana root].

Investigations on Arabidopsis thaliana--GFP- MAP4 and A. thaliana--GFP-ABD2 plants treated with tubulin and actin inhibitors has shown that stability of cell growth in the distal elongation zone of plant root is achieved by structural interrelation of microtubules and microfilaments. Use of clinorotation and application of inhibitor analysis allowed us to show that gravity force defines the type of such interrelation.

Role of cytoskeleton in gravisensing of the root elongation zone in Arabidopsis thaliana plants.

In order to reveal the involvement of tubulin microtubules and actin microfilaments in gravisensing reactions in the distal elongation zone of root, Arabidopsis thaliana plants stably transformed with MAP4-GFP construct were grown under slow clinorotation. Experiments have shown that stabilization of cell growth in the distal elongation zone of Arabidopsis seedling root is provided by common structural organization of microtubules and microfilaments, and interrelations between microtubules and microfilaments is highly dependent upon the type of cell differential growth. Less pronounced effect of microfilament disruption on microtubule organization has been observed under clinorotation and it suggests the existence of complex mechanism of cooperation between microtubules and microfilaments which is probably, masked on earth.

Organization of cytoskeleton during differentiation of gravisensitive root sites under clinorotation.

Key role in cell gravisensing is attributed to the actin cytoskeleton which acts as a mediator in signaling reactions, including graviperception. Despite of increased attention to the actin cytoskeleton, major gaps in our understanding of its functioning in plant gravisensing still remain. To fill these gaps, we propose a novel approach focused on the investigation of actin involvement in the development of columella cells and cells in the transition zone of roots submitted to clinorotation. Both statocytes and cells in the transition zone represent the postmitotic cells which take origin in root meristems and are specified into graviperceptive (root cap) and gravireacting (transition zone) root tissues. The aim of the research was to investigate and compare the microfilament arrangements in root cap statocytes and peripheral root tissues (epidermis and cortex cells) in the transition zone and to find out how the actin cytoskeleton is involved in their specification under clinostat conditions. So far, our experiments have shown that under clinorotation the cytoplasmic microfilament network in the cortex cells in the transition zone is significantly enhanced. It is suggested that more abundant cytoplasmic microfilaments could strengthen the cortical actin cytoskeleton arranged parallel with the cortical microtubules, which are found to be partially disorganized in this area. Due to microtubule disorganization, the functioning of cellulose-synthesizing machinery and proper deposition of cell wall might be affected and could cause the alterations in the growth mode. But, in our case growth of the cells in the transition zone under clinorotation was rather stable. Due to our opinion, general stability of cell growth under clinorotation is promoted by mutual functional interrelation between actin and tubulin cytoskeletons. It is suggested that a strengthened cortical actin cytoskeleton restricts the cell growth instead of disorganized microtubules.

Application of GFP technique for cytoskeleton visualization onboard the International Space Station.

Cytoskeleton recently attracted wide attention of cell and molecular biologists due to its crucial role in gravity sensing and trunsduction. Most of cytoskeletal research is conducted by the means of immunohistochemical reactions, different modifications of which are beneficial for the ground-based experiments. But for the performance onboard the space vehicles, they represent quite complicated technique which requires time and special skills for astronauts. In addition, immunocytochemistry provides only static images of the cytoskeleton arrangement in fixed cells while its localization in living cells is needed for the better understanding of cytoskeletal function. In this connection, we propose a new approach for cytoskeletal visualization onboard the ISS, namely, application of green fluorescent protein (GFP) from Aequorea victoria, which has the unique properties as a marker for protein localization in vivo. The creation of chimerical protein-GFP gene constructs, obtaining the transformed plant cells possessed protein-GFP in their cytoskeletal composition will allow receiving a simple and efficient model for screening of the cytoskeleton functional status in microgravity.

[Current issues of medical informatics]. / Tekushchie zadachi informatizatsii meditsinskoi nauki.

Due to the modern high standards of information technologies it is only natural to promote our medical care and to ensure a new quality of clinical services. Information technologies should be introduced into the medical field with due respect to clearly predetermined principles. Analyzed in the paper are the key reasons for a huge number of problems occurring in the sphere of medical informatics; optimal methods of medical-informatics introduction are defined.

[Infrastructure of information support to clinical medicine]. / Ob infrastrukture informatsionnoi podderzhki klinicheskoi meditsiny.

Issues are discussed related with designing an effective infrastructure of the information-and-computer support to clinical medicine with regard for the recent achievements in informatics, automated systems, medical equipment and robotics. It is suggested to focus on creating a module-type infrastructure meeting evenly both the trends towards integration and towards autonomy. A scheme of an information infrastructure constructed for Bakulev's National Center for Cardiovascular Surgery of the Russian Academy of Medical Sciences is described.

[Role of cytoskeleton in gravisensitivity of a plant cell: experimental data and hypotheses]. / Rol' tsitoskeleta v gravichuvstvitel'nosti rastitel'noi kletki: éksperimental'nye dannye i gipotezy.

The article highlights the hypotheses of plant gravisensing, including those where cytoskeleton is involved. Data concerning arrangement of microfilaments and microtubules in specified and non-specified to graviperception cells of higher and low plants and concepts of the role of cytoskeleton in plant gravisensing are considered.

Orientation of root hair growth is influenced by simulated microgravity.

We have tried to investigate the mechanisms supporting the plagiotropic growth (growth in parallel to the Earth) of root hairs in simulated microgravity. Our strategy to understand the regulation of such type of growth depends upon the study of cytoskeleton topography and calcium ions distribution in root hairs both in control and simulated microgravity.

[The actin cytoskeletal characteristics of the root meristem cells in Beta vulgaris L. at different mitotic stages]. / Osobennosti aktinovogo tsitoskeleta kletok kornevoi meristemy Beta vulgaris L. na raznykh stadiiakh mitoza.

For the first time the actin cytoskeleton in meristem cells of B. vulgaris root has been studied. It was established that all discovered earlier patterns of microfilament arrangement were also specific for meristem of present species. Specific rearrangements of actin in the sites of spindle (prophase) and phragmoplast formation (telophase) were revealed. It is supposed, that actin cytoskeleton determines certain position of spindle in cells and takes part in the cell plate formation.

Patterns of cortical microtubules formed in epidermis of Beta vulgaris L. roots under clinorotation.

Changes of cortical microtubules (MTs) from the normal transverse arrangement were observed in epidermal cells of Beta vulgaris roots under clinorotation. We hypothesize that the epidermis is sensitive to clinorotation and that the microtubular cytoskeleton plays a key role in the ensuing growth response.

[The distribution of glial fibrillary acidic protein and protein S-100 in experimental brain tumors in rats]. / Raspredelenie glial'nogo fibrilliarnogo kislogo belka i belka S-100 v éksperimental'nykh opukholiakh mozga krys.

The methods of quantitative immunoelectrophoresis and indirect immunofluorescence were used study the content of glial fibrillary acid protein in 10 serially reinoculated rat gliomas induced primarily by ethylnitrosourea (a total of 135 tumors). It was found that the GFAP content reduced with increase of malignancy. However, wide scattering of the GFAP content in some of the tumors was characteristic of all strains. In the group of slowly growing glial tumors (2 malignant astrocytomas and one malignant oligoastrocytoma) the GFAP content ranged from 50 to 600% and exceeded the normal content two-to threefold on the average. In the group of highly malignant gliomas (4 malignant ependymomas, 2 malignant gliomas, and one glioblastoma) the GFAP content was within the limits of 65-120%. In most cases the GFAP level was below normal or could not be determined at all. At the same time, tumors with a high GFAP content were encountered. The GFAP-positive cells were unevenly distributed in the gliomas: separately, in foci, and around the vessels. Their number increased in the direction of the periphery of the tumor. Intensive fluorescence was noted on the tumor--brain borderline. The content of protein S-100 in the experimental gliomas was always below normal.

[Characteristics of concanavalin A-binding neurospecific glycoproteins from various regions of the rat brain during experimental neurosis]. / Kharakteristika konkanavalin-A-sviazyvaiushchikh neirospetsificheskikh glikoproteinov v razlichnykh otdelakh golovnogo mozga krys pri éksperimental'nom nevroze.

The polypeptide composition of neurospecific glycoproteins in different areas of the rat brain under experimental neurosis is characterized using SDS-PAG-electrophoresis followed by electroblot and immunofixation on nitrocellulose membranes. The soluble and membrane-bound glycoproteins are purified by Con A-Sepharose column chromatography. Changes in the glycoprotein polypeptide composition in different areas of the rat brain under experimental neurosis are qualitative. Soluble glycopolypeptide 27 kDa and membrane glycopolypeptide 32 kDa are not revealed in the midbrain and corpus striatum. Soluble polypeptide 47 kDa is absent in the cerebral cortex and hippocampus. It is suggested that the above mentioned glycopolypeptides are important for the CNS physiological functioning.

[Identification and characteristics of concanavalin A-binding neurospecific glycoproteins in human brain and brain tumors]. / Identifikatsiia i kharakteristika konkanavalin A-sviazyvaiushchikh neirospetsificheskikh glikoproteinov v mozge i opukholiakh mozga cheloveka.

Soluble and membrane-bound neurospecific Con A-binding glycoproteins from human brain and tumours were identified and characterized, using a procedure which included stepwise extraction with low and high ionic strength buffers, buffered. Triton X-100 and sodium deoxycholate followed by ConA-Sepharose column chromatography, SDS-PAAG electrophoresis and immunoblotting. Adsorbed antisera against different types of neurospecific glycoproteins were used. The bulk of neurospecific glycoproteins (11 and 13) were revealed in protein fractions extracted with low ionic strength buffers and Triton X-100. In astrocytomas and glyoblastomas, some neurospecific glycoproteins were absent. Some glycoproteins were found in tumours, but were absent in brain tissue. Soluble, 77 kD glycoprotein, 11 and 16 kD glycoproteins solubilized with high ionic strength buffers and intrinsic membrane-bound 51, 57, 61, 74 and 77 kD glycoproteins can be viewed as stable neurospecific markers in malignant brain tumours.

[Neurospecific cytoskeletal proteins]. / Neirospetsificheskie belki tsitoskeleta.

Data available in literature on neurospecific proteins of cytoskeletal structures--microtubules, microfilaments and intermediate filaments are generalized. Properties of tissue-specific cytoskeletal proteins which are typical of nerve cells are summarized. The structure, physicochemical properties, cell localization, metabolism and function of cytoskeletal proteins are characterized. The coexpression and interaction of different cytoskeletal structures are considered. An analysis of neurospecific cytoskeletal proteins is of great practical importance for neurobiology, neurooncology, neurosurgery. The proteins can be used as markers of different pathologies in the nervous system.