Efficient enrichment of synchronized mouse spermatocytes suitable for genome-wide analysis.

Chromatin-based mechanisms regulating developmental transitions during meiosis are fundamental but understudied aspects of male gametogenesis. Indeed, chromatin undergoes extensive remodeling dur-ing meiosis, leading to specific patterns of gene expression and chromosome organization, which ulti-mately controls fundamental meiotic processes such as recombination and homologous chromosome associations. Recent game-changing advances have been made by analysis of chromatin binding sites of meiotic specific proteins genome-wide in mouse spermatocytes. However, further progress is still highly dependent on the reliable isolation of sufficient quantities of spermatocytes at specific stages of prophase I. Here, we describe a combination of methodologies adapted for rapid and reliable isolation of synchronized fixed mouse spermatocytes. We show that chromatin isolated from these cells can be used to study chromatin binding sites by ChIP-seq. High quality data we obtained from INO80 ChIP-seq in zygotene cells was used for functional analysis of chromatin binding sites.

Prevalence of non-O157 Shiga Toxin-producing E. Coli in Children and Calves in Al- Muthanna Province, Iraq

The present study focus on non-O157 Shiga toxin-producing E. Coli (STEC), included a bacteriological study was subjected to provide additional information for non-O157 STEC prevalence in children and calves. Isolation by using selective culturing media (CHROMagar STEC and CHROMagar O157) from 127 children suffering from diarrhea and 133 calves in Al- Muthanna province. Characterization depends on culturing positive colony on MacConkey agar and Levin’s Eosin Methylene blue agar, staining single colony from the growth by gram stain, biochemical tests; Indole, the Methyl Red, Voges-Proskauer, Citrate test, Oxidase, Catalase, Urease, Motility, Kligler Iron and Api-20E, were done to confirm a diagnosis of non-O157 STEC, The reliable isolation as non-O157 STEC serotyping by specific latex agglutination test for the target non-O157 STEC (big six) serogroup (O26, O45, O103, O111, O121 and O145). The current study showed the prevalence of non-O157 STEC was 20 of out 127 (15.73%) in samples collected from children and 27 / 133 (20.30%) in calves samples in conclusion the Non-O157 STEC is an important cause of diarrhea in children, and calves; finally, the calves play an important reservoir for Non-O157 STEC.

Development of a Specific Monoclonal Antibody to Detect Male Cells Expressing the RPS4Y1 Protein

Hemophilia is an X-linked recessive bleeding disorder. In pregnant women carrier of hemophilia, the fetal sex can be determined by non-invasive analysis of fetal DNA circulating in the maternal blood. However, in case of a male fetus, conventional invasive procedures are required for the diagnosis of hemophilia. Fetal cells, circulating in the maternal bloodstream, are an ideal target for a safe non-invasive prenatal diagnosis. Nevertheless, the small number of cells and the lack of specific fetal markers have been the most limiting factors for their isolation. We aimed to develop monoclonal antibodies (mAbs) against the ribosomal protein RPS4Y1 expressed in male cells. By Western blotting, immunoprecipitation and immunofluorescence analyses performed on cell lysates from male human hepatoma (HepG2) and female human embryonic kidney (HEK293) we developed and characterized a specific monoclonal antibody against the native form of the male RPS4Y1 protein that can distinguish male from female cells. The availability of the RPS4Y1-targeting monoclonal antibody should facilitate the development of novel methods for the reliable isolation of male fetal cells from the maternal blood and their future use for non-invasive prenatal diagnosis of X-linked inherited disease such as hemophilia.

Return to neptunism: review article

The article tries to revive the traditions of neptunizm. The neptunist Cuvier believed that «the fossil is the key to the past. The past is characterized by a radical break with the present, perhaps it even proceeded according to another physical laws. This is evidenced by the fossils. With the modern action of physical forces, all the deceased remnants decompose. Fossils could have formed during catastrophically rapid burial with reliable isolation from oxygen. The present is the key to the past according to the lawyer Lyell. The past is like the present. It contains an incredible amount of time and years. All processes were going with the same energy as they do now and were not overshadowed by any gigantic disaster. Lyell's statements entered science under the name of the principle of actualism. He gave a paradigmal form not only to the earth sciences, but to all of science, and formed the basis of the evolutionary doctrine. The fundamental dogma of randomness of both natural and all historical processes is the basis of modern scientific ideology. This randomness has no purpose and cannot have. Nevertheless, paradoxically, it predetermines the evolutionary progress of all things. With the appearance of the anthropic principle the teleologism regains its completeness and direction. The anthropic principle is teleological. Since the entire Universe and the biosphere are attuned to human nature, the more such attunement can be expected from earthly nature. There are opportunities for harmonizing human with natural landscapes for health-improving purposes. Nothing was known about the subtlest adjustment of the Universe for humans, or about the incredible complexity of the biosphere even 50-70 years ago. The universe could be explained by evolution and actualism. The discovered incredible complexity of the world brings religion and science closer together. A catastrophic and probably anti-random picture of the planet's history is emerging more and more clear. In this light, the power of science is seen again not in confrontation with religion, but in harmonization with it.

CLINICAL AND LABORATORY ANALYSIS OF THE EFFECTIVENESS OF TREATMENT OF DEEP CARIES WITH CALCIUM-CONTAINING PADS

The task of a dentist in the treatment of deep caries (pulp hyperemia) is not only to restore the integrity of the tooth, but also to stimulate the production of replacement dentin with the help of a medical pad, to ensure reliable isolation of the pulp from the toxic effects of materials. Having a wide range of different therapeutic pads, but not having methods to objectively assess the condition of the pulp, doctors have to «blindly» give preference to a particular manufacturer. In this work, based on a clinical and laboratory study, an analysis of the effectiveness of calcium-containing pads «Life» and «Dycal» – was performed. The clinical stage was performed on patients in the treatment of deep caries. Changes in the pulp microcirculation were evaluated using laser Doppler flowmetry. During histological examination, samples of tooth cuts were studied for analysis of micro-flows of the dye at the «seal-tooth» border and the depth of penetration of the dye. The clinical results are consistent with laboratory results, which can be used to draw conclusions about the benefits of the «Life» treatment pad in the treatment of deep caries.

DEVELOPMENT OF A MATHEMATICAL APPARATUS FOR THE CHOICE OF PARAMETERS OF AIR-INFLATED SLEEVE OF A MINING VENTILATION WALL OF A NEW TYPE

The results of mathematical modeling are presented, on the basis of which a finite-element model of air-inflated sleeve of a shaft ventilation wall of a new type is developed, which is the main element of its design and provides reliable isolation of the excavation from air leaks. If necessary, the developed model can be adapted to any section of the excavation.

Analysis of technical solutions for preventing admission of formation fluids in the annular space of the well during the waiting on cement time

The creation of a sealed support, providing reliable isolation of layers from each other, is one of the most important tasks in the construction of oil and gas wells. The article presents the most common in recent years in our country and abroad special technological and technical measures used to improve the quality of isolation of oil and gas reservoirs at the stage of cementing and waiting on cement with different gradients of reservoir pressures. Attention is drawn to technical and technological solutions for preventing admission of formation fluids in the annular space of the well during cementing and waiting on cement. The solution to this problem is based on creating the design pressure at the wellhead in the annulus at a rate proportional to the rate of reduction of the hydrostatic pressure column of cement slurry (drilling mud) due to the structure hardening cement slurry.