Transcriptional analysis of biopsies derived from in vitro produced bovine blastocysts in relation to pregnancy success after transfer to recipients

This study was carried out to address the relationship between the transcriptional profile of embryos and the pregnancy success based on gene expression analysis of blastocyst biopsies taken prior to transfer to recipients. For this, biopsies (30-40% of the intact embryo) were taken from day 7. Blastocysts (n=118) and 60-70% part were transferred to recipients after re-expansion. Based on the success of pregnancy, biopsies were pooled in three groups (each 10 biopsies) namely: those resulting in no pregnancy (G1), resorption (G2) and those resulting in delivery of calf (G3). Gene expression analysis of these groups was performed using a home made bovine preimplantation specific cDNA array (with 219 clones) and BlueChip (with ~2000 clones). Data analysis using Significant Analysis for Microarray (SAM) software revealed that a total of 52 and 58 genes were differentially regulated during comparison between G1 versus G3 and G2 versus G3 respectively. Quantitative real-time PCR was used to validate the results of the microarray experiments. G3-Biopsies are enriched with genes necessary for implantation (COX-2 and CDX2), carbohydrate metabolism (ALOX15), growth factor (BMP15), response to oxidative stress (TXN), signal transduction (PLAU) and placenta-specific 8 (PLAC8). G2-Biopsies are enriched with transcripts involved protein phosphorylation (KRT8), plasma membrane (OCLN) and glucose metabolism (PGK, AKR1B1). G1-Biopsies are enriched with transcripts involved inflammatory cytokines (TNF), and factors relevant for protein amino acid binding (EEF1A1), transcription factors (MSX1, PTTG1), glucose metabolism (PGK1, AKR1B1) and CD9 which is an inhibitor of implantation. The bovine MSX1 protein detected by immunohistochemistry was localized in the cytoplasm of immature oocytes and distributed at periphery of the cytoplasm of matured oocytes. Throughout the preimplantation period the staining was apparently more concentrated around the nuclei, whereas the ICM in blastocyst showed weaker labelling for MSX1 than the trophectoderm. In conclusion, we generated direct candidates of genes which may play an important role in determining the fate of the embryo after transfer. Genexpressionsprofile von in vitro produzierten Rinderembryobiopsien in Relation zur in vivo Entwicklung nach Transfer In dieser Arbeit wurde der Zusammenhang zwischen Genexpression in Embryonen und Trächtigkeitserfolg, basierend auf der Expressionsanalyse von vor dem Transfer an den Rezipienten gewonnenen Blastozystenbiopsien, untersucht. Hierzu wurden Biopsien (30- 40% des intakten Embryos) von Tag 7 Blastozysten genommen (n=118) und der 60-70% Restteil den Rezipienten nach Re-expansion transferiert. Basierend auf dem Erfolg der Trächtigkeit wurden die Biopsien in 3 Gruppen (je 10 Biopsien) gepoolt, nämlich: jene, die in keiner Trächtigkeit resultierten (G1), resorbierte Embryonen (G2) und jene, die in Geburt eines Kalbes resultierten (G3). Die Genexpressionsanalyse dieser Gruppen wurde mit einem selbsthergestellten bovinen präimplantationsspezifischen cDNA Array (mit 219 Clonen) und mit BlueChip (mit ~2000 Clonen) durchgeführt. Die Datenanalyse mittels Significant Analysis for Microarray (SAM) Software zeigte insgesamt 52 bzw. 58 unterschiedlich regulierte Gene im Vergleich zwischen G1 versus G3 und G2 versus G3. Quantitative real-time PCR wurde zur Bestätigung der durch das Microarray-Experiment entdeckten unterschiedlich exprimierten Gene eingesetzt. G3-Biopsien exprimierten herstäarkt solche Gene, die notwendig für Implantation (COX-2 und CDX2), Kohlenhydratmetabolismus (ALOX15), Wachstum (BMP15), oxidative Stressantwort (TXN), Signalübermittlung (PLAU) und Plazentafunktion-8 (PLAC8), sind die Biopsien der resorbierten Embryonen zeigten vermehrt Transkripte, die in Protein-phosphorylation (KRT8), Plasmamembranaufbau (OCLN) und Glucosemetabolismus (PGK1, AKR1B1). Involvirt sind die Biopsien von G1-Embryonen, exprimierten vermehrt Transkripte von Zytokinen (TNF), Proteinaminoacidbindung (EEF1A1), Transkriptionfaktoren (MSX1, PTTG1), Enzymen des Glucosemetabolismus (PGK1, AKR1B1) und CD9, einem Inhibitor der Implantation. Zusammengefasst lässt sich sagen, dass wir direkte Kandidatengene identifiziert haben, die eine wichtige Rolle in der Bestimmung der Enwicklungsfähigheit des Embryos nach dem Transfer spielen könnten

Sprayer for Quantitative Application of Odor Stimuli

A novel device is described for the quantitative application of chemical stimuli. The device uses ultrasound to disperse a solution of volatile chemicals as an aerosol. A motor-driven syringe controls the rate at which the solution is released from a glass capillary. Vibration of the capillary disperses the released solution into microdroplets that evaporate completely within a few centimeters of the tip. The ratio of chemical stimulus to solvent is maintained until the liquid is dispersed and therefore the release rate of the chemical stimulus can be set and calculated straightforwardly from the dilution factor and the dynamically controllable speed of the syringe plunger. The sprayer permits the delivery of chemical stimulus independent of relative vapor pressures of the components and of environmental factors such as temperature. The sprayer is easy to operate, can be constructed from inexpensive materials, and can be used to emit odor stimuli in the wind tunnel or any other bioassay for pheromones and plant volatile

Morphological and molecular characterization of L-methioninase producing Aspergillus species

Six species of L-methioninase producing Aspergillus species, isolated from Egyptian soil, were selected for comprehensive morphotypic and molecular characterization. Based on morphological and physiological features, these isolates were identified as Aspergillus flavipes, Aspergillus carneus, Aspergillus flavus, Aspergillus tamari, Aspergillus oryzae, and Aspergillus parasiticus. Regarding to the maximum enzyme productivity by A. flavipes, it was selected as authentic strain for ribosomal ribonucleic acid (rRNA) primer design. Using primer combinations for 18S rRNA and internal transcribed spacers (ITS)1 amplification, these isolates gave the same polymerase chain reaction (PCR) amplicon size, revealing the relative molecular identity. Moreover, using ITS2 primers, among the six isolates, Aspergillus flavipes EK and A. carneus displayed PCR products on agarose gel, approving the actual morphological and biochemical similarities of these two isolates, A. flavipes group. By sequencing of ITS1-5.8S-ITS2 region, blasting and alignment from the data base, A. flavipes EK showed a typical identity to gene bank deposited A. flavipes isolates. The rRNA sequence of A. flavipes EK was deposited to genbank under accession number JF831014.Key words: Aspergillus, morphological descriptions, 18 S rRNA, internal transcribed spacers (ITS) regions

Experimental behavior based on effective slab width acting as a flange with supporting steel beams in composite floors with openings

An experimental study with many variables was carried out to investigate the effective width of simply supported composite beams (T&L sec. beams) with & without slab opening. Three full-scale composite slab models with six different carrier simply supported composite beams (three with T-sec. & three with L-sec.) with constant slab rectangularity was tested.  Each model loaded by uniform load (in the elastic zone) then loaded by means of two-concentrated load system acting at equal distances from supports till failure. One of them without slab opening and the others with two symmetric slab openings and constant opening horizontal position. All standard tests were performed and all instructions and precautions were followed as perfect as the laboratory instruments permitted.  All experimental steps were executed in a manner such that meeting practical conditions as exactly as could. In this research, the tested model features, materials properties and measurement instrumentations are described in brief. The maximum load carrying capacity; crack patterns, generated strains and deflections at different positions were experimentally investigated. For the sake of analysis generalization and error minimization, all results were analyzed by means of dimensionless relations. The results were recorded and analyzed guided by their mechanical behavior. Interesting gained results, conclusions and recommendations are introduced

Screening, morphological and molecular characterization of fungi producing cystathionine γ-lyase

The potency for production of cystathionine γ-lyase (CGL) by the fungal isolates was screened. Among the tested twenty-two isolates, Aspergillus carneus was the potent CGL producer (6.29 U/mg), followed by A. ochraceous (6.03 U/mg), A. versicolor (2.51 U/mg), A. candidus (2.12 U/mg), A. niveus and Penicillium notatum (2.0 U/mg). The potent six isolates producing CGL was characterized morphologically, A. carneus KF723837 was further molecularly characterized based on the sequence of 18S–28S rDNA. Upon sulfur starvation, the yield of A. carneus extracellular CGL was increased by about 1.7- and 4.1-fold comparing to non-sulfur starved and L-methionine free medium, respectively. Also, the uptake of L-methionine was duplicated upon sulfur starvation, assuming the activation of specific transporters for L-methionine and efflux of CGL. Also, the intracellular thiols and GDH activity of A. carneus was strongly increased by S starvation, revealing the activation of in vivo metabolic antioxidant systems. Upon irradiation of A. carneus by 2.0 kGy of γ-rays, the activity of CGL was increased by two-fold, regarding to control, with an obvious decreases on its yield upon further doses. Practically, CGL activity from the solid A. carneus cultures, using rice bran as substrate, was increased by 1.2-fold, comparing to submerged cultures, under optimum conditions

Simulation: Early Detection of Brain Vessels Stroke by Applying Electromagnetic Waves Non-Invasively

Introduction: Early recognition of stroke with its two types Ischemic and Hemorrhagic, is one of the most crucial research points, commonly used methods are CT- (computerized tomography), and MRI- (Magnetic resonance imaging). These techniques cause a delay in the detection of the condition, which causes permanent disability. The main reason behind the fatal consequences of stroke is the delay of detection. Therefore, this research paper aims to early detection of the type of stroke without delay until the appropriate diagnosis of each type is made, and then the appropriate treatment without delay. Method: Using a non-invasive and fast technique to determine the stroke type by wave, we simulate and design a vessel containing a liquid as a laminar flow with the same density and velocity of blood, and it was surrounded by a Homogenized multi-turn coil consisting of (n) turns to represent the magnetic field, using specific frequency (HZ) with Electrical field in coil current (A) to see the changing in magnetic flux density (MFD), Depending on the changes in MFD, the flow of blood in laminar flow can be affected by clotting (Ischemic) or Hemorrhagic (cutting) in our vessel designed. We have built three different scenarios to apply the technique which are: First: Normal Scenario (where the blood in vessel has no problem), second: clotting (ischemic, where the vessel blocked in specific three position) and Third: Cutting (Hemorrhagic, where the vessel cut in certain nine positions). Results: This paper presents-through our own design-the studying of applying the electromagnetic waves on blood inside the vessel to detect the stroke type in our three scenarios (normal, ischemic three positions or hemorrhagic nine positions), Studying the magnetic field and laminar flow. This study covered in three areas. First: coil geometry analysis, Second: stationary, and Third: frequency domain. through the changes in Magnetic Flux Density -MFD- waves. The results were promising and distinct for distinguishing between the three scenarios which are normal, ischemic (3 positions) and hemorrhagic (9 positions) the results of MFD are: 0.09 to 3.3*10^-3, 0.08 to 3.15*10^-4, 0.15 to 6.2*10^-3 respectively

The impact of information/decision support systems (I/DSS) in debt management: The Egyptian experience.

The introduction of technology-based tools into developing countries is usually impeded by a number of potential problems. The problems become murkier when the context of the issue, such as external debt management, where technology is to be used, does not have a standard textbook of rules and procedures to follow. The principle problems encountered during the course of our research for this thesis hinged the issue of possible inapplicability of information technology (IT), especially information and decision support systems (I/DSS). This is also the case for other tools and techniques built and used by developed countries in the developing world. The great deal of difference in the context of use; content of the systems; and the attitudes of the parties involved in the process forms the basis for our argument. The research was intended to draw on the lessons learned by the Cabinet Information and Decision Support Center (IDSC) during the introduction I/DSS for improving the decision making process in developing countries. Moreover, the research intended to tackle the issue of using I/DSS in a totally new context, other than for very structured purposes such as manufacturing and the like, and a challenging environment such as those in developing countries where difficulties of implementation and use are more in context, content and cultural issues than technological ones. The research would then show implications, identify problems and challenges and try to develop generalizations and recommendations. The research, since its initial phases, had to consider the forces of centralization versus decentralization and the organizational structure of decision making especially at the top level. Other issues such as I/DSS project planning and implementation; organizational dynamics and the research at the organizational level related to national policy; and technical information systems development were taken into consideration. This experience could be viewed as a documentation phase where the Egyptian Cabinet IDSC and the Central Bank of Egypt (CBE) together hove built, implemented and sustained state-of-the-art I/DSS in the process of establishing a powerful Egyptian debt management office. The analysis of these experiences displays many lessons for the implementation of sophisticated systems under conditions of extreme difficulty. It offers insight into a number of problems that concern designers, implementors, users, and researchers in I/DSS use in managing development planning and socio-economic change especially in developing countries. It also delves into important aspects related to project planning and implementation, organizational dynamics and the effective use of accurate, timely and relevant information in development planning. We conclude this thesis with on analytical framework that gives detailed methods and guidelines for future implementation of similar programs in developing countries that might wish to benefit from the experience of the Cabinet of Egypt IDSC

The impact of national culture on knowledge management processes in the context of an economic transition to a knowledge-based economy: the case of Qatar.

In recent years, a global movement towards a knowledge-based economy through the provision of adequate and appropriate infrastructure and development programs for leveraging the potential of human capital has started. There has been tremendous cognitive development in the developed countries including the Arab countries, but in the developing countries, things are still slowly progressing towards a level of knowledge capture, creation, and dissemination that will allow them to rise to the level of developed countries. The central administration derived from the authoritative patriarchal structure of the Arab society significantly affects the motives of their qualified workforce in their quest to utilize their knowledge for the desired development towards a knowledge society. The research investigates the influence of national culture on the progress of knowledge economy four pillars ranked by World Bank Knowledge Economy index, education, information and communication technology infrastructure, innovation, and adequacy of supporting institutional framework. The research adopts Nonaka & Takeuchi's SECI knowledge management model to study the impact of national culture upon knowledge acquisition, transfer, creation, and dissemination as facilitated through Nonaka and Takeuchi's (1995) cyclical knowledge management model by People, Process, and Technology. Furthermore, the research introduces a conceptual framework and detailed analytical framework illustrating relationships and mapping governing principles to the research aim and objective. The research keeps a specific focus on the influence of Qatar's national culture on the knowledge management process prevalent in the country as the research aims to explore elements that may affect Qatar's transition towards a knowledge-based economy. The research surveys a sample of Qatari nationals community to profiles Qatar National culture, interviews a sample of Qatari professionals to investigate the impact of cultural dimensions on knowledge management process, and validate the concluded analysis by identifying Qatar's ranking in relevant Knowledge Economy global indexes. The research assert the possible influence of national culture on knowledge conversion processes as critical to the success of country's economic transition to knowledge economy