Enhanced Differentiation of Stromal Cells and Embryonic Stem Cells with Vitamin D3

In-vitro mouse co-culture of E14 embryonic stem cells (ESCs) and OP9 stromal cells can recapitulate the earliest stages of haematopoietic development, not accessible in human embryos, supporting both haemogenic precursors and their primitive haematopoietic progeny. 1α, 25-Dihydroxy-vitamin D3 (VD3) has been demonstrated to be a powerful differentiation inducer for a wide variety of neoplastic cells, and could enhance early differentiation of ESCs into blood cells in E14/OP9 co-culture. This study aims to ascertain whether VD3 is key in promoting differentiation and suppressing proliferation, by separately investigating the effects of VD3 on the proliferation phase of the E14 cell line and on stromal OP9 cells.The results showed that VD3 inhibited the proliferation of the cells in a dose-dependent manner, quantitatively by decreased cell number, and qualitatively by alkaline-phosphatase staining that revealed significant differences between VD3-treated and untreated cells, characterised by decreased enzyme expression (colourless cells). Propidium-iodide cell-cycle analyses showed no significant percentage change in VD3-treated E14 and OP9 cells within their G and S-phases, compared to the untreated controls, despite the increased percentage of G-phase compared to the S-phase in a dosedependent manner. These results with E14 and OP9 cells indicate that adequate VD3 concentration enhances cellular differentiation and inhibits proliferation. The results also suggest that if E14 and OP9 cells were co-cultured andVD3-treated, there would be furtherenhanced differentiation of ESCs into blood cells.

The First Prevalence Report of Direct Identification and Differentiation of B. abortus and B. melitensis using Real Time PCR in House Mouse of Iran

Brucellosis is a zoonotic disease; its symptoms and appearances are not exclusive in human and its traditional diagnosis is based on culture, serological methods and conventional PCR. For more sensitive, specific detection and differentiation of Brucella spp., the real time PCR method is recommended. This research has performed to determine the presence and prevalence of Brucella spp. and differentiation of Brucella abortus and Brucella melitensis in house mouse (Mus musculus) in west of Iran. A TaqMan analysis and single-step PCR was carried out in total 326 DNA of Mouse's spleen samples. From the total number of 326 samples, 128 (39.27%) gave positive results for Brucella spp. by conventional PCR, also 65 and 32 out of the 128 specimens were positive for B. melitensis, B. abortus, respectively. These results indicate a high presence of this pathogen in this area and that real time PCR is considerably faster than current standard methods for identification and differentiation of Brucella species. To our knowledge, this study is the first prevalence report of direct identification and differentiation of B. abortus and B. melitensis by real time PCR in mouse tissue samples in Iran.

Methodology Issues and Design Approach of VLE on Mathematical Concepts Acquisition within Secondary Education in England

This study used positivist quantitative approach to examine the mathematical concepts acquisition of- KS4 (14-16) Special Education Needs (SENs) students within the school sector education in England. The research is based on a pilot study and the design is completely holistic in its approach with mixing methodologies. The study combines the qualitative and quantitative methods of approach in gathering formative data for the design process. Although, the approach could best be described as a mix method, fundamentally with a strong positivist paradigm, hence my earlier understanding of the differentiation of the students, student – teacher body and the various elements of indicators that is being measured which will require an attenuated description of individual research subjects. The design process involves four phases with five key stages which are; literature review and document analysis, the survey, interview, and observation; then finally the analysis of data set. The research identified the need for triangulation with Reid-s phases of data management providing scaffold for the study. The study clearly identified the ideological and philosophical aspects of educational research design for the study of mathematics by the special education needs (SENs) students in England using the virtual learning environment (VLE) platform.

A Model to Study the Effect of Excess Buffers and Na+ Ions on Ca2+ Diffusion in Neuron Cell

Calcium is a vital second messenger used in signal transduction. Calcium controls secretion, cell movement, muscular contraction, cell differentiation, ciliary beating and so on. Two theories have been used to simplify the system of reaction-diffusion equations of calcium into a single equation. One is excess buffer approximation (EBA) which assumes that mobile buffer is present in excess and cannot be saturated. The other is rapid buffer approximation (RBA), which assumes that calcium binding to buffer is rapid compared to calcium diffusion rate. In the present work, attempt has been made to develop a model for calcium diffusion under excess buffer approximation in neuron cells. This model incorporates the effect of [Na+] influx on [Ca2+] diffusion,variable calcium and sodium sources, sodium-calcium exchange protein, Sarcolemmal Calcium ATPase pump, sodium and calcium channels. The proposed mathematical model leads to a system of partial differential equations which have been solved numerically using Forward Time Centered Space (FTCS) approach. The numerical results have been used to study the relationships among different types of parameters such as buffer concentration, association rate, calcium permeability.

A Novel Nucleus-Based Classifier for Discrimination of Osteoclasts and Mesenchymal Precursor Cells in Mouse Bone Marrow Cultures

Bone remodeling occurs by the balanced action of bone resorbing osteoclasts (OC) and bone-building osteoblasts. Increased bone resorption by excessive OC activity contributes to malignant and non-malignant diseases including osteoporosis. To study OC differentiation and function, OC formed in in vitro cultures are currently counted manually, a tedious procedure which is prone to inter-observer differences. Aiming for an automated OC-quantification system, classification of OC and precursor cells was done on fluorescence microscope images based on the distinct appearance of fluorescent nuclei. Following ellipse fitting to nuclei, a combination of eight features enabled clustering of OC and precursor cell nuclei. After evaluating different machine-learning techniques, LOGREG achieved 74% correctly classified OC and precursor cell nuclei, outperforming human experts (best expert: 55%). In combination with the automated detection of total cell areas, this system allows to measure various cell parameters and most importantly to quantify proteins involved in osteoclastogenesis.

Improving the Flexibility of Employment in Polish Economic Practice

Modern organizations operate under the pressure of dynamic and often unpredictable changes, both in external and internal environment. Market success, in this context, requires a particular competence in the form of flexibility, interpreted here both on the level of individuals and on the level of organization. This paper addresses the changes taking place in the sphere of employment, as observed in economic entities operating on Polish market. Based on own empirical studies, the authors focus on the progressing trend of ‘flexibilization’ of employment, particularly in the context of transformations in organizational structure, designed to facilitate the transition into management by projects and differentiation of labor forms.