Competitiveness as a function of local and regional growth and development

Each economic entity, institution and individual has the responsibility of contributing to the economic development in its region. Creating conditions for the development and empowerment of the business sector are activities that in the long run lead to the strengthening of not only certain economic sectors, but the entire region. In the recent period sources from EU funds for co-financing of capital projects have become available to investors. Given the uncertainty in business conditions, investors’ poor capitalization, lack of business profitability and, in terms of profitability and risk, lack of high-quality capital projects, the benefits of these resources are insufficient and/or inadequately used. The aim of this paper is to analyse the strength and capabilities of Croatian companies for financing and implementation of high-quality capital projects. For this purpose, this paper will present the results of research of financial position of selected companies in 2012. Also, it presents the results of research from 2011 that examined the reality of projections of later activated investment projects. These results are the basis for a conclusion about the ability of management, in the analysed region, to make realistic plans and carry out high-quality capital projects

A new estimation method for continuous threshold expectile model

<p>The continuous threshold expectile regression model could capture the effect of a covariate on the response variable with two different straight lines, while intersecting an unknown threshold needed be estimated. This article proposes a new estimation method via a linearization technique to estimate the regression coefficients and the threshold simultaneously. Statistical inferences of the proposed estimators are easily derived from the existing theory. Moreover, the estimation procedure is readily implemented by the current software. Simulation studies and an application on GDP per capita and quality of electricity supply data illustrate the proposed method.</p

Down-regulation of PlGF in central marrow ECs diminishes their ability to support primitive hematopoietic cells.

<p>(A) Total RNA was obtained from central marrow ECs transduced with shRNA clones targeting PlGF. The mRNA expression level of PlGF was measured using quantitative RT-PCR and normalized to <i>hprt</i> levels (*p<0.05, ***p<0.001; n = 6 from 3 independent experiments; error bars represent standard deviation). (B) Uptake of DiI-Ac-LDL (red) in control-transduced and PlGF knockdown central marrow ECs. Nuclei were visualized with DAPI (Scale bar: 50 µm). (C) The formation of endothelial capillary like tube was evaluated under a phase-contrast microscope on Matrigel following 10 hours of culture (Scale bar: 200 μm). (D, E) LSK cells were seeded in serial dilutions on central marrow ECs, central marrow ECs transduced with E8, E12 or E8 and E12 lentiviral shRNA-PlGF vectors or shRNA-GFP vector and cultured at 33°C and 5% CO₂. CAFCs were scored on week 2 and 5 (*p<0.05, **p<0.01; n = 3 from 3 independent experiments; error bars represent standard deviation).</p

Transplanted central marrow ECs incorporate into the adult BM vasculature.

<p>(A) Central marrow ECs, PlGF knockdown central marrow ECs, mock transduced central marrow ECs or central marrow SCs were labeled with CFDA-SE and transplanted into Balb/c mice following 550cGy total body irradiation (1×10⁶ cells per mouse; intravenous injection on day 0 and intraperitoneal injection on day 1 to day 4). The percentage of labeled cells present in the BM and spleen on day 5 post irradiation was determined by flow cytometry. Representative flow plots (left) and percentage of labeled cells present in the BM and spleen (right) are shown (for central marrow ECs and SCs, n = 6 from 2 independent experiments; for PlGF knockdown central marrow ECs and mock transduced central marrow ECs, n = 2). (B) Lodgment of the central marrow ECs in the BM, classified as contiguous with the vascular endothelium (top image) or within the BM space (bottom image). The BM vessel is indicated by an asterix. Bone is outlined by the dashed line. The yellow arrows represent the transplanted central marrow EC labeled with CFDA-SE. Nuclei were visualized with DAPI (blue) present in Vectashield (scale bar: 50 μm; 40 sections per mouse, n = 6 from 2 independent experiments).</p

Effects of miR-204 on SOX4 expression by luciferase reporter assay.

<p>The potential miR-204 binding site at the 3′-UTR of SOX4 mRNA was computationally predicted by Targetscan. Cells were co-transfected with miR-204 mimics (or negative control) with pGL3-SOX4 (or pGL3-SOX4-mut) vector. Luciferase activity was normalized by the ratio of firefly and Renilla luciferase signals. * indicates P<0.05.</p

Reduced PlGF expression diminishes hematopoietic recovery <i>in vivo</i>.

<p>(A) BM MNCs from Balb/c mice following treatment with central marrow ECs, PlGF knockdown central marrow ECs, central marrow SCs or PBS were evaluated for LT-HSC, ST-HSC and HPC frequencies using flow cytometry (**p<0.01; n = 9 from 3 independent experiments; error bars represent standard deviation). (B) BM MNCs from the same mice were assessed for CAFC frequencies at weeks 2 and 5 (*p<0.05; n = 9 from 3 independent experiments; error bars represent standard deviation).</p

Decreased miR-204 in H. pylori-Associated Gastric Cancer Promotes Cancer Cell Proliferation and Invasion by Targeting SOX4

<div><p>Background</p><p>The molecular mechanism between Helicobacter pylori (H. pylori) infection and gastric cancer remained largely unknown. In this study, we determined the role of miRNA in H. pylori induced gastric cancer.</p><p>Methods and Results</p><p>We found that miR-204 was decreased in H. pylori positive tissues by qRT-PCR. Knockdown of miR-204 enhanced the invasion and proliferation ability of gastric cancer cells in vitro. Luciferase assay revealed that SOX4 was target gene of miR-204, which was found up-regulated in H. pylori positive tissues. Down-regulation of miR-204 and over-expression of SOX4 promoted epithelial-mesenchymal transition process.</p><p>Conclusion</p><p>Taken together, our findings demonstrated that miR-204 may act as a tumor suppressor in H. pylori induced gastric cancer by targeting SOX4.</p></div

Reduced PlGF expression diminishes vascular recovery <i>in vivo</i>.

<p>(A) Central marrow ECs, PlGF knockdown central marrow ECs, central marrow SCs or PBS were transplanted into Balb/c mice following 550cGy total body irradiation. On day 20 post irradiation, the tibias were harvested and assessed by immunohistochemistry for VE-cadherin expression (scale bar: 50 μm). (B) Representative images of normal and pathologic vessels (scale bar: 50 μm). (C) Quantitation of normal or pathologic vessels in the BM of the irradiated mice transplanted with the different cell types. Vessels were counted in three 200X fields per section. (*p<0.05, **p<0.01; 40 sections per mouse, n = 9 from 3 independent experiments). (D) BM MNCs from the treated mice were harvested and stained with the antibody to CD31, CD45 and Ter-119. The percentage of BM ECs was determined by CD31⁺CD45⁻Ter119⁻ cells using flow cytometric analysis (*p<0.05; n = 9 from 3 independent experiments; error bars represent standard deviation).</p

Central marrow ECs demonstrate enhance support of primitive hematopoietic cells.

<p>(A) BM MNCs were seeded in serial dilutions on central marrow EC, central marrow SC, endosteal marrow EC, endosteal marrow SC and spleen EC and cultured at 33°C and 5% CO₂. CAFCs were scored on day 14, 28, 35, 42 and 49. (B) BM LSK cells were seeded in serial dilutions on central marrow EC and central marrow SC and cultured at 33°C and 5% CO2. CAFCs were scored on day 14, 21, 28 and 35 (**p<0.01, *p<0.05 n = 4 from 3 independent experiments; error bars represent standard deviation). (C) 1000 LSK cells were co-cultured with the five supportive cell layers and the number of CD45⁺ cells on day 7 was assessed by flow cytometry (**P<0.01; n = 5 from 2 independent experiments; error bars represent standard deviation). (D) 100 LSK cells were co-cultured with the five supportive cell layers and the number of CFU-Cs was assessed on day 7 (*P<0.05, **P<0.01; n = 5 from 2 independent experiments; error bars represent standard deviation).</p

Characteristics of cells grown in endothelial or stromal cell culture conditions.

<p>(A, B) Total RNA was extracted from central marrow EC, endosteal marrow EC, spleen EC, central marrow SC and endosteal marrow SC. The mRNA expression level of Tie-2 and VE-Cadherin was measured using quantitative RT-PCR. The relative expression was normalized to Hypoxanthine-guanine phosphoribosyltransferase (HPRT) levels and calculated from standard curves. (*p<0.05, **p<0.01; n = 6 from 3 independent experiments; error bars represent standard deviation). (C) Cells cultured in endothelial or stromal culture conditions were stained with antibodies to CD31 and VE-Cadherin. Positive signals were visualized with FITC conjugated secondary antibody. Nuclei were visualized with 4,6-diamidino-2-phenylindole (DAPI). (D) Central marrow EC, endosteal marrow EC, spleen EC, central marrow SC and endosteal marrow SC were harvested by enzyme-free cell dissociation solution. The expression of CD31 and CD45 was analyzed by flow cytometry with PE conjugated anti-CD31 and APC-Cy7 conjugated anti-CD45. (E) Cells cultured in endothelial or stromal culture conditions were stained with alkaline phosphatase activity. Positive reaction was observed as dark blue violet in the cells.</p