Entrepreneurship as a potential driving force for the further development of rural areas

Entrepreneurial human capital and entrepreneurship are crucial elements for the further develop - ment of rural areas. In order to understand the role of rural enterprises it is essential to learn the characte - ristics of successful companies in local economies. The main aim of this paper is to demonstrate through the introduction of the activity, everyday practice and production procedure how non-agricultural and non-tourism enterprises can foster the development of rural and peripheral areas in the Visegrad countries. Nine local enter - prises of different size, profile and development pathway were examined in order to highlight main experiences and lessons being vital for and adaptable in other disadvantaged areas. The results and company descriptions provided in the article are based on in-depth interviews, online data collection and statistical analys

Neutral Red Assay Modification to PreventCytotoxicity and Improve Repeatability Using E-63 Rat Skeletal Muscle Cells

Cellular uptake of neutral red dye (NR) is currently used as an indirect measure of viable cells in cultures. We used E-63 rat skeletal muscle cells to identify causes of NR assay variability and to develop modifications that substantially reduce it. Three methods of NR preparation and/or addition to cells were used. When NR medium was prepared, incubated overnight, and filtered to remove precipitates, the amount of dye precipitated varied greatly. Coefficients of variation (CVs) in NR uptake were greater than 25% between assays. Higher NR concentrations, longer incubation times, increased pH, and decreased temperature promoted NR precipitation in media. NR media prepared and filtered just prior to use or direct addition of prefiltered NR stock solution to cell cultures resulted in much smaller CVs between assays. NR was cytotoxic to E-63 rat muscle and primary quail myoblasts in a time-and concentration-dependent manner. NR exposure to E-63 cells for greater than 1.25 and 2 hr at 157 or 127 μg/ml, respectively, was associated with swelling and rupture of lysosomes. By contrast, there was no evidence of cytotoxicity when E-63 cells were exposed to NR for 1 hr at either 127 or 157 μg/ml. Primary quail myoblasts developed lysosomal swelling and ruptured more rapidly than E-63 cells when exposed to NR at either 127 or 157 μg/ml. For confluent 10-day cultures of E-63 cells exposed to NR at 127 μg/ml for 1 hr, the CVs within assay and between assays were 3.3-3.9% and 5.1%, respectively. For similarly exposed, actively replicating 3-day cultures of E-63 cells, the CVs within and between assays were 6.2-9.6% and 2.4%, respectively. NR uptake by the E-63 cells was linear with respect to viable cell number