The embryonic cardiomyocyte cell line H9C2 is commonly used in numerous in vitro studies, including cardiotoxicity analyses of new drugs. So far no results were published for studies on cell parameters variability during the cell line ageing process. For this reason the aim of the study was to evaluate the effect of a number of H9C2 rat embryonic cardiomyocytes passages on repeatability of study results for selected cytotoxicity parameters, with doxorubicin as a model toxic agent. The cultures were passaged twenty-five times. Cells from passage 1, 5, 10, 15, 20 and 25 were treated with doxorubicin for 24 h. Then drug cytotoxicity was evaluated with the MTT test and additionally the nuclear factor erythroid 2–related factor (Nrf2) gene expression was examined. The analysis of oxidative stress intensity and cell morphology was also assessed. The microscopic appearance of cells indicates that untreated cardiomyocytes morphology changes as well as sensitivity to toxic effects increases with the number of passages. Also an increase in oxidative stress in cells occurs with further passaging of cardiomyocytes. Statistical significance of differences in conducted tests results depended on doxorubicin concentration but in many cases the H9C2 line was found to be a reliable in vitro model only for the first five passages. For this reason it is important to take into consideration that further culturing of cardiomyocytes may not ensure repeatability of study results due to the culture ageing.

No abstract

Acta Technologica Agriculturae 1/2016Dušan Páleš et al.The most effective way for determination of curves for practical use is to use a set of control points. These control points can be accompanied by other restriction for the curve, for example boundary conditions or conditions for curve continuity (Sederberg, 2012). When a smooth curve runs only through some control points, we refer to curve approximation. The B-spline curve is one of such approximation curves and is addressed in this contribution. A special case of the B-spline curve is the Bézier curve Rédl et al., 2014). The B-spline curve is applied to a set of control points in a space, which were obtained by measurement of real vehicle movement on a slope (Rédl, 2007(Rédl, , 2008. Data were processed into the resulting trajectory (Rédl, 2012;Rédl and Kučera, 2008). Except for this, the movement of the vehicle was simulated using motion equations (Rédl, 2003;Rédl and Kročko, 2007). B-spline basis functionsBézier basis functions known as Bernstein polynomials are used in a formula as a weighting function for parametric representation of the curve (Shene, 2014). B-spline basis functions are applied similarly, although they are more complicated. They have two different properties in comparison with Bézier basis functions and these are: 1) solitary curve is divided by knots, 2) basis functions are not nonzero on the whole area. Every B-spline basis function is nonzero only on several neighbouring subintervals and thereby it is changed only locally, so the change of one control point influences only the near region around it and not the whole curve.These numbers are called knots, the set U is called the knot vector, and the half-opened interval 〈u i , u i + 1 ) is the i-th knot span. Seeing that knots u i may be equal, some knot spans may not exist, thus they are zero. If the knot u i appears p times, hence u i = u i + 1 = ... = u i + p -1 , where p >1, u i is a multiple knot of multiplicity p, written as u i (p). If u i is only a solitary knot, it is also called a simple knot. If the knots are equally spaced, i.e. (u i + 1 -u i ) = constant, for every 0 ≤ i ≤ (m -1), the knot vector or knot sequence is said uniform, otherwise it is non-uniform.Knots can be considered as division points that subdivide the interval 〈u 0 , u m 〉 into knot spans. All B-spline basis functions are supposed to have their domain on 〈u 0 , u m 〉. We will use u 0 = 0 and u m = 1.To define B-spline basis functions, we need one more parameter k, which gives the degree of these basis functions. Recursive formula is defined as follows:This definition is usually referred to as the Cox-de Boor recursion formula. If the degree is zero, i.e. k = 0, these basis functions are all step functions that follows from Eq. (1). N i, 0 (u) = 1 is only in the i-th knot span 〈u i , u i + 1 ). For example, if we have four knots u 0 = 0, u 1 = 1, u 2 = 2 and u 3 = 3, knot spans 0, 1 and 2 are 〈0, 1), 〈1, 2) and 〈2, 3), and the basis functions of degree 0 are N 0, 0 (u) = 1 on interval 〈0, 1) Acta In this co...