The EPR spectra of spin labels in lipid bilayers are well known to contain proteins sometimes composed of two spectral components. The more restricted component is associated with boundary lipids where the spin labels surround the hydrophobic regions of proteins, whereas the more mobile component arises from the spin labels located in the bulk bilayer phase, away from the protein [13]. The fitting program provides the τ c and population of each component. Thus, the mean of the rotational correlation time was selleck products calculated as τ c = N 1 *τ c1 + N 2
*τ c2 , in which N 1 and N 2 are the fractions of the population in components 1 and 2, respectively, and τ c1 and τ c2 are the corresponding rotational time correlations. Figure 6 Experimental EPR spectra (black line) and theoretical fits (red line) of spin-label 5-DSA in this website Leishmania membrane. The experiment was conducted at 26°C for samples untreated and treated with parthenolide at the indicated concentrations. EPR spectra were simulated with the NLLS fitting program, and the values of the parameter rotational correlation time, τ C , obtained from the fit for each spectrum are indicated on a
nanosecond scale. The EPR parameter 2A//is the separation in magnetic-field units between the first and last resonance lines of the spectrum. The vertical lines indicate the 2A//for the control samples, and the smaller vertical lines illustrate the increase in 2A//for the sample treated with 9 × 109 molecules/cell. The measured 2A//values and τC values indicate that the presence of parthenolide selleck significantly reduced lipid fluidity. The estimated
experimental errors for the 2A//and τC parameters are 0.5 G and 1.0 ns, respectively. Discussion For many years, parasites of the genus Leishmania have displayed extraordinary plasticity to face modifications in their environment [14]. The expansion Celastrol of risk factors related to environmental changes and man-made transformations are making leishmaniasis a growing public health concern in many countries worldwide [15]. Leishmaniasis urgently needs novel drugs with improved features, and many compounds primarily derived from plants are promising leads for the development of novel chemotherapeutics [16]. The development of axenic cultures of amastigotes of Leishmania species yielded new opportunities to investigate the antileishmanial activities of new compounds directly at the mammalian stage of the parasite [17]. Assays that use intracellular amastigote cell cultures are relevant because this life cycle stage of the parasite is important to its pathogenicity, and data obtained exclusively from promastigote cell lines are insufficient [16]. Therefore, in the present study, we determined the leishmanicidal activity of parthenolide, which is naturally occurring, in both axenic and intracellular amastigotes.