Proapoptotic activity of Ukrain is based on Chelidonium majus L. alkaloids and mediated via a mitochondrial death pathway

BACKGROUND: The anticancer drug Ukrain (NSC-631570) which has been specified by the manufacturer as semisynthetic derivative of the Chelidonium majus L. alkaloid chelidonine and the alkylans thiotepa was reported to exert selective cytotoxic effects on human tumour cell lines in vitro. Few clinical trials suggest beneficial effects in the treatment of human cancer. Aim of the present study was to elucidate the importance of apoptosis induction for the antineoplastic activity of Ukrain, to define the molecular mechanism of its cytotoxic effects and to identify its active constituents by mass spectrometry. METHODS: Apoptosis induction was analysed in a Jurkat T-lymphoma cell model by fluorescence microscopy (chromatin condensation and nuclear fragmentation), flow cytometry (cellular shrinkage, depolarisation of the mitochondrial membrane potential, caspase-activation) and Western blot analysis (caspase-activation). Composition of Ukrain was analysed by mass spectrometry and LC-MS coupling. RESULTS: Ukrain turned out to be a potent inducer of apoptosis. Mechanistic analyses revealed that Ukrain induced depolarisation of the mitochondrial membrane potential and activation of caspases. Lack of caspase-8, expression of cFLIP-L and resistance to death receptor ligand-induced apoptosis failed to inhibit Ukrain-induced apoptosis while lack of FADD caused a delay but not abrogation of Ukrain-induced apoptosis pointing to a death receptor independent signalling pathway. In contrast, the broad spectrum caspase-inhibitor zVAD-fmk blocked Ukrain-induced cell death. Moreover, over-expression of Bcl-2 or Bcl-x(L )and expression of dominant negative caspase-9 partially reduced Ukrain-induced apoptosis pointing to Bcl-2 controlled mitochondrial signalling events. However, mass spectrometric analysis of Ukrain failed to detect the suggested trimeric chelidonine thiophosphortriamide or putative dimeric or monomeric chelidonine thiophosphortriamide intermediates from chemical synthesis. Instead, the Chelidonium majus L. alkaloids chelidonine, sanguinarine, chelerythrine, protopine and allocryptopine were identified as major components of Ukrain. Apart from sanguinarine and chelerythrine, chelidonine turned out to be a potent inducer of apoptosis triggering cell death at concentrations of 0.001 mM, while protopine and allocryptopine were less effective. Similar to Ukrain, apoptosis signalling of chelidonine involved Bcl-2 controlled mitochondrial alterations and caspase-activation. CONCLUSION: The potent proapoptotic effects of Ukrain are not due to the suggested "Ukrain-molecule" but to the cytotoxic efficacy of Chelidonium majus L. alkaloids including chelidonine

Effects of auditory distraction on voluntary movements: exploring the underlying mechanisms associated with parallel processing

Highly demanding cognitive-motor tasks can be negatively influenced by the presence of auditory stimuli. The human brain attempts to partially suppress the processing of potential distractors in order that motor tasks can be completed successfully. The present study sought to further understand the attentional neural systems that activate in response to potential distractors during the execution of movements. Nineteen participants (9 women and 10 men) were administered isometric ankle-dorsiflexion tasks for 10 s at a light intensity. Electroencephalography was used to assess the electrical activity in the brain, and a music excerpt was used to distract participants. Three conditions were administered: auditory distraction during the execution of movement (auditory distraction; AD), movement execution in the absence of auditory distraction (control; CO), and auditory distraction in the absence of movement (stimulus-only; SO). AD was compared with SO to identify the mechanisms underlying the attentional processing associated with attentional shifts from internal association (task-related) to external (task-unrelated) sensory cues. The results of the present study indicated that the EMG amplitude was not compromised when the auditory stimulus was administered. Accordingly, EEG activity was upregulated at 0.368 s in AD when compared to SO. Source reconstruction analysis indicated that right and central parietal regions of the cortex activated at 0.368 s in order to reduce the processing of task-irrelevant stimuli during the execution of movements. The brain mechanisms that underlie the control of potential distractors during exercise were possibly associated with the activity of the frontoparietal network.This research was supported, in part, by grants from the Coordination for the Improvement of Higher Education Personnel (CAPES)