Mitochondrial genome acquisition restores respiratory function and tumorigenic potential of cancer cells without mitochondrial DNA

We report that tumor cells devoid of their mitochondrial genome (mtDNA) show delayed tumor growth and that tumor formation is associated with acquisition of mtDNA from host cells. This leads to partial recovery of mitochondrial function in cells derived from primary tumors grown from cells without mtDNA and a shorter lag in tumor growth. Cell lines from circulating tumor cells showed further recovery of mitochondrial respiration and an intermediate lag to tumor growth, while cells from lung metastases exhibited full restoration of respiratory function and no lag in tumor growth. Stepwise assembly of mitochondrial respiratory supercomplexes was correlated with acquisition of respiratory function. Our findings indicate horizontal transfer of mtDNA from host cells in the tumor microenvironment to tumor cells with compromised respiratory function to re-establish respiration and tumor-initiating efficacy. These results suggest a novel pathophysiological process for overcoming mtDNA damage and support the notion of high plasticity of malignant cells.

Mitochondrial membrane potential in a small subset of artemisinin-induced dormant plasmodium falciparum parasites in vitro

Artemisinin induced dormancy is a proposed mechanism for failures of mono-therapy and is linked with artemisinin resistance in Plasmodium falciparum. The biological characterization and dynamics of dormant parasites are not well understood. Here we report that following dihydroartemisinin (DHA) treatment in vitro, a small subset of morphologically dormant parasites was stained with rhodamine 123 (RH), a mitochondrial membrane potential (MMP) marker, and persisted to recovery. FACS sorted RH-positive parasites resumed growth at 10,000/well while RH-negative parasites failed to recover at 5 million/well. Furthermore, transcriptional activity for mitochondrial enzymes was only detected in RH-positive dormant parasites. Importantly, after treating dormant parasites with different concentrations of atovaquone, a mitochondrial inhibitor, the recovery of dormant parasites was delayed or stopped. This demonstrates that mitochondrial activity is critical for survival and regrowth of dormant parasites and that RH staining provides a means of identifying these parasites. These findings provide novel paths for studying and eradicating this dormant stage.

Disrupted sensory gating in pathological gambling

Background Some neurochemical evidence as well as recent studies on molecular genetics suggest that pathologic gambling may be related to dysregulated dopamine neurotransmission. Methods The current study examined sensory (motor) gating in pathologic gamblers as a putative measure of endogenous brain dopamine activity with prepulse inhibition of the acoustic startle eye-blink response and the auditory P300 event-related potential. Seventeen pathologic gamblers and 21 age- and gender-matched healthy control subjects were assessed. Both prepulse inhibition measures were recorded under passive listening and two-tone prepulse discrimination conditions. Results Compared to the control group, pathologic gamblers exhibited disrupted sensory (motor) gating on all measures of prepulse inhibition. Sensory motor gating deficits of eye-blink responses were most profound at 120-millisecond prepulse lead intervals in the passive listening task and at 240-millisecond prepulse lead intervals in the two-tone prepulse discrimination task. Sensory gating of P300 was also impaired in pathologic gamblers, particularly at 500-millisecond lead intervals, when performing the discrimination task on the prepulse. Conclusions In the context of preclinical studies on the disruptive effects of dopamine agonists on prepulse inhibition, our findings suggest increased endogenous brain dopamine activity in pathologic gambling in line with previous neurobiological findings.

Teaching and Assessing Reflective Writing in a Large Undergraduate Core Substantive Law Unit

The focus of higher education has shifted towards building students’ skills and self-awareness for future employment, in addition to developing substantive discipline knowledge. This means that there is an increasing need for embedding approaches to teaching and learning which provide a context for skills development and opportunities for students to prepare for the transition from legal education to professional practice. This chapter reports on a large (500-600 students) core undergraduate Equity law unit in an Australian University. ePortfolio has been embedded in Equity as a means of enabling students to document their reflections on their skill development in that unit. Students are taught, practice and are assessed on their teamwork and letter writing skills in the context of writing a letter of advice to a fictional client in response to a real world problem. Following submission of the team letter, students are asked to reflect on their skill development and document their reflections in ePortfolio. A scaffolded approach to teaching reflective writing is adopted using a blended model of delivery which combines face to face lectures and online resources, including an online module, facts sheets designed to guide students through the process of reflection by following the TARL model of reflection, and exemplars of reflective writing. Although students have engaged in the process of reflective writing in Equity for some years, in semester one 2011 assessment criteria were developed and the ePortfolio reflections were summatively assessed for the first time. The model of teaching and assessing reflective practice was evaluated in a range of ways by seeking feedback from students and academic staff responsible for implementing the model and asking them to reflect on their experiences. This chapter describes why skill development and reflective writing were embedded in the undergraduate law unit Equity; identify the teaching and learning approaches which were implemented to teach reflective writing to online and internal Equity students; explain the assessment processes; analyse the empirical evidence from evaluations; document the lessons learnt and discuss planned future improvements to the teaching and assessment strategies.

Tactics for teaching evidence-based practice : improving self-efficacy in finding and appraising evidence in a master's evidence-based practice unit

Universal application of evidence-based practice (EBP) is far from a reality with many clinicians feeling ill equipped to adopt this approach in their clinical practice (Melnyk Fineout- Overholt, Feinstein, Sadler, & Green-Hernandez, 2008; Sherriff, Wallis, & Chaboyer, 2007) and, thus, to be an intelligent consumer of evidence (Ciliska, 2005). While recognizing the benefit of EBP, many health professionals have low confidence in their skills for using evidence in clinical settings (Nagy, Lumby, McKinley, &Macfarlane, 2001). Educational initiatives are often recommended for promoting adoption of EBP with much of the focus being on providing knowledge of associated processes. Levin, Melnyk, Fineout-Overholt, Barnes, and Vetter (2011) demonstrated that providing knowledge of EBP process alone does not increase clinicians’ confidence in their ability to apply EBP to their practice...

The harnessing of peptide-monolith constructs for single step plasmid DNA purification

The availability of synthetic peptides has paved the way for their use in tailor-made interactions with biomolecules. In this study, a 16mer LacI-based peptide was used as an affinity ligand to examine the scale up feasibility for plasmid DNA purification. First, the peptide was designed and characterized for the affinity purification of lacO containing plasmid DNA, to be employed as a high affinity ligand for the potential capturing of plasmid DNA in a single unit operation. It was found there were no discernible interactions with a control plasmid that did not encode the lacO nucleotide sequence. The dissociation equilibrium constant of the binding between the 16mer peptide and target pUC19 was 5.0 ± 0.5 × 10-8 M as assessed by surface plasmon resonance. This selectivity and moderated affinity indicate that the 16mer is suitable for the adsorption and chromatographic purification of plasmid DNA. The suitability of this peptide was then evaluated using a chromatography system with the 16mer peptide immobilized to a customized monolith to purify plasmid DNA, obtaining preferential purification of supercoiled pUC19. The results demonstrate the applicability of peptide-monolith supports to scale up the purification process for plasmid DNA using designed ligands via a biomimetic approach.

Potential role of bioactive lipids in the development of non-antibody mediated transfusion-related acute lung injury (TRALI)

Transfusion-related acute lung injury (TRALI) has been the leading cause of transfusion-related morbidity and mortality in the UK and the USA in recent years. A threshold mechanism of TRALI has been proposed in which both patient factors (type and/or severity of clinical insult) and blood product factors (strength and/or concentration of antibodies or biological response modifiers) interact to surpass a threshold for TRALI development (Bux et al. Br J Haematol; 2007; 136: 788-99). The risk of developing antibody-mediated TRALI has been minimised by the introduction of risk-reduction strategies such as limiting the use of plasma from female donors. In contrast, there are no strategies currently in place to mitigate the development of non-antibody mediated TRALI as the mechanisms remain largely undefined. Previous studies have implicated non-polar lipids such as arachidonic acid and various species of hydroxyeicosatetranoic acid (HETE) in the development of non-antibody mediated TRALI (Silliman et al. Transfusion; 2011; 51: 2549-54), however the contribution of these lipids to the development of an inflammatory response in TRALI is poorly understood.

An improved robust field weakening algorithm for direct torque controlled synchronous reluctance motor drives

This paper presents an improved field weakening algorithm for synchronous reluctance motor (RSMs) drives. The proposed algorithm is robust to the variations in the machine d- and q-axes inductances. The transition between the maximum torque per ampere (MTPA), current and voltage limits as well as the maximum torque per flux (MTPF) trajectories is smooth. The proposed technique is combined with the direct torque control method to attain a high performance drive in the field weakening region. Simulation and experimental results are supplemented to verify the effectiveness of the proposed approach.