The Development of Novel Vorapaxar Analogues as Topical Protease Activated Receptor-1 Antagonists for the Treatment of Idiopathic Pulmonary Fibrosis

The overall aim of this project was to design and synthesise novel Protease Activated Receptor-1 (PAR-1) inhibitors in order to develop a novel treatment for Idiopathic Pulmonary Fibrosis (IPF). After finding the commercially available, allosteric PAR-1 inhibitor, “Q94” inactive and insoluble, focus switched to making novel vorapaxar analogues. A newly published synthetic route towards himbacine was successfully adapted to give vorapaxar analogues with a C-9-keto group. These were shown to have moderate potency against PAR-1 in a FLIPR calcium flux assay. After further synthetic route optimisation and investigation another novel vorapaxar analogue was identified. Compound (-)-94 was shown to be the most potent of the series of compounds tested, achieving an IC50 of 27 nM. In the same assay vorapaxar gave an IC50 of 13 nM. This makes (-)-94 a novel, potent, PAR-1 inhibitor worthy of further investigation and development as a potential IPF drug

Emerging technologies in physics education

Three emerging technologies in physics education are evaluated from the interdisciplinary perspective of cognitive science and physics education research. The technologies - Physlet Physics, the Andes Intelligent Tutoring System (ITS), and Microcomputer-Based Laboratory (MBL) Tools - are assessed particularly in terms of their potential at promoting conceptual change, developing expert-like problem-solving skills, and achieving the goals of the traditional physics laboratory. Pedagogical methods to maximize the potential of each educational technology are suggested.Comment: Accepted for publication in the Journal of Science Education and Technology; 20 page

Using a disciplinary discourse lens to explore how representations afford meaning making in a typical wave physics course

We carried out a case study in a wave physics course at a Swedish university in order to investigate the relations between the representations used in the lessons and the experience of meaning making in interview–discussions. The grounding of these interview–discussions also included obtaining a rich description of the lesson environment in terms of the communicative approaches used and the students’ preferences for modes of representations that best enable meaning making. The background for this grounding was the first two lessons of a 5-week course on wave physics (70 students). The data collection for both the grounding and the principal research questions consisted of video recordings from the first two lessons: a student questionnaire of student preferences for representations (given before and after the course) and video-recorded interview–discussions with students (seven pairs and one on their own). The results characterize the use of communicative approaches, what modes of representation were used in the lectures, and the trend in what representations students’ preferred for meaning making, all in order to illustrate how students engage with these representations with respect to their experienced meaning making. Interesting aspects that emerged from the study are discussed in terms of how representations do not, in themselves, necessarily enable a range of meaning making; that meaning making from representations is critically related to how the representations get situated in the learning environment; and how constellations of modes of disciplinary discourse may be necessary but not always sufficient. Finally, pedagogical comments and further research possibilities are presented.Web of Scienc

Low-frequency cortical activity is a neuromodulatory target that tracks recovery after stroke.

Recent work has highlighted the importance of transient low-frequency oscillatory (LFO; <4 Hz) activity in the healthy primary motor cortex during skilled upper-limb tasks. These brief bouts of oscillatory activity may establish the timing or sequencing of motor actions. Here, we show that LFOs track motor recovery post-stroke and can be a physiological target for neuromodulation. In rodents, we found that reach-related LFOs, as measured in both the local field potential and the related spiking activity, were diminished after stroke and that spontaneous recovery was closely correlated with their restoration in the perilesional cortex. Sensorimotor LFOs were also diminished in a human subject with chronic disability after stroke in contrast to two non-stroke subjects who demonstrated robust LFOs. Therapeutic delivery of electrical stimulation time-locked to the expected onset of LFOs was found to significantly improve skilled reaching in stroke animals. Together, our results suggest that restoration or modulation of cortical oscillatory dynamics is important for the recovery of upper-limb function and that they may serve as a novel target for clinical neuromodulation

A two-domain elevator mechanism for sodium/proton antiport

Sodium/proton (Na+/H+) antiporters, located at the plasma membrane in every cell, are vital for cell homeostasis1. In humans, their dysfunction has been linked to diseases, such as hypertension, heart failure and epilepsy, and they are well-established drug targets2. The best understood model system for Na+/H+ antiport is NhaA from Escherichia coli1, 3, for which both electron microscopy and crystal structures are available4, 5, 6. NhaA is made up of two distinct domains: a core domain and a dimerization domain. In the NhaA crystal structure a cavity is located between the two domains, providing access to the ion-binding site from the inward-facing surface of the protein1, 4. Like many Na+/H+ antiporters, the activity of NhaA is regulated by pH, only becoming active above pH 6.5, at which point a conformational change is thought to occur7. The only reported NhaA crystal structure so far is of the low pH inactivated form4. Here we describe the active-state structure of a Na+/H+ antiporter, NapA from Thermus thermophilus, at 3 Å resolution, solved from crystals grown at pH 7.8. In the NapA structure, the core and dimerization domains are in different positions to those seen in NhaA, and a negatively charged cavity has now opened to the outside. The extracellular cavity allows access to a strictly conserved aspartate residue thought to coordinate ion binding1, 8, 9 directly, a role supported here by molecular dynamics simulations. To alternate access to this ion-binding site, however, requires a surprisingly large rotation of the core domain, some 20° against the dimerization interface. We conclude that despite their fast transport rates of up to 1,500 ions per second3, Na+/H+ antiporters operate by a two-domain rocking bundle model, revealing themes relevant to secondary-active transporters in general

A new framework for designing programmes of assessment

Research on assessment in medical education has strongly focused on individual measurement instruments and their psychometric quality. Without detracting from the value of this research, such an approach is not sufficient to high quality assessment of competence as a whole. A programmatic approach is advocated which presupposes criteria for designing comprehensive assessment programmes and for assuring their quality. The paucity of research with relevance to programmatic assessment, and especially its development, prompted us to embark on a research project to develop design principles for programmes of assessment. We conducted focus group interviews to explore the experiences and views of nine assessment experts concerning good practices and new ideas about theoretical and practical issues in programmes of assessment. The discussion was analysed, mapping all aspects relevant for design onto a framework, which was iteratively adjusted to fit the data until saturation was reached. The overarching framework for designing programmes of assessment consists of six assessment programme dimensions: Goals, Programme in Action, Support, Documenting, Improving and Accounting. The model described in this paper can help to frame programmes of assessment; it not only provides a common language, but also a comprehensive picture of the dimensions to be covered when formulating design principles. It helps identifying areas concerning assessment in which ample research and development has been done. But, more importantly, it also helps to detect underserved areas. A guiding principle in design of assessment programmes is fitness for purpose. High quality assessment can only be defined in terms of its goals

Characterization of long and stable de novo single alpha-helix domains provides novel insight into their stability

Naturally-occurring single α-helices (SAHs), are rich in Arg (R), Glu (E) and Lys (K) residues, and stabilized by multiple salt bridges. Understanding how salt bridges promote their stability is challenging as SAHs are long and their sequences highly variable. Thus, we designed and tested simple de novo 98-residue polypeptides containing 7-residue repeats (AEEEXXX, where X is K or R) expected to promote salt-bridge formation between Glu and Lys/Arg. Lys-rich sequences (EK3 (AEEEKKK) and EK2R1 (AEEEKRK)) both form SAHs, of which EK2R1 is more helical and thermo-stable suggesting Arg increases stability. Substituting Lys with Arg (or vice versa) in the naturally-occurring myosin-6 SAH similarly increased (or decreased) its stability. However, Arg-rich de novo sequences (ER3 (AEEERRR) and EK1R2 (AEEEKRR)) aggregated. Combining a PDB analysis with molecular modelling provides a rational explanation, demonstrating that Glu and Arg form salt bridges more commonly, utilize a wider range of rotamer conformations, and are more dynamic than Glu–Lys. This promiscuous nature of Arg helps explain the increased propensity of de novo Arg-rich SAHs to aggregate. Importantly, the specific K:R ratio is likely to be important in determining helical stability in de-novo and naturally-occurring polypeptides, giving new insight into how single α-helices are stabilized

Downregulation of MIP-1α/CCL3 with praziquantel treatment in Schistosoma haematobium and HIV-1 co-infected individuals in a rural community in Zimbabwe

The results of our study show that the MIP-1alpha/CCL3 levels were positively associated with S. haematobium egg counts at baseline but not with HIV-1 infection status. MIP-1alpha/CCL3 levels were significantly reduced at three months post treatment with praziquantel. We therefore conclude that MIP-1alpha/CCL3 is produced during infection with S haematobium. S. haematobium infection is associated with increased MIP-1alpha/CCL3 levels in an egg intensity-dependent manner and treatment of S. haematobium is associated with a reduction in MIP-1alpha/CCL3

Rapamycin Pharmacokinetic and Pharmacodynamic Relationships in Osteosarcoma: A Comparative Oncology Study in Dogs

Signaling through the mTOR pathway contributes to growth, progression and chemoresistance of several cancers. Accordingly, inhibitors have been developed as potentially valuable therapeutics. Their optimal development requires consideration of dose, regimen, biomarkers and a rationale for their use in combination with other agents. Using the infrastructure of the Comparative Oncology Trials Consortium many of these complex questions were asked within a relevant population of dogs with osteosarcoma to inform the development of mTOR inhibitors for future use in pediatric osteosarcoma patients.This prospective dose escalation study of a parenteral formulation of rapamycin sought to define a safe, pharmacokinetically relevant, and pharmacodynamically active dose of rapamycin in dogs with appendicular osteosarcoma. Dogs entered into dose cohorts consisting of 3 dogs/cohort. Dogs underwent a pre-treatment tumor biopsy and collection of baseline PBMC. Dogs received a single intramuscular dose of rapamycin and underwent 48-hour whole blood pharmacokinetic sampling. Additionally, daily intramuscular doses of rapamycin were administered for 7 days with blood rapamycin trough levels collected on Day 8, 9 and 15. At Day 8 post-treatment collection of tumor and PBMC were obtained. No maximally tolerated dose of rapamycin was attained through escalation to the maximal planned dose of 0.08 mg/kg (2.5 mg/30 kg dog). Pharmacokinetic analysis revealed a dose-dependent exposure. In all cohorts modulation of the mTOR pathway in tumor and PBMC (pS6RP/S6RP) was demonstrated. No change in pAKT/AKT was seen in tumor samples following rapamycin therapy.Rapamycin may be safely administered to dogs and can yield therapeutic exposures. Modulation pS6RP/S6RP in tumor tissue and PBMCs was not dependent on dose. Results from this study confirm that the dog may be included in the translational development of rapamycin and potentially other mTOR inhibitors. Ongoing studies of rapamycin in dogs will define optimal schedules for their use in cancer and evaluate the role of rapamycin use in the setting of minimal residual disease