Sarah Etherington

Poor numeracy/quantitative skills (QS) development is a widespread issue across Australian tertiary education. Lack of fundamental QS can impede students' progression in STEM degrees, and disadvantage individual students across other domains of life (e.g., financial literacy and active citizenship). Our ACDS-funded Diversity in Numbers (DiN) project seeks to evaluate a targeted, course-wide, just-in-time model for undergraduate development of QS. Digital numeracy modules will be designed to scaffold QS development through embedded interactive content and rich automated feedback. Each module targets a core QS concept (e.g., statistical testing, unit conversions, mathematical relationships) and is framed around a published article relevant to unit content, to expand student awareness of numbers as a tool across diverse fields of science. Given the ongoing under-representation of women, LGBTIQA+ people and other minorities in STEM, the selection of journal articles aims to increase students' appreciation of diversity from many different viewpoints, while developing their QS. At a broader level, the project aims to address the ongoing lack of diversity among STEM graduates and within the STEM workforce by enabling students to "see themselves" within published research. Here we will present the design of our research project to assess the success of our pilot DiN modules.

Background Pulsed Magnetic Fields (PMFs) are currently being used to treat a range of neurological conditions such as depression. However, little is known about the cellular mechanisms behind their therapeutic effects. Objective To investigate the influence of PMFs on neuronal biochemical processes. Method B50 rat neuroblastoma cells were seeded on to 6-well plates, grown to confluence and stimulated with PMFs (~10 mT) at 1 or 10 Hz for 10 minutes (controls were unstimulated). Cells were immediately quenched post-stimulation with ice-cold PBS and then freeze-dried. Cells were subsequently lysed, derivatised and analysed using untargeted gas chromatography-mass spectrometry (GC-MS). Key Findings Initial principal component analysis revealed 3 distinct groups (control, 1 and 10 Hz). Significant differences were found in 12 metabolites (PMF stimulated vs unstimulated controls, ANOVA, n= 4-6). PMF stimulated cells had significantly lower levels (p≤0.05) of GABA precursors (succinate, aspartate, proline and glutamate). These changes may be due to increased inhibitory neurotransmitter release during PMF stimulation. Two metabolites involved in calcium signaling (inositol and serine) were also significantly lower in PMF treated cells. In addition, PMF stimulation at 1Hz reduced metabolite levels to a greater extent than 10 Hz, an effect which reached significance for glycine (p≤0.05). Conclusion PMFs at either 1 or 10 Hz significantly reduced intracellular metabolites involved with inhibitory neurotransmission and calcium signaling in neuronal cell cultures. The changes to neuronal metabolism were frequency dependent. Our data suggest that acute PMF stimulation induce biochemical changes that may modulate inhibitory neurotransmission and calcium signaling.

Purpose: Complex regional pain syndrome (CRPS) is a neuropathic pain condition that can be acquired after minor trauma or surgery to soft tissues and nerves. Biopsies from affected tissue show an increased density of α1-adrenergic receptors (AR1) compared to controls. In order to further study the role of AR1 in CRPS, we wish to use an immortalized cortical neuron cell line, NIE115. The present study aimed to (i) characterise the expression of AR1 on NIE115 cells and (ii) document how the metabolic profile of NIE115 cells is affected by adrenergic pharmacological intervention. Methods: NIE115 cultures (n = 15) were dual-or triple-labelled with antibodies directed against AR1 as well as specific neuronal markers (neurofilament, TRPV1, TUJ1, CGRP). Cells were then exposed to an AR1 agonist (phenylephrine, n = 18), an antagonist (prazosin, n = 12) or a combination of both (n = 12) and the biochemical effects studied using gas chromatography-mass spectrometry-based metabolomics. Results: Immunohistochemistry confirmed the presence of AR1 on the NIE115 cells. Treatment of the cells with phenylephrine led to changes in both carbon and nitrogen metabolism consistent with stimulation of AR1. It was expected that prazosin would block the metabolic effects of phenylephrine, but a different set of changes to carbon and nitrogen metabolism were observed. This provides further evidence to the observations that prazosin may in fact be acting as an inverse agonist. Conclusion: These data indicate that metabolomics is a powerful technology in the study of receptor signaling, and have provided us with a new tool to investigate CRPS.

Introduction. Small Leucine Rich Proteins (SLRPs) are a family of 17 proteoglycans and glycoproteins found within the extracellular matrix (ECM). The roles of these proteins in osteoblast development have not been explored in human. Problem Statement. This study investigates the gene expression of six members of the SLRP family during osteogenesis in donor matched human subcutaneous adipose tissue-derived stem cells (ADSC) and bone marrow derived stem cells (BMSC). Procedures/Data/Observations. Global gene expression data was collected from two mouse osteoblast cell lines. Donor matched subcutaneous adipose tissue and bone marrow tissue was collected from 5 female patients (mean age 72.8 ± 6.9) during abdominal or orthopaedic surgery. ADSC and BMSC were isolated and differentiated using osteogenic media (OSM). mRNA was extracted at days 0, 7 and 28. Gene expression of SLRPs was analysed using qRT-PCR. Results. Six out of 17 SLRP family members were up-regulated in mouse osteoblasts thus they have been selected. In ADSC, Osteomodulin (OMD) was up-regulated by 26.6-fold at day 7 and 17.9-fold at day 28. In BMSC, higher OMD up-regulation was seen at day 7 (125.4-fold) and a smaller one at day 28 (47.5-fold). Epiphycan (EPYC) was also up-regulated at both time points in both cells (2.6-fold at day 7 and 3.3-fold at day 28 in ADSC; 1.6-fold at day 7 and 5.9-fold at day 28 in BMSC). On the contrary, Lumican (LUM) was down –regulated by 0.6-fold at day 7 and 0.2-fold at day 28 in ADSC; 0.6-fold at day 7 and 0.04-fold at day 28 in BMSC. Other SLRPS were not changed by osteogenic stimulation. Conclusions. Previously unrecognized roles of SLRP members were demonstrated in two human MSC models undergoing osteogenesis. OMD and EPYC are likely to be most involved. Further study on their protein expressions and locations in ECM niche is needed.

Internationalisation of the curriculum is central to the core values and strategic direction of many modern universities. Among science-based academics, a global perspective is often considered to be inevitable, given the multinational nature of modern scientific collaborations and the widespread international mobility of professional scientists. Unfortunately, these realities are often distant from the undergraduate science classroom, particularly for non-mobile students. More worryingly, explicit instruction in cross-cultural engagement and communication skills, essential for working effectively in a multinational team upon graduation, is commonly absent from tertiary science courses. This study addressed one major challenge to promoting cross-cultural competence among undergraduate science students: finding time to scaffold such learning within the context of content-heavy, time-poor units. Utilising the ‘Interaction for Learning Framework’ developed by Arkoudis et al. (2012), small changes aimed at enhancing global and cross-cultural awareness were incorporated into existing assessments and teaching activities within a second-year biomedical physiology unit. In student surveys, 40% of domestic and 60% international student respondents articulated specific learning about interaction in cross cultural groups resulting from unit activities. Many students also identified specific examples of how cultural beliefs would impact on the place of biomedical physiology within the global community. In addition, staff observed more widespread benefits for student engagement and learning. It is concluded that significant development of cross-cultural awareness and a more global perspective on scientific understanding can be supported among science undergraduates with relatively minor adaptations to course content.

Internationalisation of the curriculum is central to the core values and strategic direction of many modern universities. However, academic staff at the coalface in the ‘hard’ sciences are frequently resistant to implementing internationalization of the curriculum (Clifford, 2009). Science-based academics commonly consider development of a global perspective as inevitable, citing the intrinsic cultural neutrality of core scientific principles. A worrying consequence of this belief is that explicit instruction in intercultural engagement and communication skills is commonly absent from tertiary science courses. Given the multinational nature of modern scientific collaborations and the widespread international mobility of professional scientists, these are likely to be essential skills for effective science graduates. This study addressed one major challenge to promoting intercultural competence among undergraduate science students: finding time to scaffold such learning within the context of content-heavy, time-poor units. Utilising the ‘Interaction for Learning Framework’ developed by Arkoudis et al. (2010), small changes to enhance global and intercultural awareness were incorporated into existing teaching activities within a second-year biomedical physiology unit. Interventions included a short orientation session to promote intercultural exchange and awareness, incorporation of a global perspective into an assessment item and structuring increased opportunities for interaction in tutorial classes. The project was approved by the Murdoch University Human Ethics Committee (Permit 2011/175). In student surveys, 40% of domestic and 60% of international student respondents articulated specific learning about interaction in intercultural groups as a result of unit activities. Themes among student responses were the logistics of working in intercultural groups (40%), insight into other perspectives (33%) and the experience of finding common ground (20%). 60% of students indicated that “interacting with students from other cultures [had] helped them develop a deeper or more informed understanding of biomedical physiology”. Many students identified specific examples of how cultural factors would impact on the study or application of biomedical physiology within the global community. In addition, staff observed more widespread benefits for student engagement and mastery of physiology content. It is concluded that significant development of cross-cultural awareness and a more global perspective on scientific understanding can be supported among physiology undergraduates with relatively minor adaptations to course content. Clifford VA (2009). Engaging the disciplines in internationalising the curriculum. International Journal for Academic Development 14, 133-143. Arkoudis S, Yu X, Baik C, Borland H, Chang S, Lang I, Lang J, Pearce A & Watty K. (2010). Finding Common Ground: Enhancing interaction between domestic and international students. Melbourne, ALTC.

Purpose: The somatic neuromuscular junction (NMJ) is a valuable experimental model of synapse formation. Such experiments are most commonly performed on placental mammals (e.g. mouse), where all four limbs develop on a similar timescale. By contrast, some Australian marsupials have very segmented limb development, where forelimbs are precociously developed to assist the journey to the pouch, while hindlimbs are comparatively unformed. We have compared the morphological development of the NMJ in kangaroo hindlimbs and forelimbs, to investigate how NMJ development proceeds in an animal with this novel developmental pattern. Methods: Three Western Grey Kangaroo neonates aged postnatal day (P) 0, 26 and 100 were ethically sourced from licensed shooters in South-Western Australia. 5 forelimb and 4 hindlimb muscles from each specimen were embedded, cryosectioned, and stained immunohistochemically for NMJ compartment proteins. Results: Confocal micrographs of adult kangaroo NMJs revealed an unusual oval endplate with a internal perforation at its centre. When markers of NMJ development described in rodents (e.g. neurofilament distribution, ACh receptor clustering) were analysed in P0 neonates, forelimb NMJs were considerably more developed than hindlimb junctions. However, between P0 and 100, NMJs in kangaroo forelimbs developed at a slower rate than hindlimb NMJs, so that all limbs were similarly developed by P100. Conclusions: Our preliminary data indicates that the segmented limb development in Western Grey Kangaroos is reflected at the level of the NMJ. This segmented NMJ development, combined with the relative accessibility of kangaroo neonates (which undergo a large degree of maturation in the pouch), present novel opportunities for studying synaptic development compared with placental mammals.

The principles of synapse formation have been largely unraveled by investigations at the somatic neuromuscular junction (NMJ). Such experiments are commonly performed on placental mammals (e.g. mouse), where all four limbs develop on a similar timescale. By contrast, some Australian marsupials have very segmented limb development, where forelimbs are precociously developed to assist the journey to the pouch, while hindlimbs are comparatively unformed. We explored the implications of this novel developmental pattern for the morphological development of marsupial NMJs. Western Grey Kangaroo neonates (n = 8) aged postnatal day (P) 0-100 were ethically sourced from licensed shooters. Multiple (5-9) muscles from each specimen were embedded, cryosectioned, and stained immunohistochemically for NMJ compartment proteins. Confocal micrographs of adult kangaroo NMJs revealed small oval endplates with a central internal perforation. Standard indices of synapse maturation (e.g. ACh receptor clustering) confirmed significant NMJ development from P0-P100. Notably, forelimb NMJs were considerably more developed than hindlimb junctions at P0. From P0-100, kangaroo forelimb NMJs developed at a slower rate than hindlimb NMJs, so that all limbs were similarly developed by P100. Interestingly, within the hindlimb, a detailed comparison of 5 muscles suggested a single, homogenous developmental profile, in contrast with recent studies in mouse suggesting both fast and delayed synapsing muscle populations (Pun et al., 2002). The segmented nature of NMJ development in Western Grey Kangaroos combined with the relative accessibility provided by a protracted maturation in the pouch, suggest that marsupials may present novel opportunities for studying synapse maturation compared with placental mammals.