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Organelle heterogeneity and inter-organelle contacts within a single cell contribute to the limited sensitivity of current organelle separation techniques, thus hindering organelle subpopulation characterization. Here, we use direct current insulator-based dielectrophoresis (DC-iDEP) as an unbiased separation method and demonstrate its capability by identifying distinct distribution patterns of insulin vesicles from INS-1E insulinoma cells. A multiple voltage DC-iDEP strategy with increased range and sensitivity has been applied, and a differentiation factor (ratio of electrokinetic to dielectrophoretic mobility) has been used to characterize features of insulin vesicle distribution patterns. We observed a significant difference in the distribution pattern of insulin vesicles isolated from glucose-stimulated cells relative to unstimulated cells, in accordance with maturation of vesicles upon glucose stimulation. We interpret the difference in distribution pattern to be indicative of high-resolution separation of vesicle subpopulations. DC-iDEP provides a path for future characterization of subtle biochemical differences of organelle subpopulations within any biological system.

For many years, a key driver for Higher Education Institutions (HEIs) has been ensuring that students are prepared for employment post-graduation. This has often resulted in HEIs providing specific modules of study focussing on employability and soft skills. The existing research literature presents a variety of strategies for embedding employability within curricula, often focusing on frameworks to review the provision of employability opportunities within a degree programme. The authors have previously investigated an employability module within an Undergraduate Chemistry Programme at their UK HEI, particularly focusing on students' perceptions -- these coalesced around the themes of personal progression, relevance of the module and interactions. However, before using these findings to further develop the module, it was deemed necessary to investigate the module in a different context. This study investigates the employability module in an Australian HEI and its delivery to undergraduate and postgraduate students from a range of different disciplines. Free text comments from questionnaires answered by 85 participants were subjected to thematic analysis in order to understand students' perceptions of the module in this new context. The new thematic map was compared to that from the original study to reveal several key similarities and differences between students' perceptions. However, the most important finding is thast employability modules should be tailored to a specific cohort to ensure they are aligned with their needs. This suggests that Central University employability module providers should work closely with individual departments to maximise the impact of these for students.

Gender issues, and specifically the lack of women in the physical sciences, has been a subject of intense debate for decades. The problem is so acute, that national initiatives have been developed to analyse and address the issues, with some success in STEM, particularly in higher education and also in industry. However, despite this progress, there is little understanding as to why women are less likely to study the chemical sciences in particular. In this research, a survey and interviews were used to find out why female A-level chemistry students choose, or do not choose, to study chemistry at higher education level. Two distinct phases were identified. Firstly, intelligence gathering to understand the location, content, entry requirements, and career options for potential course and institution combinations. Secondly, self-reflection to establish whether, knowing themselves, students feel as though they would be successful on a particular course at a particular institution. These findings align with research into gender imbalance in STEM and Higher Education more broadly, but go beyond this to broaden current debates with a focus on chemistry in particular.

The "mathematics problem" is a well-known source of difficulty for students attempting numerical problem solving questions in the context of science education. This paper illuminates this problem from a biology education perspective by invoking Hogan's numeracy framework. In doing so, this study has revealed that the contextualisation of mathematics within the domain of biology is not the main source of difficulty for students but rather more fundamental mathematical skills.

School pupils often perceive STEM subjects to be challenging; thus, in this project, an interactive, nationwide STEM investigation was designed to stimulate their interest. The "Soils in Scottish Schools" project provided teachers with a flexible package of resources to allow them to enhance the profile and understanding of soil science of their pupils. This package included presentations, experimental procedures, activities, and monthly newsletters which could be adapted to the particular learning environment of their pupils. In the focal point of the project, teachers were supported to encourage their pupils to collect samples from their school ground which were then analyzed for copper, lead, and zinc levels. This year-long project also afforded teachers with the opportunity to develop the pupils' investigative skills through engagement with experiments in their own classrooms where they could engage with real data.

Recognising the importance of graduates being equipped with appropriate employability skills alongside their subject-specific skills, we have had transferable skills training embedded throughout our degree programmes for 30 years. More recently, a specific employability skills module for final-year honours students has been created. This module consists of a programme of activities supporting employability skills, which was delivered to final-year undergraduate students from 2012 to 2015. A key feature in the development and delivery of these activities was the involvement of external experts. Detailed questionnaires have captured student perceptions and thematic analysis has revealed key themes. The module has been perceived to be highly useful, resulting in significant increases in students' confidence across key areas of employability skills. Furthermore, students may hold skewed perceptions of the relevance of generic employability skills to their chosen career path. This fact should be considered when delivering employability skills programmes.

Plants exploit phenotypic plasticity to adapt their growth and development to prevailing environmental conditions. Interpretation of light and temperature signals is aided by the circadian system, which provides a temporal context. Phenotypic plasticity provides a selective and competitive advantage in nature but is obstructive during large-scale, intensive agricultural practices since economically important traits (including vegetative growth and flowering time) can vary widely depending on local environmental conditions. This prevents accurate prediction of harvesting times and produces a variable crop. In this study, we sought to restrict phenotypic plasticity and circadian regulation by manipulating signaling systems that govern plants' responses to environmental signals. Mathematical modeling of plant growth and development predicted reduced plant responses to changing environments when circadian and light signaling pathways were manipulated. We tested this prediction by utilizing a constitutively active allele of the plant photoreceptor phytochrome B, along with disruption of the circadian system via mutation of EARLY FLOWERING3. We found that these manipulations produced plants that are less responsive to light and temperature cues and thus fail to anticipate dawn. These engineered plants have uniform vegetative growth and flowering time, demonstrating how phenotypic plasticity can be limited while maintaining plant productivity. This has significant implications for future agriculture in both open fields and controlled environments. Plants utilize phenotypic plasticity to adapt to prevailing environmental conditions. This work demonstrates that manipulation of phytochrome B- and EARLY FLOWERING3-regulated signaling pathways can limit phenotypic plasticity with respect to light and temperature signals. This has implications for future crop development by enabling consistent plant growth and development despite the effects of climate change. [ABSTRACT FROM AUTHOR]

Background Acanthamoeba spp. is the causative agent of Acanthamoeba keratitis and granulomatous amoebic encephalitis. Strathclyde minor groove binders (S-MGBs) are a promising new class of anti-infective agent that have been shown to be effective against many infectious organisms. Objectives To synthesize and evaluate the anti- Acanthamoeba activity of a panel of S-MGBs, and therefore determine the potential of this class for further development. Methods A panel of 12 S-MGBs was synthesized and anti- Acanthamoeba activity was determined using an alamarBlue™-based trophocidal assay against Acanthamoeba castellanii. Cross-screening against Trypanosoma brucei brucei , Staphylococcus aureus and Escherichia coli was used to investigate selective potency. Cytotoxicity against HEK293 cells allowed for selective toxicity to be measured. DNA binding studies were carried out using native mass spectrometry and DNA thermal shift assays. Results and discussion S-MGB-241 has an IC50 of 6.6 µM against A. castellanii , comparable to the clinically used miltefosine (5.6 µM) and negligible activity against the other organisms. It was also found to have an IC50 > 100 µM against HEK293 cells, demonstrating low cytotoxicity. S-MGB-241 binds to DNA as a dimer, albeit weakly compared to other S-MGBs previously studied. This was confirmed by DNA thermal shift assay with a Δ T m = 1 ± 0.1°C. Conclusions Together, these data provide confidence that S-MGBs can be further optimized to generate new, potent treatments for Acanthameoba spp. infections. In particular, S-MGB-241, has been identified as a 'hit' compound that is selectively active against A. castellanii , providing a starting point from which to begin optimization of DNA binding and potency. [ABSTRACT FROM AUTHOR]

The avian egg is a closed system that protects the growing embryo from external factors but prevents direct observation of embryo development. Various culture systems exist in the literature to study the development of the embryo for short periods of incubation (from 12 h up to a maximum of 60 h of egg incubation). A common flaw to these culture techniques is the inability to culture the unincubated avian blastoderm with intact tissue tensions on its native yolk. The goal of this work is to create a unique novel egg-in-cube system that can be used for long-term quail embryo culture initiated from its unincubated blastoderm stage. The egg-in-cube acts as an artificial transparent eggshell system that holds the growing embryo, making it amenable to microscopy. With the egg-in-cube system, quail embryos can be grown up to 9 days from the unincubated blastoderm (incubated in air, 20.9% O2), which improves to 15 days on switching to a hyperoxic environment of 60% O2. Using transgenic fluorescent quail embryos in the egg-in-cube system, cell movements in the unincubated blastoderm are imaged dynamically using inverted confocal microscopy, which has been challenging to achieve with other culture systems. Apart from these observations, several other imaging applications of the system are described in this work using transgenic fluorescent quail embryos with upright confocal or epifluorescence microscopy. To demonstrate the usefulness of the egg-in-cube system in perturbation experiments, the quail neural tube is electroporated with fluorescent mRNA “in cubo”, followed by the incubation of the electroporated embryo and microscopy of the electroporated region with the embryo in the cube. The egg-in-cube culture system in combination with the “in cubo” electroporation and dynamic imaging capabilities described here will enable researchers to investigate several fundamental questions in early embryogenesis with the avian (quail) embryo on its native yolk.

Introduction: There are approximately 90 Aboriginal and Torres Strait Islander community controlled art centres across Australia, the majority in geographically remote locations. This survey explored how these centres are supporting older people, including people living with dementia, if and how they are collaborating with aged care services and what challenges and opportunities they identify in these arrangements. Methods: An online survey was developed by a team of Aboriginal and non-Aboriginal researchers, art centre staff and art centre consultants. The survey was distributed in 2018 to art centres across Australia via their four art centre peak bodies: Desart; the Association of Northern, Kimberley and Arnhem Aboriginal Artists; the Indigenous Art Centre Alliance - Far North Queensland and Torres Strait Islands; and the Aboriginal Art Centre Hub - Western Australia. The survey was also conducted face-to-face with participants at art centre annual events; and on field trips to North Western and Central Australia, conducted as part of an overarching study. Results: There were 53 completed surveys, with the highest proportion of responses (43%) from art centre managers. The survey generated 330 qualitative responses to the 13 questions that provided this option. The results showed that art centres play a wide ranging and vital role in supporting the health and wellbeing of older artists, many of whom are considered Elders within their communities, and that this reaches far beyond the production of art. The results showed that art centres are a safe place providing older people with the purpose and means to generate income, to enact governance, and to share cultural knowledge through intergenerational connection. Additionally, the results indicate that art centres provide a significant amount of direct care for older people, and that relationships are fundamental to delivering this social, emotional, spiritual and physical care. Furthermore, they showed a great deal of collaboration between art centres and aged care services, although little of this is formally documented or resourced. Conclusion: The survey results demonstrate that art centres play a significant and previously unexplored role in supporting the wellbeing of older people and people living with dementia in remote Aboriginal and Torres Strait Islander communities across Australia. Respondents shared diverse examples of providing physical, social, emotional, spiritual and cultural care, assistance with navigating health and aged care systems, as well as examples of collaborations with aged care and health providers. The results demonstrate opportunities to recognise and resource this vital work. These findings are particularly important in the context of Australia's recent Royal Commission into Aged Care Quality and Safety, which found that systemic change is urgently required. Additionally, Indigenous scholars have called for a swell of system reform to address inequities in health and aged care systems. They advocate for a fundamental shift from biomedical and siloed models of care to integrated models that centralise culture, intergenerational connection and the cultural determinants of health. The results show that art centres could bring their expertise to this conversation.

Like most university learners, students in the Department of Pure and Applied Chemistry at the University of Strathclyde, experienced hybrid learning environments from March 2020 transitioning into hybrid approaches for the full academic year 21/22. As students who were in the early years of their programme at this time, and experienced a shift from traditional education practices are soon to graduate, a short research project was undertaken to better understand the perception of employers on the impact of remote and hybrid learning on current, and future, graduate skills, and competencies. Several employers in the chemical industries were surveyed to rate changes in their perception of, and importance of, key graduate skills and competencies after the pandemic. In general, it was shown that the employers responding to this survey did not show concern about the level of personal, professional, and higherlevel cognitive skills developed by students working in remote/hybrid environments during the pandemic. A gap in practical skills was acknowledged as indicated by employers' viewing students as being less skilled in this area, however the skills gap was not thought to be important with regards to employability. Additionally, employers indicated an increased importance on a student's ability to develop and demonstrate basic personal and professional skills including communication, collaboration, interpersonal/networking, autonomy, self-management, and resilience. The research suggests programme leaders creating or modifying curricula should place attention on the development of these skills and competencies to ensure students are self-led to successfully transition into the workplace. [ABSTRACT FROM AUTHOR]

Basic mathematical fluency is a prerequisite for success in a wide array of areas in biology and it has been noted that many students are deficient in this skill. In this paper, the use of a "simulated" peer-assessment activity is investigated as a method to improve performance in numerical problem-solving questions in high school biology. Additionally, the benefits of using "simulated," rather than real, student's answers in peer-assessment is discussed. The study involved a small cohort of students who carried out a "simulated" peer-assessment as a classroom activity and their improvement in performance and attitude towards the activity was measured. The results demonstrate that a "simulated" peer-assessment activity is suitable as a replacement for standard peer-assessment and that students' attitudes favour the simulated approach.

Summary: Metabolism is a key regulator of hematopoietic stem cell (HSC) functions. There is a lack of real-time, non-invasive approaches to evaluate metabolism in single HSCs. Using fluorescence lifetime imaging microscopy, we developed a set of metabolic optical biomarkers (MOBs) from the auto-fluorescent properties of metabolic coenzymes NAD(P)H and FAD. The MOBs revealed the enhanced glycolysis, low oxidative metabolism, and distinct mitochondrial localization of HSCs. Importantly, the fluorescence lifetime of enzyme-bound NAD(P)H (τbound) can non-invasively monitor the glycolytic/lactate dehydrogenase activity in single HSCs. As a proof of concept for metabolism-based cell sorting, we further identified HSCs within the Lineage-cKit+Sca1+ (KLS) hematopoietic stem/progenitor population using MOBs and a machine-learning algorithm. Moreover, we revealed the dynamic changes of MOBs, and the association of longer τbound with enhanced glycolysis under HSC stemness-maintaining conditions during HSC culture. Our work thus provides a new paradigm to identify and track the metabolism of single HSCs non-invasively and in real time. : Optical Imaging; Biological Sciences; Cellular Physiology; Stem Cells Research Subject Areas: Optical Imaging, Biological Sciences, Cellular Physiology, Stem Cells Research

Objective Advance care planning (ACP) assists people to identify their goals, values and treatment preferences for future care. Ideally, preferences are documented in an advance care directive (ACD) and used by doctors to guide medical decision-making should the patient subsequently lose their decision-making capacity. However, studies demonstrate that ACDs are not always adhered to by doctors in clinical practice. We aim to describe the attitudes and perspectives of doctors regarding ACD adherence and the utility of ACDs in clinical practice.Design Face-to-face semistructured interviews were conducted using three case-based vignettes to explore doctors’ decision-making and attitudes towards ACDs. Transcripts were analysed using a thematic analysis.Setting Doctors from a variety of medical specialties and with varying experience levels were recruited from a large tertiary hospital in Melbourne, Australia.Participants A total of 21 doctors were interviewed, 48% female (10/21). Most (19/21) reported having experience using ACDs.Results Four themes were identified: aligning with patient preferences (avoiding unwanted care, prioritising autonomy and anticipating family opposition), advocating best interests (defining futile care, relying on clinical judgement, rejecting unreasonable decisions and disregarding legal consequences), establishing validity (doubting rigour of the decision-making process, questioning patients’ ability to understand treatment decisions, distrusting outdated preferences and seeking confirmation) and translating written preferences into practice (contextualising patient preferences, applying subjective terminology and prioritising emergency medical treatment).Conclusions ACDs provide doctors with opportunities to align patient preferences with treatment and uphold patient autonomy. However, doctors experience decisional conflict when attempting to adhere to ACDs in practice, especially when they believe that adhering to the ACD is not in the patients’ best interests, or if they doubt the validity of the ACD. Future ACP programmes should consider approaches to improve the validity and applicability of ACDs. In addition, there is a need for ethical and legal education to support doctors’ knowledge and confidence in ACP and enacting ACDs.

Gold nanoparticles continue to generate interest for use in several biomedical applications. Recently, researchers have been focusing on exploiting their dual diagnostic/therapeutic theranostic capabilities. Before clinical translation can occur, regulatory agencies will require a greater understanding of their biodistribution and safety profiles post administration. Previously, the real-time identification and tracking of gold nanoparticles in free-flowing vasculature had not been possible without extrinsic labels such as fluorophores. Here, we present a label-free imaging approach to examine gold nanoparticle (AuNP) activity within the vasculature by utilizing multiphoton intravital microscopy. This method employs a commercially available multiphoton microscopy system to visualize the intrinsic luminescent signal produced by a multiphoton absorption-induced luminescence effect observed in single gold nanoparticles at frame rates necessary for capturing real-time blood flow. This is the first demonstration of visualizing unlabeled gold nanoparticles in an unperturbed vascular environment with frame rates fast enough to achieve particle tracking. Nanoparticle blood concentration curves were also evaluated by the tracking of gold nanoparticle flow in vasculature and verified against known pre-injection concentrations. Half-lives of these gold nanoparticle injections ranged between 67 and 140 s. This label-free imaging approach could provide important structural and functional information in real time to aid in the development and effective analysis of new metallic nanoparticles for various clinical applications in an unperturbed environment, while providing further insight into their complex uptake and clearance pathways.