Redefining online biology education: a study on interactive branched video utilisation and student learning experiences.

This study evaluated the use of interactive, branched videos compared with traditional passive linear delivery for enhancing student engagement and learning in online courses. Undergraduate biology students were provided with either branched decision-based or linear videos on cell biology and protein purification as self-guided or consolidation activities. While the interactive branched videos did not improve learning gains, thematic analysis revealed that students found them more enjoyable and preferable for revision. However, most students felt linear passive videos were more logically structured for core content delivery. In a revised format, with clearer scaffolding, the interactive branched videos were perceived as significantly more engaging and useful when utilised for a problem-solving activity. Students welcomed the autonomy of directing their learning path but desired support to avoid missing critical information. Overall, thoughtfully designed branched videos can increase student motivation, but their utility depends on context. Our findings indicate the importance of balancing interactivity, clear organisation and purpose when incorporating these innovative formats into online learning. Branched videos show promise for increasing engagement but require intentional instructional design tailored to learning objectives.

Embedding retrieval practice in undergraduate biochemistry teaching using PeerWise.

Retrieval practice is an evidence-based approach to teaching; here, we evaluate the use of PeerWise for embedding retrieval practice into summative assessment. PeerWise allows anonymous authoring, sharing, answering, rating, and feedback on peer-authored multiple choice questions. PeerWise was embedded as a summative assessment in a large first-year introductory biochemistry module. Engagement with five aspects of the tool was evaluated against student performance in coursework, exam, and overall module outcome. Results indicated a weak-to-moderate positive but significant correlation between engagement with PeerWise and assessment performance. Student feedback showed PeerWise had a polarizing effect; the majority recognized the benefits as a learning and revision tool, but a minority strongly disliked it, complaining of a lack of academic moderation and irrelevant questions unrelated to the module. PeerWise can be considered a helpful learning tool for some students and a means of embedding retrieval practice into summative assessment.

Engagement with video content in the blended classroom.

Blended learning is becoming the expected norm for core content delivery in many institutions. Pre-recorded videos in the form of screencasts are the primary delivery method, with students being asked to engage with the content in this medium. Usage is only likely to increase into the future as delivery moves away from traditional lectures and seminars. In this perspective, we look at the use of video material as a means of content delivery and how to help students engage with it. Theoretical literature around cognitive loading and active learning, alongside personal experience of delivery, is drawn on to give a framework for creating engaging recordings and learning activities.

Reimagining laboratory-based immunology education in the time of COVID-19.

The pandemic has brought challenges to teaching lab and research skills. Here Nigel Francis and colleagues explore the diverse approaches taken to replace lab-based immunology teaching, explain how networks of educators have driven this innovation and discuss the importance of retaining best practice into the future.

Adopting a flipped classroom approach for teaching molar calculations to biochemistry and genetics students.

The flipped classroom is a relatively new active learning pedagogical intervention, gaining popularity as a blended learning methodology. The flipped classroom comprises two distinct parts, directed learning carried out at the student's own pace away from the classroom and an interactive, class-based activity encouraging problem-solving and experiential learning. This research presents a 1-year study to measure student performance and perception toward a flipped classroom approach to teaching core biochemical calculations to first-year undergraduate biochemistry and genetics students. A post-task questionnaire showed an overall positive student perception with an associated significant improvement in the end of module summative assessment. These results suggest that this teaching approach offers some advantages over more traditional teaching pedagogies.

Akt and STAT5 mediate naïve human CD4+ T-cell early metabolic response to TCR stimulation.

Metabolic pathways that regulate T-cell function show promise as therapeutic targets in diverse diseases. Here, we show that at rest cultured human effector memory and central memory CD4+ T-cells have elevated levels of glycolysis and oxidative phosphorylation (OXPHOS), in comparison to naïve T-cells. Despite having low resting metabolic rates, naive T-cells respond to TCR stimulation with robust and rapid increases in glycolysis and OXPHOS. This early metabolic switch requires Akt activity to support increased rates of glycolysis and STAT5 activity for amino acid biosynthesis and TCA cycle anaplerosis. Importantly, both STAT5 inhibition and disruption of TCA cycle anaplerosis are associated with reduced IL-2 production, demonstrating the functional importance of this early metabolic program. Our results define STAT5 as a key node in modulating the early metabolic program following activation in naive CD4+ T-cells and in turn provide greater understanding of how cellular metabolism shapes T-cell responses.

Metabolic Adaptation of Human CD4+ and CD8+ T-Cells to T-Cell Receptor-Mediated Stimulation.

Linking immunometabolic adaptation to T-cell function provides insight for the development of new therapeutic approaches in multiple disease settings. T-cell activation and downstream effector functions of CD4+ and CD8+ T-cells are controlled by the strength of interaction between the T-cell receptor (TCR) and peptides presented by human leukocyte antigens (pHLA). The role of TCR-pHLA interactions in modulating T-cell metabolism is unknown. Here, for the first time, we explore the relative contributions of the main metabolic pathways to functional responses in human CD4+ and CD8+ T-cells. Increased expression of hexokinase II accompanied by higher basal glycolysis is demonstrated in CD4+ T-cells; cytokine production in CD8+ T-cells is more reliant on oxidative phosphorylation. Using antigen-specific CD4+ and CD8+ T-cell clones and altered peptide ligands, we demonstrate that binding affinity tunes the underlying metabolic shift. Overall, this study provides important new insight into how metabolic pathways are controlled during antigen-specific activation of human T-cells.

Bioenergetic analysis of human peripheral blood mononuclear cells.

Leucocytes respond rapidly to pathogenic and other insults, with responses ranging from cytokine production to migration and phagocytosis. These are bioenergetically expensive, and increased glycolytic flux provides adenosine triphosphate (ATP) rapidly to support these essential functions. However, much of this work is from animal studies. To understand more clearly the relative role of glycolysis and oxidative phosphorylation in human leucocytes, especially their utility in a translational research setting, we undertook a study of human peripheral blood mononuclear cells (MNCs) bioenergetics. Glycolysis was essential during lipopolysaccharide (LPS)-mediated interleukin (IL)-1ß, IL-6 and tumour necrosis factor (TNF)-α production, as 2-deoxy-D-glucose decreased significantly the output of all three cytokines. After optimizing cell numbers and the concentrations of all activators and inhibitors, oxidative phosphorylation and glycolysis profiles of fresh and cryopreserved/resuscitated MNCs were determined to explore the utility of MNCs for determining the bioenergetics health profile in multiple clinical settings. While the LPS-induced cytokine response did not differ significantly between fresh and resuscitated cells from the same donors, cryopreservation/resuscitation significantly affected mainly some measures of oxidative phosphorylation, but also glycolysis. Bioenergetics analysis of human MNCs provides a quick, effective means to measure the bioenergetics health index of many individuals, but cryopreserved cells are not suitable for such an analysis. The translational utility of this approach was tested by comparing MNCs of pregnant and non-pregnant women to reveal increased bioenergetics health index with pregnancy but significantly reduced basal glycolysis and glycolytic capacity. More detailed analysis of discrete leucocyte populations would be required to understand the relative roles of glycolysis and oxidative phosphorylation during inflammation and other immune responses.

A novel hybrid CFH/CFHR3 gene generated by a microhomology-mediated deletion in familial atypical hemolytic uremic syndrome.

Genomic disorders affecting the genes encoding factor H (fH) and the 5 factor H related proteins have been described in association with atypical hemolytic uremic syndrome. These include deletions of CFHR3, CFHR1, and CFHR4 in association with fH autoantibodies and the formation of a hybrid CFH/CFHR1 gene. These occur through nonallelic homologous recombination secondary to the presence of large segmental duplications (macrohomology) in this region. Using multiplex ligation-dependent probe amplification to screen for such genomic disorders, we have identified a large atypical hemolytic uremic syndrome family where a deletion has occurred through microhomology-mediated end joining rather than nonallelic homologous recombination. In the 3 affected persons of this family, we have shown that the deletion results in formation of a CFH/CFHR3 gene. We have shown that the protein product of this is a 24 SCR protein that is secreted with normal fluid-phase activity but marked loss of complement regulation at cell surfaces despite increased heparin binding. In this study, we have therefore shown that microhomology in this area of chromosome 1 predisposes to disease associated genomic disorders and that the complement regulatory function of fH at the cell surface is critically dependent on the structural integrity of the whole molecule.

Lactoferrin inhibits neutrophil apoptosis via blockade of proximal apoptotic signaling events.

Neutrophils are the most abundant leukocyte and have a short lifespan, dying by apoptosis approximately five days after leaving the bone marrow. Their apoptosis can be delayed at sites of inflammation to extend their functional lifespan, but inappropriate inhibition of apoptosis contributes to chronic inflammatory disease. Levels of the physiological iron chelator lactoferrin are raised at sites of inflammation and we have shown previously that iron-unsaturated lactoferrin inhibited human neutrophil apoptosis, but the mechanisms involved were not determined. Here we report that the anti-apoptotic effect of lactoferrin is dependent upon its iron saturation status as iron-saturated lactoferrin did not affect neutrophil apoptosis. We also show that the effect of lactoferrin is mediated at an early stage in apoptosis as it inhibited activation of sphingomyelinase, generation of ceramide, activation of caspase 8 and Bax and cleavage of Bid. Lactoferrin did not inhibit apoptosis induced by exogenous ceramide, supporting the proposal that it acts upstream of ceramide generation. We therefore conclude that raised lactoferrin levels are likely to contribute to chronic inflammation by delaying neutrophil apoptosis and that this is achieved by inhibiting proximal apoptotic signaling events.

Common polymorphisms in C3, factor B, and factor H collaborate to determine systemic complement activity and disease risk.

Common polymorphisms in complement alternative pathway (AP) proteins C3 (C3(R102G)), factor B (fB(R32Q)), and factor H (fH(V62I)) are associated with age-related macular degeneration (AMD) and other pathologies. Our published work showed that fB(R32Q) influences C3 convertase formation, whereas fH(V62I) affects factor I cofactor activity. Here we show how C3(R102G) (C3S/F) influences AP activity. In hemolysis assays, C3(102G) activated AP more efficiently (EC(50) C3(102G): 157 nM; C3(102R): 191 nM; P < 0.0001). fB binding kinetics and convertase stability were identical, but native and recombinant fH bound more strongly to C3b(102R) (K(D) C3b(102R): 1.0 µM; C3b(102G): 1.4 µM; P < 0.0001). Accelerated decay was unaltered, but fH cofactor activity was reduced for C3b(102G), favoring AP amplification. Combining disease "risk" variants (C3(102G), fB(32R), and fH(62V)) in add-back assays yielded sixfold higher hemolytic activity compared with "protective" variants (C3(102R), fB(32Q), and fH(62I); P < 0.0001). These data introduce the concept of a functional complotype (combination of polymorphisms) defining complement activity in an individual, thereby influencing susceptibility to AP-driven disease.