All "wrapped" up in reflection: supporting metacognitive awareness to promote students' self-regulated learning.

Self-regulated learning (SRL) is the process of utilizing effective strategies to acquire knowledge or skills and is influenced by motivation, metacognitive processing, and study-related behaviors. We hypothesized that by using survey tools that allow reflection on and refinement of students' study strategies, we could nurture metacognitive skill development, encourage positive motivation and study-related behaviors, and hence promote academic success. Undergraduate students in a semester-long, second-year biology course were provided with resources to promote SRL and three survey instruments that encouraged them to create study plans and reflect on the effectiveness of their study strategies. Using a student-partnered approach, we sought to investigate the role of metacognition, motivation, and study-related behaviors on academic performance by (i) identifying the self-regulated learning strategies most utilized by students, (ii) investigating the role of reflection in enhancing metacognitive processing and academic performance, and (iii) understanding whether students created and/or modified their study strategies as an outcome of self-regulation. Survey responses allowed us to understand the repertoire of study strategies used by students. Our analyses suggest that students demonstrated metacognitive skill development through the use of the resources and reflection instruments, as they accurately reported on the effectiveness of their study strategies and indicated future plans to shift study-related behaviors from passive to active reviewing techniques. Students across the grade spectrum perceived the reflection instruments as beneficial in identifying areas of improvement and developing long-term study habits, suggesting that these instruments were effective in promoting metacognitive skill development for a variety of student learners. We conclude that supporting students with resources that promote SRL and providing opportunities for timely reflection can promote metacognitive skill development, a key feature of academic success.

Investigating Anti-Vaccination Stances on Social Media: an Assignment To Promote Science Literacy.

In this digital age in which social media use among young adults continues to rise, consideration of the impact of these platforms on our students and on science literacy pedagogy is essential. This has been highlighted during the 2019 coronavirus disease pandemic, when mis- and disinformation surrounding the pandemic and vaccinations were so prevalent on social media platforms that it provoked a cautionary announcement from the World Health Organization. We describe here the structure of an assignment aimed to promote science literacy by encouraging students to explore antivaccination stances on social media and evaluate the scientific validity of such claims using scientific literature. To comprehensively analyze these antivaccination sentiments, we encouraged students to develop succinct arguments to demonstrate the social, economic, or other cultural influences likely contributing to antivaccination stances. In alignment with the philosophical-educational concept of Bildung, we hope to nurture an understanding of scientific literacy that focuses on both evidence-based critical thinking as well as empathetic understanding of the socio-political circumstances that influence public opinion on scientific matters. Student work provided compelling evidence for the success of our field-tested assignment in fostering students to be authoritative voices of science in everyday life and highlighted the importance of efforts to explicitly focus on science literacy within biology curricula.

COL1A1 C-propeptide mutations cause ER mislocalization of procollagen and impair C-terminal procollagen processing.

Mutations in the type I procollagen C-propeptide occur in ~6.5% of Osteogenesis Imperfecta (OI) patients. They are of special interest because this region of procollagen is involved in α chain selection and folding, but is processed prior to fibril assembly and is absent in mature collagen fibrils in tissue. We investigated the consequences of seven COL1A1 C-propeptide mutations for collagen biochemistry in comparison to three probands with classical glycine substitutions in the collagen helix near the C-propeptide and a normal control. Procollagens with C-propeptide defects showed the expected delayed chain incorporation, slow folding and overmodification. Immunofluorescence microscopy indicated that procollagen with C-propeptide defects was mislocalized to the ER lumen, in contrast to the ER membrane localization of normal procollagen and procollagen with helical substitutions. Notably, pericellular processing of procollagen with C-propeptide mutations was defective, with accumulation of pC-collagen and/or reduced production of mature collagen. In vitro cleavage assays with BMP-1 ±â€¯PCPE-1 confirmed impaired C-propeptide processing of procollagens containing mutant proα1(I) chains. Overmodified collagens were incorporated into the matrix in culture. Dermal fibrils showed alterations in average diameter and diameter variability and bone fibrils were disorganized. Altered ER-localization and reduced pericellular processing of defective C-propeptides are expected to contribute to abnormal osteoblast differentiation and matrix function, respectively.

Glucose transporter expression in an avian nectarivore: the ruby-throated hummingbird (Archilochus colubris).

Glucose transporter (GLUT) proteins play a key role in the transport of monosaccharides across cellular membranes, and thus, blood sugar regulation and tissue metabolism. Patterns of GLUT expression, including the insulin-responsive GLUT4, have been well characterized in mammals. However, relatively little is known about patterns of GLUT expression in birds with existing data limited to the granivorous or herbivorous chicken, duck and sparrow. The smallest avian taxa, hummingbirds, exhibit some of the highest fasted and fed blood glucose levels and display an unusual ability to switch rapidly and completely between endogenous fat and exogenous sugar to fuel energetically expensive hovering flight. Despite this, nothing is known about the GLUT transporters that enable observed rapid rates of carbohydrate flux. We examined GLUT (GLUT1, 2, 3, & 4) expression in pectoralis, leg muscle, heart, liver, kidney, intestine and brain from both zebra finches (Taeniopygia guttata) and ruby-throated hummingbirds (Archilochus colubris). mRNA expression of all four transporters was probed using reverse-transcription PCR (RT-PCR). In addition, GLUT1 and 4 protein expression were assayed by western blot and immunostaining. Patterns of RNA and protein expression of GLUT1-3 in both species agree closely with published reports from other birds and mammals. As in other birds, and unlike in mammals, we did not detect GLUT4. A lack of GLUT4 correlates with hyperglycemia and an uncoupling of exercise intensity and relative oxidation of carbohydrates in hummingbirds. The function of GLUTs present in hummingbird muscle tissue (e.g. GLUT1 and 3) remain undescribed. Thus, further work is necessary to determine if high capillary density, and thus surface area across which cellular-mediated transport of sugars into active tissues (e.g. muscle) occurs, rather than taxon-specific differences in GLUT density or kinetics, can account for observed rapid rates of sugar flux into these tissues.

Selective processing and metabolism of disease-causing mutant prion proteins.

Prion diseases are fatal neurodegenerative disorders caused by aberrant metabolism of the cellular prion protein (PrP(C)). In genetic forms of these diseases, mutations in the globular C-terminal domain are hypothesized to favor the spontaneous generation of misfolded PrP conformers (including the transmissible PrP(Sc) form) that trigger downstream pathways leading to neuronal death. A mechanistic understanding of these diseases therefore requires knowledge of the quality control pathways that recognize and degrade aberrant PrPs. Here, we present comparative analyses of the biosynthesis, trafficking, and metabolism of a panel of genetic disease-causing prion protein mutants in the C-terminal domain. Using quantitative imaging and biochemistry, we identify a misfolded subpopulation of each mutant PrP characterized by relative detergent insolubility, inaccessibility to the cell surface, and incomplete glycan modifications. The misfolded populations of mutant PrPs were neither recognized by ER quality control pathways nor routed to ER-associated degradation despite demonstrable misfolding in the ER. Instead, mutant PrPs trafficked to the Golgi, from where the misfolded subpopulation was selectively trafficked for degradation in acidic compartments. Surprisingly, selective re-routing was dependent not only on a mutant globular domain, but on an additional lysine-based motif in the highly conserved unstructured N-terminus. These results define a specific trafficking and degradation pathway shared by many disease-causing PrP mutants. As the acidic lysosomal environment has been implicated in facilitating the conversion of PrP(C) to PrP(Sc), our identification of a mutant-selective trafficking pathway to this compartment may provide a cell biological basis for spontaneous generation of PrP(Sc) in familial prion disease.

Prion protein biosynthesis and its emerging role in neurodegeneration.

Various fatal neurodegenerative disorders are caused by altered metabolism of the prion protein (PrP). These diseases are typically transmissible by an unusual 'protein-only' mechanism in which a misfolded isomer, PrP(Sc), confers its aberrant conformation onto normal cellular PrP. An impressive range of studies has investigated nearly every aspect of this fascinating event; yet, our understanding of how PrP(Sc) accumulation might lead to cellular dysfunction and neurodegeneration is trifling. Recent advances in our understanding of normal PrP biosynthesis and degradation might have unexpectedly shed new light on this complex problem. Indeed, our current understanding of normal PrP cell biology, coupled with a growing appreciation of its complex metabolism, is providing new hypotheses for PrP-mediated neurodegeneration.

Retrotranslocation of prion proteins from the endoplasmic reticulum by preventing GPI signal transamidation.

Neurodegeneration in diseases caused by altered metabolism of mammalian prion protein (PrP) can be averted by reducing PrP expression. To identify novel pathways for PrP down-regulation, we analyzed cells that had adapted to the negative selection pressure of stable overexpression of a disease-causing PrP mutant. A mutant cell line was isolated that selectively and quantitatively routes wild-type and various mutant PrPs for ER retrotranslocation and proteasomal degradation. Biochemical analyses of the mutant cells revealed that a defect in glycosylphosphatidylinositol (GPI) anchor synthesis leads to an unprocessed GPI-anchoring signal sequence that directs both ER retention and efficient retrotranslocation of PrP. An unprocessed GPI signal was sufficient to impart ER retention, but not retrotranslocation, to a heterologous protein, revealing an unexpected role for the mature domain in the metabolism of misprocessed GPI-anchored proteins. Our results provide new insights into the quality control pathways for unprocessed GPI-anchored proteins and identify transamidation of the GPI signal sequence as a step in PrP biosynthesis that is absolutely required for its surface expression. As each GPI signal sequence is unique, these results also identify signal recognition by the GPI-transamidase as a potential step for selective small molecule perturbation of PrP expression.

Deficiency of cartilage-associated protein in recessive lethal osteogenesis imperfecta.

Classic osteogenesis imperfecta, an autosomal dominant disorder associated with osteoporosis and bone fragility, is caused by mutations in the genes for type I collagen. A recessive form of the disorder has long been suspected. Since the loss of cartilage-associated protein (CRTAP), which is required for post-translational prolyl 3-hydroxylation of collagen, causes severe osteoporosis in mice, we investigated whether CRTAP deficiency is associated with recessive osteogenesis imperfecta. Three of 10 children with lethal or severe osteogenesis imperfecta, who did not have a primary collagen defect yet had excess post-translational modification of collagen, were found to have a recessive condition resulting in CRTAP deficiency, suggesting that prolyl 3-hydroxylation of type I collagen is important for bone formation.

Virus receptors and tropism.

Polyomaviruses are small, tumorigenic, nonenveloped viruses that infect several different species. Interaction of these viruses with cell surface receptors represents the initial step during infection of host cells. This interaction can be a major determinant of viral host and tissue tropism. This chapter reviews what is currently known about the cellular receptors for each of five polyomavirus family members: Mouse polyomavirus (PyV), JC virus (JCV), BK virus (BKV), Lymphotropic papovavirus (LPV) and Simian virus 40 (SV40). These polyomaviruses serve to illustrate the enormous diversity of virus-cell surface interactions and allow us to closely evaluate the role of receptors in their life cycles. The contribution of other factors such as transcriptional regulators and signaling pathways are also summarized.

Contrasting roles of endosomal pH and the cytoskeleton in infection of human glial cells by JC virus and simian virus 40.

Infection of eukaryotic cells by pathogens requires the efficient use of host cell endocytic and cytoplasmic transport mechanisms. Understanding how these cellular functions are exploited by microorganisms allows us to better define the basic biology of pathogenesis while providing better insight into normal cellular functions. In this report we compare and contrast intracellular transport and trafficking of the human polyomavirus JC virus (JCV) with that of simian virus 40 (SV40). We have previously shown that infection of human glial cells by JCV requires clathrin-dependent endocytosis. In contrast, infection of cells by SV40 proceeds by caveola-dependent endocytosis. We now examine the roles of endosomal pH and the cellular cytoskeleton during infection of glial cells by both viruses. Our results demonstrate that JCV infection is sensitive to disruption of endosomal pH, whereas SV40 infection is pH independent. Infection by JCV is inhibited by treatment of glial cells with cytochalasin D, nocodazole, and acrylamide, whereas SV40 infection is affected only by nocodazole. These data point to critical differences between JCV and SV40 in terms of endocytosis and intracellular trafficking of their DNA genomes to the nucleus. These data also suggest a unique sequential involvement of cytoskeletal elements during infection of glial cells by JCV.