Automated Writing Assessments Measure Undergraduate Learning after Completion of a Computer-Based Cellular Respiration Tutorial.

The focus of biology education has shifted from memorization to conceptual understanding of core biological concepts such as matter and energy relationships. To examine undergraduate learning about matter and energy, we incorporated constructed-response (CR) questions into an interactive computer-based tutorial. The objective of this tutorial is to teach students about matter and energy and help dispel common misconceptions through the context of cellular respiration. We used a constructed-response classifier (CRC) tool to categorize ideas in responses to three CR questions and measure changes in student thinking about cellular respiration. Our data set includes 841 undergraduates from 19 geographically diverse institutions including two-year colleges, primarily undergraduate institutions, and research-intensive colleges and universities. We found students from all institution types included more scientific ideas in CRs post-tutorial. Students used an average of 2.1 ideas in CRs and frequently used both scientific and developing ideas. We found this mixed thinking persisted after the tutorial regardless of institution type. Students' multiple-choice (MC) selections were correlated with their CRs, but CRs revealed more mixed thinking than would be inferred from MC responses. Our study shows a CRC tool can measure student learning after a computer-based tutorial and provides more complete information than MC responses.

Introductory biology undergraduate students' mixed ideas about genetic information flow.

The core concept of genetic information flow was identified in recent calls to improve undergraduate biology education. Previous work shows that students have difficulty differentiating between the three processes of the Central Dogma (CD; replication, transcription, and translation). We built upon this work by developing and applying an analytic coding rubric to 1050 student written responses to a three-question item about the CD. Each response was previously coded only for correctness using a holistic rubric. Our rubric captures subtleties of student conceptual understanding of each process that previous work has not yet captured at a large scale. Regardless of holistic correctness scores, student responses included five or six distinct ideas. By analyzing common co-occurring rubric categories in student responses, we found a common pair representing two normative ideas about the molecules produced by each CD process. By applying analytic coding to student responses preinstruction and postinstruction, we found student thinking about the processes involved was most prone to change. The combined strengths of analytic and holistic rubrics allow us to reveal mixed ideas about the CD processes and provide a detailed picture of which conceptual ideas students draw upon when explaining each CD process.

Mixed Student Ideas about Mechanisms of Human Weight Loss.

Recent calls for college biology education reform have identified "pathways and transformations of matter and energy" as a big idea in biology crucial for students to learn. Previous work has been conducted on how college students think about such matter-transforming processes; however, little research has investigated how students connect these ideas. Here, we probe student thinking about matter transformations in the familiar context of human weight loss. Our analysis of 1192 student constructed responses revealed three scientific (which we label "Normative") and five less scientific (which we label "Developing") ideas that students use to explain weight loss. Additionally, students combine these ideas in their responses, with an average number of 2.19 ± 1.07 ideas per response, and 74.4% of responses containing two or more ideas. These results highlight the extent to which students hold multiple (both correct and incorrect) ideas about complex biological processes. We described student responses as conforming to either Scientific, Mixed, or Developing descriptive models, which had an average of 1.9 ± 0.6, 3.1 ± 0.9, and 1.7 ± 0.8 ideas per response, respectively. Such heterogeneous student thinking is characteristic of difficulties in both conceptual change and early expertise development and will require careful instructional intervention for lasting learning gains.

Wobble pairs of the HDV ribozyme play specific roles in stabilization of active site dynamics.

The hepatitis delta virus (HDV) is the only known human pathogen whose genome contains a catalytic RNA motif (ribozyme). The overall architecture of the HDV ribozyme is that of a double-nested pseudoknot, with two GU pairs flanking the active site. Although extensive studies have shown that mutation of either wobble results in decreased catalytic activity, little work has focused on linking these mutations to specific structural effects on catalytic fitness. Here we use molecular dynamics simulations based on an activated structure to probe the active site dynamics as a result of wobble pair mutations. In both wild-type and mutant ribozymes, the in-line fitness of the active site (as a measure of catalytic proficiency) strongly depends on the presence of a C75(N3H3+)N1(O5') hydrogen bond, which positions C75 as the general acid for the reaction. Our mutational analyses show that each GU wobble supports catalytically fit conformations in distinct ways; the reverse G25U20 wobble promotes high in-line fitness, high occupancy of the C75(N3H3+)G1(O5') general-acid hydrogen bond and stabilization of the G1U37 wobble, while the G1U37 wobble acts more locally by stabilizing high in-line fitness and the C75(N3H3+)G1(O5') hydrogen bond. We also find that stable type I A-minor and P1.1 hydrogen bonding above and below the active site, respectively, prevent local structural disorder from spreading and disrupting global conformation. Taken together, our results define specific, often redundant architectural roles for several structural motifs of the HDV ribozyme active site, expanding the known roles of these motifs within all HDV-like ribozymes and other structured RNAs.

Disparate HDV ribozyme crystal structures represent intermediates on a rugged free-energy landscape.

The hepatitis delta virus (HDV) ribozyme is a member of the class of small, self-cleaving catalytic RNAs found in a wide range of genomes from HDV to human. Both pre- and post-catalysis (precursor and product) crystal structures of the cis-acting genomic HDV ribozyme have been determined. These structures, together with extensive solution probing, have suggested that a significant conformational change accompanies catalysis. A recent crystal structure of a trans-acting precursor, obtained at low pH and by molecular replacement from the previous product conformation, conforms to the product, raising the possibility that it represents an activated conformer past the conformational change. Here, using fluorescence resonance energy transfer (FRET), we discovered that cleavage of this ribozyme at physiological pH is accompanied by a structural lengthening in magnitude comparable to previous trans-acting HDV ribozymes. Conformational heterogeneity observed by FRET in solution appears to have been removed upon crystallization. Analysis of a total of 1.8 µsec of molecular dynamics (MD) simulations showed that the crystallographically unresolved cleavage site conformation is likely correctly modeled after the hammerhead ribozyme, but that crystal contacts and the removal of several 2'-oxygens near the scissile phosphate compromise catalytic in-line fitness. A cis-acting version of the ribozyme exhibits a more dynamic active site, while a G-1 residue upstream of the scissile phosphate favors poor fitness, allowing us to rationalize corresponding changes in catalytic activity. Based on these data, we propose that the available crystal structures of the HDV ribozyme represent intermediates on an overall rugged RNA folding free-energy landscape.