Crystallization of Escherichia coli Catabolite Gene Activator Protein with its DNA Binding Site: The use of Modular DNA

To obtain crystals of the Escherichia coli catabolite gene activator protein (CAP) complexed with its DNA-binding site, we have searched for crystallization conditions with 26 different DNA segments ≥28 base-pairs in length that explore a variety of nucleotide sequences, lengths, and extended 5′ or 3′ termini. In addition to utilizing uninterrupted asymmetric lac site sequences, we devised a novel approach of synthesizing half-sites that allowed us to efficiently generate symmetric DNA segments with a wide variety of extended termini and lengths in the large size range (≥28 bp) required by this protein. We report three crystal forms that are suitable for X-ray analysis, one of which (crystal form III) gives measurable diffraction amplitudes to 3 Å resolution. Additives such as calcium, n-octyl-β-d-glucopyranoside and spermine produce modest improvements in the quality of diffraction from crystal form III. Adequate stabilization of crystal form III is unexpectedly complex, requiring a greater than tenfold reduction in the salt concentration followed by addition of 2-methyl-2,4-pentanediol and then an increase in the concentration of polyethylene glycol.

Development of the Heat and Energy Concept Inventory: Preliminary Results on the Prevalence and Persistence of Engineering Students' Misconceptions

BACKGROUND Students frequently hold a number of misconceptions related to temperature, heat and energy. There is not currently a concept inventory with sufficiently high internal reliability to assess these concept areas for research purposes. Consequently, there is little data on the prevalence of these misconceptions amongst undergraduate engineering students. PURPOSE (HYPOTHESIS) This work presents the Heat and Energy Concept Inventory (HECI) to assess prevalent misconceptions related to: (1) Temperature vs. Energy, (2) Temperature vs. Perceptions of Hot and Cold, (3) Factors that affect the Rate vs. Amount of Heat Transfer and (4) Thermal Radiation. The HECI is also used to document the prevalence of misconceptions amongst undergraduate engineering students. DESIGN/METHOD Item analysis, guided by classical test theory, was used to refine individual questions on the HECI. The HECI was used in a one group, pre-test-post-test design to assess the prevalence and persistence of targeted misconceptions amongst a population of undergraduate engineering students at diverse institutions. RESULTS Internal consistency reliability was assessed using Kuder-Richardson Formula 20; values were 0.85 for the entire instrument and ranged from 0.59 to 0.76 for the four subcategories of the HECI. Student performance on the HECI went from 49.2% to 54.5% after instruction. Gains on each of the individual subscales of the HECI, while generally statistically significant, were similarly modest. CONCLUSIONS The HECI provides sufficiently high estimates of internal consistency reliability to be used as a research tool to assess students' understanding of the targeted concepts. Use of the instrument demonstrates that student misconceptions are both prevalent and resistant to change through standard instruction.

Investigating the Role of System Justification in Mental Illness Stigma

The current study integrates system justification theory with research on mental illness stigma. Stereotypes of both low- and high-status groups in society can be a means of satisfying the system justification motive, or the motive to view societal inequalities as justified (as reviewed in Jost, Banaji, & Nosek, 2004). Corrigan, Watson, and Ottati (2003) proposed that system justification theory may be able to explain the origins of particular stereotypes of people with mental illness, such as dangerousness and incompetence. The primary goal of the present study was to investigate whether the stigmatization of people with mental illness – a specific form of stigmatization of a lowstatus group – can be at least partially attributed to a broader motive to justify societal inequalities. To test this, the current study included both an experimental manipulation of the perceived legitimacy of the social system and a measure of system-justifying beliefs. Stigmatization of individuals with mental illness was measured with both explicit selfreport measures (semantic differentials and the Attribution Questionnaire) and an implicit measure (a computer-based Implicit Association Test). The relationships between participant characteristics, such as personal experience with mental illness, and stigma were also investigated. Consistent with past research demonstrating only modest correlations between explicit and implicit stigma, greater self-reported fear toward a person with a chronic mental illness was weakly associated with increased implicit bias against mental illness in favor of physical disability. There was little support for the involvement of system justification in explicit stigma. Participants with personal experience with mental illness were less likely to self-report fear and avoidance of a person with a chronic mental illness. These findings have implications for stigmareduction efforts.