Separation and enrichment of sodium-motile bacteria using cost-effective microfluidics

Many motile bacteria are propelled by the rotation of flagellar filaments. This rotation is driven by a membrane protein known as the stator-complex, which drives the rotor of the bacterial flagellar motor. Torque generation is powered in most cases by proton transit through membrane protein complexes known as stators, with the next most common ionic power source being sodium. Sodium-powered stators can be studied through the use of synthetic chimeric stators that combine parts of sodium- and proton-powered stator proteins. The most well studied example is the use of the sodium-powered PomA-PotB chimeric stator unit in the naturally proton-powered Escherichia coli. Here we designed a fluidics system at low cost for rapid prototyping to separate motile and non-motile populations of bacteria while varying the ionic composition of the media and thus the sodium-motive force available to drive this chimeric flagellar motor. We measured separation efficiencies at varying ionic concentrations and confirmed using fluorescence that our device delivered eightfold enrichment of the motile proportion of a mixed population. Furthermore, our results showed that we could select bacteria from reservoirs where sodium was not initially present. Overall, this technique can be used to implement the selection of highly motile fractions from mixed liquid cultures, with applications in directed evolution to investigate the adaptation of motility in bacterial ecosystems.     

Anatomy integration blueprint: A fourth‐year musculoskeletal anatomy elective model

Current undergraduate medical school curricular trends focus on both vertical integration of clinical knowledge into the traditionally basic science‐dedicated curricula and increasing basic science education in the clinical years. This latter type of integration is more difficult and less reported on than the former. Here, we present an outline of a course wherein the primary learning and teaching objective is to integrate basic science anatomy knowledge with clinical education. The course was developed through collaboration by a multi‐specialist course development team (composed of both basic scientists and physicians) and was founded in current adult learning theories. The course was designed to be widely applicable to multiple future specialties, using current published reports regarding the topics and clinical care areas relying heavily on anatomical knowledge regardless of specialist focus. To this end, the course focuses on the role of anatomy in the diagnosis and treatment of frequently encountered musculoskeletal conditions. Our iterative implementation and action research approach to this course development has yielded a curricular template for anatomy integration into clinical years. Key components for successful implementation of these types of courses, including content topic sequence, the faculty development team, learning approaches, and hidden curricula, were developed. We also report preliminary feedback from course stakeholders and lessons learned through the process. The purpose of this report is to enhance the current literature regarding basic science integration in the clinical years of medical school. Anat Sci Educ 7: 379–388. © 2014 American Association of Anatomists.     

Promoting Inquiry-Based Teaching in Laboratory Courses: Are We Meeting the Grade?

Over the past decade, repeated calls have been made to incorporate more active teaching and learning in undergraduate biology courses. The emphasis on inquiry-based teaching is especially important in laboratory courses, as these are the courses in which students are applying the process of science. To determine the current state of research on inquiry-based teaching in undergraduate biology laboratory courses, we reviewed the recent published literature on inquiry-based exercises. The majority of studies in our data set were in the subdisciplines of biochemistry, cell biology, developmental biology, genetics, and molecular biology. In addition, most exercises were guided inquiry, rather than open ended or research based. Almost 75% of the studies included assessment data, with two-thirds of these studies including multiple types of assessment data. However, few exercises were assessed in multiple courses or at multiple institutions. Furthermore, assessments were rarely based on published instruments. Although the results of the studies in our data set show a positive effect of inquiry-based teaching in biology laboratory courses on student learning gains, research that uses the same instrument across a range of courses and institutions is needed to determine whether these results can be generalized.     

Teacher behavior, student interest and affective learning: putting theory to practice

Students of Project Adapt (PA), an instructional program for at-risk middle school students in Brooklyn NY, have displayed startling improvement on cognitive outcome measures. Teacher given grades and standardized tests illuminate the progress students have made since entering the program. The current investigation attempts to analyze PA on students' affective responses in a pre/post-test experimental design. Upon entering PA, students completed measures of interest, affect, and their previous teachers' behavior (pre). These ratings were then compared to end of year ratings on the same instruments (post). Results indicate students' perceive PA teachers utilizing different behaviors than their previous teachers. Additionally, student affect and interest increased significantly. Results of this study have implications for both instructional researchers and educational practitioners.     

Multitrait-multimethod investigation of a novel body image measurement technique

A multitrait-multimethod (MTMM) matrix was used to evaluate validity evidence for a digital image manipulation (DIM) body image measurement technique in young women. One hundred one young women completed the DIM procedure and the Thompson and Gray (1995) Contour Drawing Rating Scale to measure self-ideal discrepancy and size perception accuracy components of body image. Seven-day test-retest reliability was acceptable (R = .81-.95). Convergent validity for self-ideal discrepancy was higher (r = .74) than the corresponding heterotrait, monomethod coefficients (r = .46, r = .23) and heterotrait-heteromethod coefficients (r = .18, r = .12). However, the convergent validity coefficient for size perception accuracy was r = .12. The pattern of correlations in the MTMM matrix met the criteria of Campbell and Fiske (1959) for validity of these procedures to measure self-ideal discrepancy but not size perception accuracy. The DIM procedure addresses some of the criticisms associated with figure-rating scales, such as unrepresentativeness of the figures, scale coarseness, and restriction of range in responses. DIM, therefore, represents a realistic, valid alternative to figure-rating scales for measuring self-ideal discrepancy.     

Analysis of a laminar-flow diffusional mixer for directed self-assembly of liposomes

The present work describes the operation and simulation of a microfluidic laminar-flow mixer. Diffusive mixing takes place between a core solution containing lipids in ethanol and a sheath solution containing aqueous buffer, leading to self assembly of liposomes. Present device architecture hydrodynamically focuses the lipid solution into a cylindrical core positioned at the center of a microfluidic channel of 125 × 125-μm² cross-section. Use of the device produces liposomes in the size range of 100–300 nm, with larger liposomes forming at greater ionic strength in the sheath solution and at lower lipid concentration in the core solution. Finite element simulations compute the concentration distributions of solutes at axial distances of greater than 100 channel widths. These simulations reduce computation time and enable computation at long axial distances by utilizing long hexahedral elements in the axial flow region and fine tetrahedral elements in the hydrodynamic focusing region. Present meshing technique is generally useful for simulation of long microfluidic channels and is fully implementable using comsol Multiphysics. Confocal microscopy provides experimental validation of the simulations using fluorescent solutions containing fluorescein or enhanced green fluorescent protein.     

Validity and reliability of GPS for measuring instantaneous velocity during acceleration, deceleration, and constant motion

In this study, we assessed the validity and reliability of 5 and 10 Hz global positioning systems (GPS) for measuring instantaneous velocity during acceleration, deceleration, and constant velocity while straight-line running. Three participants performed 80 running trials while wearing two GPS units each (5 Hz, V2.0 and 10 Hz, V4.0; MinimaxX, Catapult Innovations, Scoresby, VIC, Australia). The criterion measure used to assess GPS validity was instantaneous velocity recorded using a tripod-mounted laser. Validity was established using the standard error of the estimate (± 90% confidence limits). Reliability was determined using typical error (± 90% confidence limits, expressed as coefficient of variation) and Pearson's correlation. The 10 Hz GPS devices were two to three times more accurate than the 5 Hz devices when compared with a criterion value for instantaneous velocity during tasks completed at a range of velocities (coefficient of variation 3.1-11.3%). Similarly, the 10 Hz GPS units were up to six-fold more reliable for measuring instantaneous velocity than the 5 Hz units (coefficient of variation 1.9-6.0%). Newer GPS may provide an acceptable tool for the measurement of constant velocity, acceleration, and deceleration during straight-line running and have sufficient sensitivity for detecting changes in performance in team sport. However, researchers must account for the inherent match-to-match variation reported when using these devices.     

The effects of exercise in hypoxic and normoxic conditions on endothelin-1 and arterial compliance

The purpose of this study was to determine the effects of short-term normoxic and hypoxic exercise on plasma endothelin-1 and nitric oxide levels, and the relationship of arterial compliance and pulmonary artery pressure to endothelin-1. Seven endurance-trained males completed two incremental and two steady-state exercise tests performed at ventilatory threshold in normoxia and hypoxia (fraction of inspired oxygen = 0.14). Plasma endothelin-1was measured throughout steady-state tests. Arterial compliance using applanation tonometry, plasma nitric oxide and pulmonary artery pressure using Doppler echocardiography were measured before and after exercise. Small arterial compliance and pulmonary artery pressure significantly increased following exercise. There were no main effects of condition or time for plasma endothelin-1and nitric oxide levels. There were no significant relationships between plasma endothelin-1 and arterial compliance or pulmonary artery pressure. In conclusion, mechanisms other than the endothelial system may play a role in the exercise-induced changes in small artery compliance in this study population. Moderate hypoxia and a 30-minute steady-state exercise have limited effects on plasma endothelin-1 in endurance-trained males.