Single-cell attachment and culture method using a photochemical reaction in a closed microfluidic system

Recently, interest in single cell analysis has increased because of its potential for improving our understanding of cellular processes. Single cell operation and attachment is indispensable to realize this task. In this paper, we employed a simple and direct method for single-cell attachment and culture in a closed microchannel. The microchannel surface was modified by applying a nonbiofouling polymer, 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer, and a nitrobenzyl photocleavable linker. Using ultraviolet (UV) light irradiation, the MPC polymer was selectively removed by a photochemical reaction that adjusted the cell adherence inside the microchannel. To obtain the desired single endothelial cell patterning in the microchannel, cell-adhesive regions were controlled by use of round photomasks with diameters of 10, 20, 30, or 50 μm. Single-cell adherence patterns were formed after 12 h of incubation, only when 20 and 30 μm photomasks were used, and the proportions of adherent and nonadherent cells among the entire UV-illuminated areas were 21.3%±0.3% and 7.9%±0.3%, respectively. The frequency of single-cell adherence in the case of the 20 μm photomask was 2.7 times greater than that in the case of the 30 μm photomask. We found that the 20 μm photomask was optimal for the formation of single-cell adherence patterns in the microchannel. This technique can be a powerful tool for analyzing environmental factors like cell-surface and cell-extracellular matrix contact.     

Novel mathematical model to estimate ball impact force in soccer

To assess ball impact force during soccer kicking is important to quantify from both performance and chronic injury prevention perspectives. We aimed to verify the appropriateness of previous models used to estimate ball impact force and to propose an improved model to better capture the time history of ball impact force. A soccer ball was fired directly onto a force platform (10 kHz) at five realistic kicking ball velocities and ball behaviour was captured by a high-speed camera (5,000 Hz). The time history of ball impact force was estimated using three existing models and two new models. A new mathematical model that took into account a rapid change in ball surface area and heterogeneous ball deformation showed a distinctive advantage to estimate the peak forces and its occurrence times and to reproduce time history of ball impact forces more precisely, thereby reinforcing the possible mechanics of ‘footballer’s ankle’. Ball impact time was also systematically shortened when ball velocity increases in contrast to practical understanding for producing faster ball velocity, however, the aspect of ball contact time must be considered carefully from practical point of view.     

Numerical and experimental investigations of human swimming motions

This paper reviews unsteady flow conditions in human swimming and identifies the limitations and future potential of the current methods of analysing unsteady flow. The capability of computational fluid dynamics (CFD) has been extended from approaches assuming steady-state conditions to consideration of unsteady/transient conditions associated with the body motion of a swimmer. However, to predict hydrodynamic forces and the swimmer’s potential speeds accurately, more robust and efficient numerical methods are necessary, coupled with validation procedures, requiring detailed experimental data reflecting local flow. Experimental data obtained by particle image velocimetry (PIV) in this area are limited, because at present observations are restricted to a two-dimensional 1.0 m² area, though this could be improved if the output range of the associated laser sheet increased. Simulations of human swimming are expected to improve competitive swimming, and our review has identified two important advances relating to understanding the flow conditions affecting performance in front crawl swimming: one is a mechanism for generating unsteady fluid forces, and the other is a theory relating to increased speed and efficiency.     

DEVELOPING CURRICULA FOR GRANDPARENTS IN JAPAN

The purpose of this study was to determine how three generations view the contribution and needs of grandparents in Japan. A sample consisting of 239 grandparents, 266 parents, and 274 grandchildren from, urban and rural areas completed the Grandparent Strengths and Needs Inventory (Strom & Strom, 1993). Grandparents reported more satisfaction, success, and involvement in teaching than was observed by parents or grandchildren. Grandparents experienced greater difficulty as well as frustration and saw themselves as less informed about their role than was reported by younger people. Significant main effects were generation, grandchild gender, grandchild age, generations living together, frequency of grandchild care by the grandparent, and amount of time the generations spent together. Implications for building curricula and guidelines for program development are identified.     

Reconsideration of the Paradox of Inquiry

It is well known that the Meno presents the argument called “the paradox of inquiry.” This paper has two purposes. First, I analyze the paradox of inquiry and reformulate the argument as the “renewed paradox of inquiry.” Second, I clarify that the problem of inquiry posed by this paradox concerns the necessary conditions for a novice learner to learn to begin and advance an inquiry on her own. This problem of inquiry will be important to address even today in considering scientific and educational practice related to inquiry.     

Selective cell capture and analysis using shallow antibody-coated microchannels

Demand for analysis of rare cells such as circulating tumor cells in blood at the single molecule level has recently grown. For this purpose, several cell separation methods based on antibody-coated micropillars have been developed (e.g., Nagrath et al., Nature 450, 1235–1239 (2007)). However, it is difficult to ensure capture of targeted cells by these methods because capture depends on the probability of cell-micropillar collisions. We developed a new structure that actively exploits cellular flexibility for more efficient capture of a small number of cells in a target area. The depth of the sandwiching channel was slightly smaller than the diameter of the cells to ensure contact with the channel wall. For cell selection, we used anti-epithelial cell adhesion molecule antibodies, which specifically bind epithelial cells. First, we demonstrated cell capture with human promyelocytic leukemia (HL-60) cells, which are relatively homogeneous in size; in situ single molecule analysis was verified by our rolling circle amplification (RCA) method. Then, we used breast cancer cells (SK-BR-3) in blood, and demonstrated selective capture and cancer marker (HER2) detection by RCA. Cell capture by antibody-coated microchannels was greater than with negative control cells (RPMI-1788 lymphocytes) and non-coated microchannels. This system can be used to analyze small numbers of target cells in large quantities of mixed samples.     

Practice Makes Pretty Good: Assessment of Primary Literature Reading Abilities across Multiple Large-Enrollment Biology Laboratory Courses

Primary literature is essential for scientific communication and is commonly utilized in undergraduate biology education. Despite this, there is often little time spent training our students how to critically analyze a paper. To address this, we introduced a primary literature module in multiple upper-division laboratory courses. In this module, instructors conduct classroom discussions that dissect a paper as researchers do. While previous work has identified classroom interventions that improve primary literature comprehension within a single course, our goal was to determine whether including a scientific paper module in our classes could produce long-term benefits. On the basis of performance in an assessment exam, we found that our module resulted in longitudinal gains, including increased comprehension and critical-thinking abilities in subsequent lab courses. These learning gains were specific to courses utilizing our module, as no longitudinal gains were seen in students who had taken other upper-division labs that lacked extensive primary literature discussion. In addition, we assessed whether performance on our assessment correlated with a variety of factors, including grade point average, course performance, research background, and self-reported confidence in understanding of the article. Furthermore, all of the study conclusions are independent of biology disciplines, as we observe similar trends within each course.