A more sensitive MS detector does not obviously lead to a more sensitive assay: experiences with ES-285

In this paper the transfer of an existing method for the quantitative determination of the anticancer agent ES-285 in human plasma using liquid chromatography tandem mass spectrometry on an API 365 to an API 3000 mass spectrometer is described. The transfer appeared not to be straightforward. Problems arose resulting from carry-over and interferences. In addition, due to the expansion of the calibration range, data ought to be weighted with a different factor to increase the accuracy of the lower concentrations. After finding appropriate solutions for these problems, the lower limit of quantitation could be lowered from 10 to 1 ng/mL for ES-285 in human plasma. The usefulness and necessity of the modified assay was demonstrated by analysis of plasma samples from a patient receiving a low dosage of the drug.     

Vocal Tract Resonance Analysis of Aging Voice Using Long-Term Average Spectra

This study is the first to use long-term average spectra (LTAS) to investigate resonance characteristics of dynamic speech in young adulthood and old age. A total of 80 speakers participated, divided equally by age group and gender. All elderly speakers were healthy, active members of the community. Measurement of the first three spectral peaks in LTAS from the first paragraph of the Rainbow Passage revealed significant lowering of peak 1 from young adulthood to old age in both men and women. Peaks 2 and 3 also lowered significantly across the adult lifespan in women and showed a tendency to lower in men. These acoustic findings are consistent with anatomic data suggesting that aging results in lengthening of the supraglottic vocal tract. Findings that women demonstrate more substantial lowering of spectral peaks with aging than men suggest that women may undergo more pronounced age-related lengthening of the supraglottic vocal tract. Alternatively, it is possible that elderly men systematically alter tongue position during vowel articulation while elderly women are less inclined to do so. Taken in conjunction with previous research, these findings suggest a "mixed model" of vocal tract resonance changes with aging in which an interaction exists between gender, the resonance effects of laryngeal lowering, and vowel articulatory patterns.     

How Students “Unpack” the Structure of a Word Problem: Graphic Representations and Problem Solving

This research investigated how fourth and fifth grade students spontaneously ‘unpacked’ a word problem when generating a graphic representation to aid in problem solution. Relationships among the type of graphic representation produced, spatial visualization, drawing ability, gender, and problem solving also were examined and described. Instrumentation developed for the study included several math challenge tasks, a spatial visualization task, and a drawing task. For one of the math challenge tasks, students were instructed to draw a picture to assist them with problem solution. These graphic representations generated by students were rated as pictorial or as displaying some level of schematic representation. Schematic representations included germane information from the problem supportive of problem solution. Pictorial representations included expressive and extraneous elements not necessary for problem solution, with no schematic elements. Findings indicated that the majority of students rendered schematic representations, with girls more likely than boys to use schematic representations at a statistically significant level. Students who used schematic visual representations were more successful problem solvers than those pictorially representing problem elements. The more “schematic‐like” the visual representation, the more successful students were at problem solution. Drawing a pictorial representation in the math challenge task also was negatively correlated to drawing skill.     

Spontaneous Assembly of Magnetic LEGO Bricks

This paper describes simple methods of demonstrating macroscale spontaneous assembly. These demonstrations can illustrate topics in college or high school chemistry courses, such as the thermodynamics of crystallization and the chelate effect. The assembling units are built from a combination of magnetic and conventional LEGO building blocks. Unlike many other spontaneous-assembly experiments, these assemblies do not require surface flotation to form.     

Efficient protocol for quantum Monte Carlo calculations of hydrogen abstraction barriers: Application to methanol

Accurate calculation of hydrogen abstraction reaction barriers is a challenging problem, often requiring high level quantum chemistry methods that scale poorly with system size. Quantum Monte Carlo (QMC) methods provide an alternative approach that exhibit much better scaling, but these methods are still computationally expensive. We describe approaches that can significantly reduce the cost of QMC calculations of barrier heights, using the hydrogen abstraction of methanol by a hydrogen atom as an illustrative example. By analysing the combined influence of trial wavefunctions and pseudopotential quadrature settings on the barrier heights, variance, and time‐step errors, we devise a simple protocol that minimizes the cost of the QMC calculations while retaining accuracy comparable to large‐basis coupled cluster theory. We demonstrate that this protocol is transferable to other hydrogen abstraction reactions.     

Llama-derived single-domain antibodies for the detection of botulinum A neurotoxin

Single-domain antibodies (sdAb) specific for botulinum neurotoxin serotype A (BoNT A) were selected from an immune llama phage display library derived from a llama that was immunized with BoNT A toxoid. The constructed phage library was panned using two methods: panning on plates coated with BoNT A toxoid (BoNT A Td) and BoNT A complex toxoid (BoNT Ac Td) and panning on microspheres coupled to BoNT A Td and BoNT A toxin (BoNT A Tx). Both panning methods selected for binders that had identical sequences, suggesting that panning on toxoided material may be as effective as panning on bead-immobilized toxin for isolating specific binders. All of the isolated binders tested were observed to recognize bead-immobilized BoNT A Tx in direct binding assays, and showed very little cross-reactivity towards other BoNT serotypes and unrelated protein. Sandwich assays that incorporated selected sdAb as capture and tracer elements demonstrated that all of the sdAb were able to recognize soluble (“live”) BoNT A Tx and BoNT Ac Tx with virtually no cross-reactivity with other BoNT serotypes. The isolated sdAb did not exhibit the high degree of thermal stability often associated with these reagents; after the first heating cycle most of the binding activity was lost, but the portion of the protein that did refold and recover antigen-binding activity showed only minimal loss on subsequent heating and cooling cycles. The binding kinetics of selected binders, assessed by both an equilibrium fluid array assay as well as surface plasmon resonance (SPR) using toxoided material, gave dissociation constants (K _D ) in the range 2.2 × 10⁻¹¹ to 1.6 × 10⁻¹⁰ M. These high-affinity binders may prove beneficial to the development of recombinant reagents for the rapid detection of BoNT A, particularly in field screening and monitoring applications.