A Biosilification Fusion Protein for a ‘Self‐immobilising’ Sarcosine Oxidase Amperometric Enzyme Biosensor

Monomeric sarcosine oxidase (mSOx) fusion with the silaffin peptide, R5, designed previously for easy protein production in low resource areas, was used in a biosilification process to form an enzyme layer electrode biosensor. mSOx is a low activity enzyme (10–20 U/mg) requiring high amounts of enzyme to obtain an amperometric biosensor signal, in the clinically useful range <1 mM sarcosine, especially since the K_m is >10 mM. An amperometric biosensor model was fitted to experimental data to investigate dynamic range. mSOx constructs were designed with 6H (6×histidine) and R5 (silaffin) peptide tags and compared with native mSOx. Glutaraldehyde (GA) cross‐linked proteins retained ～5 % activity for mSOx and mSOx‐6H and only 0.5 % for mSOx‐R5. In contrast R5 catalysed biosilification on (3‐mercaptopropyl) trimethoxysilane (MPTMS) and tetramethyl orthosilicate (TMOS) particles created a ‘self‐immobilisation’ matrix retaining 40 % and 76 % activity respectively. The TMOS matrix produced a thick layer (>500 μm) on a glassy carbon electrode with a mediated current due to sarcosine in the clinical range for sarcosinemia (0–1 mM). The mSOx‐R5 fusion protein was also used to catalyse biosilification in the presence of creatinase and creatininase, entrapping all three enzymes. A mediated GC enzyme linked current was obtained with dynamic range available for creatinine determination of 0.1–2 mM for an enzyme layer ～800 nm.     

Student-Invented Numeration Systems: Pattern-Analysis and Mathematical Understanding

Thirteen fifth graders were given an assignment to invent their own numeration systems, following a unit on bases and a look at early events in the history of numbers. The task presented options that required the students to make decisions (such as whether to use a base, which base to use, design of symbols, etc.), and build a rationale for the elements of their system. Analyses of patterns embedded in their invented systems provided an assessment of student understanding of numeration. The progression of more and less complex thinking related to the student's choice of a base other than 10, consistency of logic throughout the system in words and symbols, rationale for change, and perception of real life examples that would change if the system was adopted. The invention task is presented as another way to make connections.     

Truncated Newton method for sparse unconstrained optimization using automatic differentiation

When solving large complex optimization problems, the user is faced with three major problems. These are (i) the cost in human time in obtaining accurate expressions for the derivatives involved; (ii) the need to store second derivative information; and (iii), of lessening importance, the time taken to solve the problem on the computer. For many problems, a significant part of the latter can be attributed to solving Newton-like equations. In the algorithm described, the equations are solved using a conjugate direction method that only needs the Hessian at the current point when it is multiplied by a trial vector. In this paper, we present a method that finds this product using automatic differentiation while only requiring vector storage. The method takes advantage of any sparsity in the Hessian matrix and computes exact derivatives. It avoids the complexity of symbolic differentiation, the inaccuracy of numerical differentiation, the labor of finding analytic derivatives, and the need for matrix store. When far from a minimum, an accurate solution to the Newton equations is not justified, so an approximate solution is obtained by using a version of Dembo and Steihaug's truncated Newton algorithm (Ref. 1).This paper was presented at the SIAM National Meeting, Boston, Massachusetts, 1986.     

Announcement of Proficiency Testing Schemes

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Products of residue currents of Cauchy–Fantappiè–Leray type

With a given holomorphic section of a Hermitian vector bundle, one can associate a residue current by means of Cauchy–Fantappiè–Leray type formulas. In this paper we define products of such residue currents. We prove that, in the case of a complete intersection, the product of the residue currents of a tuple of sections coincides with the residue current of the direct sum of the sections.     

Ring expansion of 5- to 6-member zirconacycles by carbenoid insertion

A wide range of carbenoids (1-lithio-1-halo species), including those with α-SiR₃, OEt, SPh, SO₂Ph, P(O)(OEt)₂, and CN substituents, insert into 5-member zirconacycles (saturated and unsaturated, mono- and bi-cyclic) to afford functionalized 6-member zirconacycles. 1-Lithio-1-haloalkenes insert to afford 6-member zirconacycles with an alkylidene substituent next to the metal. Unexpected double insertion of some carbenoids, and evidence for endocylic β-hydride transfer processes provide additional mechanistic interest.     

The Central Limit Problem for Random Vectors with Symmetries

Motivated by the central limit problem for convex bodies, we study normal approximation of linear functionals of high-dimensional random vectors with various types of symmetries. In particular, we obtain results for distributions which are coordinatewise symmetric, uniform in a regular simplex, or spherically symmetric. Our proofs are based on Stein’s method of exchangeable pairs; as far as we know, this approach has not previously been used in convex geometry. The spherically symmetric case is treated by a variation of Stein’s method which is adapted for continuous symmetries. This work was done while at Stanford University.     

Efficient N- and C-functionalisation of cyclam macrocycles utilising bisaminal methodology

An efficient synthesis of C-functionalised cyclam macrocycles that employs bisaminal intermediates and allows N-substitution to be controlled is reported.     

Expert consensus in solo voice performance evaluation

Experts were interviewed to identify criteria for evaluation of vocal performance. A scale was then constructed and inter- and intrajudge reliability assessed. Experts listened to 19 different performances, plus 6 presented a second time. Interjudge reliability for one judge was modest, but increased dramatically as the size of the judge panel increased. The most reliable items were overall score and intonation accuracy. Diction was less reliable than other items. Intrajudge reliability was higher for overall score than for any other item. A factor analysis on the test items yielded factors labelled intrinsic quality, execution, and diction. Another factor analysis, using the experts as variables, revealed two underlying evaluative dimensions. It was found that 13 experts were primarily influenced by execution, and that 8 were mainly affected by intrinsic quality. Interjudge and intrajudge reliabilities of these two groups differed.