Rapid characterization of protein productivity and production stability of CHO cells by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

The biopharmaceutical industry has been in pursuit of strategies which can isolate stable and high-producing cell lines. The whole cell mass spectrometry method by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) is a rapid and simple method for cell characterization based on the differences in the fingerprints of the mass spectra. This work describes how the method was evaluated for the application of screening for stable and high-producing clones from a panel of recombinant Chinese hamster ovary (CHO) cell lines. Detectable m/z values and their relative intensities were collected and processed by partial least squares (PLS). To reduce the errors introduced by the preparation method and spectra noise, high intensity preliminary data was selected and the number of variables introduced was validated by leave-one-out cross-validation. The differences in recombinant protein productivity and titer were revealed by PLS regression with promising results. Partial least-squares discriminant analysis (PLS-DA) was applied to differentiate stable and unstable cell lines as traditional stability testing would require several months involving numerous continuous passages. Results confirmed that the whole cell MALDI-TOF method can be a powerful method for routine monitoring of bioprocesses and study can be further developed by extending the number of the cell lines tested to establish a recombinant cell line database.     

Squaramides: physical properties, synthesis and applications

Squaramides are remarkable four-membered ring systems derived from squaric acid that are able to form up to four hydrogen bonds. A high affinity for hydrogen bonding is driven through a concomitant increase in aromaticity of the ring. This hydrogen bonding and aromatic switching, in combination with structural rigidity, have been exploited in many of the applications of squaramides. Substituted squaramides can be accessed via modular synthesis under relatively mild or aqueous conditions, making them ideal units for bioconjugation and supramolecular chemistry. In this tutorial review the fundamental electronic and structural properties of squaramides are explored to rationalise the geometry, conformation, reactivity and biological activity.     

Promoting interdisciplinarity through mathematical modelling

This article presents one approach to addressing the important issue of interdisciplinarity in the primary school mathematics curriculum, namely, through realistic mathematical modelling problems. Such problems draw upon other disciplines for their contexts and data. The article initially considers the nature of modelling with complex systems and discusses how such experiences differ from existing problem-solving activities in the primary mathematics curriculum. Principles for designing interdisciplinary modelling problems are then addressed, with reference to two mathematical modelling problems—one based in the scientific domain and the other in the literary domain. Examples of the models children have created in solving these problems follow. A reflection on the differences in the diversity and sophistication of these models raises issues regarding the design of interdisciplinary modelling problems. The article concludes with suggested opportunities for generating multidisciplinary projects within the regular mathematics curriculum.     

How useful is commonality? Inventory and production decisions to maximize survival probability in start-ups

This paper deals with component commonality in start-up manufacturingfirms. We present a two-product Markov decision model that examinesthe implications of the inventory and production strategiesfor the survival probability of the firm. The advantage of usingcomponent commonality is studied for varying costs, demand correlationsand order replenishment lead times. Optimal policies are derived,and minimum stock levels for survival are obtained. Moreover,we state the conditions under which simplified production decisionscan be made. It is shown that commonality is not only usefulas a way of dealing with demand uncertainty, but that its increaseduse is preferred for strongly substitutable products, and shorterreplenishment lead times.     

Cis-UROCANIC ACID SYNERGIZES WITH HISTAMINE FOR INCREASED PGE2 PRODUCTION BY HUMAN KERATINOCYTES: LINK TO INDOMETHACIN-INHIBITABLE UVB-INDUCED IMMUNOSUPPRESSION

Abstract— There is considerable evidence that suppression of the immune system by UVB (280–320 nm UV) irradiation is initiated by UVB-dependent isomerization of a specific skin photoreceptor, urocanic acid (UCA), from the trans to the cis form. Previous studies have confirmed that cis -UCA administration to mice 3–5 days prior to hapten sensitization at a distant site, suppresses the contact hypersensitivity (CHS) response upon challenge. This study demonstrates in mice that cis -UCA, like UVB, suppresses CHS to trinitrochlorobenzene by a mechanism partly dependent on prostanoid production. In vitro experimentation showed that human keratinocytes, isolated from neonatal foreskin, increased prostaglandin E₂ (PGE₂) production in response to histamine but not UCA alone. However, cis -UCA synergized with histamine for increased PGE₂ production by keratinocytes. cis -urocanic acid also increased the sensitivity of keratinocytes for PGE₂ production in response to histamine. Prostaglandin E₂ from keratinocytes exposed to cis -UCA and histamine may contribute directly, or indirectly, to the regulation of CHS responses by UVB irradiation.     

The binding of hafnium to nitrilotriacetate

Using the TDPAC method the formation of Hf-Nitrilotriacetate (Hf-NTA) was investigated. The dependence of the complex formation on the molar ratios of Hf to NTA and upon the pH value was studied. With additional data taken from the chemical analogue Zr-NTA the structure of the complex can be given.     

Modifying uncertainty from sampling to achieve fitness for purpose: a case study on nitrate in lettuce

Existing methods have been applied to estimate the uncertainty of measurement, caused by both sampling and analysis, and fitness-for-purpose of these measurements. A new approach has been taken to modify the measurement uncertainty by changing the contribution made by the sampling process. A case study on nitrate in lettuce has been used to demonstrate the applicability of this new generic approach. The sampling theory of Gy was used to predict the alterations in the sampling protocol required to achieve the necessary change in sampling uncertainty. An experimental application of this altered sampling protocol demonstrated that the predicted change in sampling uncertainty was achieved in practice. For the lettuce case study, this approach showed that composite samples containing 40 heads, rather than the usual ten heads, produced measurements of nitrate that where more fit-for-purpose.     

Antifungal metabolites from Blumea balsamifera

The leaves of Blumea balsamifera afforded icthyothereol acetate, cryptomeridiol, lutein, and beta-carotene. The structures of icthyothereol acetate (1) and cryptomeridiol (2) were elucidated by extensive 1D and 2D NMR spectroscopy, while those of lutein and beta-carotene were identified by comparison with literature data. Antimicrobial tests indicated that 1 has moderate activity against the fungi Aspergillus niger, Trichophyton mentagrophytes, and Candida albicans, while 2 has low activity against A. niger, T. mentagrophytes, and C. albicans. Both compounds have no antimicrobial activity against Psuedomonas aeruginosa, Staphylococcus aureus, Bacillus subtilis, and Escherichia coli.     

Optimised uncertainty in food analysis: application and comparison between four contrasting 'analyte-commodity' combinations

The optimised uncertainty (OU) methodology is applied across a range of analyte-commodity combinations. The commodities and respective analytes under investigation were chosen to encompass a range of input factors: measurement costs (sampling and analytical), sampling uncertainties, analytical uncertainties and potential consequence costs which may be incurred as a result of misclassification. Two types of misclassification are identified-false compliance and false non-compliance. These terms can be used across a wide range of foodstuffs that have regulations requiring either minimum compositional requirements, maximum contaminant allowances or compositional specifications. The latter refers to foodstuffs with regulations that state an allowable tolerance around the compositional specification, i.e. the upper specification limit (USL) and the lower specification limit (LSL). The traditional OU methodology has been adapted so that it is applicable in these cases and has been successfully applied in practice. The Newton-Raphson method has been used to determine the optimal uncertainty value for the two case studies in which analyte concentration is assessed against a 'single threshold' regulatory requirement. This numerical method was shown to give a value of the optimal uncertainty that is practically identical to that given by the previously used method of visual inspection. The expectation of financial loss was reduced by an average of 65% over the four commodities by the application of the OU methodology, showing the benefit of the method.     

TDPAD studies on19F implanted into diamond

¹⁹F has been used to probe the electric field gradients in the diamond lattice. The measurements were performed via the time differential perturbed angular distribution (TDPAD) technique applied to the 197 keV state (=128 ns, Q=0.10 b) of¹⁹F. The nuclei were excited and recoil implanted into the diamond crystal using a (p,p)-reaction initiated by a pulsed 4 MeV proton beam. Spin rotation spectra were generated to study the dependence of the electric field gradients on the orientation of the crystal planes, the diamond type, and temperature.We are greatly indebted to Dr. Thies for invaluable help in evaluation of the data.