Construction of kinetic models for metabolic reaction networks: Lessons learned in analysing short-term stimulus response data

Construction of dynamic models of large-scale metabolic networks is one of the central issues in the engineering of living cells. However, construction of such models is often hampered by a number of challenges, for example, data availability, compartmentalization and parameter identification coupled with design of in vivo perturbations. As a solution to the latter, short-term perturbation experiments are proposed and are proven to be a useful experimental method to obtain insights into the in vivo kinetic properties of the metabolic pathways.

The aim of this work is to construct a kinetic model using the available experimental data obtained by short-term perturbation experiments, where the steady state of a glucose-limited anaerobic chemostat culture of Saccharomyces cerevisiae was perturbed. In constructing the model, we first determined the steady-state flux distribution using the data before the glucose pulse and the known stoichiometry. For the rate expressions, we used approximative linlog kinetics, which allows the enzyme–metabolite kinetic interactions to be represented by an elasticity matrix. We performed a priori model reduction based on timescale analysis and parameter identifiability analysis allowing the information content of the experimental data to be assessed. The final values of the elasticities are estimated by fitting the model to the available short-term kinetic response data.

The final model consists of 16 metabolites and 14 reactions. With 25 parameters, the model adequately describes the short-term response of the cells to the glucose perturbation, pointing to the fact that the assumed kinetic interactions in the model are sufficient to account for the observed response.     

A novel polyphosphazene with nitroxide radical side groups as cathode‐active material in Li‐ion batteries

A novel polyphosphazene carrying stable nitroxide aromatic radical groups as a pendant with four electrons involvement per repeating unit is synthesized. To do so, series of macromolecular substitution reactions of poly (dichlorophosphazene) with 3,5‐dibromophenol, 2‐methyl‐2‐nitrosopropane, and lead oxide, respectively, are performed. After characterization of the newly synthesized polymers by standard spectroscopic techniques (such as Fourier transform infrared [FT‐IR], nuclear magnetic resonance [NMR], or electron paramagnetic resonance [EPR]), the targeted polymer is further investigated as a cathode‐active material for rechargeable lithium‐ion batteries (LIBs). The cell delivered a good rate performance with a discharge capacity of 100 mAh/g at a C/2 current density over 500 cycles.     

Determination of flurbiprofen in pharmaceutical preparations by GC–MS

This article describes a gas chromatography–mass spectrometry (GC–MS) method for the determination of flurbiprofen in pharmaceutical preparations. The method is based on the derivatization of flurbiprofen with N-methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA). For GC–MS, electron ionization mode (EI = 70 eV) and selected ion monitoring (SIM) mode were used for quantitative analysis (m/z 180 for flurbiprofen). Calibration curve was linear between the concentration range of 0.25–5.0 μg/mL. Intra- and inter-day precision values for flurbiprofen were less than 3.64, and accuracy (relative error) was better than 2.67%. The mean recovery of flurbiprofen was 99.4% for pharmaceutical preparations. The limits of detection and quantification of flurbiprofen were 0.05 and 0.15 μg/mL, respectively. No interference was found from tablet excipients at the selected assay conditions. Also, the method was applied for the quality control of five commercial flurbiprofen dosage forms to quantify the drug and to check the formulation content uniformity.     

NO2 adsorption on ultrathin theta-Al2O3 films: formation of nitrite and nitrate species

Interaction of NO2 with an ordered theta-Al2O3/NiAl(100) model catalyst surface was investigated using temperature programmed desorption (TPD) and X-ray photoelectron spectroscopy (XPS). The origin of the NO(x) uptake of the catalytic support (i.e., Al2O3) in a NO(x) storage catalyst is identified. Adsorbed NO2 is converted to strongly bound nitrites and nitrates that are stable on the model catalyst surface at temperatures as high as 300 and 650 K, respectively. The results show that alumina is not completely inert and may stabilize some form of NO(x) under certain catalytic conditions. The stability of the NO(x) formed by exposing the theta-Al2O3 model catalyst to NO2 adsorption increases in the order NO2 (physisorbed or N2O4) < NO2 (chemisorbed) < NO2- < NO3-.     

Quantification and identification of components released from dental composites using different chromatographic techniques

This study aimed to review recent chromatographic methods for quantifying and identifying components released from dental composites. Resin-based dental restorative materials are extensively used in dentistry today. Although composite materials are known to be highly stable structures, they are susceptible to degradation because of the incomplete polymerization. Several components may be released from resin composite restorations into the oral environment. The elution of components from composite resins may affect the biocompatibility of the restorations. Therefore, it is essential to understand the nature and quantity of substances that are segregated into the oral cavity.     

Some Gaussian binomial sum formulæ with applications

We introduce and compute some Gaussian q-binomial sums formulæ. In order to prove these sums, our approach is to use q-analysis, in particular a formula of Rothe, and computer algebra. We present some applications of our results.     

Scanning Ion Conductance Microscopic Study for Cellular Uptake of Cationic Conjugated Polymer Nanoparticles

Positively charged conjugated polymer nanoparticles (CPNs) are emerging biomaterials exhibiting high levels of cellular entry. High rate of cellular entry efficiency is believed that the amphiphilic CPNs interact efficiently with the negatively charged hydrophobic cellular membranes. For the first time, the cell surface morphological changes of human cervical cancer cells treated with CPNs using a scanning probe microscopy technique, scanning ion conductance microscopy (SICM) are imaged. After 1 h of CPN incubation, distinct changes are observed in cell surface morphology such as interconnected protrusions and pits with sub‐micrometer sizes, which are not observed from cells treated with positively charged polyethyleneimine (PEI) under the same treatment conditions. The change on cell surface morphology is quantified by surface roughness ratio, which is increased as CPN concentration increases, while the ratio first increases and then decreases as the incubation time increases. These results suggest that cells respond actively toward CPN with both positive charges on the side chain and the hydrophobicity from rigid aromatic backbone, which leads to subsequent endocytosis. In conclusion, it is demonstrated that SICM is a suitable imaging technique to reveal the dynamic alternations on the cell surface morphology at the early stage of nanoparticles endocytosis with high resolution.

     

Testing the durability of copper based preservative treated bamboos in ground-contact for six years

Cellulose - In this study, durability of bamboo samples in terms of the variability of location along culm height (top, middle and bottom) were evaluated in a ground-contact field test for six...     

Divisibility of L-polynomials for a family of Artin–Schreier curves

In this paper we consider the curves $C_k^{(p,a)}:y^p?y=x^{p^k+1}+ax$ defined over ${\mathbb{F}}_p$ and give a positive answer to a conjecture about a divisibility condition on L-polynomials of the curves $C_k^{(p,a)}$. Our proof involves finding an exact formula for the number of ${\mathbb{F}}_{p^n}$-rational points on $C_k^{(p,a)}$ for all n, and uses a result we proved elsewhere about the number of rational points on supersingular curves.     

Physical and Covalent Immobilization of <Emphasis Type="Italic">Lipase</Emphasis> onto Amine Groups Bearing Thiol-Ene Photocured Coatings

In this study, amine groups containing thiol-ene photocurable coating material for lipase immobilization were prepared. Lipase (EC 3.1.1.3) from Candida rugosa was immobilized onto the photocured coatings by physical adsorption and glutaraldehyde-activated covalent bonding methods, respectively. The catalytic efficiency of the immobilized and free enzymes was determined for the hydrolysis of p-nitrophenyl palmitate and also for the synthesis of p-nitrophenyl linoleate. The storage stability and the reusability of the immobilized enzyme and the effect of temperature and pH on the catalytic activities were also investigated. The optimum pH for free lipase and physically immobilized lipase was determined as 7.0, while it was found as 7.5 for the covalent immobilization. After immobilization, the optimum temperature increased from 37 °C (free lipase) to 50–55 °C. In the end of 15 repeated cycles, covalently bounded enzyme retained 60 and 70 % of its initial activities for hydrolytic and synthetic assays, respectively. While the physically bounded enzyme retained only 56 % of its hydrolytic activity and 67 % of its synthetic activity in the same cycle period. In the case of hydrolysis V _max values slightly decreased after immobilization. For synthetic assay, the V _max value for the covalently immobilized lipase was found as same as free lipase while it decreased dramatically for the physically immobilized lipase. Physically immobilized enzyme was found to be superior over covalent bonding in terms of enzyme loading capacity and optimum temperature and exhibited comparable re-use values and storage stability. Thus, a fast, easy, and less laborious method for lipase immobilization was developed.