CYP154C5 Regioselectivity in Steroid Hydroxylation Explored by Substrate Modifications and Protein Engineering**

CYP154C5 from Nocardia farcinica is a P450 monooxygenase able to hydroxylate a range of steroids with high regio- and stereoselectivity at the 16α-position. Using protein engineering and substrate modifications based on the crystal structure of CYP154C5, an altered regioselectivity of the enzyme in steroid hydroxylation had been achieved. Thus, conversion of progesterone by mutant CYP154C5 F92A resulted in formation of the corresponding 21-hydroxylated product 11-deoxycorticosterone in addition to 16α-hydroxylation. Using MD simulation, this altered regioselectivity appeared to result from an alternative binding mode of the steroid in the active site of mutant F92A. MD simulation further suggested that the entrance of water to the active site caused higher uncoupling in this mutant. Moreover, exclusive 15α-hydroxylation was observed for wild-type CYP154C5 in the conversion of 5α-androstan-3-one, lacking an oxy-functional group at C17. Overall, our data give valuable insight into the structure–function relationship of this cytochrome P450 monooxygenase for steroid hydroxylation.     

Quantification of secondary structure prediction improvement using multiple alignments

The use of multiple sequence alignments for secondary structurepredictions is analysed. Seven different protein families, containingonly sequences of known structure, were considered to providea range of alignment and prediction conditions. Using alignmentsobtained by spatial superposition of main chain atoms in knowntertiary protein structures allowed a mean of 8% in secondarystructure prediction accuracy, when compared to those obtainedfrom the individual sequences. Substitution of these alignmentsby those determined directly from an automated sequence alignmentalgorithm showed variations in the prediction accuracy whichcorrelated with the quality of the multiple alignments and distanceof the primary sequence. Secondary structure predictions canbe reliably improved using alignments from an automatic alignmentprocedure with a mean increase of 6.87percnt;, giving an overallprediction accuracy of 68.5%, if there is a minimum of 25% sequenceidentity between all sequences in a family.     

Some aspects of the role of coal thermoplasticity and coke structure in coal gasification. 2. The effects of pressure and heating rate on dilatometric parameters and optical anisotropy development

An examination of the influence of pressure and heating rate on the thermoplastic properties of a weakly coking coal and of coal with various pitch-like additives was carried out using a high pressure dilatometer. Pressure up to 4 MPa markedly increased the swelling properties and increased the plastic range by decreasing the softening temperature, but the effects of pressure were strongly influenced by rate of heating with high heating rates enhancing the effects. Additions of tar or pitch also enhanced swelling at low pressure. The solid carbonization residues from the dilatometer were examined by polarized light microscopy to determine the content and composition of optically anisotropic species. The anisotropic content was increased by increases in pressure and heating rate and pitch additives enhanced the anisotropic content, especially at low pressure, without affecting the composition of the anisotropy. No direct correlations exist between the dilatometric parameters and the optical anisotropy but their dependence on the conditions emphasizes that when considering high pressure gasification of coal, it is necessary to obtain data under appropriate conditions.     

Determinants of users’ continuance of social networking sites: A self-regulation perspective

Social networking sites (SNS) have transformed how individuals interact, build and maintain social relationships. We proposed a research model on the determinants of user continuance using Bagozzi's framework of self-regulation as the theoretical foundation. Following the process of appraisal → emotional reactions → coping responses, we developed the model by leveraging findings from social presence and IS continuance research. Based on survey data from Facebook users, we found that appraisal factors (pleasure, awareness, connectedness, and system quality) were strong determinants of emotional reaction (user satisfaction and sense of belonging). User satisfaction and sense of belonging together positively influenced continuance intention.     

Evaluation of four steering wheels to determine driver hand placement in a static environment

While much research exists on occupant packaging both proprietary and in the literature, more detailed research regarding user preferences for subjective ratings of steering wheel designs is sparse in published literature. This study aimed to explore the driver interactions with production steering wheels in four vehicles by using anthropometric data, driver hand placement, and driver grip design preferences for Generation-Y and Baby Boomers. In this study, participants selected their preferred grip diameter, responded to a series of questions about the steering wheel grip as they sat in four vehicles, and rank ordered their preferred grip design. Thirty-two male participants (16 Baby Boomers between ages 47 and 65 and 16 Generation-Y between ages 18 and 29) participated in the study. Drivers demonstrated different gripping behavior between vehicles and between groups. Recommendations for future work in steering wheel grip design and naturalistic driver hand positioning are discussed.     

Factors influencing the efficacy of an interfacial modifier for the interface in an immiscible polymer blend

The modification of the interface in immiscible polymer blends is critical in order to optimize physical properties. Despite the great commercial importance of polymer blends, many aspects of the emulsification process remain unclear In this paper, it will be shown that an emulsification curve can be used to estimate the influence of the architecture molar mass of styrene/hydrogenated butadiene block copolymer interfacial modifiers on the emulsifion of polystyrene/ethylene-propylene rubber blends under melt processing conditions. An expression for eing the critical amount of copolymer to saturate the interface, as well as aspects concerning the localisation of the modifier in the blend system will also be addressed.     

Crystallization behavior of oxyethylene/oxybutylene diblock and triblock copolymers

The crystallization behavior of poly(oxyethylene)-b-poly(oxybutylene) block copolymers with different compositions, morphologies and architectures (E_mB_n diblock copolymers and E_mB_nE_m, B_nE_mB_n triblock copolymers) were investigated and the effect of volume fraction and architecture on the crystallization temperature (T_c) in non-isothermal crystallization was determined. It is found that the E_mB_n diblock copolymers having long E blocks exhibit similar crystallization temperatures, irrespective of volume fraction and morphology, but for the block copolymers with shorter E blocks the crystallization temperature increases with both the volume fraction, φ_E, and the length, m, of the E block. Some block copolymers with extremely low T_c, which fall into the temperature range normally associated with homogenous nucleation, were chosen for time-resolved small-angle X-ray scattering (SAXS) and isothermal crystallization kinetics experiments. The results show that breakout crystallization occurs in all these block copolymers. Therefore, unlike E_mB_n/B_h blends, there is no obvious relationship between T_c and crystallization behavior in neat block copolymers and homogeneous nucleation does not definitely lead to confined crystallization. The values of χ_c/χ_ODT for all the block copolymers with hex and bcc morphology were also calculated. It is found that all the block copolymers have χ_c/χ_ODT<3, in agreement with the previously reported critical value and consistent with their breakout crystallization behavior.     

Substrate-mediated interactions and intermolecular forces between molecules adsorbed on surfaces

Adsorbate interactions and reactions on metal surfaces have been investigated using scanning tunneling microscopy. The manners in which adsorbates perturb the surface electronic structure in their vicinity are discussed. The effects these perturbations have on other molecules are shown to be important in overlayer growth. Interactions of molecules with surface steps are addressed, and each molecule's electron affinity is shown to dictate its adsorption sites at step edges. Standing waves emanating from steps are demonstrated to effect transient molecular adsorption up to 40 A away from the step edge. Halobenzene derivatives are used to demonstrate how the surface is important in aligning reactive intermediates.     

Étude expérimentale du mélange réactif au sein d'un réacteur torique ondulé (RTO)

Reactive mixing in a wavy torus reactor (WTR) has been examined experimentally. Its performances are compared to these of two other reactors: a plane torus reactor (PTR) and a baffled stirred tank reactor (BSTR). hydrodynamics of mixing has been studied using a plug flow model with axial dispersion for the WTR and PTR. The effect of axial dispersion on the kinetic of a chemical reaction was then analysed for the WTR, PTR and BSTR. Mixing times were slightly longer for the WTR due to a weaker axial dispersion. We also noticed an improvement of the convertion rate for low dissipated power.