Diastereomeric Drug Glucuronides: Enzymatic Glucuronidation and Analysis by Capillary Electrophoresis and Liquid Chromatography-Mass Spectrometry

Foreign compounds, such as drugs are metabolised in the body in numerous reactions. Metabolic reactions are divided into phase I (functionalisation) and phase II (conjugation) reactions. Uridine diphosphoglucuronosyltransferase enzymes (UGTs) are important catalysts of phase II metabolic system. They catalyse the transfer of glucuronic acid to small lipophilic molecules and convert them to hydrophilic and polar glucuronides that are readily excreted from the body. Liver is the main site of drug metabolism. Many drugs are racemic mixtures of two enantiomers. Glucuronidation of a racemic compound yields a pair of diastereomeric glucuronides. Stereoisomers are interesting substrates in glucuronidation studies since some UGTs display stereoselectivity. Diastereomeric glucuronides of O-desmethyltramadol (M1) and entacapone were selected as model compounds in this work. The investigations of the thesis deal with enzymatic glucuronidation and the development of analytical methods for drug metabolites, particularly diastereomeric glucuronides. The glucuronides were analysed from complex biological matrices, such as urine or from in vitro incubation matrices. Various pretreatment techniques were needed to purify, concentrate and isolate the analytes of interest. Analyses were carried out by liquid chromatography (LC) with ultraviolet (UV) or mass spectrometric (MS) detection or with capillary electromigration techniques. Commercial glucuronide standards were not available for the studies. Enzyme-assisted synthesis with rat liver microsomes was therefore used to produce M1 glucuronides as reference compounds. The glucuronides were isolated by LC/UV and ultra performance liquid chromatography (UPLC)/MS, while tandem mass spectrometry (MS/MS) and nuclear magnetic resonance (NMR) spectroscopy were employed in structural characterisation. The glucuronides were identified as phenolic O-glucuronides of M1. To identify the active UGT enzymes in (±)-M1 glucuronidation recombinant human UGTs and human tissue microsomes were incubated with (±)-M1. The study revealed that several UGTs can catalyse (±)-M1 glucuronidation. Glucuronidation in human liver microsomes like in rat liver microsomes is stereoselective. The results of the studies showed that UGT2B7, most probably, is the main UGT responsible for (±)-M1 glucuronidation in human liver. Large variation in stereoselectivity of UGTs toward (±)-M1 enantiomers was observed. Formation of M1 glucuronides was monitored with a fast and selective UPLC/MS method. Capillary electromigration techniques are known for their high resolution power. A method that relied on capillary electrophoresis (CE) with UV detection was developed for the separation of tramadol and its free and glucuronidated metabolites. The suitability of the method to identify tramadol metabolites in an authentic urine samples was tested. Unaltered tramadol and four of its main metabolites were detected in the electropherogram. A micellar electrokinetic chromatography (MEKC) /UV method was developed for the separation of the glucuronides of entacapone in human urine. The validated method was tested in the analysis of urine samples of patients. The glucuronides of entacapone could be quantified after oral entacapone dosing.

Visualization of enantiomers and determination of homo- and hetero-nuclear residual dipolar and scalar couplings: The natural abundant C-13 edited J/D-resolved NMR techniques

The simple two dimensional C-13-satellite J/D-resolved experiments have been proposed for the visualization of enantiomers, extraction of homo- and hetero-nuclear residual dipolar couplings and also H-1 chemical shift differences between the enantiomers in the anisotropic medium. The significant advantages of the techniques are in the determination of scalar couplings of bigger organic molecules. The scalar couplings specific to a second abundant spin such as F-19 can be selectively extracted from the severely overlapped spectrum. The methodologies are demonstrated on a chiral molecule aligned in the chiral liquid crystal medium and two different organic molecules in the isotropic solutions. (C) 2010 Elsevier B.V. All rights reserved.

Nature inspired optimization techniques for the design optimization of laminated composite structures using failure criteria

The design optimization of laminated composites using naturally inspired optimization techniques such as vector evaluated particle swarm optimization (VEPSO) and genetic algorithms (GA) are used in this paper. The design optimization of minimum weight of the laminated composite is evaluated using different failure criteria. The failure criteria considered are maximum stress (MS), Tsai-Wu (TW) and failure mechanism based (FMB) failure criteria. Minimum weight of the laminates are obtained for different failure criteria using VEPSO and GA for different combinations of loading. From the study it is evident that VEPSO and GA predict almost the same minimum weight of the laminate for the given loading. Comparison of minimum weight of the laminates by different failure criteria differ for some loading combinations. The comparison shows that FMBFC provide better results for all combinations of loading. (C) 2010 Elsevier Ltd. All rights reserved.

An investigation of well-characterized small gold clusters by photoelectron spectroscopy, tunneling spectroscopy, and cognate techniques

After microscopic characterization of the size distributions of gold clusters, deposited on carbon substrates by vacuum evaporation or by soft landing, Au(4f') binding energy of the clusters has been measured as a function of the mean cluster size. Similar measurements have been carried out on Au clusters prepared from sols by chemical means and high-nuclearity cluster compounds. In general, small clusters with a mean diameter of $2 nm show significantly larger binding energies than the bulk metal value, due to the onset of nonmetallicity. Nonmetallicity manifests itself in terms of a tunneling conductance gap only in clusters of diameter ;5 1 nm containing 40 atoms or fewer.

Investigation of the (PEG)xLiCl system using conductivity, DSC and NMR techniques

Electrolytes based on polyethylene glycol (PEG, mol.wt.8000) and LiCl of compositions, (PEG)(x)LiCl, x=4, 6, 8, 10, 12, 40, 60, where x is the O/Li ratio, were prepared by solution casting from methanol solutions. FTIR studies indicate that the ether oxygens of the polymer chain participate in Li+ ion conduction. The presence of a salt-polymer complex that melts around 190 degrees C was evidenced by DSC measurements for the electrolytes with compositions x<12. The highest conductivity was obtained at the composition x=10 which was attributed to the presence of a mostly amorphous compound. NMR measurements indicated two regions of motional narrowing, one attributable to the glass transition and another to translational diffusion.

Sport, body and power: Reassessing the myth of President Kekkonen

Former President of Finland Urho Kekkonen was not only a powerful politician but also a well-known sportsman and keep-fit enthusiast. The president’s sports hobbies were covered and celebrated in the media and thus became an integral part of his public persona. This paper looks at Kekkonen’s athletic and able-bodied image and its significance for his power from the perspective of gender. In his exercise activities, Kekkonen was able to display his bodily prowess and demonstrate his version of masculinity, which emphasized both physical and mental strength. The union of mind and muscle in turn buttressed his political ascendancy. Kekkonen’s athletic body served as a cornerstone of his dominance over his country and, simultaneously, as a shield protecting Finland from both internal and external threats. Furthermore, Kekkonen’s sports performances were essential elements in the myth that was created around the president during his term and which was carefully conserved after his fall from power. Drawing upon scholarship on men and masculinities, this paper reassesses the still-effective mythical image of Kekkonen as an invincible superman. The article reveals the performative nature of his athletic activities and shows that in part, his pre-eminence in them was nothing more than theatre enacted by him and his entourage. Thus, Kekkonen’s superior and super-masculine image was actually surprisingly vulnerable and dependent on the success of the performance. The president’s ageing, in particular, demonstrates the fragility of his displays of prowess, strength and masculinity, and shows how fragile the entanglement of body and power can be.

Schlieren visualization of shock wave phenomena over a missile-shaped body at hypersonic Mach numbers

The flow over a missile-shaped configuration is investigated by means of Schlieren visualization in short-duration facility producing free stream Mach numbers of 5.75 and 8. This visualization technique is demonstrated with a 41 degrees full apex angle blunt cone missile-shaped body mounted with and without cavity. Experiments are carried out with air as the test gas to visualize the flow field. The experimental results show a strong intensity variation in the deflection of light in a flow field, due to the flow compressibility. Shock stand-off distance measured with the Schlieren method is in good agreement with theory and computational fluid dynamic study for both the configurations. Magnitude of the shock oscillation for a cavity model may be greater than the case of a model without cavity. The picture of visualization shows that there is an outgoing and incoming flow closer to the cavity. Cavity flow oscillation was found to subside to steady flow with a decrease in the free stream Mach number.

Understanding changes in the Himalayan cryosphere using remote sensing techniques

In the Himalayas, a large area is covered by glaciers and seasonal snow and changes in its extent can influence availability of water in the Himalayan Rivers. In this paper, changes in glacial extent, glacial mass balance and seasonal snow cover are discussed. Glacial retreat was estimated for 1868 glaciers in 11 basins distributed in the Indian Himalaya since 1962. The investigation has shown an overall reduction in glacier area from 6332 to 5329km2 from 1962 to 2001/2 - an overall deglaciation of 16%. Snow line at the end of ablation season on the Chhota Shigri glacier observed using field and satellite methods suggests a change in altitude from 4900 to 5200m from the late 1970s to present. Seasonal snow cover was monitored in the 28 river sub-basins using normalized difference snow index (NDSI) technique in Central and Western Himalaya. The investigation has shown that in the early part of winter, i.e. from October to December, a large amount of snow retreat was observed. For many basins located in lower altitude and in the south of the Pir Panjal range, snow ablation was observed throughout the winter season. In addition, average stream runoff of the Baspa basin for the month of December increased by 75%. This combination of glacial retreat, negative mass balance, early melting of seasonal snow cover and winter-time increase in stream runoff might suggest an influence of global warming on the Himalayan cryosphere.

Ring Resonators and Micro Fluidics: Novel Design for Drug/Disease Detection

MEMS systems are technologically developed from integrated circuit industry to create miniature sensors and actuators. Originally these semiconductor processes and materials were used to build electrical and mechanical systems, but expanded to include biological, optical fluidic magnetic and other systems 12]. Here a novel approach is suggested where in two different fields are integrated via moems, micro fluidics and ring resonators. It is well known at any preliminary stage of disease onset, many physiological changes occur in the body fluids like saliva, blood, urine etc. The drawback till now was that current calibrations are not sensitive enough to detect the minor physiological changes. This is overcome using optical detector techniques 1]. The basic concepts of ring resonators, with slight variations can be used for optical detection of these minute disease markers. A well known fact of ring resonators is that a change in refractive index will trigger a shift in the resonant wavelength 5]. The trigger for the wavelength shift in the case discussed will be the presence of disease agents. To trap the disease agents specific antibody has to be used (e. g. BSA).

3-D Warping in Four-Bar Laminated Linkages

This paper deals with the evaluation of the component-laminate load-carrying capacity, i.e., to calculate the loads that cause the failure of the individual layers and the component-laminate as a whole in four-bar mechanism. The component-laminate load-carrying capacity is evaluated using the Tsai-Wu-Hahn failure criterion for various layups. The reserve factor of each ply in the component-laminate is calculated by using the maximum resultant force and the maximum resultant moment occurring at different time steps at the joints of the mechanism. Here, all component bars of the mechanism are made of fiber reinforced laminates and have thin rectangular cross-sections. They could, in general, be pre-twisted and/or possess initial curvature, either by design or by defect. They are linked to each other by means of revolute joints. We restrict ourselves to linear materials with small strains within each elastic body (beam). Each component of the mechanism is modeled as a beam based on geometrically nonlinear 3-D elasticity theory. The component problems are thus split into 2-D analyses of reference beam cross-sections and nonlinear 1-D analyses along the three beam reference curves. For the thin rectangular cross-sections considered here, the 2-D cross-sectional nonlinearity is also overwhelming. This can be perceived from the fact that such sections constitute a limiting case between thin-walled open and closed sections, thus inviting the nonlinear phenomena observed in both. The strong elastic couplings of anisotropic composite laminates complicate the model further. However, a powerful mathematical tool called the Variational Asymptotic Method (VAM) not only enables such a dimensional reduction, but also provides asymptotically correct analytical solutions to the nonlinear cross-sectional analysis. Such closed-form solutions are used here in conjunction with numerical techniques for the rest of the problem to predict more quickly and accurately than would otherwise be possible. Local 3-D stress, strain and displacement fields for representative sections in the component-bars are recovered, based on the stress resultants from the 1-D global beam analysis. A numerical example is presented which illustrates the failure of each component-laminate and the mechanism as a whole.

Reduced Order Suboptimal Control Design for a Class of Nonlinear Distributed Parameter Systems Using POD and /spl thetas/-D Techniques

A new computational tool is presented in this paper for suboptimal control design of a class of nonlinear distributed parameter systems. First proper orthogonal decomposition based problem-oriented basis functions are designed, which are then used in a Galerkin projection to come up with a low-order lumped parameter approximation. Next, a suboptimal controller is designed using the emerging /spl thetas/-D technique for lumped parameter systems. This time domain sub-optimal control solution is then mapped back to the distributed domain using the same basis functions, which essentially leads to a closed form solution for the controller in a state feedback form. Numerical results for a real-life nonlinear temperature control problem indicate that the proposed method holds promise as a good suboptimal control design technique for distributed parameter systems.