SELECTIVE INACTIVATION OF MICROSOMAL DRUG METABOLIZING PROTEINS BY VISIBLE LIGHT

Abstract— Rat liver microsomes treated with heterochromatic visible light (λ > 400 nm) and O₂ showed a preferential inactivation of NADPH-cytochrome P₄₅₀ reductase and cytochrome P₄₅₀, proteins involved in drug metabolism. Cytochrome P₄₅₀ destruction correlates with lipid peroxidation; both are oxygen dependent and affected to the same extent by antioxidants and radical scavengers. Under anaerobic conditions. NADPH-cytochrome P₄₅₀ reductase is the only component which is inactivated; this is accompanied by FMN loss and activity can be restored by reconstitution.     

Insights into the Ultrafast Photodissociation Dynamics of Isoprene-Derived Criegee Intermediates

Isoprene is the most abundant nonmethane volatile organic compound emitted into the troposphere by terrestrial vegetation. Reaction with ozone represents an important isoprene removal process from the troposphere and is a well-known source of Criegee intermediates (CIs), which are reactive carbonyl oxides. Three CIs, formaldehyde oxide (CH₂OO), methyl vinyl ketone oxide (MVK-oxide) and methacrolein oxide (MACR-oxide) are formed during isoprene ozonolysis. All three CIs contain strongly absorbing ππ* states, electronic excitation, which leads to dissociation to form aldehyde/ketone + oxygen products. Here, we compare the excited state chemistry of CH₂OO, MVK-oxide and MACR-oxide in order to ascertain how increasing molecular complexity affects their photodynamics. In CH₂OO, vertical excitation to the S₂ state leads to prompt O-O bond fission with a unity quantum yield. Branching into both the O (¹D) + H₂CO (S₀) and O (³P) + H₂CO (T₁) product channels is predicted, with 80% of trajectories dissociating to form the former product pair. Analogous vertical excitation of the lowest energy conformers of MVK-oxide and MACR-oxide also undergoes O-O bond fission to form O + MVK/MACR products—albeit with a nonunity quantum yield. In the latter case, ca. 10% and 25% of trajectories remain as the parent MVK-oxide and MACR-oxide molecules, respectively. Additionally, at most only 5% of the dissociating trajectories form O (³P) + MVK/MACR (T₁) products, with a greater fraction forming O (¹D) + MVK/MACR (S₀) products (cf. CH₂OO). This latter observation coupled with the greater fraction of undissociated trajectories aligns with the bathochromic shift in the electronic absorption of the MACR-oxide and MVK-oxide (cf. CH₂OO). We discuss the implications of the results in a broader context, including those that are relevant to the atmosphere.     

Structural characterisation of the Ce1−xLaxNbO4+δ solid solution series: In-situ high-temperature powder diffraction studies

High-resolution neutron powder diffraction has been utilised to investigate the effect of lanthanum substitution on the structure of cerium niobate, CeNbO_4+δ, as a function of temperature. Two members of the Ce_1−xLa_xNbO_4+δ solid solution series, Ce_0.8La_0.2NbO_4+δ and Ce_0.2La_0.8NbO_4+δ, were examined over a temperature range of 293-923 K under a positive pressure of O₂ (500 mbar). From this data it was found that on increasing lanthanum substitution there was an associated reduction in the temperature of the monoclinic-to-tetragonal phase transition. The data also suggested that increasing lanthanum substitution caused an associated decrease in the excess oxygen content. In addition, high-temperature X-ray powder diffraction data recorded in static air were also examined for four compositions of the Ce_1−xLa_xNbO_4+δ series (x=0.2, 0.4, 0.6 and 0.8). These data corroborated the results of the neutron diffraction experiments and also suggested that there was formation of an intermediate phase, analogous to the CeNbO_4.08 phase of the parent material, during the phase transitions of the x=0.8 and 0.6 compositions.     

A simple model for a two-stage chemical reaction with diffusion

We analyse the evolution of a two-stage chemical reaction betweentwo neighbouring plumes of reactants. Under the assumption thatthe plumes are approximately Gaussian we derive a system ofordinary differential equations for the total amount, the centroidand the variance of each reactant. We compare the solution ofthese equations with full numerical simulation of the reaction.Excellent agreement is obtained, with solution of the near-Gaussianmodel requiring considerably less computational effort thanthe full simulations. Of key importance is the yield of thereaction, and we discuss this feature in particular.     

Two-dimensional propagators and spatio-temporal correlations for flow in porous media: A comparative study

The two-dimensional displacement joint probability densityPΔ(X,Z) and the two-time probability density W₂(Z₁,Δ₁;Z₂,A₂) for water flowing through several porous systems have been measured by means of pulsed field-gradient nuclear magnetic resonance (PFG-NMR). The simultaneous particle displacementsX and Z perpendicular and parallel to the pressure gradient, respectively, at a given encoding time Δ are obtained from an experiment employing orthogonal magnetic field gradients. Time-correlated propagators which relate the displacement spectra at two consecutive times Δ_l, and Δ₂ with each other were derived by applying rephasing gradients in two steps. Flow through a random packing of glass beads and through natural sandstone is compared to flow through arrays of either oriented or unoriented fibers with different solid volume fractions. The dependence of the dispersion tensor D^* as a function of time is discussed and related to a characteristic length ξ_tT transverse to the flow direction. Within a certain range of Z values, displacements inX and Z are related by a power law <X²(Z)> ∝Z _γ. The spreading exponent γ is found to increase with growing orientational order in the porous system and is largest for fiber bundles being twisted with respect to the mean pressure gradient axis. The evolution of the correlation coefficient px²,z with time gives a measure for the typical correlation length of the system parallel to the flow axis, ξ_‖. Analyzing the shape ofW ₂(Z₁,Δ₁Z₂,Δ₂) allows one to investigate the loss of coherence in flow by an alternative approach. The decay of the two-time correlation coefficient,pZ ₁,Z₂, is sensitive to the change of the z-component of the particle velocity and probes a different lengthscale thanpx ₂z^.     

Site-specific characterisation of densely O-glycosylated mucin-type peptides using electron transfer dissociation ESI-MS/MS

Site-specific characterisation of mucin-type O-linked glycosylation is an analytical challenge due to glycan heterogeneity, lack of glycosylation site consensus sequence and high density of occupied glycosylation sites. Here, we report the use of electron transfer dissociation (ETD) for the site-specific characterisation of densely glycosylated mucin-type O-linked glycopeptides using ESI-IT-MS/MS. Synthetic glycopeptides from the human mucin-1 (MUC-1) tandem repeat region containing a range of O-linked, tumour-associated carbohydrate antigens, namely Tn, T and sialyl T, with different glycosylation site occupancies and an increasing number of tandem repeats were studied. In addition, a glycopeptide from the anti-freeze glycoprotein of Antarctic and Arctic notothenoids, bearing four O-linked, per-acetylated T antigens was characterised. ETD MS/MS of infused or capillary LC-separated glycopeptides provided broad peptide sequence coverage (c/z·-type fragment ions) with intact glycans still attached to the Ser/Thr residues. Thus, the glycosylation sites were unambiguously determined, while simultaneously obtaining information about the attached glycan mass and peptide identity. Highly sialylated O-glycopeptides showed less efficient peptide fragmentation, but some sequence and glycosylation site information was still obtained. This study demonstrates the capabilities of ETD MS/MS for site-specific characterisation of mucin-type glycopeptides containing high-density O-linked glycan clusters, using accessible and relative low-resolution/low-mass accuracy IT MS instrumentation.     

Proton spin relaxation and self-diffusion in some ‘isotropic’ nematogens

Angular distributions of reactive scattering arising from a long-lived collision complex dissociating via a transition state that approximates to a linear rotator are predicted for H atom displacement occurring over a distribution of bending angles β with respect to the rotator axis. A conical angular distribution may be generated when only a narrow range of bending angles contributes to the reactive scattering. However, the angular distribution may take on quite different character as the distribution of bending angles becomes broader. Thus, an isotropic angular distribution is generated by a bending angle distribution which follows the spherical polar weighting P(β) ∝ sin β. Forward and backward scattering is generated by bending angle distributions favouring less strongly bent displacement, while sideways scattering is generated by a bending angle distribution favouring more strongly bent displacement. The experimental angular distribution for OH + CO reactive scattering which shows mild forward and backward peaking, corresponds to a range of bending angles for H atom displacement from the H-OCO transition state which is broadened about the nominal preferred direction β = 45° but avoids strongly bent directions with β ≈ 90°.     

Imaging cytoplasmic cAMP in mouse brainstem neurons

Background  

cAMP is an ubiquitous second messenger mediating various neuronal functions, often as a consequence of increased intracellular Ca²⁺ levels. While imaging of calcium is commonly used in neuroscience applications, probing for cAMP levels has not yet been performed in living vertebrate neuronal tissue before.     

Time Correlations in Fluid Transport Obtained by Sequential Rephasing Gradient Pulses

We present a basic experiment by which the evolution of the displacement probability density (propagator) of static or flowing fluid inNsuccessive time intervals is obtained by single labeling, coupled with multiple rephasing events during the course of a pulsed field-gradient sequence. We term this type of sequence SERPENT: SEquential Rephasing by Pulsed field-gradients Encoding N Time-intervals. Realizations of the SERPENT experiment for the caseN= 2 which include spin echo, stimulated echo, and Carr–Purcell pulse sequences are suggested. They have in common a spatial spin-labeling of the initial magnetization by a gradient of area q₀, followed by successive rephasing via gradients q₁and q₂at timest= Δ₁andt= Δ₂, respectively, where q₀+ q₁+ q₂= 0. A two-dimensional Fourier transform with respect to q₁and q₂gives directly the joint probability densityW₂(R₁, Δ₁; R₂, Δ₂) for displacements R₁and R₂in times Δ₁and Δ₂, respectively. q₁and q₂may be in arbitrary directions. Assuming R₁R₂, the correlation coefficient ρ_R1,R2then reflects the time-history of the fluctuating velocities. The behavior of the cross moment R₁(Δ₁) · R₂(Δ₂) can be obtained from either a full two-dimensional or a set of one-dimensional SERPENT measurements. Experimental results are presented for water flowing through a bed of packed glass beads. While Δ₁is appropriately chosen to sample the short-time velocity field within the system, increasing Δ₂clearly shows the loss of correlation when the average fluid element displacement exceeds the bead diameter.