The role of phenylalanine 4-monooxygenase (PAH) in the S-oxidation of S-carboxymethyl-L-cysteine (SCMC) in the rat has now been well established in rat cytosolic fractions in vitro. However, the role of PAH in the S-oxidation of SCMC in human cytosolic fractions or hepatocytes has yet to be investigated. The aim of this investigation was to analyse the kinetic parameters of PAH oxidation of both L-phenylalanine (Phe) and SCMC in the human HepG2 cell line in order to investigate the use of these cells as a model for the cellular regulation of SCMC S-oxidation. The experimentally determined Km and V(max) were 7.14 +/- 0.32 mM and 0.85 +/- 0.32 nmole Tyr formed min(-1) x mg protein(-1) using Phe as substrate. For SCMC the values were 25.24 +/- 5.91 mM and 0.79 +/- 0.09 nmole SCMC (RIS) S-oxides formed min(-1) x mg protein(-1). The experimentally determined Km and V(max) for the cofactor BH4 were 6.81 +/- 0.21 microM and 0.41 +/- 0.004 nmole Tyr formed min(-1) x mg protein(-1) for Phe and 7.24 +/- 0.19 microM and 0.42 +/- 0.002 nmole SCMC (R/S) S-oxides formed min(-1) x mg protein(-1) for SCMC. The use of various PAH inhibitors confirmed that HepG2 cells contained PAH and that the enzyme was capable of converting SCMC to its (R) and (S) S-oxide metabolites in an in vitro PAH assay. Thus HepG2 cells have become a useful additional tool for the investigation of the cellular regulation of PAH in the S-oxidation of SCMC.

We study multi-frequency transitions in the transient dynamics of a viscously damped dispersive finite rod with an essentially nonlinear end attachment. The attachment consists of a small mass connected to the rod by means of an essentially nonlinear stiffness in parallel to a viscous damper. First, the periodic orbits of the underlying hamiltonian system with no damping are computed, and depicted in a frequency–energy plot (FEP). This representation enables one to clearly distinguish between the different types of periodic motions, forming back bone curves and subharmonic tongues. Then the damped dynamics of the system is computed; the rod and attachment responses are initially analyzed by the numerical Morlet wavelet transform (WT), and then by the empirical mode decomposition (EMD) or Hilbert–Huang transform (HTT), whereby, the time series are decomposed in terms of intrinsic mode functions (IMFs) at different characteristic time scales (or, equivalently, frequency scales). Comparisons of the evolutions of the instantaneous frequencies of the IMFs to the WT spectra of the time series enables one to identify the dominant IMFs of the signals, as well as, the time scales at which the dominant dynamics evolve at different time windows of the responses; hence, it is possible to reconstruct complex transient responses as superposition of the dominant IMFs involving different time scales of the dynamical response.\ud Moreover, by superimposing the WT spectra and the instantaneous frequencies of the IMFs to the FEPs of the underlying hamiltonian system, one is able to clearly identify the multi-scaled transitions that occur in the transient damped dynamics, and to interpret them as ‘jumps’ between different branches of periodic orbits of the underlying hamiltonian system. As a result, this work develops a physics-based, multi-scaled framework and provides the necessary computational tools for multi-scaled analysis of complex multi-frequency transitions of essentially nonlinear dynamical systems

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Abstract. In this article, the problem of model reduction of 2-D systems is studied via orthogonal series. The algorithm proposed reduces the problem to an overdetermined linear algebraic system of equations, which may readily be solved to yield the simplified model. When this model approximates adequately the original system, it has many important advantages, e.g., it simplifies the analysis and simulation of the original system, it reduces the computational effort in design procedures, it reduces the hardware complexity of the system, etc. Several examples are included which illustrate the efficiency of the proposed method and gives some comparison with other model reduction techniques.

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1. Incubation of beagle hepatic cytosol, under conditions promoting phenylalanine hydroxylase activity, led to the formation of the sulfoxide derivatives of S-carboxymethyl-L-cysteine, N-acetyl-S-carboxymethyl-L-cysteine, S-methyl-L-cysteine and N-acetyl-S-methyl-L-cysteine. Thiodiglycolic acid was not a substrate. Enzyme kinetic parameters (Km, Vmax) were derived indicating S-carboxymethyl-L-cysteine had the greatest clearance; no enantioselective preference was observed for this S-oxygenation reaction. 2. Following oral administration of S-carboxymethyl-L-cysteine to beagle dogs, the parent substance and its sulfoxide were the only compounds identified in the plasma. Pharmacokinetic data have been obtained indicating that the small amount of sulfoxide formed persisted within the body for longer than the parent material, but that the majority of the ingested dose remained in the administered sulfide form. 3. The sulfide moiety within the muco-regulatory drug, S-carboxymethyl-L-cysteine, is thought to be vital as it acts as a free radical scavenger, resulting in the inactive sulfoxide. Additional extensive enyzme-mediated sulfoxidation would decrease the amount of active sulfide available. In the dog this appears to not be an issue, signalling possible exploitation for therapeutic benefit in treating airway disease.