Landslides and tsunamis predicted by incompressible smoothed particle hydrodynamics (SPH) with application to the 1958 Lituya Bay event and idealized experiment.

Tsunamis caused by landslides may result in significant destruction of the surroundings with both societal and industrial impact. The 1958 Lituya Bay landslide and tsunami is a recent and well-documented terrestrial landslide generating a tsunami with a run-up of 524 m. Although recent computational techniques have shown good performance in the estimation of the run-up height, they fail to capture all the physical processes, in particular, the landslide-entry profile and interaction with the water. Smoothed particle hydrodynamics (SPH) is a versatile numerical technique for describing free-surface and multi-phase flows, particularly those that exhibit highly nonlinear deformation in landslide-generated tsunamis. In the current work, the novel multi-phase incompressible SPH method with shifting is applied to the Lituya Bay tsunami and landslide and is the first methodology able to reproduce realistically both the run-up and landslide-entry as documented in a benchmark experiment. The method is the first paper to develop a realistic implementation of the physics that in addition to the non-Newtonian rheology of the landslide includes turbulence in the water phase and soil saturation. Sensitivity to the experimental initial conditions is also considered. This work demonstrates the ability of the proposed method in modelling challenging environmental multi-phase, non-Newtonian and turbulent flows.

Computational estimation of the influence of the main body-to-iliac limb length ratio on the displacement forces acting on an aortic endograft. Theoretical application to Bolton Treovance® Abdominal Stent-Graft.

AIM: The influence of the relative iliac limb length of an endograft (EG) on the displacements forces (DF) predisposing to adverse effects are under-appreciated in the literature. Therefore, we conducted a computational study to estimate the magnitude of the DF acting over an entire reconstructed EG and its counterparts for a range of main body-to-iliac limb length (L1/L2) ratios. METHODS: A customary bifurcated 3D model was computationally created and meshed using the commercially available ANSYS ICEM (Ansys Inc., Canonsburg, PA, USA) software. Accordingly, Fluid Structure Interaction was used to estimate the DF. The total length of the EG was kept constant, while the L1/L2 ratio ranged from 0.3 to 1.5. RESULTS: The increase in L1/L2 slightly affected the DF on the EG (ranging from 3.8 to 4.1 N) and its bifurcation (4.0 to 4.6 N). However, the forces exerted at the iliac sites were strongly affected by the L1/L2 values (ranging from 0.9 to 2.2 N), showing a parabolic pattern with a minimum for 0.6 ratio. CONCLUSION: It is suggested that the hemodynamic effect of the relative limb lengths should not be considered negligible. A high main body-to-iliac limb length ratio seems to favor hemodynamically a low bifurcation but it attenuates the main body-iliac limbs modular stability. Further clinical studies should investigate the relevant value of these findings. The Bolton Treovance(®) device is presented as a representative, improved stent-graft design that takes into account these hemodynamic parameters in order to achieve a promising, improved clinical performance.

The hemodynamic impact of misalignment of fenestrated endografts: a computational study.

OBJECTIVE: The hemodynamic consequences of misaligned stent-grafts (SG) in fenestrated endografts (EG) have not been adequately studied. Our aim was to study the hemodynamic effects of positional variations of SG, investigating the potential influence on the total displacement forces acting on the EG and the shear stress values at the stented segments. METHODS: This was a computational study. An idealized EG model with two renal fenestrations was computationally reconstructed and centrally extended up to the suprarenal level to treat a suprarenal aneurysm. The misalignment of SG was represented by a variable take-off angle between the SG and the EG centerline axis, corresponding to angles of 90°, 176°, 142°, 38°, and 4°, respectively. Accordingly, the maximum EG displacement forces and the shear stress within the stented segments were calculated, using commercially available software. RESULTS: The variable positions of the SG caused no effect on the maximum displacement force acting on the EG, being quite steady and equal to 5.55 N. On the contrary, the values of maximum shear stress acting on the stented segments were influenced by their orientation. The narrow transition zone between the distal end of the mating stent and the target artery showed higher stresses than any other segment. The right-angle take off SG position (90°) was associated with the lowest stresses (12.5 Pa), whereas the highest values were detected at 38° and 142° (16.5 and 16.1 Pa, respectively). The vessel segments distal to the SG exhibited constantly lower stress values (1.9-2.2 Pa) than any other segment. CONCLUSION: We detected differences in the values of shear stress exerted on the stented arteries, depending on different positions that SG can adapt after the deployment of fenestrated EG. The pathophysiologic implication of our findings and their potential association with clinical events deserve further investigation and clinical validation.

Esterification of phenolic acids catalyzed by lipases immobilized in organogels.

Lipases from Rhizomucor miehei and Candida antarctica B were immobilized in hydroxypropylmethyl cellulose organogels based on surfactant-free microemulsions consisting of n-hexane, 1-propanol and water. Both lipases kept their catalytic activity, catalyzing the esterification reactions of various phenolic acids including cinnamic acid derivatives. High reaction rates and yields (up to 94%) were obtained when lipase from C. antarctica was used. Kinetic studies have been performed and apparent kinetic constants were determined showing that ester synthesis catalyzed by immobilized lipases occurs via the Michaelis-Menten mechanism.

Esterification reactions catalyzed by lipases immobilized in organogels: effect of temperature and substrate diffusion.

Rhizomucor miehei lipase was immobilized in hydroxy(propylmethyl) cellulose or agar gels containing lecithin or AOT microemulsions. The effect of the diffusion of substrates and products to this catalyst was studied, as well as the effect of temperature on the initial rate of ester synthesis. The composition of the gel affects the reaction rate due to mass transport phenomena. The apparent activation energies were higher for the systems based on agar, independently of the microemulsion used, and lower for the systems based on AOT microemulsions, independently of the polymer used.

Oxidative stability and radical scavenging activity of extra virgin olive oils: an electron paramagnetic resonance spectroscopy study.

The oxidative stability of extra virgin olive oils (EVOO) from the Greek island of Crete was evaluated by electron paramagnetic resonance (EPR) spectroscopy and the spin trapping technique. The spin trap N-t-butyl-alpha-phenylnitrone (PBN) was added to the olive oil samples and the production of free radicals was monitored during heating at 70 degrees C. Induction time for the accelerated oxidation of virgin olive oils at 70 degrees C was determined. The EPR results were compared with the oxidative stability values provided by the Rancimat method at 110 degrees C and high linear correlations were found (r=0.922). EPR spin trapping provides a sensitive and rapid method for evaluating the oxidative stability of EVOO. The same samples of Greek extra virgin olive oils were also examined for their radical scavenging activity (RSA) toward the stable galvinoxyl radical by EPR spectroscopy. The decrease of the intensity of the EPR signal upon incubation time was followed. Both oxidative stability and radical scavenging activity of EVOO samples were correlated to their content in polyphenols and tocopherols.

Unexpected reaction profile observed in the synthesis of propyl laurate when using Candida rugosa lipases immobilized in microemulsions based organogels.

1-Propyl laurate synthesis should not be used as standard reaction test of immobilized enzymes in microemulsion-based organogels (MBGs) prepared using lecithin/1-propanol as surfactant when extremely active enzymes with high load are used. In these cases, an anomalous kinetic reaction constant value is observed over short reaction times. Such an anomalous profile is strongly dependent on the concentration of catalyst in the crude powder and, consequently, is not appreciated when either commercial or low activity lipase samples are employed.

A new homogeneous enzyme immunoassay for thyroxine using glycogen phosphorylase b-thyroxine conjugates.

BACKGROUND: [corrected] Measurement of serum thyroxine (T(4)) concentration is important for diagnosis of thyroid gland diseases. We developed a practical homogeneous enzyme immunoassay for thyroxine analysis in unextracted sera. METHODS: A thyroxine derivative conjugated to a reactive sulfhydryl group of glycogen phosphorylase b (GPb). Conjugation caused inhibition of enzyme activity and the enzyme conjugate was re-activated upon the binding of a polyclonal anti-T(4) antibody. Antibody-activation was blocked by the presence of free T(4). RESULTS: Conjugation affected the allosteric character of the enzyme and the K(m) for the allosteric activator AMP was increased 28 times, while anti-T(4) antibody partially reversed this effect. The optimum concentration ratio of enzyme conjugate to anti-T(4) antibody was determined, and T(4) was measured with desired sensitivity and accuracy in the range between 10 and 240 microg/l. Furosemide was used to inhibit the interaction of thyroxine with serum T(4)-binding sites. Human serum T(4) values obtained by this method correlated well with those obtained by a radioimmunoassay (y=1.9+1.0x, r=0.97, N=72). CONCLUSIONS: Chemical modification of glycogen phosphorylase b with a T(4) derivative led to the development of a simple homogenous enzyme immunoassay for T(4) analysis with the desired sensitivity and accuracy.

Homogeneous enzyme immunoassay for triiodothyronine in serum.

BACKGROUND: The concentration of triiodothyronine (T3) in human serum is extremely low and can be determined only by very sensitive methods. We developed a homogeneous enzyme immunoassay for T3 analysis in unextracted serum. METHODS: A T(3) derivative was conjugated to the -SH groups of glycogen phosphorylase b (GPb) from rabbit muscle. Conjugation caused inhibition of enzyme activity, and the enzyme conjugate was reactivated upon binding of anti-T3 antibody. Activation was blocked by the presence of non-antibody-bound T3; this was the basis for the development of the homogeneous enzyme immunoassay for T3 by determining GPb activity fluorometrically. RESULTS: We used furosemide to block the interaction of T3 with serum proteins with T3-binding sites, avoiding any serum treatment step. T3 was measured in the range 0.3-8 microg/L. T3 values obtained by this assay correlated well with those obtained by a RIA (y = 0.97x - 0.07 microg/L; r = 0.96; n = 92). Within- and between-run imprecision (CV) was 5-9% for normal and high concentrations and 16-20% for low concentrations. CONCLUSIONS: Chemical modification of GPb with a T3 derivative allows the development of a simple homogeneous enzyme immunoassay for T3 in unextracted serum.

The effect of proteolytic enzymes on hair follicles of transgenic mice expressing the lac Z-protein in cells of the bulge region.

OBJECTIVE: To study the effects of proteolytic enzymes on mice hair follicles, particularly on cells of the bulge area regarded as follicle stem cells. BACKGROUND: Previous application by iontophoresis of proteolytic enzymes on guinea pig skin resulted in degenerative effects on hair follicles and the hypothesis was proposed that some of the affected cells could be stem cells. METHODS: To mark putative stem cells transgenic mice were produced carrying the lac-Z gene fused to the Upstream Regulatory Region (URR) of Human Papilloma Virus 11 (HPV11), as they express this gene specifically in the cells of the bulge area. Chymotrypsin and papain were applied on skin by iontophoresis, trypsin in the form of liposomes. RESULTS: Enzyme application, both by electrophoresis and as liposomes, led to intense degenerative effects of the hair follicle, such as detachment of the inner root sheath, cystic dilation of the hair shaft and presence of epithelial cells within the lumen. Some of these cells represent hair follicle stem cells expressing beta-galactosidase (beta-gal), having been detached from the bulge area as a result of enzyme treatment, implying impairment of their function.

Bioorganic reactions in microemulsions: the case of lipases.

Water-in-oil microemulsions, or reverse micelles, are being evaluated as a reaction medium for a variety of enzymatic reactions. These systems have many potential biotechnological applications. Important examples are the use of various lipase microemulsion systems for hydrolytic or synthetic reactions. This review illustrates the biotechnological applications of microemulsions as media for bioorganic reactions. The principal focus is on lipase catalyzed processes.

Structural and Dynamic Properties of Lecithin-Alcohol Based w/o Microemulsions: A Luminescence Quenching Study

Water-in-oil microemulsions have been made with lecithin in the presence of some alcohols. The structure of the microemulsions has been studied by steady-state and time-resolved analysis of the luminescence quenching of Ru(bipy)32+ by Fe(CN)63-. We found that well-defined microemulsions can be made only in the presence of short-chain alcohols such as propanol-1 and butanol-1. There exists a threshold for water content in order to obtain typical reverse micelles. Thus in the case of propanol-1, water/surfactant ratio wo should be above 20. By varying water content in the range 20 < wo

Lipase catalyzed esterification in AOT reverse micelles: a structural study.

AOT reverse micelles are used to cosolubilize hydrophilic and hydrophobic reactants of lipase catalysed esterification. Depending on the nature of the alcohol, a drastic change of the initial rate of the esterification is observed. A structural study of the micellar system with and without reactant is undertaken to explain the change in the activity with the various alcohols.

Catalytic behavior of Pseudomonas cepacia lipase in w/o microemulsions.

The activity of purified Pseudomonas cepacia lipase has been investigated in esterification reactions of various aliphatic alcohols with natural fatty acids. The reactions were carried out in microemulsions formed in isooctane by bis-(2-ethylhexyl)sulfosuccinate sodium salt (AOT). Kinetic studies showed that the reaction follows a ping-pong bi-bi mechanism with inhibition by both substrates. The apparent kinetic parameters of the reaction were found to be K(m octanol) = 310 mM, K(m lauric acid) = 78 mM, and V(max) = 250 mumol min(-1) mg(-1). The same system was used for the synthesis of mono- and diglycerides from glycerol and lauric acid, which was successful at very low w(o) values. The catalytic behavior of P. cepacia lipase was also studied in esterification reactions performed in a nonionic microemulsion system formulated by tetraethyleneglycoldodecylether (C(12)E(4)). The optimum activity was found at about w(o) = 8. The apparent values of V(max app) and K(m app) for octanol were calculated and found to be 100 mumol min(-1) mg(-1) and 76 mM, respectively. (c) 1995 John Wiley & Sons, Inc.

Kinetic study of lipase catalyzed esterification reactions in water-in-oil microemulsions.

The kinetics of the esterification of lauric acid by (-)menthol, catalyzed by Penicillium simplicissimum lipase, was studied in water/bis-(2-ethylhexyl)sulfosuccinate sodium salt (AOT)/isooctane microemulsions. Due to their low water content, microemulsions assist in reversing the direction of lipase activity, favoring synthetic reactions. The kinetics of this synthesis follows a Ping-Pong Bi--Bi mechanism. The values of all apparent kinetic parameters were determined. The theoretical model for the expression of enzymic activity in reverse micelles, proposed by Verhaert et al. (Verhaert, R., Hilhorst, R., Vermüe, M., Schaafsma, T. J., Veeger, C. 1990. Eur. J. Biochem. 187: 59-72) was extended to express the lipase activity in an esterification reaction involving two hydrophobic substrates in microemulsion systems. The model takes into account the partitioning of the substrates between the various phases and allows the calculation of the intrinsic kinetic constants. The experimental results showing the dependence of the initial velocity on the hydration ratio, W(o) = [H(2)O]/[AOT], of the reverse micelles, were in accordance with the theoretically predicted pattern.

Esterification reactions catalyzed by lipases in microemulsions: the role of enzyme localization in relation to its selectivity.

The activity of lipases from Rhizopus delemar, Rhizopus arrhizus, and Penicillium simplicissimum entrapped in microemulsions formulated by bis-(2-ethylhexyl)sulfo-succinate sodium salt (AOT) in isooctane has been studied in esterification reactions of various aliphatic alcohols with fatty acids. The effect of the nature of the fatty acids (chain length) and of the alcohols (primary, secondary, or tertiary; chain length; cyclic structures) on the lipase activities was investigated in relation to the reverse micellar structure. The lipases tested showed a selectivity regarding the structure of the substrates used when hosted in the AOT/isooctane microemulsion systems. Penicillium simplicissimum lipase showed higher reaction rates in the esterification of long chain alcohols as well as secondary alcohols. Primary alcohols had a low reaction rate and tertiary a very slow rate of esterification. Long chain fatty acids were better catalyzed as compared to the shorter ones. Rhizopus delemar and R. arrhizus lipases showed a preference for the esterification of short chain primary alcohols, while the secondary alcohols had a low rate of esterification and the tertiary ones could not be converted. The reaction of medium chain length fatty acids was also better catalyzed than in the case of the long ones. The observed lipase selectivity appeared to be related to the localization of the enzyme molecule within the micellar microstructure due to the hydrophobic/hydrophilic character of the protein. The reverse micellar structural characteristics, as well as the localization of the enzyme, were examined by fluorescence quenching measurements and spectroscopical studies.

Pest sequences present in the subunits of phosphorylase kinase.

We report that the amino acid sequences of all four subunits of rabbit skeletal muscle phosphorylase kinase possess one or more regions rich in proline (P), glutamic acid (E), serine (S) and threonine (T). alpha and beta subunits contain strong PEST sequences, showing PEST scores greater than 0 (Rogers et al. (1986) Science 234, 364-368), while gamma and delta subunits contain weak PEST regions (negative PEST scores greater than -5.3). In addition to PEST sequences, alpha, beta and gamma subunits contain clusters of arginine pairs. The above sequence characteristics may serve to signal rapid turnover of phosphorylase kinase.

The ammonium sulfate activation of phosphorylase b.

The ammonium sulfate activation of phosphorylase b has been studied. Ammonium sulfate, when present in high concentrations, induces properties of phosphorylase a in phosphorylase b, such as an enhanced affinity for AMP, a reversal of the glucose-6-P inhibition and enzyme tetramerization. The data are consistent with the interpretation that sulfates bind to the Ser-14 site and the sulfate-protein interactions at this site are responsible for activation of phosphorylase b.

Spin-label studies of glycogen phosphorylase hosted in microemulsion droplets.

ESR spectra of Phosphorylase b labelled with two different paramagnetic probes were recorded in microemulsions of various compositions, formulated with AOT and isooctane. Measurements of the hyperfine coupling constant aN and of the degree of anisotropy H1/H0, showed that: i) the polarity of the microenvironment of both spin-labels bound to the enzyme, expressed in terms of aN, is increased with increasing the hydration ratio of the reversed micelles, R; ii) a transition of the polarity between bound and free water is observed at R = 10; iii) the mobilities of the spin-labelled moieties of the enzyme are different, depending on the hydrophobicity of the site where the probe is bound and on the size of the microdroplets.