Biomedical subjects

E J Woo

Publications and source records attributed to E J Woo.

18 recordsLinked to original sources

PubMed2001

Crystal structure of mannanase 26A from Pseudomonas cellulosa and analysis of residues involved in substrate binding.

The crystal structure of Pseudomonas cellulosa mannanase 26A has been solved by multiple isomorphous replacement and refined at 1.85 A resolution to an R-factor of 0.182 (R-free = 0.211). The enzyme comprises (beta/alpha)(8)-barrel architecture with two catalytic glutamates at the ends of beta-strands 4 and 7 in precisely the same location as the corresponding glutamates in other 4/7-superfamily glycoside hydrolase enzymes (clan GH-A glycoside hydrolases). The family 26 glycoside hydrolases are therefore members of clan GH-A. Functional analyses of mannanase 26A, informed by the crystal structure of the enzyme, provided important insights into the role of residues close to the catalytic glutamates. These data showed that Trp-360 played a critical role in binding substrate at the -1 subsite, whereas Tyr-285 was important to the function of the nucleophile catalyst. His-211 in mannanase 26A does not have the same function as the equivalent asparagine in the other GH-A enzymes. The data also suggest that Trp-217 and Trp-162 are important for the activity of mannanase 26A against mannooligosaccharides but are less important for activity against polysaccharides.

Amino Acid Sequence↗

PubMed2001

Phenol block of peripheral nerve conduction: Titrating for optimum effect.

OBJECTIVES: To verify the dose-response relationship in phenol nerve block and to determine the concentration and volume of phenol injectate required for effective nerve conduction block. DESIGN: Before-after, experimental study. SETTING: A research institute laboratory. ANIMALS: Seventy-one New Zealand white rabbits. INTERVENTIONS: Group I (n = 48) received tibial nerve block by perineural injection (phenol, n = 40; saline, n = 8), group II (n = 21) by submerging the nerve in phenol solution. The 6 subgroups of group I each received different concentrations (3%, 4%, 5%) and volumes (0.1mL, 0.2mL, 0.3mL). The 2 subgroups of group II received 3% (n = 8) and 5% (n = 13) phenol. MAIN OUTCOME MEASURES: Compound muscle action potential (CMAP) and tension of triceps surae muscles by electric stimulation of the sciatic nerve were measured preintervention and at day 1, and weeks 1, 2, 4, and 8 postblock. Histologic studies were performed on 2 animals from group I. RESULTS: Two rabbits in group I died before results were obtained. In the remaining animals, CMAP amplitude reduced significantly (p <.05) as the volume of 5% phenol solution increased from 0.1mL, 0.2mL, to 0.3mL. A high concentration of phenol produced a more pronounced conduction block; however, no significant (p =.0589) difference existed among the 3 concentrations. Submerged tibial nerve had a greater degree of conduction block than perineurally injected nerve. Depth of the degeneration area in nerve fascicle varied with distance from the injection point. CONCLUSIONS: The nerve block effect of phenol can be titrated by adjusting the concentration and volume of phenol solution if the technique of application and localization of a block site are standardized.

Action Potentials↗

PubMed2000

Thermal--electrical finite element modelling for radio frequency cardiac ablation: effects of changes in myocardial properties.

Finite element (FE) analysis has been utilised as a numerical tool to determine the temperature distribution in studies of radio frequency (RF) cardiac ablation. However, none of the previous FE analyses clarified such computational aspects as software requirements, computation time or convergence test. In addition, myocardial properties included in the previous models vary greatly. A process of FE modelling of a system that included blood, myocardium, and an ablation catheter with a thermistor embedded at the tip is described. The bio-heat equation is solved to determine the temperature distribution in myocardium using a commercial software application (ABAQUS). A Cauchy convergence test (epsilon = 0.1 degree C) was performed and it is concluded that the optimal number of elements for the proposed system is 24610. The effects of changes in myocardial properties (+/- 50% electric conductivity, +100%/-50% thermal conductivity, and +100%/-50% specific heat capacity) in both power-controlled (PCRFA) and temperature-controlled RF ablation (TCRFA) were studied. Changes in myocardial properties affect the results of the FE analyses of PCRFA more than those of TCRFA, and the maximum changes in lesion volumes were -58.6% (-50% electric conductivity), -60.7% (+100% thermal conductivity), and +43.2% (-50% specific heat).

Arrhythmias, Cardiac↗

PubMed2000

Germin is a manganese containing homohexamer with oxalate oxidase and superoxide dismutase activities.

Germin is a hydrogen peroxide generating oxalate oxidase with extreme thermal stability; it is involved in the defense against biotic and abiotic stress in plants. The structure, determined at 1.6 A resolution, comprises beta-jellyroll monomers locked into a homohexamer (a trimer of dimers), with extensive surface burial accounting for its remarkable stability. The germin dimer is structurally equivalent to the monomer of the 7S seed storage proteins (vicilins), indicating evolution from a common ancestral protein. A single manganese ion is bound per germin monomer by ligands similar to those of manganese superoxide dismutase (MnSOD). Germin is also shown to have SOD activity and we propose that the defense against extracellular superoxide radicals is an important additional role for germin and related proteins.

Binding Sites↗

PubMed2000

Crystallization and preliminary X-ray analysis of the auxin receptor ABP1.

Auxin-binding protein (ABP1) is an important receptor for the plant hormone auxin that is involved with many growth and developmental responses in plants. The maize ABP1 gene has been expressed in insect cells, purified and crystallized. Type II crystals are monoclinic, with two glycosylated homodimers in the asymmetric unit, and diffract to 1.9 A using synchrotron radiation.

Crystallization↗

PubMed2000

A new catheter design using needle electrode for subendocardial RF ablation of ventricular muscles: finite element analysis and in vitro experiments.

Radio-frequency (RF) cardiac ablation has been very successful for treating arrhythmias related with atrioventricular junction and accessory pathways with successful cure rates of more than 90%. Even though ventricular tachycardia (VT) is a more serious problem, it is known to be rather difficult to cure VT using RF ablation. In order to apply RF ablation to VT, we usually need to create a deeper and wider lesion. Conventional RF ablation electrodes often fail to produce such a lesion. We propose a catheter-electrode design including one or more needle electrodes with a diameter of 0.5-1.0 mm and length of 2.0-10 mm to create a lesion large enough to treat VT. One temperature sensor could be placed at the middle of the needle electrode for temperature-controlled RF ablation. From finite element analyses and in vitro experiments, we found that the depth of a lesion is 1-2 mm deeper than the insertion depth of the needle and the width increases as we increase the diameter of the needle and the time duration. We showed that a single needle electrode can produce a lesion with about 10-mm width and any required depth. If a wider lesion is required, more than one needle with suggested structures can be used. Or, repeated RF ablations around a certain area using one needle could produce a cluster of lesions. In some cases, a catheter with both conventional electrode and needle electrode at its tip may be beneficial to take advantage of both types of electrode.

Animals↗

PubMed2000

Finite element analyses of uniform current density electrodes for radio-frequency cardiac ablation.

The high current density at the edge of a metal electrode causes hot spots, which can lead to charring or blood coagulation formation during radio-frequency (RF) cardiac ablation. We used finite element analysis to predict the current density distribution created by several electrode designs for RF ablation. The numerical results demonstrated that there were hot spots at the edge of the conventional tip electrode and the insulating catheter. By modifying the shape of the edge of the 5-mm tip electrode, we could significantly reduce the high current density at the electrode-insulator interface. We also studied the current density distribution produced by a cylindrically shaped electrode. We modified the shape of a cylindrical electrode by recessing the edge and filled in a coating material so that the overall structure was still cylindrical. We analyzed the effects of depth of recess and the electrical conductivity of the added material. The results show that more uniform current density can be accomplished by recessing the electrode, adding a curvature to the electrode, and by coating the electrode with a resistive material.

Catheter Ablation↗

PubMed2000

Dependence of apparent resistance of four-electrode probes on insertion depth.

The apparent resistance of a finite-thickness layer measured with a four-electrode plunge probe depends on the electrode insertion depth, electrode spacing, and layer thickness, as well as the resistivity ratio of an underlying layer. A physical model consisting of air, a saline solution layer, and an agar layer simulates the real situation of resistivity measurement. The saline layer represents the finite-thickness layer whose resistivity is to be measured by a plunge electrode probe, and the agar layer represents an underlying perturbing layer. A micropositioner controls the insertion depth of the four electrodes into the saline solution. With the apparent resistance measured on a semi-infinite-thickness layer of saline solution as standard, measurement results show decreasing apparent resistance and increasing error with increasing electrode insertion depth. This information is important for correct measurement of myocardial resistivity in vivo and in vitro.

Calibration↗

PubMed2000

Temperature measurement within myocardium during in vitro RF catheter ablation.

While most commercial ablation units and research systems can provide catheter tip temperature during ablation, they do not provide information about the temperature change inside the myocardium, which determines the lesion size. We present the details of a flow simulation and temperature measurement system, which allows the monitoring of the temperature change inside the myocardium during in vitro radio frequency (RF) cardiac catheter ablation at different blood flow rates to which the catheter site may be exposed. We set up a circulation system that simulated different blood flow rates of 0 to 5 L/min at 37 degrees C. We continuously measured the temperature at the catheter tip using the built-in thermistor and inside the myocardium using a three-thermocouple probe. The system provides a means for further study of the temperature inside myocardium during RF catheter ablation under different flow conditions and at different penetration depths.

Animals↗

PubMed2000

Functional inactivation of the IGF-I receptor delays differentiation of skeletal muscle cells.

Skeletal myoblasts are inherently programmed to leave the cell cycle and begin the differentiation process following removal of exogenous growth factors. Serum withdrawal results in a marked induction of IGF production which is essential for skeletal muscle differentiation in vitro. However, the potential role of the tyrosine kinase IGF-I receptor (thought to be the principal mediator of both IGF-I and II signaling in skeletal muscle) in the decision of myoblasts to begin differentiation following serum withdrawal is unknown. To explore the role of the IGF-I receptor in this decision by skeletal myoblasts, we functionally inactivated endogenous IGF-I receptors in mouse C2C12 cells using a dominant negative, kinase-inactive IGF-I receptor in which the ATP-binding site lysine (K) at residue 1003 has been mutated to alanine (A). Cell lines with the greatest degree of mutant IGF-I receptor expression (A/K cells) demonstrated functional inactivation of endogenous IGF-I receptors as determined by their impaired ability to phosphorylate the principal substrate of the IGF-I receptor, IRS-1, in response to treatment with IGF-I. In addition, the proliferative response of myoblasts to IGF-I was completely abolished in A/K cells. Following withdrawal of exogenous growth factors, A/K cells demonstrated a marked delay in the induction of the gene expression of myogenin, a skeletal muscle-specific transcription factor essential for differentiation, and a subsequent delay in the induction of muscle creatine kinase activity. Delayed differentiation in A/K cells was associated with prolonged phosphorylation of the cell cycle regulatory retinoblastoma (Rb) protein; it is the un- (or hypo-) phosphorylated form of Rb which is known to promote differentiation in skeletal myoblasts. Thus, the IGF-I receptor regulates the timing of myoblast differentiation induced by serum withdrawal. The delayed differentiation of skeletal myoblasts with functionally inactive IGF-I receptors may result, at least in part, from delayed induction of myogenin gene expression and prolonged phosphorylation of the Rb protein.

Animals↗

PubMed1998

Barley oxalate oxidase is a hexameric protein related to seed storage proteins: evidence from X-ray crystallography.

The oxalate oxidase enzyme expressed in barley roots is a thermostable, protease-resistant enzyme that generates H2O2. It has great medical importance because of its use to assay plasma and urinary oxalate, and it has also been used to generate transgenic, pathogen-resistant crops. This protein has now been purified and three types of crystals grown. X-ray analysis shows that the symmetry present in these crystals is consistent with a hexameric arrangement of subunits, probably a trimer of dimers. This structure may be similar to that found in the related seed storage proteins.

Crystallography, X-Ray↗

PubMed1997

Differential regulation of insulin-like growth factor-I (IGF-I) receptor gene expression by IGF-I and basic fibroblastic growth factor.

Insulin-like growth factor-I receptor (IGF-IR) gene expression is regulated by various stimuli, including hormones, growth factors, and nutritional status. We have investigated the molecular mechanism by which two growth factors, insulin-like growth factor-I (IGF-I) and basic fibroblast growth factor (bFGF) regulate IGF-IR gene expression. bFGF increases the endogenous IGF-IR mRNA levels and IGF-IR promoter activity. This effect is mediated by a region of the IGF-IR promoter located between nucleotides -476 and -188 in the 5'-flanking region. In contrast, IGF-I decreases the IGF-IR mRNA levels. IGF-I down-regulates IGF-IR transcriptional activity as deduced from experiments in which the levels of pre-mRNA and mRNA were measured. IGF-I reduced pre-mRNA and mRNA levels in parallel, while the mRNA stability was found to be unchanged by IGF-I treatment. While these results strongly suggest an effect of IGF-I on IGF-IR transcriptional activity, no specific IGF-I response element was demonstrated in the 5'-untranslated region or 5'-flanking region studied. Thus, bFGF and IGF-I have differential effects on IGF-IR gene transcription, with the IGF-I response region as yet unidentified.

Animals↗

PubMed1994

Finite-element method in electrical impedance tomography.

In electrical impedance tomography (EIT), current patterns are injected into a subject and boundary voltages are measured to reconstruct a cross-sectional image of resistivity distribution. Static EIT image reconstruction requires a computer model of a subject, an efficient data-collection method and robust and fast reconstruction algorithms. The finite-element method is used as the computer model. The paper describes the finite-element analysis software package developed, including an interactive graphical mesh generator and fast algorithms for solving linear systems of equations using sparse-matrix and vector techniques. Various models of irregularly shaped subjects are developed using mesh-design tools, including automatic mesh generation and optimisation using the Delaunay algorithm. Even though the software package is customised for use in electrical impedance tomography, it can be used for other biomedical research areas, such as impedance cardiography, cardiac defibrillation and impedance pneumography.

Algorithms↗

PubMed1993

Using compound electrodes in electrical impedance tomography.

In electrical impedance tomography, we inject currents and measure voltages to estimate an object's resistivity distribution. The electrode configuration affects measured voltage data because the electrode-skin contact impedance is high and varies with electrode location. We developed a compound electrode which is composed of two electrodes: a large outer electrode to inject current and a small inner electrode to sense voltage. We used these compound electrodes to measure voltages from a physical phantom. We showed that the measured voltages from the compound electrodes are smaller in amplitude than those from conventional electrodes. This demonstrates that the compound electrode can minimize contact impedance voltage drop from the measured data. We used a finite element model for the compound electrode and incorporated the model into the regularized Newton-Raphson reconstruction algorithm. We performed a sensitivity study and showed that the reconstructed resistivity distributions are less dependent on the unknown contact resistance values for a compound electrode than a conventional electrode and that the use of a compound electrode results in improved images for the reconstruction algorithm.

Algorithms↗

PubMed1993

Finite element modeling of electrode-skin contact impedance in electrical impedance tomography.

In electrical impedance tomography (EIT), we inject currents through and measure voltages from an array of surface electrodes. The measured voltages are sensitive to electrode-skin contact impedance because the contact impedance and the current density through this contact impedance are both high. We used large electrodes to provide a more uniform current distribution and reduce the contact impedance. A large electrode differs from a point electrode in that it has shunting and edge effects which cannot be modeled by a single resistor. We used the finite element method (FEM) to study the electric field distributions underneath an electrode, and developed three models: a FEM model, a simplified FEM model and a weighted load model. We showed that the FEM models considered both shunting and edge effects and matched closely the experimental measurements. FEM models for electrodes can be used to improve the performance of an electrical impedance tomography reconstruction algorithm.

Algorithms↗

PubMed1992

Skin impedance measurements using simple and compound electrodes.

We have studied the effect of the electrode configuration on the measurement of body impedance and found that the electrode configuration greatly affects the impedance measurement using the four-electrode method. We studied the characteristics of the compound electrode and found that the compound electrode provides the four-electrode method in a compact form. A new method of measuring the skin impedance using simple electrodes at low frequencies was developed. At high frequencies where the effect of internal tissue impedance is not negligible, we used the compensation method using compound electrodes, because they measure the voltage right under the skin. At 50 kHz, we measured the real part of the skin impedance of less than 80 omega on the thorax. We propose a simple instrument which can measure accurate skin impedance at various frequencies.

Electric Conductivity↗

PubMed1992

Measuring lung resistivity using electrical impedance tomography.

We propose the use of electrical impedance tomography (EIT) imaging techniques in the measurement of lung resistivity for detection and monitoring of apnea and edema. In EIT, we inject currents into a subject using multiple electrodes and measure boundary voltages to reconstruct a cross-sectional image of internal resistivity distribution. We found that a simplified, therefore fast, version of the impedance imaging method can be used for detection and monitoring of apnea and edema. We have showed the feasibility of this method through computer simulations and human experiments. We speculate that the EIT imaging technique will be more reliable than the current impedance apnea monitoring method, since we are monitoring the change of internal lung resistivity. However, more study is required to verify that this method performs better in the presence of motion artifact than the conventional two-electrode impedance apnea monitoring method. Future work should include experiments which carefully simulate different kinds of motion artifacts.

Adult↗

PubMed1985

Separation of physiological factors influencing glucose-insulin kinetics in diabetic patients.

A new quantitative method was developed for separation of physiological factors influencing glucose intolerance in diabetes mellitus using a three-compartmental model and the intravenous glucose tolerance test (IVGTT) in humans. The present model includes the physiologic factors of the hepatic glucose balance function, the peripheral tissue's glucose utilization rate, and the insulin secretion rate. The insulin sensitivity parameter and hepatic glucose sensitivity parameter were estimated in optimal fitting of the model-based data of glucose and insulin concentrations to the measured IVGTT data in 9 normal and 11 diabetic subjects. The results show that these sensitivity parameters are important for separation of the effects of the interactive physiologic factors, and, also useful in evaluating different glucose-insulin kinetics in 3 clinical groups of normal and diabetic subjects.

Adult↗