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Differential diagnosis of lung nematode parasites from livestock by electrophoretic techniques.

The protein profile, determined by SDS-PAGE, from different geographical strains (Slovakia and Spain) of Dictyocaulus filaria and Protostrongylus rufescens parasitizing Capra hircus and Ovis aries has been assayed. This protein profile has also been comparatively determined in D. viviparus isolated from lungs of Bos taurus killed in Slovakia, by SDS-PAGE. Protein profiles of D. viviparus and both strains of D. filaria were very similar while in P. rufescens a quite different protein profile was found. Furthermore, the isoenzymatic pattern of Lactate Dehydrogenase (LDH) has been studied in two different geographical strains of D. filaria and P. rufescens and in D. viviparus by starch gel electrophoresis. From both strains of D. filaria and from both sexes, the isoenzymatic pattern of LDH was characterized by the presence of five isoenzymes, four anodical and one cathodical. In contrast, males and females of D. viviparus showed a different LDH isoenzymatic pattern; males presenting two isoenzymes with anodical and cathodical migration, respectively, and females showed only a single isoenzyme with anodical migration. Moreover, the electrophoretic mobility of the D. viviparus isoenzymes was different to that of D. filaria. Therefore, LDH has been designated as an important diagnostic tool to differentiate between species of genus Dictyocaulus. Finally, the LDH isoenzymatic pattern in P. rufescens (Slovakian and Spanish strain) was identical in both sexes appearing as a single band with cathodical migration.

Animals↗

Sensitivity and selectivity of the electrochemical detection of the copper(II) complexes of bioactive peptides, and comparison to model studies by rotating ring-disc electrode.

Post-column reaction of peptides with Cu(II) can be used for the electrochemical detection of peptides as their biuret complexes. Understanding of the behavior (sensitivity at the anode and cathode in the dual-series electrochemical detector) of the system is facilitated through the observation of the rotating ring disc voltammetry of some model compounds. In operation, the anodic signal from the oxidation of the Cu(II)-peptide to the Cu(III) form can be used to detect peptides, or the downstream cathode can be used to detect the Cu(III) form. The signals appear at about 0.4 V (anode) for tetra- and longer peptides, 0.65 V for tripeptides. The anode signal is augmented by tyrosine (oxidation at 0.4-0.5 V) and tryptophan (0.5-0.6 V). If the cathode is used as the detector in a two working electrode cell, the sensitivity depends on the stability of the Cu(III) product. This is peptide dependent, but the signal is significant and useful analytically. Twenty-three bioactive peptides in two groups, naturally electrochemically active and naturally electrochemically silent, and several model compounds have been studied. Both naturally electrochemically active peptides (contain tyrosine and/or tryptophan) and naturally electrochemically silent peptides have been studied. Chromatography with an acetonitrile gradient has been used to separate the peptides in each group. Detection limits are for non-electroactive peptides in the range of 16-100 fmol (10- microliters injection 1.6-10 nM, 100 microliters injection 0.16-1.0 nM), and for electroactive peptides in the range of 6-40 fmol (0.6-4.0 nM for a 10- microliters injection and 60-400 pM for a 100- microliters injection). A tryptic digest of bovine cytochrome c is easily seen at 100 nM.

Amino Acid Sequence↗

The accuracy and spatial resolution of the measurement of tissue oxygen removal rate.

An analysis is presented of factors determining the accuracy and spatial resolution of a method for the estimation of oxygen removal rate in tissues using a multi-cathode electrode placed on the tissue surface. The effect of oxygen consumption by the cathodes is shown to be small for 25-micrometer diameter cathodes, but covering the tissue surface with a thin oxygen oxygen-permeable plastic film can introduce a systematic error into the estimate of removal rate, the size of which depends upon the oxygen diffusion coefficient and solubility in the plastic compared with those in the tissue. Spatial resolution is shown to be adequate for distinguishing changes in removal rate over distances of a few hundred microns, and the averaging of removal rate in the direction normal to the tissue surface is weighted heavily towards the superficial tissue layer.

Oxygen Consumption↗

Electrochemical studies on the influence of proteins on the corrosion of implant alloys.

The effect of proteins on corrosion rates of 316L stainless steel, commercially pure titanium and titanium 6-aluminium 4-vanadium was studied in the static and fretting modes. The static mode was studied using cylindrical specimens as per ASTM F-746, and static fracture fixation plates. The fretting mode was studied using a two-hole plate fretting machine which caused a cyclic rocking motion between the plate and the screws, as per ASTM F-897. Electrochemical techniques of polarization resistance and Tafel slope measurements were used to study effects of proteins on the anodic and cathodic corrosion reactions. It was found that proteins increased the corrosion rate of the stainless steel and C.P. titanium cylindrical specimens, but did not have an effect on the Ti-6AI-4V cylinders. In the fretting mode proteins decreased the corrosion rate of the stainless steel plates, but did not have an appreciable effect on either of the titanium alloys. The presence of proteins appeared to cause an increase in the anodic Tafel constant and a decrease in the cathodic Tafel constant of stainless steel specimens. Significant differences in the shapes of the cathodic Tafel slopes were also seen with cylinders with different surface conditions, and static versus fretting plates.

Alloys↗

Historical perspective and current trends in emission microscopy, mirror electron microscopy and low-energy electron microscopy. An introduction to the proceedings of the Second International Symposium and Workshop on Emission microscopy and Related Techniques.

Emission microscopes and related instruments comprise a specialized class of electron microscopes that have in common an acceleration field in combination with the first stage of imaging (i.e., an immersion objective lens, also called a cathode lens or emission lens). These imaging techniques include photoelectron emission microscopy (PEEM or PEM), electron emission induced by heat, ions, or neutral particles, mirror electron microscopy (MEM), and low-energy electron microscopy (LEEM), among others. In these instruments the specimen is placed on a flat cathode or is the cathode itself. The low-energy electrons that are emitted, reflected, or backscattered from the specimen are first accelerated and then imaged by means of an electron lens system resembling that of a transmission electron microscope. The image is formed in a parallel mode in all of the above instruments, in contrast to the image in scanning electron microscopes, where the information is collected sequentially by scanning the specimen. A brief history and introduction to emission microscopy, MEM, and LEEM is presented as a background for the Proceedings of the Second International Symposium and Workshop on this subject, held in Seattle, Washington, August 16-17, 1990. Current trends in this field gleaned from the presentations at that meeting are discussed.

China↗

The occurrence of calcium salt deposition on dermal collagen fibres following electrical injury to porcine skin.

Deposition of calcium salts on collagen fibres has been shown to occur in cathode areas from 2 days to 2 months after exposure to direct current (d.c.) via contact knobs measuring 12 mm in diameter using energy level from 0.5 to 96 J and on day 7 after exposure to alternating current (a.c.) via pointed electrodes using energy level from 30 to 50 J. In order to determine the statistical relation of this histological observation to the type of energy applied 1095 biopsies from 49 pigs including biopsies from skin areas exposed to heat, 50 Hz a.c., 100 kHz a.c. and d.c. as well as from unexposed skin were examined. The specificity was 1.0 using calcium deposition as the test criterium. The sensitivity for cathode areas was found to vary from 0.52 to 1.0 depending on the biopsy technique and the number of days after exposure. Calcium salts on collagen fibres seem uniformly to be present in the cathode area from day 4 to 7, the positive test answer being depending on the biopsy technique. For 50 Hz a.c. the sensitivity using a pointed electrode was found to vary from 0.08 to 0.27 dependent on the number of days after exposure. For all other types of energy the sensitivity was 0.

Animals↗

Importance of electrode design, lead configuration and impedance for successful low energy transcatheter atrial defibrillation in dogs.

OBJECTIVES: We assessed the feasibility of low energy endocardial defibrillation in a canine model of atrial fibrillation, comparing catheters with large surface area electrodes and standard electrode catheters, and evaluated the effects of lead configuration and circuit impedance on defibrillation energy requirements. BACKGROUND: Although recent animal studies have demonstrated the feasibility of low energy endocardial atrial defibrillation, their results have been conflicting with regard to important methodologic aspects. METHODS: In 14 anesthetized greyhounds, atrial fibrillation was induced by rapid atrial pacing and maintained by vagal stimulation. Two large surface area braided electrode catheters and two standard electrode catheters were introduced percutaneously, one of each, in the right atrial appendage and right ventricular apex. A cutaneous patch electrode was placed on the left thorax. Biphasic shocks synchronized to the ventricular electrogram were used to terminate atrial fibrillation. Seven configurations were evaluated. Three used standard electrodes: proximal atrial cathode to distal atrial, ventricular or cutaneous anode. Four used braided electrodes: three with atrial cathode to ventricular, cutaneous or combined anode; one with ventricular cathode to atrial anode. RESULTS: Defibrillation with standard electrode catheters was associated with high impedance (576 +/- 112 omega) and low success rates for all configurations (28% success at < or = 40 J, no successes at 10 J). Low energy defibrillation was readily achieved with the braided electrodes with significantly lower impedance (75 +/- 13 omega, p < 0.0001). Ventricular fibrillation did not occur. The success rate of cardioversion increased in a dose-response manner, allowing fitting of a sigmoid curve and calculation of energy associated with 50% (ED50) and 90% (ED90) success. The most successful configuration was ventricular cathode/atrial anode (ED50 1.5 +/- 0.4 J), and the least successful was atrial anode/cutaneous patch (ED50 6.5 +/- 3.2 J, p = 0.0001). CONCLUSIONS: Low energy atrial defibrillation is feasible using large surface area electrodes but not with standard electrode catheters owing to high impedance. An intracardiac anode provides lower impedance and higher success rates than are provided by a cutaneous anode.

Analysis of Variance↗

Effect of first-phase polarity of biphasic shocks on defibrillation threshold with a single transvenous lead system.

OBJECTIVES: The purpose of this study was to determine whether the polarity of the first phase of a biphasic shock affects the defibrillation threshold. BACKGROUND: The polarity of a monophasic shock has been shown to affect the defibrillation threshold. METHODS: A transvenous defibrillation lead with distal and proximal shocking electrodes was used in this study. In 15 consecutive patients, the defibrillation threshold was determined twice using a step-down protocol, in random order: with the distal coil as the anode for the initial phase (anodal biphasic shock) and with the polarity reversed (cathodal biphasic shock). The power to detect a 5.0-J difference in this study is 0.96. These patients were 61 +/- 11 years old (mean +/- SD), and the mean left ventricular ejection fraction was 0.32 +/- 0.10. RESULTS: Mean defibrillation threshold using anodal biphasic shocks was 9.9 +/- 4.8 J, compared with 9.5 +/- 4.2 J using cathodal biphasic shocks (p = 0.8). In three patients the defibrillation threshold was lower by a mean of 6.3 +/- 2.9 J with the former configuration; in three patients the defibrillation threshold was lower by a mean of 6.7 +/- 2.5 J with the latter configuration; and in nine patients it was the same. Using the standard cathodal configuration, a defibrillation threshold < or = 10 J was obtained in approximately 70% of patients, and a subcutaneous patch was not required in any patient. CONCLUSIONS: The polarity of the first phase of a biphasic shock used with a single transvenous lead does not affect the defibrillation threshold.

Defibrillators, Implantable↗

Deactivation of bilirubin oxidase by a product of the reaction of urate and O2.

The "wired" bilirubin oxidase (BOD) bioelectrocatalyst is superior to pure platinum as an electrocatalyst of the four-electron electroreduction of O(2) to water. Not only is its overpotential for O(2) reduction lower, but unlike platinum, it is not affected by organic compounds like glucose. The "wired" BOD-coated carbon cathode operates for >1 week at 37 degrees C in a glucose-containing physiological buffer solution. One of its key applications would be in a glucose-O(2) biofuel cell, which would operate in living tissues. The cathode is, however, short-lived in serum, losing its electrocatalytic activity in a few hours. Here we show that the damaging serum component is a product of the reaction of urate and dissolved oxygen. Exclusion of urate, by application of Nafion film on the cathode, improves the stability in serum.

Catalysis↗

A composite coating by electrolysis-induced collagen self-assembly and calcium phosphate mineralization.

A composite coating that is composed of collagen protein and calcium phosphate minerals is considered to be bioactive and may enhance bone growth and fixation of metallic orthopedic implants. In this study, we have successfully developed a uniform collagen fibril/octacalcium phosphate composite coating on silicon substrate by electrolytic deposition (ELD). The coating deposition was done through applying a constant potential to the cathode in a three-electrode electrochemistry cell that contain a mild acidic (pH 4.8-5.3) aqueous solution of collagen molecules, calcium and phosphate ions. The coating process involved self-assembly of collagen fibrils and the deposition of calcium phosphate minerals as a result of cathode reaction and local pH increase. The two steps could be synchronized to form a bone-like composite at nanometer scale through proper adjustment of the solution and deposition parameters. Coating morphology, crystal structure and compositions were analyzed by optical and fluorescence microscopy, scanning and transmission electron microscopy, energy dispersive X-ray analysis, inductively coupled argon plasma optical emission spectrophotometry, and Fourier-transformed infrared spectroscopy. Under typical deposition conditions, the cathode (Si) surface formed a thin (100 nm) layer of calcium phosphate coating, on top of which a thick (approximately 100 microm) composite layer formed. The porous composite layer consists of a collagen fibril network on which clusters of octacalcium phosphate crystals nucleate and grow. By combining photolithography and ELD, we were also able to pattern the composite coating into regular arrays of squares. Preliminary results by nanoindentation tests showed that properly prepared composite coating may have higher elastic modulus and scratch resistance than monolithic porous calcium phosphate coating. The results not only provide a novel bioactive coating for biomedical implants, but also establish a new experimental protocol for studying biomineralization mechanisms of collagen based biological tissues.

Bone Substitutes↗

Urea biosensor based on PANi(urease)-Nafion/Au composite electrode.

The polyaniline (PANi)-Nafion composite film was prepared onto the ceramic plate by the cyclic voltammetry (CV) method with the various cycle numbers. When the PANi-Nafion/Au/ceramic plate with the preparing cycle number of 5 was as working electrode, the cathodic peak current was achieved as 84.0 microA in 60 mg dl(-1) NH4Cl buffer solution. On the other hand, the small cathodic peak currents for buffer solution in the presence of 60 mg dl(-1) LiOH, NaCl and KCl, respectively, were found with the same composite electrode as working electrode. The cathodic peak current decreased from 84.0 to 16.3 microA in the 60 mg dl(-1) NH4Cl buffer solution when the cycle number for preparing PANi-Nafion/Au/ceramic plate composite electrode with the CV method increased from 5 to 15. The enzyme of urease was immobilized onto the PANi-Nafion/Au/ceramic plate composite film by the electrochemical immobilization and the casting methods and used as sensing electrode to detect the concentration of urea in the buffer solution. The sensitivity of composite electrode immobilized with the casting method was greater than that of electrochemical immobilization method. The sensitivity and the detecting limit of the urea sensor were found to be 0.7 and 5.27 microA (mg dl(-1))(-1)cm(-2), as well as 6 and 0.3 mg dl(-1), respectively, when urease was immobilized by glutaraldehyde (GA) cross-linker and Nafion network, respectively.

Biosensing Techniques↗

Input-output relationship in galvanotactic response of Dictyostelium cells.

Under a direct current electric field, Dictyostelium cells exhibit migration towards the cathode. To determine the input-output relationship of the cell's galvanotactic response, we developed an experimental instrument in which electric signals applied to the cells are highly reproducible and the motile response are analyzed quantitatively. With no electric field, the cells moved randomly in all directions. Upon applying an electric field, cell migration speeds became about 1.3 times faster than those in the absence of an electric field. Such kinetic effects of electric fields on the migration were observed for cells stimulated between 0.25 and 10 V/cm of the field strength. The directions of cell migrations were biased toward the cathode in a positive manner with field strength, showing galvanotactic response in a dose-dependent manner. Quantitative analysis of the relationship between field strengths and directional movements revealed that the biased movements of the cells depend on the square of electric field strength, which can be described by one simple phenomenological equation. The threshold strength for the galvanotaxis was between 0.25 and 1 V/cm. Galvanotactic efficiency reached to half-maximum at 2.6 V/cm, which corresponds to an approximate 8 mV voltage difference between the cathode and anode direction of 10 microm wide, round cells. Based on these results, possible mechanisms of galvanotaxis in Dictyostelium cells were discussed. This development of experimental system, together with its good microscopic accessibility for intracellular signaling molecules, makes Dictyostelium cells attractive as a model organism for elucidating stochastic processes in the signaling systems responsible for cell motility and its regulations.

Animals↗

Shortening of distal motor latency in anode distal stimulation.

OBJECTIVE: Distal motor latency (DML) is shortened when the anode is held distally instead of the cathode by increasing the stimulus intensity. The objective of this study was to clarify the mechanism responsible for this shortening. METHODS: In seven healthy subjects, compound muscle action potential (CMAP) was obtained from the thenar muscle by bipolar stimulation of the median nerve at the wrist, and the intensity at which the first motor units were stimulated was defined as the threshold. Bipolar stimulation with extended interpole distance was employed to identify the generating site of the CMAP and F-wave. RESULTS: The shortening of DML was dependent on the stimulus intensity and threshold. For the low threshold condition, the CMAP generating site was replaced from the proximal cathodal pole to the distal anodal pole by increasing the stimulus intensity. The generating site of the F-wave remained at the proximal cathodal pole irrespective of stimulus intensity. CONCLUSIONS: Replacement of the generating site results in the shortening of DML. When the F-wave is recorded after being induced by anode distal stimulation, CMAP should not be simultaneously evaluated. SIGNIFICANCE: This study clarified the generation sites of CMAP and the F-wave when induced by anode distal stimulation.

Action Potentials↗

Transcranial direct current stimulation applied over the somatosensory cortex - differential effect on low and high frequency SEPs.

OBJECTIVE: Transcranial direct current stimulation (tDCS) has an influence on the excitability of the human motor cortex measured by motor evoked potentials (MEPs) after transcranial magnetic stimulation. Low and high frequency (HFOs) components of somatosensory evoked potentials (SEPs) were studied questioning whether a comparable effect can be observed after applying tDCS to the human somatosensory cortex. METHODS: Multichannel median nerve SEPs were recorded before and after applying tDCS of 1mA over a period of 9min with the cathode placed over the somatosensory cortex and the anode over the contralateral forehead and vice versa in a second session. The source activity of the N20, N30 and HFOs was evaluated before and after application of tDCS. RESULTS: After cathodal tDCS to the somatosensory cortex we found a significant reduction of the N20 source amplitude while there was no effect after anodal stimulation. For the N30 component and HFOs no change in source activity was observed. CONCLUSIONS: Corresponding to the results for the motor cortex a sustained reduction of the excitability of the somatosensory cortex after cathodal tDCS was shown. SIGNIFICANCE: We demonstrated differential effects of tDCS on the high and low frequency components of SEPs confirming the hypothesis of locally and functionally distinct generators of these two components.

Adult↗

Effects of transcranial direct current stimulation coupled with repetitive electrical stimulation on cortical spreading depression.

We have recently shown that two techniques of brain stimulation - repetitive electrical stimulation (ES) (that mimics transcranial magnetic stimulation) and transcranial direct current stimulation (tDCS) - modify the velocity of cortical spreading depression (CSD) significantly. Herein we aimed to study the effects of these two techniques combined on CSD. Thirty-two Wistar rats were divided into four groups according to the treatment: sham tDCS/sham ES, sham tDCS/1 Hz ES, anodal tDCS/1 Hz ES, cathodal tDCS/1 Hz ES. Our findings show that 1 Hz ES reduced CSD velocity, and this effect was modified by either anodal or cathodal tDCS. Anodal tDCS induced larger effects than cathodal tDCS. Hereby CSD velocity was actually increased significantly after anodal tDCS/1 Hz ES. Our results show that combining two techniques of brain stimulation can modify significantly the effects of ES alone on cortical excitability as measured by the neurophysiological parameter of cortical spreading depression and therefore provide important insights into the effects of this new approach of brain stimulation on cortical activity.

Animals↗

Role of intramural virtual electrodes in shock-induced activation of left ventricle: optical measurements from the intact epicardial surface.

BACKGROUND: According to one hypothesized mechanism of defibrillation, shocks directly excite the bulk of ventricular myocardium in the excitable state due to intramural virtual electrodes; however, this hypothesis has not been examined in intact myocardium. OBJECTIVES: The purpose of this study was examine the role of intramural virtual electrodes in shock-induced activation of intact left ventricular (LV) tissue. METHODS: Twelve isolated porcine LV preparations were stained with a transmembrane potential (V(m))-sensitive dye by two methods: (1) surface staining and (2) global staining via coronary perfusion. Shocks (E approximately 0.8-48 V/cm, duration = 10 ms) were applied across the wall from epicardium to endocardium during diastole via transparent electrodes. Shock-induced V(m) responses were measured optically from the intact epicardial surface after surface staining and global staining. RESULTS: Surface-staining recordings demonstrated different V(m) responses to cathodal and anodal shocks. Whereas cathodal shocks caused depolarization and rapid activation of the epicardial surface, anodal shocks induced hyperpolarization and delayed surface activation. In contrast, global-staining V(m) responses to cathodal and anodal shocks were qualitatively similar. Both responses were characterized by activation with small latency and rapid propagation. Weak shocks of both polarities induced monotonic action potential upstrokes; stronger shocks induced nonmonotonic upstrokes with two rising phases at shock onset and end. Such features of global-staining V(m) responses as make activation of the epicardium by anodal shocks and the nonmonotonic action potential upstrokes can be explained by the presence of subepicardial intramural virtual electrodes. CONCLUSION: These data suggest that shocks induce intramural virtual electrodes that directly excite LV tissue and account for the shape of optical V(m) responses recorded from the epicardial surface.

Animals↗

Theoretical study of the surface energy and electronic structure of pyrite FeS2 (100) using a total-energy pseudopotential method, CASTEP.

The geometric and electronic structures of FeS(2) (100) surface have been studied by a quantum-mechanical calculation using a total-energy pseudopotential code, CASTEP. The (100) surface is very stable and does not give any significant geometric relaxation. The electronic structure of FeS(2) (100) surface is characterized by the appearance of new native surface states in the bulk band gap, which correspond to antibonding mixed Fea-Ssp(3) states. These surface states play an important role as mediators of electron transfer on both anodic and cathodic sites in the incipient oxidation of pyrite. Moreover, the (100) surface has small band gaps and shows some metallic character. It is predicted that the rate of cathodic reductive reaction of O(2) in the incipient oxidation of pyrite is much faster than previously considered. The transport of electrons from the anodic sites to the cathodic sites on the (100) surface is faster and hole injection of anodic sites is not the rate-determining step. So we can deduce that the rate-determining step of incipient oxidation for pyrite consists of both electron transfer of pyrite/aqueous O(2) interface and the splitting of H(2)O.

Journal Article↗

Effect of electroporation on the electroosmosis across hairless mouse skin in vitro.

The effect of electroporation on the iontophoresis-produced electroosmosis across the skin was evaluated by measuring the permeability of hairless mouse skin, to mannitol, a non-electrolyte, in vitro. Immediately after electroporation by squared pulses (10 times/s) at 100, 150 or 200 V for 1 ms, anodal iontophoretic permeations were determined at 0.4 mA/cm2 for 4 h. The observed iontophoretic permeability of mannitol was higher with electroporation pretreatment than without pretreatment. The enhanced flux of mannitol induced by electroporation, however, was due to increased passive diffusion. The contribution of convective or osmotic flow caused by anodal iontophoresis on skin permeation of mannitol was decreased by the pretreatment. In addition, osmotic flow was decreased with an increase in the applied voltage for electroporation. In contrast, mannitol flux during cathodal iontophoresis at 0.4 mA/cm2 after 150 or 200 V electroporation was higher than without electroporation as well as anodal iontophoresis, but cathodal iontophoretic flux after electroporation was lower than without iontophoresis. The neutral high-molecular compound dextran rhodamine B was also used as a second model. Anodal iontophoresis alone did not increase skin permeability of the compound. However, electroporation pretreatment before anodal iontophoresis enhanced the skin permeation of dextran rhodamine B, which was due to increased osmotic flow induced by this combination. These results suggest that electroporation decreases the electroosmosis produced by iontophoresis, and that electroporation increases skin permeability to neutral low and high model compounds (mannitol and dextran rhodamine B) probably due to an enlarged permeation pathway. Thus, electroporation affects osmotic flow from the anode to cathode during iontophoresis. Therefore, one has to pay attention to the change in electroosmosis produced by iontophoresis for the combined use of electroporation and iontophoresis to attain a high skin-penetration enhancing effect.

Administration, Topical↗