A controlled trial to investigate whether the orientation of the bevel and angle of approach determine the side of endobronchial intubation in an adult manikin.

One of the commonest complications of endotracheal intubation occurs when the tip of the endotracheal tube passes distal to the carina and enters one of the main bronchi. The perioperative practitioner may observe high airway pressures, hypoxia or even pneumothorax. The most common reason given for the high incidence of right endobronchial intubation is that the right main bronchus comes off the trachea at a more acute angle from the midline. We sought, however, to explore two other factors which may explain this phenomenon ­ the angle of the tube's bevel and its trajectory of approach. We conducted a prospective controlled trial in which doctors from our department intubated the trachea of an adult manikin in three distinct sets using standard tube, reversed tubes and reversed laryngoscope blades. We found that the angle of the bevel and trajectory of approach determines the side of endobronchial intubation in an adult manikin.

Nitrous oxide emission from riparian buffers in relation to vegetation and flood frequency.

The nitrate (NO(3)(-)) removal capacity of riparian zones is well documented, but information is lacking with regard to N(2)O emission from riparian ecosystems and factors controlling temporal dynamics of this potent greenhouse gas. We monitored N(2)O fluxes (static chambers) and measured denitrification (C(2)H(2) block using soil cores) at six riparian sites along a fourth-order stretch of the White River (Indiana, USA) to assess the effect of flood regime, vegetation type, and forest maturity on these processes. The study sites included shrub/grass, aggrading (<15 yr-old), and mature (>80 yr) forests that were flooded either frequently (more than four to six times per year), occasionally (two to three times per year), or rarely (every 20 yr). While the effect of forest maturity and vegetation type (0.52 and 0.65 mg N(2)O-m(-2) d(-1) in adjacent grassed and forested sites) was not significant, analysis of variance (ANOVA) revealed a significant effect ( < 0.01) of flood regime on N(2)O emission. Among the mature forests, mean N(2)O flux was in this order: rarely flooded (0.33) < occasionally flooded (0.99) < frequently flooded (1.72). Large pulses of N(2)O emission (up to 80 mg N(2)O-m(-2) d(-1)) occurred after flood events, but the magnitude of the flux enhancement varied with flood event, being higher after short-duration than after long-duration floods. This pattern was consistent with the inverse relationship between soil moisture and mole fraction of N(2)O, and instances of N(2)O uptake near the river margin after flood events. These results highlight the complexity of N(2)O dynamics in riparian zones and suggest that detailed flood analysis (frequency and duration) is required to determine the contribution of riparian ecosystems to regional N(2)O budget.

Electromobility of plasmid DNA in tumor tissues during electric field-mediated gene delivery.

Interstitial transport is a crucial step in plasmid DNA-based gene therapy. However, interstitial diffusion of large nucleic acids is prohibitively slow. Therefore, we proposed to facilitate interstitial transport of DNA via pulsed electric fields. To test the feasibility of this approach to gene delivery, we developed an ex vivo technique to quantify the magnitude of DNA movement due to pulsed electric fields in two tumor tissues: B16.F10 (a mouse melanoma) and 4T1 (a mouse mammary carcinoma). When the pulse duration and strength were 50 ms and 233 V/cm, respectively, we found that the average plasmid DNA movements per 10 pulses were 1.47 microm and 0.35 microm in B16.F10 and 4T1 tumors, respectively. The average plasmid DNA movements could be approximately tripled, ie to reach 3.69 microm and 1.01 microm, respectively, when the pulse strength was increased to 465 V/cm. The plasmid DNA mobility was correlated with the tumor collagen content, which was approximately eight times greater in 4T1 than in B16.F10 tumors. These data suggest that electric field can be a powerful driving force for improving interstitial transport of DNA during gene delivery.

Changes in anisotropic conduction caused by remodeling cell size and the cellular distribution of gap junctions and Na(+) channels.

Because gene therapy presents a new frontier in the treatment of arrhythmias, it has become important to know how manipulation of the cellular distribution of proteins changes electrical events within individual cells, and whether these cellular changes affect conduction at the larger macroscopic size scale. However, experimental limitations in cardiac bundles prevent measurement of conduction delays across specific gap junctions, as well as the intracellular distribution of the maximum rate of rise of the action potential (V(max)). In view of these limitations, we used immunohistochemical morphological results as a basis to develop two-dimensional cellular models of neonatal and mature canine ventricular muscle in order to obtain insight into the electrophysiological effects of changes in the cellular distribution of proteins; eg, the major protein of cardiac gap junctions, connexin43. Morphological results showed that when the cells enlarged after birth, the gap junctions shifted from the sides to the ends of ventricular myocytes. At birth, V(max) was not different during longitudinal and transverse propagation. However, growth hypertrophy produced a selective increase in mean transverse V(max) with no significant change in longitudinal V(max). Two-dimensional cellular computational models of neonatal and mature ventricular muscle showed that the observed changes in the cellular distribution of the gap junctions and change in cell size accounted for the experimental results. The results unexpectedly showed that cellular scaling (cell size) is as important (or more so) as changes in gap junction distribution in determining the properties of transverse propagation. The results suggest that in pathological states that are arrhythmogenic, maintenance of cell size during remodeling the distribution of gap junctions is important in sustaining a maximum rate of rise of the action potential.

Electrophysiological effects of remodeling cardiac gap junctions and cell size: experimental and model studies of normal cardiac growth.

The increased incidence of arrhythmias in structural heart disease is accompanied by remodeling of the cellular distribution of gap junctions to a diffuse pattern like that of neonatal cardiomyocytes. Accordingly, it has become important to know how remodeling of gap junctions due to normal growth hypertrophy alters anisotropic propagation at a cellular level (V(max)) in relation to conduction velocities measured at a macroscopic level. To this end, morphological studies of gap junctions (connexin43) and in vitro electrical measurements were performed in neonatal and adult canine ventricular muscle. When cells enlarged, gap junctions shifted from the sides to the ends of ventricular myocytes. Electrically, normal growth produced different patterns of change at a macroscopic and microscopic level. Although the longitudinal and transverse conduction velocities were greater in adult than neonatal muscle, the anisotropic velocity ratios were the same. In the neonate, mean V(max) was not different during longitudinal (LP) and transverse (TP) propagation. However, growth hypertrophy produced a selective increase in mean TP V(max) (P<0.001), with no significant change in mean LP V(max). Two-dimensional neonatal and adult cellular computational models show that the observed increases in cell size and changes in the distribution of gap junctions are sufficient to account for the experimental results. Unexpectedly, the results show that cellular scaling (cell size) is as important (or more so) as changes in gap junction distribution in determining TP properties. As the cells enlarged, both mean TP V(max) and lateral cell-to-cell delay increased. V(max) increased because increases in cell-to-cell delay reduced the electric current flowing downstream up to the time of V(max), thus enhancing V(max). The results suggest that in pathological substrates that are arrhythmogenic, maintaining cell size during remodeling of gap junctions is important in sustaining a maximum rate of depolarization.

Extracellular discontinuities in cardiac muscle: evidence for capillary effects on the action potential foot.

It has become of fundamental importance to understand variations in the shape of the upstroke of the action potential in order to identify structural loading effects. One component of this goal is a detailed experimental analysis of the time course of the foot of the cardiac action potential (Vm foot) during propagation in different directions in anisotropic cardiac muscle. To this end, we performed phase-plane analysis of transmembrane action potentials during anisotropic propagation in adult working myocardium. The results showed that during longitudinal propagation there was initial slowing of Vm foot that resulted in deviations from a simple exponential; corollary changes occurred at numerous sites during transverse propagation. We hypothesized that the effect on Vm foot observed in the experimental data was created by the microscopic structure, especially the capillaries. This hypothesis predicts that the phase-plane trajectory of Vm foot will deviate from linearity in the presence of a high density of capillaries, and that a linear trajectory will occur in the absence of capillaries. Comparison of the results of Fast and Kléber (Circ Res. 1993;73:914-925) in a monolayer of neonatal cardiac myocytes, which is devoid of capillaries, and our results in newborn ventricular muscle, which is rich in capillaries, showed drastic differences in Vm foot as predicted. Because this comparison provided experimental support for the capillary hypothesis, we explored the underlying biophysical mechanisms due to interstitial electrical field effects, using a "2-domain" model of myocytes and capillaries separated by interstitial space. The model results show that a propagating interstitial electrical field induces an inward capacitive current in the inactive capillaries that causes a feedback effect on the active membrane (source) that slows the initial rise of its action potential. The results show unexpected mechanisms related to extracellular structural loading that may play a role in selected conduction disturbances, such as in a reperfused ischemic region surrounded by normal myocardium.

Threshold variability in fibers with field stimulation of excitable membranes.

The central focus of this report is the evolution of transmembrane potentials following initiation of a point-source field stimulus, particularly when the stimulus is short and the stimulating electrode is close to the fiber. The transmembrane voltage threshold in response to a point-source field stimulus was determined in a numerical model of a single unmyelinated fiber. Both nerve (Hodgkin-Huxley) and cardiac (Ebihara-Johnson [1]) models of the fiber membrane were evaluated. A central question is whether it is possible to know in advance whether a stimulus of specific magnitude, duration, and location will result in a subsequent action potential. Such determination can be based on the membrane's "voltage threshold." In contrast to the commonly held view, the voltage threshold was found to vary markedly depending on the duration and location of the field stimulus. Voltage thresholds ranged from about 8 mV above baseline to more than 100 mV above baseline, the higher thresholds occurring with shorter stimuli and electrode locations closer to the membrane. A related question is whether the passive membrane response can be used as a tool in determining whether a subsequent action potential is elicited. If the answer is affirmative, this finding can be very useful, since passive properties are linear and thereby much simpler to evaluate than active ones. The results show that the passive response tracks active responses long enough to be a good estimator of subsequent action potential development. Examples show that the evaluation of Vm at 0.2-0.5 msec after stimulus initiation, times chosen on the basis of membrane characteristics, was a better predictor of subsequent excitation than was either initial transmembrane current or Vm at the time when the stimulus ends. Most of the circumstances analyzed here with electric field stimulation also appear likely to be valid with magnetic field stimulation.

Electric field stimulation of excitable tissue.

This paper examines the transmembrane voltage response of an unmyelinated fiber to a stimulating electric field from a point current source. For subthreshold conditions, analytic expressions for the transmembrane potential, vm, are developed that include the specific effects of fiber-source distance, h, and time from the onset of the stimulus, T. Suprathreshold effects are determined for two examples by extending the analytical results with a numerical model. The vm response is a complex evolution in time, especially for small h, that differs markedly from the "activating function." In general, the subthreshold response is a good predictor of the wave shape of the suprathreshold vm, but a poor predictor of its magnitude. The subthreshold response also is a good (but not a precise) predictor of the region where excitation begins.