Excitability of muscle afferent neurones determined by depolarizing current injection in control and OA animals.

<p>2nA direct current was injected into neurones at 4 weeks after surgery in control and in OA animals. (<b>A and B</b>) show repetitive firing in a control and an OA muscle afferent neurone, respectively. In both recordings, the upper trace indicates the 2 nA depolarizing current, and the lower trace is the intracellular recording signal. (<b>C</b>) Shows the histogram showing the number of neurones with various evoked APs following depolarizing current injection in both control and OA muscle afferent neurones. (<b>D</b>) Shows the comparison of the number of APs evoked by 2 nA direct current injection between OA (<i>N</i> = 37) and control (<i>N</i> = 25) muscle afferent neurones. The Mann-Whitney <i>U</i>-test was used.</p

Scatter plots of conduction velocity (A), AP duration (B), AP rise time (C), maximum rising rate (D), fall time (E) and maximum falling rate (F) of individual muscle afferent neurones in control and OA animals.

<p>In each case the median (horizontal line) is superimposed. Student’s <i>t</i>-tests were used in the comparisons between OA (<i>N</i> = 40) and control (<i>N</i> = 35) muscle afferent neurones, except that Mann-Whitney <i>U</i>-tests were used in the comparison for the AP fall time and maximum falling rate, because the control AP fall time and OA maximum falling rate data failed the D'Agostino and Pearson omnibus normality test. The data indicate slower axonal conduction velocities and slower dynamics of AP generation particularly depolarization in neurones in OA animals.</p

Activation threshold of dorsal root in control and OA animals.

<p>A 0.04 ms rectangular pulse stimulus was delivered to dorsal roots at 4 weeks after surgery in control and in OA animals. (A) Shows the comparison of minimal electrical current sufficient to evoke an AP between OA (<i>N</i> = 25) and control muscle afferent neurones (<i>N</i> = 21). The Mann-Whitney <i>U</i>-test was used. (B) Shows the number of neurones evoked at various current strengths to the dorsal root in both control and OA muscle afferent neurones.</p

Effects of knee derangement on differential hind limb weight distribution in the incapacitance test.

<p>The percentage of weight bearing of the right hind limb (ipsilateral) was compared between one day before surgery (baseline) and 4 weeks after surgery. In each scatter plot, the mean (horizontal line) is superimposed. After confirming that the data was normally distributed, paired <i>t</i>-test was used in the comparison. Significant difference in the percentage of weight bearing of the right hind limb between OA and control rats was found at 4 weeks after surgery.</p

Action potential (AP) recorded intracellularly from a muscle afferent neurone, illustrating parameters measured in each neurone studied.

<p>The AP in the upper trace was elicited in a muscle afferent neurone by electrical stimulation of the L4 dorsal root. 1, latency (by measuring the distance from the stimulating site to the centre of DRG after each experiment, the conduction velocity is calculated); 2, AP rise time; 3, AP fall time; AP duration at base (the value equals AP rise time plus AP fall time); 4, AP half width; 5, 50% afterhyperpolarization recovery time; 6, 80% afterhyperpolarization recovery time; 7, resting membrane potential; 8, AP amplitude; 9, afterhyperpolarization amplitude. Lower trace is the differentiated derivative of the upper trace recording, and plots the change of voltage over time: 10, maximum rising rate; 11, maximum falling rate.</p

Kaplan-Meier survival analysis for hospitalization for heart failure.

<p>(A) Temporal increase in QRS duration of <4ms/year (red line) versus ≥4 ms/year (blue line). (B) Development of a right bundle branch block (RBBB) pattern (red line) versus a non-RBBB [i.e., left bundle branch block (LBBB), RBBB with left anterior hemiblock, or non-specific intraventricular conduction disturbance (NSIVCD)] pattern (blue line). (C) Classification into four groups based on the temporal increase in QRS duration and the development of BBB patterns: patients with the temporal increase in QRS duration of <4 ms/year and the development of an RBBB pattern (red line), patients with the temporal increase in QRS duration of ≥4 ms/year and the development of an RBBB pattern (green line), patients with the temporal increase in QRS duration of <4 ms/year and the development of a non-RBBB pattern (blue line), and patients with the temporal increase in QRS duration of ≥4 ms/year and e development of a non-RBBB pattern.</p

Kaplan-Meier survival analysis for cardiovascular events.

<p>(A) Temporal increase in QRS duration of <5 ms/year (red line) versus ≥5 ms/year (blue line). (B) Development of an RBBB pattern (red line) versus a non-RBBB (i.e., LBBB, RBBB with left anterior hemiblock, or NSIVCD) pattern (blue line). (C) Classification into four groups based on the temporal increase in QRS duration and the development of BBB patterns: patients with the temporal increase in QRS duration of <5 ms/year and the development of an RBBB pattern (red line), patients with the temporal increase in QRS duration of ≥5 ms/year and the development of an RBBB pattern (green line), the temporal increase in QRS duration of <5 ms/year and the development of a non-RBBB pattern (blue line), and patients with the temporal increase in QRS duration of ≥5 ms/year and the development of a non-RBBB pattern (orange line). Abbreviations are as <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0157412#pone.0157412.g001" target="_blank">Fig 1</a>.</p

Comparison of ECG characteristics according to cardiovascular events.

<p>Comparison of ECG characteristics according to cardiovascular events.</p

Univariate and multivariate survival analyses of cardiovascular events.

<p>Univariate and multivariate survival analyses of cardiovascular events.</p

Comparison of ECG characteristics according to heart failure hospitalization.

<p>Comparison of ECG characteristics according to heart failure hospitalization.</p