A Novel Approach for Studying the Physiology and Pathophysiology of Myelinated and Non-Myelinated Axons in the CNS White Matter

<div><p>Advances in brain connectomics set the need for detailed knowledge of functional properties of myelinated and non-myelinated (if present) axons in specific white matter pathways. The corpus callosum (CC), a major white matter structure interconnecting brain hemispheres, is extensively used for studying CNS axonal function. Unlike another widely used CNS white matter preparation, the optic nerve where all axons are myelinated, the CC contains also a large population of non-myelinated axons, making it particularly useful for studying both types of axons. Electrophysiological studies of optic nerve use suction electrodes on nerve ends to stimulate and record compound action potentials (CAPs) that adequately represent its axonal population, whereas CC studies use microelectrodes (MEs), recording from a limited area within the CC. Here we introduce a novel robust isolated "whole" CC preparation comparable to optic nerve. Unlike ME recordings where the CC CAP peaks representing myelinated and non-myelinated axons vary broadly in size, "whole" CC CAPs show stable reproducible ratios of these two main peaks, and also reveal a third peak, suggesting a distinct group of smaller caliber non-myelinated axons. We provide detailed characterization of "whole" CC CAPs and conduction velocities of myelinated and non-myelinated axons along the rostro-caudal axis of CC body and show advantages of this preparation for comparing axonal function in wild type and dysmyelinated <i>shiverer</i> mice, studying the effects of temperature dependence, bath-applied drugs and ischemia modeled by oxygen-glucose deprivation. Due to the isolation from gray matter, our approach allows for studying CC axonal function without possible "contamination" by reverberating signals from gray matter. Our analysis of "whole" CC CAPs revealed higher complexity of myelinated and non-myelinated axonal populations, not noticed earlier. This preparation may have a broad range of applications as a robust model for studying myelinated and non-myelinated axons of the CNS in various experimental models.</p></div

PD-L1 and Survival in Solid Tumors: A Meta-Analysis

<div><p>Background</p><p>Numerous agents targeting PD-L1/PD-1 check-point are in clinical development. However, the correlation between PD-L1expression and prognosis of solid tumor is still in controversial. Here, we elicit a systematic review and meta-analysis to investigate the potential value of PD-L1 in the prognostic prediction in human solid tumors.</p><p>Methods</p><p>Electronic databases were searched for studies evaluating the expression of PD-L1 and overall survival (OS) of patients with solid tumors. Odds ratios (ORs) from individual studies were calculated and pooled by using a random-effect model, and heterogeneity and publication bias analyses were also performed.</p><p>Results</p><p>A total of 3107 patients with solid tumor from 28 published studies were included in the meta-analysis. The median percentage of solid tumors with PD-L1 overexpression was 52.5%. PD-L1 overexpression was associated with worse OS at both 3 years (OR = 2.43, 95% confidence interval (CI) = 1.60 to 3.70, P < 0.0001) and 5 years (OR = 2.23, 95% CI = 1.40 to 3.55, P = 0.0008) of solid tumors. Among the tumor types, PD-L1 was associated with worse 3 year-OS of esophageal cancer, gastric cancer, hepatocellular carcinoma, and urothelial cancer, and 5 year-OS of esophageal cancer, gastric cancer and colorectal cancer.</p><p>Conclusions</p><p>These results suggest that expression of PD-L1 is associated with worse survival in solid tumors. However, the correlations between PD-L1 and prognosis are variant among different tumor types. More studies are needed to investigate the clinical value of PD-L1 expression in prognostic prediction and treatment option.</p></div

Flow diagram of literature search and study selection.

<p>PB, peripheral blood.</p

Pronounced effects of oxygen glucose deprivation (OGD) on “whole” CC CAPs.

<p>(A) Superimposed CC CAPs recorded before (control), during OGD and during washout. (B1, B2) Time course of changes of absolute (B1) and normalized (B2) amplitudes of peaks 1 and 2 during 10-minute OGD and 20-minute washout. (C) Superimposed CAPs before, during and after OGD. (A-C) Representative data from a single experiment on CC6. The CAPs were recorded without averaging to reflect the fast changes during the test. (D) Summary of OGD–induced changes of CAP peaks after 10 min OGD and 20 min washout in CC3 and CC6. Temperature 35.5°C–36.5°C.</p

Stimulus-response relationship and refractoriness of “whole” CC compound action potential peaks.

<p>This figure shows further details on “whole” CC CAP from in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0165637#pone.0165637.g001" target="_blank">Fig 1C</a>. (A) Superimposed CAPs recorded at varying stimulus intensities. (B) stimulus-response plot of amplitudes of peaks 1 and 2 for CAPs shown in (A). (C) Superimposed CAPs evoked by paired stimuli of varying intervals, with 0.5 ms interval increments. Individual traces in (B) and (C) are averaged from 6 consecutive recordings at each stimulation intensity, taken at 10 s intervals. (D) Plot of peak amplitudes of 2^nd CAP in the pair as a function of inter-stimulus interval, normalized to peak amplitudes of the 1^st CAP in the pair. The absolute refractory period of both peaks is 1.5 ms, while the relative refractory period of both peaks exceeded 15 ms. Conduction distance 1.5 mm; room temperature.</p

Schematic showing the surgical preparations in the three groups.

<p><b>A.</b> Control Group: Both VN and PN were intact. <b>B.</b> Transection Group: Both VN and PN were transected. <b>C.</b> Bridge group: The proximal end of VN was anastomosed to the distal end of the PN.</p

Characteristics of studies included in the meta-analysis.

<p>NR: Not reported.</p><p>Characteristics of studies included in the meta-analysis.</p

Records of CMAP from Control Group (A), Transection Group (B) and Bridge Group (C) rabbit showing both distal latency (dLAT) and amplitude 20 weeks after surgery.

<p>The stimulator electrode was place on the intact VN in Control Group, the proximal end of the transected VN in the Transection Group, and the VN rostral to the anastomosed site in the Bridge Group. <b>D</b>, Comparison of dLAT. <b>E</b>, Comparison of amplitude. The error bars indicate standard deviation (SD). * p<0.001 vs. control group; # p<0.001 vs. Transected Group; ## p<0.05 vs. Transected Group</p

Comparison of “whole” CC CAPs of wild type (wt) and dysmyelinated <i>shiverer</i> (shi^-/-) mice.

<p>(A1, A2) Superimposed “whole” CC CAPs recorded at varying stimulation intensities in CC3 (A1) and CC6 (A2). Each trace in A1 and A2 represents an average of 6 consecutive recordings taken at 10 s intervals. (B1, B2) Digitally averaged “whole” CC CAPs recorded from wt (thick gray traces) and shi^-/- (thin black traces) CC3 (B1, digitally averaged from 8 experiments) and CC6 (B2, digitally averaged from 11 experiments). CAPs from wt CCs, (digitally averaged, from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0165637#pone.0165637.g004" target="_blank">Fig 4A1 and 4A2</a>; brought to same scale with shi^-/- CAPs) are shown in B1 and B2 as thick gray traces. Dashed arrows with question marks show hypothetical relationships between peaks of wt and shi^-/- CAPs (see text). (C1, C2) Statistical comparison of main parameters of wt and shi^-/- “whole” CC CAPs. ** p<0.01; *** p<0.001.</p

Detailed characteristics of “whole CC CAPs recorded at 4 mm conduction distance from CC3 and CC6.

<p>(A1, A2) Digitally averaged “whole” CC CAPs recorded from 14 CC3s (A1) and 29 CC6s (A2) from different mouse brains. The CC3 and CC6 were identified as explained in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0165637#pone.0165637.g003" target="_blank">Fig 3</a>. Note different voltage scales in A1 and A2. (B) Detailed statistical comparison of “whole” CC3 and CC6 CAP parameters: ampl.—amplitude; CV—conduction velocity; peak2/peak1 ampl.—ratio of peak amplitudes; peak2/peak1 area—ratio of peak areas. ** p<0.01; *** p<0.001. All recordings were performed at room temperature.</p