The correlation between appetite and food intake.

<p>Appetite was measured by a VAS. The correlation coefficients and corresponding <i>P</i>-values were calculated by a Spearman's test.</p

The within-subject comparisons for appetite and food intake.

<p>appetite measurements by a VAS with fasting vs. postprandial plasma (A); appetite measurements by a VAS with sad tears vs. trickled saline (B); the amount of food intake with fasting vs. postprandial plasma (C); and the amount of food intake with sad tears vs. trickled saline (D). Chi-square values and the corresponding <i>P</i>-values are shown. If the circles are on the diagonal line, there was no difference between the two conditions. If a greater number of circles are observed under the diagonal line, the values for the conditions shown in the horizontal axis were greater.</p

The study protocol.

<p>On day 1, we collected fasting and postprandial plasma samples before and during mixed meal tests in healthy male subjects (n = 20). On days 2 and 3, a sample of their own fasting or postprandial plasma at 60 min was randomly applied to the male subjects. On days 4 and 5, a sample of the fresh sad tears or the trickled saline from young healthy female volunteers was randomly applied to the male subjects. Details of the study protocol are described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042352#s2" target="_blank">methods</a> section. R denotes randomization.</p

Residual Hearing in DFNB1 Deafness and Its Clinical Implication in a Korean Population

<div><p>Introduction</p><p>The contribution of Gap junction beta-2 protein (<i>GJB2</i>) to the genetic load of deafness and its mutation spectra vary among different ethnic groups.</p><p>Objective</p><p>In this study, the mutation spectrum and audiologic features of patients with <i>GJB2</i> mutations were evaluated with a specific focus on residual hearing.</p><p>Methods</p><p>An initial cohort of 588 subjects from 304 families with varying degrees of hearing loss were collected at the otolaryngology clinics of Seoul National University Hospital and Seoul National University Bundang Hospital from September 2010 through January 2014. <i>GJB2</i> sequencing was carried out for 130 probands with sporadic or autosomal recessive non syndromic hearing loss. The audiograms were evaluated in the <i>GJB2</i> mutants.</p><p>Results</p><p>Of the 130 subjects, 22 (16.9%) were found to carry at least one mutant allele of <i>GJB2</i>. The c.235delC mutation was shown to have the most common allele frequency (39.0%) among <i>GJB2</i> mutations, followed by p.R143W (26.8%) and p.V37I (9.8%). Among those probands without the p.V37I allele in a <i>trans</i> configuration who showed some degree of residual hearing, the mean air conduction thresholds at 250 and 500 Hz were 57 dB HL and 77.8 dB HL, respectively. The c.235delC mutation showed a particularly wide spectrum of hearing loss, from mild to profound and significantly better hearing thresholds at 250 Hz and 2k Hz than in the non-p.V37I and non-235delC nonsyndromic hearing loss and deafness 1(DFNB1) subjects.</p><p>Conclusion</p><p>Despite its reputation as the cause of severe to profound deafness, c.235delC, the most frequent DFNB1 mutation in our cohort, caused a wide range of hearing loss with some residual hearing in low frequencies. This finding can be of paramount help for prediction of low frequency hearing thresholds in very young DFNB1 patients and highlights the importance of soft surgery for cochlear implantation in these patients.</p></div

<i>GJB2</i> mutations and their allele frequencies.

<p><i>GJB2</i> mutations and their allele frequencies.</p

Establishment of a Flexible Real-Time Polymerase Chain Reaction-Based Platform for Detecting Prevalent Deafness Mutations Associated with Variable Degree of Sensorineural Hearing Loss in Koreans

<div><p>Many cutting-edge technologies based on next-generation sequencing (NGS) have been employed to identify candidate variants responsible for sensorineural hearing loss (SNHL). However, these methods have limitations preventing their wide clinical use for primary screening, in that they remain costly and it is not always suitable to analyze massive amounts of data. Several different DNA chips have been developed for screening prevalent mutations at a lower cost. However, most of these platforms do not offer the flexibility to add or remove target mutations, thereby limiting their wider use in a field that requires frequent updates. Therefore, we aimed to establish a simpler and more flexible molecular diagnostic platform based on ethnicity-specific mutation spectrums of SNHL, which would enable bypassing unnecessary filtering steps in a substantial portion of cases. In addition, we expanded the screening platform to cover varying degrees of SNHL. With this aim, we selected 11 variants of 5 genes (<i>GJB2</i>, <i>SLC26A4</i>, <i>MTRNR1</i>, <i>TMPRSS3</i>, and <i>CDH23</i>) showing high prevalence with varying degrees in Koreans and developed the U-TOP™ HL Genotyping Kit, a real-time PCR-based method using the MeltingArray technique and peptide nucleic acid probes. The results of 271 DNA samples with wild type sequences or mutations in homo- or heterozygote form were compared between the U-TOP™ HL Genotyping Kit and Sanger sequencing. The positive and negative predictive values were 100%, and this method showed perfect agreement with Sanger sequencing, with a Kappa value of 1.00. The U-TOP™ HL Genotyping Kit showed excellent performance in detecting varying degrees and phenotypes of SNHL mutations in both homozygote and heterozygote forms, which are highly prevalent in the Korean population. This platform will serve as a useful and cost-effective first-line screening tool for varying degrees of genetic SNHL and facilitate genome-based personalized hearing rehabilitation for the Korean population.</p></div

Overlapping audiograms of all patients with <i>GJB2</i> mutations.

<p>All of the patients with hearing loss caused by <i>GJB2</i> mutations in this study (n = 22). (Black dots mean hearing thresholds of each subject, which may represent more than one subject with identical hearing thresholds. Red dots represent mean hearing thresholds of each frequency).</p

Overlapping audiograms of the patients with c.235delC and non-c.235delC <i>GJB2</i> mutations.

<p>A. Patients with hearing loss caused by a c.235delC mutation (excluding patients with a p.V37I mutation in the <i>trans</i> configuration) (n = 13). B. Patients with hearing loss caused by a non-c.235delC mutation (excluding patients with a p.V37I mutation in the <i>trans</i> configuration) (n = 5). Detailed lists of non-c.235delC mutations are p.R143W homozygote, p.R143W single heterozygote, p.Glu47* and p.Ala171Glufs*40 compound heterozygote, and p.T86R and p.R143W compound heterozygote. (*p = 0.05, **p = 0.03, by the Mann-Whitney U test) (Black dots mean hearing thresholds of each subject, which may represent more than one subject with identical hearing thresholds. Red dots represent mean hearing thresholds of each frequency).</p

Fenofibrate improves sensory and motor functions of the sciatic nerve in <i>db/db</i> mice with VEGF inhibition.

<p>Tactile threshold (A) and motor conduction latency of the sciatic nerves (B) in <i>db/m</i> and <i>db/db</i> mice after 12 weeks of vascular endothelial growth factor receptor (VEGFR) inhibition plus fenofibrate or VEGFR inhibition alone. *<i>P</i><0.05, **<i>P</i><0.01 and ^#<i>P</i><0.001 versus <i>db/m</i> controls, <i>db/m</i>+Feno, <i>db/m</i> VEGFR12, <i>db/m</i> VEGFR12+Feno, <i>db/db</i>+Feno, and <i>db/db</i> VEGFR12+Feno mice.</p

Fenofibrate decreases inflammatory cell infiltration, oxidative stress and apoptosis of the sciatic nerve in <i>db/db</i> mice with VEGF inhibition.

<p>Representative immunohistochemical stain for F4/80-, 8-OH-dG- and TUNEL-positive cells and quantitative analyses of the results are shown (A to F). *<i>P</i><0.05, **<i>P</i><0.01 and ^#<i>P</i><0.001 versus <i>db/m</i> controls. Representative electron microscopic images of the sciatic nerve endoneural endothelial cells (15,000×) and nerve bundles (5,000×) (G and H). Disorganized, vacuolized and thickened endothelial cells were observed in <i>db/db</i> controls and <i>db/db</i> VEGFR12 mice. These characteristics decreased after 12-week fenofibrate treatment in the <i>db/db</i>+Feno and <i>db/db</i> VEGR12+Feno mice. A marked decrease in the number of unmyelinated nerve bundles (red arrowheads) and prominent axonal degeneration (red arrows) were observed in the <i>db/db</i> controls and <i>db/db</i> VEGFR12 mice. These deficits were also improved by the 12-week fenofibrate treatment in the <i>db/db</i>+Feno and <i>db/db</i> VEGR12+Feno mice. Quantitative analyses of number of endoneurial blood vessel, area of unmyelinated fiber, axonal diameter and number of degenerated axon are shown (I to L). *<i>P</i><0.05 versus other groups, *<i>P</i><0.05, **<i>P</i><0.01, and ^#<i>P</i><0.001 versus <i>db/m</i> controls. Representative immunofluorescent stains for PECAM-1 and TUNEL-double positive cells in <i>db/db</i> mice groups and quantitative analyses of the results are shown (M and N). An increase in the number of PECAM-1 and TUNEL-double positive cells (white arrows) was observed in the <i>db/db</i> controls and <i>db/db</i> VEGFR12 mice. **<i>P</i><0.01 and ^#<i>P</i><0.001 versus <i>db/m</i> controls.</p