Comparison of variables of PN activity during normocapnia (5% CO₂) and hypercapnia (8% CO₂) (<i>n</i> = 21).

<p>Comparison of variables of PN activity during normocapnia (5% CO₂) and hypercapnia (8% CO₂) (<i>n</i> = 21).</p

Correlation between abdominal nerve (AbN) activity and CN activity.

<p>(A) Original and integrated traces of AbN, CN, and PN activity during normocapnia (5% CO₂, 95% O₂), hypercapnia (8% CO₂, 92% O₂), and recovery (5% CO₂, 95% O₂). (B) Traces averaged from consecutive integrated sweeps for 1 min in the preparation shown in (A). The filled arrowheads show the onset of nerve discharges. (C) Comparison of the duration of the AbN pre-I discharge, CN pre-I discharge, and respiratory cycle in the preparation shown in (A). Black bars show the times that the traces in (A) were recorded. The black arrow shows the time point at which the respiratory duration became shortest. The white arrow shows the time point at which the abdominal discharge appeared. (D) Correlation between the duration of the CN pre-I phase discharge and duration of the AbN pre-I discharge in the preparation shown in (A).</p

The onset of the CN, SLN, HGN and RLN discharges in relation to that of the PN discharge during normocapnia (5% CO₂) and hypercapnia (8% CO₂).

<p>The onset of the CN, SLN, HGN and RLN discharges in relation to that of the PN discharge during normocapnia (5% CO₂) and hypercapnia (8% CO₂).</p

The discharge patterns of the CN, SLN, RLN, and HGN under normocapnic conditions.

<p>Averaged traces from two preparations shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0166436#pone.0166436.g001" target="_blank">Fig 1</a> were normalized via the peak amplitude.</p

Respiratory motor activity under normocapnic conditions and hypercapnic acidosis.

<p>(A) Original and integrated traces of CN, SLN, RLN, and PN activity during normocapnia (5% CO₂, 95% O₂), hypercapnia (8% CO₂, 92% O₂), and recovery (5% CO₂, 95% O₂). (B) Traces averaged from consecutive integrated sweeps for 1 min in the preparation shown in (A). The black and gray traces show the averaged traces during hypercapnia and normocapnia, respectively. The filled arrowheads show the onset of the nerve discharges. The open arrowheads show the pre-inspiration (pre-I) discharges. ‘<i>I</i>’ and ‘<i>E</i>’ demonstrate the inspiratory and expiratory phases, respectively. (C) Original and integrated traces of the CN, SLN, HGN, and PN activity obtained from a different preparation from (A). (D) Traces averaged from consecutive integrated sweeps for 1 min in the preparation shown in (C).</p

The discharge patterns of the CN, SLN, RLN, and HGN in hypercapnic acidosis.

<p>The averaged traces from two preparations shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0166436#pone.0166436.g001" target="_blank">Fig 1</a> were normalized via the peak amplitude of the pre-I discharges.</p

Analysis of Multiple B-Value Diffusion-Weighted Imaging in Pediatric Acute Encephalopathy

<div><p>Acute encephalopathy is a disease group more commonly seen in children. It is often severe and has neurological sequelae. Imaging is important for early diagnosis and prompt treatment to ameliorate an unfavorable outcome, but insufficient sensitivity/specificity is a problem. To overcome this, a new value (fraction of high b-pair (F_H)) that could be processed from clinically acceptable MR diffusion-weighted imaging (DWI) with three different b-values was designed on the basis of a two-compartment model of water diffusion signal attenuation. The purpose of this study is to compare F_H with the apparent diffusion coefficient (ADC) regarding the detectability of pediatric acute encephalopathy. We retrospectively compared the clinical DWI of 15 children (1–10 years old, mean 2.34, 8 boys, 7 girls) of acute encephalopathy with another 16 children (1–11 years old, mean 4.89, 9 boys, 7 girls) as control. A comparison was first made visually by mapping F_H on the brain images, and then a second comparison was made on the basis of 10 regions of interest (ROIs) set on cortical and subcortical areas of each child. F_H map visually revealed diffusely elevated F_H in cortical and subcortical areas of the patients with acute encephalopathy; the changes seemed more diffuse in F_H compared to DWI. The comparison based on ROI revealed elevated mean F_H in the cortical and subcortical areas of the acute encephalopathy patients compared to control with significant difference (P<0.05). Similar findings were observed even in regions where the findings of DWI were slight. The reduction of mean ADC was significant in regions with severe findings in DWI, but it was not constant in the areas with slighter DWI findings. The detectability of slight changes of cortical and subcortical lesions in acute encephalopathy may be superior in F_H compared to ADC.</p></div

Cell Adhesion Signaling Regulates RANK Expression in Osteoclast Precursors

<div><p>Cells with monocyte/macrophage lineage expressing receptor activator of NF-κB (RANK) differentiate into osteoclasts following stimulation with the RANK ligand (RANKL). Cell adhesion signaling is also required for osteoclast differentiation from precursors. However, details of the mechanism by which cell adhesion signals induce osteoclast differentiation have not been fully elucidated. To investigate the participation of cell adhesion signaling in osteoclast differentiation, mouse bone marrow-derived macrophages (BMMs) were used as osteoclast precursors, and cultured on either plastic cell culture dishes (adherent condition) or the top surface of semisolid methylcellulose gel loaded in culture tubes (non-adherent condition). BMMs cultured under the adherent condition differentiated into osteoclasts in response to RANKL stimulation. However, under the non-adherent condition, the efficiency of osteoclast differentiation was markedly reduced even in the presence of RANKL. These BMMs retained macrophage characteristics including phagocytic function and gene expression profile. Lipopolysaccharide (LPS) and tumor necrosis factor –αTNF-α activated the NF-κB-mediated signaling pathways under both the adherent and non-adherent conditions, while RANKL activated the pathways only under the adherent condition. BMMs highly expressed RANK mRNA and protein under the adherent condition as compared to the non-adherent condition. Also, BMMs transferred from the adherent to non-adherent condition showed downregulated RANK expression within 24 hours. In contrast, transferring those from the non-adherent to adherent condition significantly increased the level of RANK expression. Moreover, interruption of cell adhesion signaling by echistatin, an RGD-containing disintegrin, decreased RANK expression in BMMs, while forced expression of either RANK or TNFR-associated factor 6 (TRAF6) in BMMs induced their differentiation into osteoclasts even under the non-adherent condition. These results suggest that cell adhesion signaling regulates RANK expression in osteoclast precursors.</p> </div

Multiple White Matter Volume Reductions in Patients with Panic Disorder: Relationships between Orbitofrontal Gyrus Volume and Symptom Severity and Social Dysfunction

<div><p>Numerous brain regions are believed to be involved in the neuropathology of panic disorder (PD) including fronto-limbic regions, thalamus, brain stem, and cerebellum. However, while several previous studies have demonstrated volumetric gray matter reductions in these brain regions, there have been no studies evaluating volumetric white matter changes in the fiber bundles connecting these regions. In addition, although patients with PD typically exhibit social, interpersonal and occupational dysfunction, the neuropathologies underlying these dysfunctions remain unclear. A voxel-based morphometry study was conducted to evaluate differences in regional white matter volume between 40 patients with PD and 40 healthy control subjects (HC). Correlation analyses were performed between the regional white matter volumes and patients' scores on the Panic Disorder Severity Scale (PDSS) and the Global Assessment of Functioning (GAF). Patients with PD demonstrated significant volumetric reductions in widespread white matter regions including fronto-limbic, thalamo-cortical and cerebellar pathways (p<0.05, FDR corrected). Furthermore, there was a significant negative relationship between right orbitofrontal gyrus (OFG) white matter volume and the severity of patients' clinical symptoms, as assessed with the PDSS. A significant positive relationship was also observed between patients' right OFG volumes and their scores on the GAF. Our results suggest that volumetric reductions in widespread white matter regions may play an important role in the pathology of PD. In particular, our results suggest that structural white matter abnormalities in the right OFG may contribute to the social, personal and occupational dysfunction typically experienced by patients with PD.</p></div

Osteoclast differentiation under adherent and non-adherent conditions.

<p><b><i>A</i></b><i>.</i> BMMs were cultured with M-CSF (50 ng/ml), TGF-β (1 ng/ml), and RANKL (150 ng/ml) under the adherent and non-adherent conditions for 24, 48, 72, or 96 hours, after which non-adherent cells were harvested and placed on plastic cell culture plates for 30 minutes, then fixed and stained for TRAP. Cells shown stained red are TRAP-positive cells. <b><i>B</i></b><i>.</i> BMMs grown under adherent and non-adherent conditions were cultured in the presence of M-CSF (50 ng/ml), TGF-β (1 ng/ml), and RANKL (150 ng/ml) for 0, 24, 48, 72, or 96 hours. Subsequently, TRAP activity was determined. Data represent mean values of three independent experiments, with error bars indicating ± SD. <b><i>C</i></b><i>.</i> FITC-conjugated zymosan particles were added to BMMs after 0 and 96 hours of culturing under the adherent and non-adherent conditions in the presence of M-CSF (50 ng/ml), TGF-β (1 ng/ml), and RANKL (150 ng/ml). One hour after zymosan addition, cells were washed with PBS and fixed, then the zymosan particles were visualized by UV illumination. Green dots indicate FITC-conjugated zymosan particles incorporated into cells. <b><i>D</i></b><i>.</i> BMMs were cultured in the presence of M-CSF (50 ng/ml), TGF-β (1 ng/ml), and RANKL (150 ng/ml) under the adherent and non-adherent conditions for 0, 12, 24, 48, and 72 hours, after which mRNA expression levels were examined by RT-PCR. <b><i>E and F</i></b><i>.</i> Relative mRNA expression levels of NFATc1 (<b><i>E</i></b>) and integrin β₃ (<b><i>F</i></b>) at the indicated times under the adherent and non-adherent conditions in the presence of M-CSF (50 ng/ml), TGF-β (1 ng/ml), and RANKL (150 ng/ml), which were quantified using real-time RT-PCR. Data represent the mean values of three independent experiments, with error bars indicating ± SD. **<i>P</i>< 0.01 vs adherent condition at same time point.</p