Time courses of the effects of CD, HFSD and HFSSD diets on blood biochemistry.

<p>The fasting plasma glucose (FPG, A), the plasma insulin (B), the free fatty acids (FFA, C) and the homeostasis model assessment insulin resistance (Homa-IR) index (D) are shown. At least five rats were used in each diet group for this statistical analysis across post diet days 0–120. ^*<i>P</i><0.05, HFSD vs. CD; # <i>P</i><0.05, HFSSD vs. CD. Vertical dashed lines indicate initial significant changes in blood chemistry caused by the diets. For abbreviations see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0057427#pone-0057427-g001" target="_blank">Fig.1</a>. Error bars: ± SEM.</p

Time courses of the effects of CD, HFSD and HFSSD diets on body weight (A) and systolic blood pressure (SBP) (B).

<p>CD, conventional diet; HFSD, high-fat and high-sucrose diets; HFSSD, high-fat, high-sucrose and high-salt diets. Vertical dashed line indicates initial significant changes in SBP. Ten rats were used in each diet group for this statistical analysis across post diet days 0–120. ^*<i>P</i><0.05, HFSD vs. CD; # <i>P</i><0.05, HFSSD vs. CD. Error bars: ± SEM.</p

DataSheet1_Interaction of Neurovascular Signals in the Degraded Condylar Cartilage.docx

Introduction: Degradation of the condylar cartilage during temporomandibular joint osteoarthritis (TMJ-OA) results in the infiltration of nerves, blood vessels and inflammatory cells from the subchondral bone into the cartilage. The interaction among innervation, angiogenesis and inflammation in the condylar cartilage of TMJ-OA remains largely unknown.Method: In the present study, microarray-based transcriptome analysis was used to detect, and quantitative real-time polymerase chain reaction was used to validate transcriptome changes in the condylar cartilage from a well-established rat TMJ-OA model. Gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG) pathway and protein-protein interaction (PPI) analyses were conducted.Result: There were 1817 differentially expressed genes (DEGs, fold change ≥2, p Conclusion: The present study demonstrated, for the first time, that intimate interactions exist among innervation, angiogenesis and inflammation in the condylar cartilage of TMJ-OA.</p

The effects of CD, HFSD and HFSSD diets on the somatic motor functions measured in conscious rats.

<p>The somatic motor function was shown by motor coordinating performance of rats on a treadmill on post diet days 120. Five to six rats were used in each diet group for this statistical analysis. ^*<i>P</i><0.05, HFSD vs. CD; # <i>P</i><0.05, HFSSD vs. CD. For abbreviations see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0057427#pone-0057427-g001" target="_blank">Fig.1</a>. Error bars: ± SEM.</p

Ion Steric Effect Induces Giant Enhancement of Thermoelectric Conversion in Electrolyte-Filled Nanochannels

Ionic thermoelectricity in nanochannels has received increasing attention because of its advantages, such as high Seebeck coefficient and low cost. However, most studies have focused on dilute simple electrolytes that neglect the effects of finite ion sizes and short-range electrostatic correlation. Here, we reveal a new thermoelectric mechanism arising from the coupling of the ion steric effect due to finite ion sizes and ion thermodiffusion in electric double layers, using both theoretical and numerical methods. We show that this mechanism can significantly enhance the thermoelectric response in nanoconfined electrolytes depending on the properties of electrolytes and nanochannels. Compared to the previously known mechanisms, the new mechanism can increase the Seebeck coefficient by 100% or even 1 order of magnitude enhancement under optimal conditions. Moreover, we demonstrate that the short-range electrostatic correlation can help preserve the Seebeck coefficient enhancement in a weaker confinement or in more concentrated electrolytes

Defective craniofacial morphogenesis and organogenesis in the NCC-<i>mTOR</i> KO mice.

<p>(A) Craniofacial morphogenesis at two successive stages. Arrowheads point to facial cleft. (B) Lateral view of E11.5 heads, NCCs are GFP-labeled. (C) SEM examination of E12.5 heads, * marks facial cleft. (D, E) Histology of E11.5 heads, showing failed midline fusion of the mutant FNs. (F, G) Histology of E14.5 heads. (H-K) Immunofluorescence for β-catenin and p-Smad1/5/8, arrowheads point to positive staining in the mandibular prominences. (L) Quantification of β-catenin and p-Smad1/5/8 levels by calculating relative fluorescence intensity. (M-Q) Whole mount in situ hybridization for <i>Alx3</i>, <i>Msx1</i>, <i>Pax3</i> and <i>Fgf8</i>. (R) Quantification of in situ hybridization staining of <i>Alx3</i>, <i>Msx1</i>, <i>Pax3</i> and <i>Fgf8</i>. *P<0.05; ** P<0.01. br: brain; ctr: control; fn: frontonasal prominence; md: mandibular prominence; mx: maxillary prominence; ns: non-significant; pal: palate; sn: snout; tb: tooth bud; ton: tongue; vs: vessel. Scale bar in (A-C): 100 μm; scale bar in (A) applies to (M-Q); scale bars in others: 200 μm.</p

The effects of CD, HFSD and HFSSD diets on the myelinated and unmyelinated fiber structures in the sciatic nerves.

<p>Electron microscopic photomicrographs show the cross-section of the sciatic nerve fibers in the CD (A and D), HFSD (B and E) and HFSSD (C and F) groups. Dramatic pathological changes characterized by myelin breakdown or disruption and axon degeneration are mainly seen in large myelinated fibers (LMF) of rats fed HFSD (B) and HFSSD (C) when comparing with CD (A). The diets-induced LMF myelin changes are often seen as myelin lamina rarefaction, focal demyelination and vacuolization (yellow arrowheads in B-C). Axon degeneration of LMFs is characterized by abnormal high electron density and axonal plasmic shrinkage (asterisks in B-C). Ultrastructures of small myelinated fibers (SMF) and unmyelinated C fibers (UMF) in three groups are also shown (D-F). The axolemma and the Schwann cell covering are well maintained in all UMFs of rats fed CD (see red arrows in D), however, the axolemma and the Schwann cell membrane are thickened and perturbed shown as high electron density in both HFSD and HFSSD (red arrows in E-F). In addition, enlarged mitochondria and lipofuscin depositions are also seen in UMF axons of high energy/salt-treated rats (see yellow arrowheads in E-F) but not in control rats (D). The ultrastructures of SMFs in HFSD and HFSSD are well preserved when compared with CD (see yellow arrows in B-F), however, broken SMF can also be seen in the diet rats (double yellow arrows in E). My, myelin sheath; SC, the nucleus of Schwann cells; other abbreviations see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0057427#pone-0057427-g001" target="_blank">Fig.1</a>. Scale bar for A-C: 5 µm, scale bar for D-F: 2 µm.</p

Craniofacial development of the NCC-<i>Rictor</i> and NCC-<i>Rptor</i> cKO mice.

<p>(A) Lateral and top view of mouse heads at the newborn stage. (B) Quantification of snout length (the most anterior point to the most anterior eye line) and width (the line at the mid-point of the length line), *p<0.05. (C, D, E) Skeleton preparation by alizarin red and alcian blue staining at the newborn stage, demonstrating hypoplasia of the craniofacial bones and an enlarged interfrontal suture in the mutant. (F, G) Immunofluorescence for PHH3 in the first PA. (H) SEM of the NCC-<i>Rptor</i> cKO mouse heads. (I) Western blots for p-S6K1,S6K1, p-4E-BP1, 4E-BP1, Cyclin D1, Vegf, c-Myc, p-Akt, Akt, p-Erk, Erk, and Gapdh of E11.5 facial primordia, * p<0.05. boc: basioccipital bone; cd: condyle of the mandible; cs: coronal suture; ctr: control; fr: frontal bone; ic: incisor; inf: interfrontal suture; is: lambdoid suture; mx (max in H): maxilla; pmx: premaxilla; nb: nasal bones; ns: non-significant; oc: occipital bone; par: parietal bone; sn: snout; jaw;l: lower jaw; sp: sphenoid bone; ss:sagittal suture; ts: tooth socket; zy: zygomatic arch. Scale bar in (A): 500μm; scale bars in (G.H): 100 μm.</p

Lowering P53 activity by <i>P53</i> copy reduction attenuates the craniofacial phenotype in the NCC-<i>mTOR</i> cKO mice.

<p>(A) Gross examination of NCC-<i>mTOR</i> cKO mice by <i>P53</i> copy deduction. (B) Alcian blue staining in parasagittal sections of E12.5 heads. (C) Histology in frontal sections of E12.5 heads. (D) Quantification of the length of snout/frontonasal prominence, which is represented by the length from the brain-nose turning point to the most anterior plane of the snout. (E) PHH3 staining in frontal sections of E11.5 FN. (F) Apoptosis in frontal sections of E11.5 heads (arrow heads). (G, H) Quantification of cell proliferation and apoptosis. * P<0.05; **P<0.01. bc: basicranium; br; brain; ctr: control; fn: frontonasal prominence; ls: length of the snout; mc: Meckel’ cartilage; md: mandibular arch; mx: maxillary arch; ne: nasal epithelium; pl: palatal shelf; sn: snout; tb: tooth bud; ton: tongue; vd: vessel dilation. Scale bar: 200μm.</p

Apoptosis and proliferation assays.

<p>(A-D) Apoptosis at E10.5 and E11.5. (E) Quantification of apoptotic cells. (F, F’) Dual staining of TUNEL and Runx2 shows that apoptotic cells are predominantly NCC descendants. (G, H) Immunofluorescence for PHH3 in E10.5 facial primordia. (I, J) Immunofluorescence for PHH3 in the FN at E11.5. (K, L) Immunofluorescence for PHH3 in the mandibular arch at E11.5. Arrowheads indicate groove in the tongue. (M) Quantification of PHH3+ cells. (N, O) O9 NCCs cultured with and without rapamycin (100nM). (P) PHH3 staining of O9 cells. (Q)Western blot for mTORC1 downstream target p-S6K1/S6K1 upon rapamycin treatment. (R) Percentage of PHH3+ cells and cell live/death assay. (S) Phase contrast images of cultured PA cells. (T) Phalloidin and Dapi staining of PA cells. (U) PHH3 and Phalloidin double staining of PA cells. br: brain; ctr: control; fn: frontonasal prominence; md: mandibular prominence; mx: maxillary prominence; rapa: rapamycin; ton: tongue. Scale bars in (A-D): 100 μm.; scale bars in (L-U): 200 μm; scale bar in (L) applies to (G-K).</p