Warm molecular gas temperature distribution in six local infrared bright Seyfert galaxies

<p><b>A, C, E and G</b>: Summarized data depicting the effect of L-NAME (100 μM) or endothelium removal (denuded) on propofol-induced (does-dependent) changes in luminal diameter in coronary microvessels obtained from control, TRPV1^-/-, TRPA1^-/- and TRPAV^-/- mice, respectively (<i>n</i> = 12). <b>B, D, F and H</b>: Summarized data depicting the effect of Pen A (50 μM) alone and in combination with L-NAME on propofol-induced changes in luminal diameter in coronary microvessels obtained from control, TRPV1^-/-, TRPA1^-/- and TRPAV^-/- mice, respectively (<i>n</i> = 12). Data are expressed as % relaxation ± SEM. *<i>P</i>< 0.05 vs. control.</p

A schematic representation showing the proposed mechanisms by which propofol causes vasodepressor responses <i>in-vivo</i>.

<p>A schematic representation showing the proposed mechanisms by which propofol causes vasodepressor responses <i>in-vivo</i>.</p

Propofol induced changes in MAP in Control and TRPV1^-/- mice: Panels A and B: Summarized data depicting the effect of L-NAME (100 mg/kg/min) on propofol-induced changes in MAP in control and TRPV1^-/- mice.

<p>Panels <b>C</b> and <b>D:</b> Summarized data depicting the effect of Pen A (50 ug/kg/min) on propofol-induced changes in MAP in control and TRPV1^-/- mice. Panels <b>E</b> and <b>F:</b> Summarized data depicting the effect of L-NAME and Pen A in combination on propofol-induced changes in MAP in control and TRPV1^-/- mice. Data are means ± SEM. *<i>P</i><0.05 compared to baseline. #<i>P</i><0.05 compared to control. <i>n</i> = 6 mice in each group.</p

Propofol induced changes in MAP in TRPA1^-/- and TRPAV^-/- mice: Panels A and B: Summarized data depicting the effect of L-NAME (100 mg/kg/min) on propofol-induced changes in MAP in TRPA1^-/- and TRPAV^-/- mice.

<p>Panels <b>C</b> and <b>D:</b> Summarized data depicting the effect of Pen A (50ug/kg/min) on propofol-induced changes in MAP in TRPA1^-/- and TRPAV^-/- mice. Panels <b>E</b> and <b>F:</b> Summarized data depicting the effect of L-NAME and Pen A in combination on propofol-induced changes in MAP in control and TRPA1^-/- and TRPAV^-/- mice. Data are means ± SEM. *<i>P</i><0.05 compared to baseline. <i>n</i> = 6 mice in each group.</p

Baseline Hemodynamics.

<p>Values are represented as Mean ± SE and were obtained after addition of hexamethonium. Body weights were recorded before anesthesia [Control: n = 26; TRPA1 ^-/-: n = 12; TRPV1^-/-: n = 12 and TRPAV^-/-: n = 12]. MAP = Mean Arterial Pressure.</p><p>(*) denotes significance from control p<0.05.</p><p>Baseline Hemodynamics.</p

Detection of TRPA1-TRPV1 complex formation in MCAECs using proximity ligation assay (PLA).

<p><b>A</b>: MCAECs stained with PLA probes only were used as negative controls. MCAECs stained with DAPI (blue), TRPA1 and TRPV1 antisera conjugated with PLA probes (red signal). <b>B</b>: Effect of propofol (10μM) or AITC (100μM) treatment on PLA signal in MCAECs. <b>C</b>: PLA of MCAECs pretreated with HC-030031 (0.5μM), then incubated with either propofol (10μM) or AITC (100μM). <b>D</b>: Lack of effect of propofol (10 μM) on aortic endothelial cells obtained from TRPA1^-/- and TRPV1^-/- mice. <i>n</i> = 4 coverslips of cells.</p

Unsuspected osteochondroma-like outgrowths in the cranial base of Hereditary Multiple Exostoses patients and modeling and treatment with a BMP antagonist in mice

<div><p>Hereditary Multiple Exostoses (HME) is a rare pediatric disorder caused by loss-of-function mutations in the genes encoding the heparan sulfate (HS)-synthesizing enzymes EXT1 or EXT2. HME is characterized by formation of cartilaginous outgrowths—called osteochondromas- next to the growth plates of many axial and appendicular skeletal elements. Surprisingly, it is not known whether such tumors also form in endochondral elements of the craniofacial skeleton. Here, we carried out a retrospective analysis of cervical spine MRI and CT scans from 50 consecutive HME patients that included cranial skeletal images. Interestingly, nearly half of the patients displayed moderate defects or osteochondroma-like outgrowths in the cranial base and specifically in the clivus. In good correlation, osteochondromas developed in the cranial base of mutant <i>Ext1</i>^<i>f/f</i><i>;Col2-CreER</i> or <i>Ext1</i>^<i>f/f</i><i>;Aggrecan-CreER</i> mouse models of HME along the synchondrosis growth plates. Osteochondroma formation was preceded by phenotypic alteration of cells at the chondro-perichondrial boundary and was accompanied by ectopic expression of major cartilage matrix genes -<i>collagen 2</i> and <i>collagen X-</i> within the growing ectopic masses. Because chondrogenesis requires bone morphogenetic protein (BMP) signaling, we asked whether osteochondroma formation could be blocked by a BMP signaling antagonist. Systemic administration with LDN-193189 effectively inhibited osteochondroma growth in conditional <i>Ext1</i>-mutant mice. In vitro studies with mouse embryo chondrogenic cells clarified the mechanisms of LDN-193189 action that turned out to include decreases in canonical BMP signaling pSMAD1/5/8 effectors but interestingly, concurrent increases in such anti-chondrogenic mechanisms as pERK1/2 and <i>Chordin</i>, <i>Fgf9</i> and <i>Fgf18</i> expression. Our study is the first to reveal that the cranial base can be affected in patients with HME and that osteochondroma formation is amenable to therapeutic drug intervention.</p></div

Osteochondroma development in long bones and ribs in juvenile mice is inhibited by systemic LDN-193189 treatment.

<p><b>(A to D)</b> μCT images (A, B and D) and X-ray image (C) of knee and ribs from mutant <i>Ext1</i>^<i>f/f</i><i>;Agr-CreER</i> mice receiving vehicle treatment and showing large osteochondromas (double arrowheads). (<b>E to H</b>) Analogous images from companion mutant mice receiving LDN treatment for 6 weeks. Note the appreciable reduction in osteochondorma size in both knee and ribs (arrowhead). Bar in (A) for A, B, E and F, 5 mm; bar in (C) for C and G, 3 mm; and bar in (D) for D and H, 1.5 mm.</p

Stereotypic osteochondromas form in a juvenile HME mouse model.

<p>(<b>A-F</b>) Bright field and fluorescence images of longitudinal sections of spheno-occipital (<i>sos</i>) synchondroses (A-C), tibia (D-E) and ribs (F) from 5 week-old control <i>R26-tdTomato;Agr-CreER</i> mice that had been injected with vehicle 3 days earlier. The fluorescence images (B and E) show absence of reporter activity in growth plates (<i>gp</i>) and perichondrium (<i>pc</i>) at any anatomical site examined and thus, lack of <i>CreER</i> leakage in this model. Note that ribs contain a large resting cartilage (<i>rc</i>) adjacent to the growth plate (F). (<b>G-H</b>) μCT images of cranial base and knee from control <i>Ext1</i>^<i>f/f</i><i>;Agr-CreER</i> mice sacrificed 6 weeks from vehicle injection and displaying normal and typical anatomical characteristics. (<b>I-N</b>) Bright field and fluorescence images from companion <i>R26-tdTomato;Agr-CreER</i> mice that were administered tamoxifen once and were sacrificed 3 days post-injection. Note that reporter activity is very strong in spheno-occipital synchondrosis (<i>sos</i>), tibia and rib growth plates and rib resting cartilage (J, K, M and N) and is also conspicuous in flanking perichondrial (<i>pc</i>) cells at each anatomical location (arrowheads in K and M). Boxed area in J is shown at higher magnification in K. (<b>O-P</b>) μCT images of cranial base and knee from mutant <i>Ext1</i>^<i>f/f</i><i>;Agr-CreER</i> mice sacrificed 6 weeks after tamoxifen injection. Note the large multiple osteochondromas (arrowheads) protruding away from the bone surfaces at each anatomical site and better appreciable when contrasted to corresponding images from companion controls (G-H). Bar in (A) for A, B, I and J: 300 μm; bar in (C for C and K, 150 μm; bar in (D) for D, E, L and M, 250 μm; bar in (F) for F and N, 150 μm; and bar in (G) for G and O, 0.8 mm; and in (H) for H and P, 5 mm.</p

LDN-193189 is a strong and direct inhibitor of chondrogenesis.

<p>(<b>A-J</b>) Alcian blue staining and optical quantification of mouse embryo limb bud cell micromass cultures on day 4 and 6 reared in control medium (A and F) or medium containing rhBMP2 (B and G), LDN (C and H) or rhBMP2 plus LDN (D and I). Note the drastic inhibition of chondrogenesis by LDN treatment at both day 4 and 6 (C and H) and partial but still major inhibition in co-treated cultures (D and I). Quantification of alcian blue staining confirms visual data and establish statistical significance (E and J). (** p < 0.005; *** p < 0.001). (<b>K-P</b>) Histograms of qPCR data showing that LDN treatment caused significant decreases in master cartilage genes Sox9 and aggrecan (Acan) on both day 4 and 6 of culture compared to levels in control or rhBMP2-stimulated cultures, while concurrently increasing the gene expression levels of the endogenous BMP antagonist Chordin. (* p < 0.05; ** p < 0.01; *** p < 0.001).</p