The aim of the study was to evaluate the impact of defoliation intensity, defoliation frequency, and interactions with N supply on N uptake, N mobilization from and N allocation to roots, adult leaves, and growing leaves. Plants of Lolium perenne were grown under two contrasted N regimes. Defoliation intensity treatments consisted of a range of percentage leaf area removal (0, 25, 50, 75, or 100%). These treatments were applied in parallel to a set of plants previously undefoliated, and to a second set of plants which had been defoliated several times at a constant height. A (15)N tracer technique was used to quantify N uptake, mobilization, and allocation over a 7 d period. A significant reduction in plant N uptake was observed with the removal of more than 75% of lamina area, but only with high N supply. As defoliation intensity increased, the amount of N taken up and subsequently allocated to growing leaves during the labelling period was maintained at the expense of N allocation to roots and adult leaves. Increasing defoliation intensity increased the relative contribution of roots supplying mobilized N to growing leaves and decreased the relative contribution of adult leaves. Defoliation frequency did not substantially alter N uptake, mobilization, and allocation between roots, adult and growing leaves on a plant basis. However, tiller number per plant was largely increased under repeated defoliation, hence indicating that allocation and mobilization of N to growing leaves, on the basis of individual tillers, was decreased by defoliation frequency.

Peroxisome proliferator-activated receptors (PPARs) regulate storage and catabolism of fats and carbohydrates. PPAR␥ activity increases insulin sensitivity and adipocyte differentiation at the expense of adipogenesis and weight gain. The goal of this study was to 1) clone the promoter of the human adipocyte fatty acid binding protein (aP2) gene, namely fatty acid-binding protein-4, 2) characterize its pharmacological regulation, and 3) determine its putative predictability for adipogenesis. Among the selected PPAR agonists, rosiglitazone and pioglitazone displayed the highest maximal efficacy (E max ) on reporter-gene assays in COS-7 cells cotransfected by either a galactosidase 4-response elementbased or a human aP2 promoter-based Luc reporter vector, along with either chimeric or full-length human PPAR expression plas- -0072), and indomethacin behaved as partial agonists relative to pioglitazone in full-length human aP2-PPAR␥2. Beyond their partial PPAR␥ agonist properties, these compounds elicited a lower maximal up-regulation of mouse aP2 mRNA in 3T3-L1 adipocytes as compared with pioglitazone; these properties paralleled a time-dependent increase in neutral lipids. By contrast, the selective PPAR␣ agonist 2,2-dichloro-12-(4-chlorophenyl)dodecanoic acid (BM-17.0744) neither stimulated the human aP2-PPAR␣ promoter reporter-gene assay, thus demonstrating a specific interaction between PPAR␥ and the aP2 promoter, nor affected lipogenesis in 3T3-L1 cells. Altogether, these data characterized a functional promoter of the human aP2 gene; its in vitro pharmacological regulation in PPAR␥-mediated reporter-gene assay may represent an interesting complement or an alternative to time-consuming procedures aiming at discriminating PPAR ligands with low lipogenic properties.Metabolic syndrome is characterized by the clustering of at least three risk factors among hypertension, certain types of dyslipidemia, impaired glucose tolerance and type II diabetes, and obesity. These metabolic abnormalities lead to atherosclerosis and related complications (Haffner and Taegtmeyer, 2003). The control of lipid and carbohydrate metabolism, including physiologic and pharmacological treatments, represents a valid rationale to reduce cardiovascular diseases in patients with metabolic syndrome (Beckman et al., 2003;Wilson and Grundy, 2003).PPARs are a subclass of the nuclear receptor superfamily acting as ligand-dependent transcription factors (Kersten et al., 2000). Three subtypes were identified; the ␣ isoform is the primary subtype expressed in liver, but also in heart and kidney, and acts as a major regulator of metabolism of fats, catabolism of fatty acids, and synthesis and catabolism of lipoproteins (Barbier et al., 2002). PPAR␣ is also involved in Parts of this work were presented as a poster at the 74th Congress of the European Atherosclerosis Society (April 17-20, 2004, Seville, Spain).Article, publication date, and citation information can be found at

The Hedgehog (HH) pathway has been linked to the formation of basal cell carcinoma (BCC), medulloblastoma, and other cancers. The recently approved orally active drugs vismodegib (GDC‐0449) and sonidegib (LDE–225) were not only efficacious for the treatment of advanced or metastatic BCC by antagonizing the smoothened (SMO) receptor, but also produced important side effects, limiting their use for less invasive BCC. Herein, we compared a large series of SMO antagonists, including GDC‐0449 and LDE‐225, the clinically tested BMS‐833923, CUR‐61414, cyclopamine, IPI‐926 (saridegib), itraconazole, LEQ‐506, LY‐2940680 (taladegib), PF‐04449913 (glasdegib), and TAK‐441 as well as preclinical candidates (PF‐5274857, MRT‐83) in two SMO‐dependent cellular assays and for G‐protein activation. We report marked differences in inhibitor potencies between compounds as well as a notable disparity between the G‐protein assay and the cellular tests, suggesting that classification of drugs is assay dependent. Furthermore, we explored topical efficacies of SMO antagonists on depilated mice using Gli1 and Ptch1 mRNA quantification in skin as biomarkers of the HH signaling inhibition. This topical model rapidly discriminated drugs in terms of efficacies and potencies for inhibition of both biomarkers. SMO antagonists showed also a large variation in their blood and skin partition, suggesting that some drugs are more favorable for topical application. Overall, our data suggested that in vitro and in vivo efficacious drugs such as LEQ‐506 and TAK‐441 may be of interest for topical treatment of less invasive BCC with minimal side effects.

It is now recognized that atherosclerosis complications are related to the unstable character of the plaque rather than its volume. Vulnerable plaques often contain a large lipid core, a reduced content of smooth muscle cells, and accumulation of inflammatory cells. Colocalization of macrophages and active matrix metalloproteinases (MMPs) is likely relevant for atherosclerotic lesion disruption. Nevertheless, MMP activity and regulation by cardiovascular drugs remains poorly defined. In this study, we evaluated the effects of avasimibe, fluvastatin, and peroxisome proliferator-activated receptor (PPAR) ligands on 92-kDa gelatinase B (MMP-9) secretion by human THP-1 macrophages. THP-1 macrophages were treated with compounds for 48 h, and secreted MMP-9 protein was quantified by immunoassay. Avasimibe, fluvastatin, and PPARalpha agonists (fenofibric acid and Wy-14643) significantly reduced, in a concentration-dependent manner, MMP-9 protein (up to 67 +/- 5% for fenofibric acid). In these assays, the PPARgamma selective agonist rosiglitazone displayed a lower efficacy than other compounds. Enzymatic activity of MMP-9 was also decreased by all cardiovascular drugs tested. MMP-9 protein/activity inhibition by cardiovascular drugs was due, at least in part, to a decrease in MMP-9 mRNA. These results show that THP-1 macrophages could be an useful cellular model to investigate effects of compounds on plaque vulnerability through MMP-9 activity.

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