Modeling lymphangiogenesis in a three-dimensional culture system

A lack of appropriate in vitro models of three-dimensional lymph vessel growth hampers the study of lymphangiogenesis. We developed a lymphatic ring assay--a potent, reproducible and quantifiable three-dimensional culture system for lymphatic endothelial cells that reproduces spreading of endothelial cells from a pre-existing vessel, cell proliferation, migration and differentiation into capillaries. In the assay, mouse thoracic duct fragments are embedded in a collagen gel, leading to the formation of lumen-containing lymphatic capillaries, which we assessed by electron microscopy and immunostaining. We developed a computerized method to quantify the lymphatic network. By applying this model to gene-deficient mice, we found evidence for involvement of the matrix metalloproteinase, MMP-2, in lymphangiogenesis. The lymphatic ring assay bridges the gap between two-dimensional in vitro models and in vivo models of lymphangiogenesis, can be used to exploit the potential of existing transgenic mouse models, and rapidly identify regulators of lymphangiogenesis.     

Novel ketooxazole based inhibitors of fatty acid amide hydrolase (FAAH)

Efforts to improve the properties of the well studied ketooxazole FAAH inhibitor OL-135 resulted in the discovery of a novel propylpiperidine series of FAAH inhibitors that has a modular design and superior properties to OL-135. The efficacy of one of these compounds was demonstrated in a rat spinal nerve ligation model of neuropathic pain in rats.     

Transient receptor potential, TRP channels: a new family of channels broadly expressed

Transient receptor potential, TRP channels are a new superfamily of functionally versatile non-selective cation channels present from yeast to mammals. On the basis of their structural homology, TRP channels are subdivided in 7 groups : TRPC 1-7 Canonical, TRPV 1-6 Vanilloid, TRPM 1-8 Melastatin, TRPP 1-3 Polycystin, TRPML Mucolipin, TRPA Ankyrin and TRPN (NO mechanotransducer potential C), the latter not expressed in mammals. Their cloning and heterologous expression allowed to demonstrating that these channels are generally weakly voltage-dependent. They are activated by various ligands involving a signal transduction cascade as well as directly by multiple compounds, heat and pH. TRP channels are found in a broad range of cell types. TRP channels are essential in allowing animals to sense the outside world and cells to sense their local environment. Following mutations or anomalous behaviour, these channels have a major role in several human diseases.     

Similar ectopic gene conversion frequencies in the backbone genome of pathogenic and nonpathogenic Escherichia coli strains

We previously showed that gene conversions were more frequent in the genomes of three Escherichia coli pathogenic strains than in the genome of the nonpathogenic K-12 E. coli strain. However, that study did not address whether the more frequent conversions observed in the genes of pathogenic strains occurred between the backbone genes common to these four strains or in the numerous horizontally transferred genes found only in pathogenic strains. Here, we show that ectopic gene conversions are equally frequent in the backbone genes of pathogenic and nonpathogenic strains, that most of these conversions are short, and that the nucleotide changes they generate are probably selectively neutral. Backbone genes are therefore under similar selective constraints in both pathogenic and nonpathogenic E. coli strains. The higher frequency of gene conversions we previously observed in pathogenic strains is therefore due to higher conversion frequencies between the numerous horizontally transferred genes found only in pathogenic strains.     

A yeast PAF acetylhydrolase ortholog suppresses oxidative death

Phospholipids containing sn-2 polyunsaturated fatty acyl residues are primary targets of oxidizing radicals, producing proapoptotic and membrane perturbing fragmented phospholipids. The only known phospholipases that specifically select these oxidized and/or short-chained phospholipids as substrates are mammalian group VII phospholipases A2s that were purified and cloned as PAF acetylhydrolases. Platelet-activating factor (PAF) is a short-chained phospholipid, and whether these enzymes actually are PAF hydrolases or evolved as oxidized phospholipid phospholipases is unknown. The fission yeast Schizosaccharomyces pombe, which does not form or use PAF as a signaling molecule, contains an open-reading frame potentially homologous to mammalian group VII phospholipase A2s. We cloned this SPBC106.11c locus and expressed it in distantly related Saccharomyces cerevisiae that lack homologous sequences. The S. pombe locus encoded a functional phospholipase A2, now renamed plg7+, that hydrolyzed PAF and a synthetic oxidized phospholipid. Expression of human type II PAF acetylhydrolase or S. pombe Plg7p enhanced the viability of S. cerevisiae subjected to oxidative stress. We conclude that a single-celled organism with an exceedingly spare genome still expresses an unusually discriminating phospholipase A2, and that selective hydrolysis of phospholipid oxidation products is an early, and critical, way to overcome oxidative membrane damage and oxidant-induced cell death.     

The role of testosterone in aggressive and non-aggressive risk-taking in adolescent boys

While there exists increasing evidence of a relationship between testosterone (T) and risk-taking (RT), many issues remain unsolved. This paper tries to address two main-questions: (i) does T influence aggressive risk-taking (ART) and/or non-aggressive risk-taking (NART) behavior and (ii) is this relationship stable throughout age and pubertal development and how is the relationship affected by affiliations with peers that are highly involved in RT, referred to as differential association (DA)? For a sample of 301 third-grade adolescent boys (mean age 14.4 years), we assessed the relationship between serum levels of T and estradiol (E2), DA and ART/NART. Significant effects of SHBG (Beta=-0.15; p<0.029) and free testosterone (FT) (Beta=0.23; p<0.003) on NART were shown. No significant effects were found with respect to ART. Further analyses showed that the FT-NART and the FT-ART relations differed as to age but not as to pubertal development (PD) and that the relationship between FT and RT is mediated by DA as such that individuals with higher levels of FT have friends that are more involved in RT and their influence contributes to increased levels of RT. Our results indicate that hormone-related interests and predispositions may influence the development of affiliations with risk-taking peers, a factor which is crucial in understanding adolescent RT.     

Monoterpene glycosides isolated from Fadogia agrestis

Six monoterpene glycosides were isolated from Fadogia agrestis. Their structures were elucidated using a combination of mass spectroscopy, 1D- and 2D-homo- and hetero-NMR spectroscopy and chemical analysis, and established as being derivatives of 2,6-dimethyl-2(E),6(Z)-octadiene-1,8-diol containing from two to four units of rhamnopyranose and, three of them, one or two additional units of glucopyranose. In three of the compounds an acyl group of 8-hydroxy-2,6-dimethyl-2(E),6(Z)-octadienoyl was found esterifying the O-2 position of one of the units of rhamnopyranose.