Magnetic resonance imaging of the vomeronasal organ

The vomeronasal organ of mice and frogs was studied by highspatial resolution magnetic resonance imaging. By this techniquethe presence of material in the vomeronasal lumen can be evidencedin vivo.     

Coumarins from Three Phebalium species

The twigs of Phebalium anceps yielded the common furocoumarins psoralen, bergapten and xanthotoxin whereas the leaves of P. coxii and P. ralstonii gare only 8-prenylated 7-methoxycoumarins. From R coxii murrangatin and murralongin were obtained whilst R ralstonii yielded both of these compounds and another coumarin, the novel 7-methoxy-8-(2-acetoxy-3-methyl-1oxobut-2-enyl)-coumarin. The chemotaxonomic implications of these results are discussed.     

X monosomy and 21 trisomy in a sibship.

The case of a sibship of 4, 2 members of which present aneuplo?dy (45,X and 47,XX,21+) is reported. The paternal grandfather and grandmother are first cousins and there is a large number of centromeric associations in the father.     

A one-step procedure for the determination of protein kinase activities and a protein kinase inhibitor in nuclear extract

An isoelectric focusing procedure has been developed that achieves the complete separation of an inhibitor protein from its target enzymes, the nuclear protein kinases, in a one-step operation. The method thus enables one to estimate the amount of inhibitory activity in nuclear extracts, an activity that can not be measured in the presence of the protein kinases. Using this procedure, we have determined the apparent and the actual protein kinase activities in nuclear extracts of rat liver and of two transplantable rat hepatomas by two independent measurements: (a) the increase in protein kinase activity upon separation of the enzyme(s) from the inhibitor, relative to the activity in the crude extracts which was taken as 100%; (b) the inhibitory activity of the pooled focused inhibitor fractions. The amount of cryptic enzyme activity closely correlated with the amount of inhibitor present in extracts from different tissues. Considerably less cryptic kinase activity was found in the tumor nuclei than in liver which may account, at least in part, for the higher phosphorylating capacity previously observed in hepatoma.     

Requirement for kasB in Mycobacterium mycolic acid biosynthesis,cell wall impermeability and intracellular survival: implications for therapy

Mycobacterium tuberculosis infects one-third of the world's population and causes two million deaths annually. The unusually low permeability of its cell wall contributes to the ability of M. tuberculosis to grow within host macrophages, a property required for pathogenesis of infection. Mycobacterium marinum is an established model for discovering genes involved in mycobacterial infection. Mycobacterium marinum mutants with transposon insertions in the beta-ketoacyl-acyl carrier protein synthase B gene (kasB) grew poorly in macrophages, although growth in vitro was unaffected. Detailed analyses by thin-layer chromatography, nuclear magnetic resonance (NMR), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, infrared spectroscopy, and chemical degradations showed that the kasB mutants synthesize mycolic acids that are 2-4 carbons shorter than wild type; the defect was localized to the proximal portion of the meromycolate chain. In addition, these mutants showed a significant (approximately 30%) reduction in the abundance of keto-mycolates, with a slight compensatory increase of both alpha- and methoxy-mycolates. Despite these small changes in mycolate length and composition, the kasB mutants exhibited strikingly altered cell wall permeability, leading to a marked increase in susceptibility to lipophilic antibiotics and the host antimicrobial molecules defensin and lysozyme. The abnormalities of the kasB mutants were fully complemented by expressing M. tuberculosis kasB, but not by the closely related gene kasA. These studies identify kasB as a novel target for therapeutic intervention in mycobacterial diseases.     

Glucocorticoid metabolism by 11-beta hydroxysteroid dehydrogenase type 2 modulates human mineralocorticoid receptor transactivation activity

The mineralocorticoid receptor (MR) binds aldosterone, but also glucocorticoid hormones (corticosterone in rodents, cortisol in humans), which largely prevail in the plasma. To prevent permanent and maximal occupancy of MR by glucocorticoid hormones in aldosterone-target cells, specific effects of aldosterone require metabolism of glucocorticoid hormones into 11-dehydroderivatives by 11-beta hydroxysteroid dehydrogenase (11-HSD2). We analyzed the effect of corticosterone or 11-dehydrocorticosterone (11-DHC) on the transactivation activity of the MR, transiently expressed in a new renal cell line expressing 11-HSD2. We show that, because of its metabolism by 11-HSD2, corticosterone is a poor activator of MR transactivation, except at micromolar concentrations, where the enzyme is saturated. We also show that high micromolar concentrations of 11 DHC are required to activate the MR. The weak antagonist property of 11-DHC on aldosterone-induced hMR transactivations is also documented. Such partial agonist activity of 11-DHC is discussed in the light of its positioning in a three-dimensional model of the MR ligand-binding domain.     

Identification of Aurora kinases as RasGAP Src homology 3 domain-binding proteins

The GTPase-activating protein RasGAP functions as both a negative regulator and an effector of Ras proteins. In tumor cells, RasGAP is no longer able to deactivate oncogenic Ras proteins, and its effector function becomes predominant. As RasGAP itself has no obvious enzymatic function that may explain this effector function, we looked for downstream RasGAP effectors that could fulfill this role. We looked for the existence of RasGAP Src homology 3 (SH3) domain partners as this domain is involved in the regulation of cell proliferation and has an anti-apoptotic effect. We report here the identification of a new RasGAP SH3 domain-binding protein, named Aurora. This Drosophila melanogaster Ser/Thr kinase has three human orthologs called Aurora/Ipl1-related kinase or HsAIRK-1, -2, and -3. Coimmunoprecipitation experiments in COS cells confirmed that HsAIRK-1 and HsAIRK-2 both interact with RasGAP. RasGAP pull-down experiments showed that it interacts with HsAIRK-1 in G(2)/M HeLa cells. We also demonstrated that RasGAP binds to the kinase domain of Aurora and that this interaction inhibits the kinase activity of HsAIRK-1 and HsAIRK-2. Finally we showed that RasGAP forms a ternary complex with HsAIRK and survivin. This complex may be involved in the regulation of the balance between cell division and apoptosis.     

Adhesion of Bacillus spores and Escherichia coli cells to inert surfaces: role of surface hydrophobicity

The ability of bacterial spores and vegetative cells to adhere to inert surfaces was investigated by means of the number of adherent spores (Bacillus cereus and Bacillus subtilis spores) and Escherichia coli cells and their resistance to cleaning or rinsing procedures (adhesion strength). Six materials (glass, stainless steel, polyethylene high density (PEHD), polyamide-6, polyvinyl chloride, and Teflon) were tested. Slight differences in the number of adherent spores (less than 1 log unit) were observed between materials, but a higher number of adherent E. coli cells was found on the hydrophobic materials PEHD and Teflon. Conversely, the resistance of both B. cereus and B. subtilis spores to a cleaning procedure was significantly affected by the material. Hydrophobic materials were harder to clean. The topography parameter derived from the Abbott-Firestone curve, RVK, and, to a lesser extent, the widely used roughness parameters RA (average roughness) and Rz (maximal roughness), were related to the number of adherent cells. Lastly, the soiling level as well as the adhesion strength were shown to depend largely on the microorganism. The number of adhering B. cereus hydrophobic spores and their resistance to a cleaning procedure were found to be 10 times greater than those of the B. subtilis hydrophilic spores. Escherichia coli was loosely bound to all the materials tested, even after 24 h biofilm formation.     

Expression analysis in plant and cell suspensions of CrCKR1, a cDNA encoding a histidine kinase receptor homologue in Catharanthus roseus (L.) G. Don

A full length cDNA (CrCKR1) encoding a hybrid histidine kinase was isolated from a Catharanthus roseus cDNA library. The kinase belongs to the subfamily of cytokinin receptors represented by CRE1/AHK4/WOL in Arabidopsis thaliana. In cell suspensions, the expression of CrCKR1 is not affected by various stress and hormonal treatments but is stimulated in cells continuously exposed to cytokinin. In plants, CrCKR1 is strongly expressed only in the petals of mature flowers. These data suggest that CrCKR1 could take part in the mechanisms leading to the production of secondary metabolites in C. roseus.     

Overexpression of Galectin-7 in Mouse Epidermis Leads to Loss of Cell Junctions and Defective Skin Repair

Background

The proteins of the galectin family are implicated in many cellular processes, including cell interactions, polarity, intracellular trafficking, and signal transduction. In human and mouse, galectin-7 is almost exclusively expressed in stratified epithelia, notably in the epidermis. Galectin-7 expression is also altered in several human tumors of epithelial origin. This study aimed at dissecting the consequences of galectin-7 overexpression on epidermis structure and functions in vivo.

Methods

We established transgenic mice specifically overexpressing galectin-7 in the basal epidermal keratinocytes and analyzed the consequences on untreated skin and after UVB irradiation or mechanical injury.

Results

The intercellular cohesion of the epidermis is impaired in transgenic animals, with gaps developing between adjacent keratinocytes, associated with loss of adherens junctions. The epidermal architecture is aberrant with perturbations in the multilayered cellular organisation of the tissue, and structural defects in the basement membrane. These transgenic animals displayed a reduced re-epithelialisation potential following superficial wound, due to a defective collective migration of keratinocytes. Finally, a single mild dose of UVB induced an abnormal apoptotic response in the transgenic epidermis.

Conclusion

These results indicate that an excess of galectin-7 leads to a destabilisation of adherens junctions associated with defects in epidermal repair. As this phenotype shares similarities with that of galectin-7 null mutant mice, we conclude that a critical level of this protein is required for maintaining proper epidermal homeostasis. This study brings new insight into the mode of action of galectins in normal and pathological situations.