Binding properties of beta-adrenergic receptors in early human fetal lung

The characteristics of beta-adrenergic receptors in human fetal lung were examined using the beta-adrenoceptor antagonist 3H-dihydroalprenolol /DHA/. Steady-state binding was reached by 15 min at 25 degrees C, and the association and dissociation rate constants were 0.0422 nM-1 min-1 and 0.0874 min-1, respectively. From saturation experiments, Bmax of 82.0 +/- 38 fmol/mg protein and KD = 1.85 +/- 0.92 nM were calculated. Inhibition of 3H-DHA binding by beta-1 /metoprolol/ and beta-2 /zinterol, IPS-339, fenoterol/ selective drugs resulted in biphasic displacement curves with slope factors less than 1.0. Analysis of these curves revealed a beta-1: beta-2 ratio of 40:60 in human fetal lung. The presence of 3H-DHA binding sites in early human lung may have a developmental importance which is not yet understood.     

Altered profile of secondary metabolites in the root exudates of Arabidopsis ATP-binding cassette transporter mutants

Following recent indirect evidence suggesting a role for ATP-binding cassette (ABC) transporters in root exudation of phytochemicals, we identified 25 ABC transporter genes highly expressed in the root cells most likely to be involved in secretion processes. Of these 25 genes, we also selected six full-length ABC transporters and a half-size transporter for in-depth molecular and biochemical analyses. We compared the exuded root phytochemical profiles of these seven ABC transporter mutants to those of the wild type. There were three nonpolar phytochemicals missing in various ABC transporter mutants compared to the wild type when the samples were analyzed by high-performance liquid chromatography-mass spectrometry. These data suggest that more than one ABC transporter can be involved in the secretion of a given phytochemical and that a transporter can be involved in the secretion of more than one secondary metabolite. The primary and secondary metabolites present in the root exudates of the mutants were also analyzed by gas chromatography-mass spectrometry, which allowed for the identification of groups of compounds differentially found in some of the mutants compared to the wild type. For instance, the mutant Atpdr6 secreted a lower level of organic acids and Atmrp2 secreted a higher level of amino acids as compared to the wild type. We conclude that the release of phytochemicals by roots is partially controlled by ABC transporters.     

The Harlequin ladybird continues to invade southeastern Europe

The ladybird Harmonia axyridis (Pallas) is considered one of the most serious invasive species around the globe. It has spread all over North America and Western Europe, while data from southeastern Europe, especially in the Mediterranean region, are scarce. In this study we present the first confirmed data of the spread of H. axyridis throughout Croatia. Specimens were sampled and identified during the period 2008–2010. The species was recorded at 18 localities in all three colour forms in various habitat types. Light trapping was found to be a satisfactory method for collecting H. axyridis. Since there is no evidence to suggest the deliberate introduction of H. axyridis in Croatia, it can be assumed that it has spread southwards from Central and Eastern Europe, and that it will probably continue to spread. Further investigations should focus on monitoring and detailed mapping of H. axyridis in Croatia and neighbouring countries, especially in the Mediterranean region, to determine whether stable populations are present.     

HADDOCK versus HADDOCK: new features and performance of HADDOCK2.0 on the CAPRI targets

Here we present version 2.0 of HADDOCK, which incorporates considerable improvements and new features. HADDOCK is now able to model not only protein-protein complexes but also other kinds of biomolecular complexes and multi-component (N > 2) systems. In the absence of any experimental and/or predicted information to drive the docking, HADDOCK now offers two additional ab initio docking modes based on either random patch definition or center-of-mass restraints. The docking protocol has been considerably improved, supporting among other solvated docking, automatic definition of semi-flexible regions, and inclusion of a desolvation energy term in the scoring scheme. The performance of HADDOCK2.0 is evaluated on the targets of rounds 4-11, run in a semi-automated mode using the original information we used in our CAPRI submissions. This enables a direct assessment of the progress made since the previous versions. Although HADDOCK performed very well in CAPRI (65% and 71% success rates, overall and for unbound targets only, respectively), a substantial improvement was achieved with HADDOCK2.0.     

LOX-1 inhibition in myocardial ischemia-reperfusion injury: modulation of MMP-1 and inflammation

A recently identified lectin-like oxidized low-density lipoprotein receptor (LOX-1) mediates endothelial cell injury and facilitates inflammatory cell adhesion. We studied the role of LOX-1 in myocardial ischemia-reperfusion (I/R) injury. Anesthetized Sprague-Dawley rats were subjected to 60 min of left coronary artery (LCA) ligation, followed by 60 min of reperfusion. Rats were treated with saline, LOX-1 blocking antibody JXT21 (10 mg/kg), or nonspecific anti-goat IgG (10 mg/kg) before I/R. Ten other rats underwent surgery without LCA ligation and served as a sham control group. LOX-1 expression was markedly increased during I/R (P < 0.01 vs. sham control group). Simultaneously, the expression of matrix metalloproteinase-1 (MMP-1) and adhesion molecules (P-selectin, VCAM-1, and ICAM-1) was also increased in the I/R area (P < 0.01 vs. sham control group). There was intense leukocyte accumulation in the I/R area in the saline-treated group. Treatment of rats with the LOX-1 antibody prevented I/R-induced upregulation of LOX-1 and reduced MMP-1 and adhesion molecule expression as well as leukocyte recruitment. LOX-1 antibody, but not nonspecific IgG, also reduced myocardial infarct size (P < 0.01 vs. saline-treated I/R group). To explore the link between LOX-1 and adhesion molecule expression, we measured expression of oxidative stress-sensitive p38 mitogen-activated protein kinase (p38 MAPK). The activity of p38 MAPK was increased during I/R (P < 0.01 vs. sham control), and use of LOX-1 antibody inhibited p38 MAPK activation (P < 0.01). These findings indicate that myocardial I/R upregulates LOX-1 expression, which through p38 MAPK activation increases the expression of MMP-1 and adhesion molecules. Inhibition of LOX-1 exerts an important protective effect against myocardial I/R injury.     

HIV-associated dementia, mitochondrial dysfunction, and oxidative stress

Over the past several decades, researchers have made large advances in unraveling the pathogenesis of HIV-related neurological disease leading to substantial benefits for affected individuals. Concomitant advances in HIV-treatment have changed the landscape of HIV care, resulting in alterations in HIV neuroepidemiolgy and potentially the neuropathogenesis of cognitive disorders. Specifically, widespread ARV medications use has heightened our awareness of mitochondrial toxicity, oxidative stress, and metabolic abnormalities and stimulated more research into the related cognitive consequences. Specific sources of oxidative stress among HIV-1-infected individuals to be discussed in this article include the direct effects of HIV-1, chronic immune activation in response to HIV-1 and other pathogens, and co-morbid factors. Continued research in this area could provide novel therapeutic targets.