Low beta diversity of ambrosia beetles (Coleoptera: Curculionidae: Scolytinae and Platypodinae) in lowland rainforests of Papua New Guinea

We assessed the effect of geographical distance on insect species turnover in a situation where other major environmental factors, including host plant species, altitude, and climate, were constant. We sampled ambrosia beetles (Coleoptera, Curculionidae: Scolytinae and Platypodinae) from four tree species: Artocarpus altilis , Ficus nodosa , Leea indica and Nauclea orientalis , at three sites forming a 1000 km transect in lowland rainforests of northern Papua New Guinea. A standardized volume of wood from trunk, branches and twigs was sampled for ambrosia beetles from three individuals of the four tree species at each site. Each tree was killed standing and left exposed to beetle colonization for 20 days prior to sampling. We obtained 12 751 individuals from 84 morphospecies of ambrosia beetles. We surveyed most of the local species richness at each site, predicted by Chao 2 species richness estimates. The similarity of ambrosia beetle communities, estimated by Chao-Sorensen index, was not correlated with their geographical distance. Likelihood analysis and Q-mode analysis using Monte Carlo-generated null distribution of beetles among sites supported the hypothesis that the assemblages of ambrosia beetles at different sites are drawn from the same species pool, regardless of their geographical distance. Tree part (trunk, branch, or twig) was more important predictor of the composition of ambrosia beetle communities than was the host species or geographical location. All three variables, however, explained only a small portion of variability in ambrosia assemblages. The distribution of ambrosia beetles among tree parts, tree species and study sites was mostly random, suggesting limited importance of host specificity or dispersal limitation.     

Ca2+ responses to thyrotropin-releasing hormone and angiotensin II: the role of plasma membrane integrity and effect of G11alpha protein overexpression on homologous and heterologous desensitization

The molecular mechanisms involved in GPCR-initiated signaling cascades where the two receptors share the same signaling cascade, such as thyrotropin-releasing hormone (TRH) and angiotensin II (ANG II), are still far from being understood. Here, we analyzed hormone-induced Ca(2+) responses and the process of desensitization in HEK-293 cells, which express endogenous ANG II receptors. These cells were transfected to express exogenously high levels of TRH receptors (clone E2) or both TRH receptors and G(11)alpha protein (clone E2M11). We observed that the characteristics of the Ca(2+) response, as well as the process of desensitization, were both strongly dependent on receptor number and G(11)alpha protein level. Whereas treatment of E2 cells with TRH or ANG II led to significant desensitization of the Ca(2+) response to subsequent addition of either hormone, the response was not desensitized in E2M11 cells expressing high levels of G(11)alpha. In addition, stimulation of both cell lines with THR elicited a clear heterologous desensitization to subsequent stimulation with ANG II. On the other hand, ANG II did not affect a subsequent response to TRH. ANG II-mediated signal transduction was strongly dependent on plasma membrane integrity modified by cholesterol depletion, but signaling through TRH receptors was altered only slightly under these conditions. It may be concluded that the level of expression of G-protein-coupled receptors and their cognate G-proteins strongly influences not only the magnitude of the Ca(2+) response but also the process of desensitization and resistance to subsequent hormone addition.     

Déjà vu in proteomics. A hit parade of repeatedly identified differentially expressed proteins

After reading many 2-DE-based articles featuring lists of the differentially expressed proteins, one starts experiencing a disturbing déjà vu. The same proteins seem to predominate regardless of the experiment, tissue or species. To quantify the occurrence of individual differentially expressed proteins in 2-DE experiment reports, we compiled the identities of differentially expressed proteins identified in human, mouse, and rat tissues published in three recent volumes of Proteomics and calculated the appearance of the most predominant proteins in the dataset. The most frequently identified protein is a highly abundant glycolytic enzyme enolase 1, differentially expressed in nearly every third experiment on both human and rodent tissues. Heat-shock protein 27 (HSP27) and heat-shock protein 60 (HSP60) were differentially expressed in about 30 percent of human and rodent samples, respectively. Considering protein families as units, keratins and peroxiredoxins are the most frequently identified molecules, with at least one member of the group being differentially expressed in about 40 percent of all experiments. We suggest that the frequent identification of these proteins must be considered in the interpretation of any 2-DE studies. We consider if these commonly observed changes represent common cellular stress responses or are a reflection of the technical limitations of 2-DE.     

Selection and screening for enzymes of nitrile metabolism

This work critically reviews the assays of nitrile-converting and nitrile-forming enzymes (nitrilases, nitrile hydratases, amidases, aldoxime dehydratases). Most of the strains producing such enzymes were obtained by selection on media with nitriles, amides or aldoximes as nitrogen sources. Activity and enantioselectivity of the enzymes was usually assayed by time-consuming chromatographic analysis of substrates and the corresponding reaction products. Attempts at introducing faster assays resulted in several spectrophotometric methods for reaction product (ammonia, hydroxamate, methacrylamide, benzamide, etc.) determination. Recently, new methods for colorimetric and fluorimetric determination of ammonia have been developed, which appear promising for high-throughput assays. Alternatively, methods consisting in determination of NADH consumed in a coupled amination reaction or pH-responsive methods are promising for this purpose. All the above selection and screening methods establish fundamental conditions for the design of hierarchical screening projects. However, the potential of these principles, in particular spectrophotometric and fluorimetric methods, will be probably further exploited and adapted to multiwell plate and robotic systems.     

VDAC2 and aldolase A identified as membrane proteins of K562 cells with increased expression under iron deprivation

We have shown previously that iron deprivation significantly stimulates the uptake of non-transferrin ferric iron from ferric citrate by erythroleukemia K562 cells and that this stimulation depends on protein synthesis. However, we have not detected increased expression of any known iron transport protein (Kovar J. et al. (2006) Blood Cells Mol Dis 37:95–99). Therefore, in order to identify membrane proteins of K562 cells with increased expression under iron deprivation, we employed the isolation of membrane proteins by two-phase partitioning system, protein separation by high-resolution 2D electrophoresis, computer differential analysis, and tandem mass spectrometry. Employing these techniques we identified two proteins with statistically significant upregulation, i.e., aldolase A (ALDA) and voltage-dependent anion channel 2 (VDAC2). The upregulation of aldolase A and VDAC2 in K562 cells under iron deprivation was also confirmed by western blot analysis. This is the first time when the control of aldolase A and VDAC2 levels by iron status of the cell is demonstrated.     

Some Aspects of PCB Metabolism by Horseradish Cells

Transformation of polychlorinated biphenyls was studied using different strains of in vitro cultured cells of horseradish (Armoracia rusticana L.). Time and concentration dependence of this process and production of intracellular and extracellular peroxidases were measured with differentiated shooty teratoma culture K54. The yield of PCB transformation and the viability of the cells were highly dependent on PCB concentration. 100 ppm PCB totally inhibited growth of the cells, and reduced their metabolism of xenobiotics. Experiments with a peroxidase (POX) inhibitor, propylgallate, and a cytochrome P450 inhibitor, aminobenztriazole, indicated the involvement of both enzymatic systems in PCB metabolism.     

The role of defective glycosylation in congenital muscular dystrophy

The dystrophin glycoprotein complex (DGC) is an assembly of proteins spanning the sarcolemma of skeletal muscle cells. Defects in the DGC appear to play critical roles in several muscular dystrophies due to disruption of basement membrane organization. O-mannosyl oligosaccharides on α-dystroglycan, a major extracellular component of the DGC, are essential for normal binding of α-dystroglycan to ligands (such as laminin) in the extracellular matrix and subsequent signal transmission to actin in the cytoskeleton of the muscle cell. Muscle-Eye-Brain disease (MEB) and Walker-Warburg Syndrome (WWS) have mutations in genes encoding glycosyltransferases needed for O-mannosyl oligosaccharide synthesis. Myodystrophic myd mice and humans with Fukuyama Congenital Muscular Dystrophy (FCMD), congenital muscular dystrophy due to defective fukutin-related protein (FKRP) and MDC1D have mutations in putative glycosyltransferases. These human congenital muscular dystrophies and the myd mouse are associated with defective glycosylation of α-dystroglycan. It is expected other congenital muscular dystrophies will prove to have mutations in genes involved in glycosylation. Published in 2004. This revised version was published online in July 2006 with corrections to the Cover Date.