Cytology of induced systemic resistance of tomato to Phytophthora infestans

The infection of tomato leaves by Phytophthora infestans was followed using cytological methods. Fungal ingress and plant reactions in untreated and induced resistant plants were studied. Systemic disease resistance was induced by a local pre-infection with the same fungus. Induction retarded fungal progress at the leaf surface, epidermis and in the mesophyll. The reduced numbers of germinated cysts indicate the presence of fungitoxic substances on the leaf surface of induced plants. Frequency of fungal penetration through the outer epidermal cell wall was reduced, but only in plants exhibiting a high level of induced resistance. Autofluor-escent material, indicating the presence of lignin-like substances, accumulated rapidly beneath some of the appressoria, but this plant response was similar in induced and non-induced plants. Staining with aniline blue indicated that callose deposition was not involved in induced resistance. Thus, none of the cytologically investigated plant reactions correlated with the reduced penetration frequency observed. In the mesophyll, however, the cytological picture corresponding to a hypersensitive reaction occurred more often in induced plants. It is concluded that reduction of disease severity by induction is the result of the combined action of several successive defence reactions.Dedicated to the memory of Professor H. Grisebach     

Contribution of a single hydroxyl group to transition-state discrimination by adenosine deaminase: evidence for an "entropy trap" mechanism

Adenosine deaminase was found to bind 6-hydroxy-1,6-dihydropurine ribonucleoside (II), formed by reversible addition of water to purine ribonucleoside (I) in a reaction analogous to formation of a tetrahedral intermediate in substrate deamination, with an apparent Ki value of 3 x 10(-13) M at 20 degrees C. 1,6-Dihydropurine ribonucleoside (IV), synthesized by photolysis of purine ribonucleoside in the presence of NaBH4, exhibited a Ki value of 5.4 x 10-6 M. After correction for differences between the relative free energies of solvation of II and IV, the 6-hydroxyl group of II was estimated to contribute more than 16 kcal to the free energy of binding, approaching the enthalapy of formation of a single hydrogen bond to charged group in the vapor phase. The relatively weak binding of IV and of substrate water suggests that entropic effects, arising from the cooperative action of binding determinants contained within these separate molecules, contribute more than 10 kcal/mol to the free energy of binding of II in which these binding determinants are contained within a single molecule. In free solution, the entropy of reversible hydration of I was evaluated by measuring the temperature dependence of equilibria of protonation of I and of pseudobase formation from I-methylpurinium ribonucleoside as -35 eu, comparable with the entropy of activation for the uncatalyzed hydrolysis of adenosine. In the active site of adenosine deaminase, this thermodynamic obstacle is evidently climbed spontaneously as a result of attractive interactions between the active site and the critical hydroxyl group at the 6-position.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of litter removal and mowing on germination and establishment of two fen-grassland species along a productivity gradient

To study the factors that limit the occurrence of species in fen grasslands, the effects of mowing and litter removal on germination, growth and establishment of two common species (Silene flos-cuculi andLotus pedunculatus) were analyzed along a productivity gradient at seven sites in Northern Germany. The sites differed in nutrient availability, vegetation composition and standing crop. In autumn 2002 a field experiment was set up at each site, including factorial combinations of two treatments (litter removal, mowing). Seeds and juvenile individuals of both species were transferred within the treatment combinations. The number of germinated and transplanted individuals was recorded during the 2003 and 2004 seasons and biomass of survived plants weighed in late summer 2004. Mowing and litter removal had positive effects on the number of germinated seeds of both species. The effect of litter removal was mostly significant at sites with a comparatively low nutrient availability and standing crop (350 g/m²), while the effect of mowing was more important at the site with the highest standing crop (1000 g/m²). In some cases the presence of litter had a positive effect on the survival of individuals, which may be caused by extremely dry weather conditions in 2003. The experiment showed that plant species reaction on mowing and litter removal differed both according to their life stage and the site conditions. These differences could be interpreted in context with Shifting Limitation Hypothesis because the abolition of seed limitation by sowing and creation of additional gaps for germination was most efficient at low productive sites, while the high standing crop reduced first of all the establishment probability, which could be increased by mowing.     

A highly pathogenic strain of Bacillus thuringiensis serovar kurstaki in lepidopteran pests

In order to detect and identify the most toxic Bacillus thuringiensis strains against pests, we isolated a B. thuringiensis strain (Bn1) from Balaninus nucum (Coleoptera: Curculionidae), the most damaging hazelnut pest. Bn1 was characterized via morphological, biochemical, and molecular techniques. The isolate was serotyped, and the results showed that Bn1 was the B. thuringiensis serovar, kurstaki (H3abc). The scanning electron microscopy indicated that Bn1 has crystals with cubic and bipyramidal shapes. The Polymerase Chain Reactions (PCRs) revealed the presence of the cry1 and cry2 genes. The presence of Cry1 and Cry2 proteins in the Bn1 isolate was confirmed via SDS-PAGE, at approximately 130 kDa and 65 kDa, respectively. The bioassays conducted to determine the insecticidal activity of the Bn1 isolate were conducted with four distinct insects, using spore-crystal mixtures. We noted that Bn1 has higher toxicity as compared with the standard B. thuringiensis subsp. kurstaki (HD-1). The highest observed mortality was 90% against Malacosoma neustria and Lymantria dispar larvae. Our results show that the B. thuringiensis isolate (Bn1) may prove valuable as a significant microbial control agent against lepidopteran pests.     

Relations of the morphological characteristic latent structure and body posture indicators in children aged seven to nine years

With the aim of determining the connection between the indicators of body posture and latent structure of morphological variables in children aged 7 and 8 years, first and second grade of primary school, a set of 17 morphological measures and 12 body posture indicators were longitudinally applied to a sample of 110 boys and 114 girls. The latent structure of morphological variables in both sexes was defined by three factors but at a different order of significance: in boys, the order was longitudinal dimensionality, voluminosity, mass and subcutaneous fat tissue and transverse dimensionality, whereas in girls the order was voluminosity, mass and subcutaneous fat tissue, longitudinal dimensionality and transverse dimensionality. The latent structure of torax body posture indicator was defined by two factors, the status of body posture of the rear part of the thorax, and status of the body posture of the front part of the thorax. The results obtained by canonical correlation analysis between predictive variables, morphological latent structure and criterion variables, latent structure of thorax body posture indicators with two posture indicators of the chest and one of the foot status, showed two important pairs of canonical roots on each measurement, suggesting a significant association between these two sets of parameters.     

A new model of selection in women's handball

The aim of the study was to assess the basic motor abilities that determine top performance in women's handball, and to identify test panel for primary selection at handball school. The study included 155 female attendants of the Split Handball School, mean age 12.5 years. Differences in the basic motor abilities between the subjects that developed into elite handball players after 7-year training process and those that abandoned handball for being unable to meet the competition criteria were evaluated by use of discriminative analysis. The former were found to have also been superior initially in all variables analyzed, and in arm coordination, overall body coordination, throw and jump explosive strength, arm movement frequency and repetitive trunk strength in particular. Motor superiority based on the abilities of coordination, explosive strength and speed determines performance in women's handball, qualifying these abilities as reliable selection criteria. Based on this study results, a new model of selection in women's handball, with fine arm coordination as the major limiting factor of performance, has been proposed.     

Beyond gastric acid reduction: proton pump inhibitors induce heme oxygenase-1 in gastric and endothelial cells

Proton pump inhibitors (PPIs) have been demonstrated to prevent gastric mucosal injury by mechanisms independent of acid inhibition. Here we demonstrate that both omeprazole and lansoprazole protect human gastric epithelial and endothelial cells against oxidative stress. This effect was abrogated in the presence of the heme oxygenase-1 (HO-1) inhibitor ZnBG. Exposure to either PPI resulted in a strong induction of HO-1 expression on mRNA and protein level, and led to an increased activity of this enzyme. Expression of cyclooxygenase isoforms 1 and 2 remained unaffected, and COX-inhibitors did not antagonize HO-1 induction by PPIs. Our results suggest that the antioxidant defense protein HO-1 is a target of PPIs in both endothelial and gastric epithelial cells. HO-1 induction might account for the gastroprotective effects of PPIs independently of acid inhibition, especially in NSAID gastropathy. Moreover, our findings provide additional perspectives for a possible but yet unexplored use of PPIs in vasoprotection.     

A fluorescence polarization-based assay for peptidyl prolyl cis/trans isomerase cyclophilin A

Peptidyl prolyl cis/trans isomerase cyclophilin A (CypA) serves as a cellular receptor for the important immunosuppressant drug, cyclosporin A. In addition, CypA and its enzyme family have been found to play critical roles in a variety of biological processes, including protein trafficking, HIV and HCV infection/replication, and Ca(2+)-mediated intracellular signaling. For these reasons, cyclophilins have emerged as potential drug targets for several diseases. Therefore, it is extremely important to screen for novel small molecule cyclophilin inhibitors. Unfortunately, the biochemical assays reported so far are not adaptable to a high-throughput screening format. Here, we report a fluorescence polarization-based assay for human CypA that can be adapted to high-throughput screening for drug discovery. The technique is based on competition and uses a fluorescein-labeled cyclosporin A analog and purified human CypA to quantitatively measure the binding capacity of unlabeled inhibitors. Detection by fluorescence polarization allows real-time measurement of binding ratios without separation steps. The results obtained demonstrated significant correlation among assay procedures, suggesting that the application of fluorescence polarization in combination with CypA is highly advantageous for the accurate assessment of inhibitor binding.     

The first study on the bacterial flora of the european spruce bark beetle, Dendroctonus micans (Coleoptera: Scolytidae)

The European spruce bark beetle, Dendroctonus micans Kugelann (Coleoptera, Scolytidae), is one of the most serious pests of oriental spruce (Picea orientalis L.) in Turkey. In this study, we investigated bacterial flora of D. micans collected from different populations of the forests of Eastern Black Sea Region of Turkey from 2002 to 2004. Seven different bacteria were isolated from healthy, diseased and dead specimens based on the color of colony and morphology. According to morphological, physiological and biochemical properties, metobolic enyzme profile by BIOLOG microtiter plate system, and total cellular fatty acid profile by Microbial Identification System (MIS), isolates were identified as Micrococcus luteus, Bacillus thuringiensis subsp. morrisoni, Serratia grimesii, Enterobacter cloaceae, Enterobacter intermedius, Streptococcus sp. and Pseudomonas putida. This is the first study on the bacterial flora of D. micans.