Acidic glycosphingolipids of cock testis elucidated by mass spectrometry and NMR spectroscopy

The main acidic glycosphingolipids (GSLs) of cock testis were identified as GalCer I³-sulfate and gangliosides GM4, GM3, GD3 and GT3. They contained N-acetylneuraminic acid as the major sialic acid, and ceramides composed mainly of sphingosine (dl8:1) and C18–24 non-hydroxy fatty acids. Appreciable amounts of hydroxy fatty acids were detected only in the GM4 preparation.     

The influence of the landscape structure within buffer zones,catchment land use and instream environmental variables on mollusc communities in a medium-sized lowland river

The world’s freshwater molluscan fauna is facing unprecedented threats from habitat loss and degradation. Declines in native populations are mostly attributed to the human impact, which results in reduced water quality. The objectives of our survey were to analyse the structure of the mollusc communities in a medium-sized lowland river and to determine the most important environmental variables at different spatial scales, including landscape structure, catchment land use and instream environmental factors that influence their structure. Our survey showed that a medium-sized river, that flows through areas included in the European Ecological Natura 2000 Network Programme of protected sites, provides diverse instream habitats and niches that support 47 mollusc species including Unio crassus, a bivalve of Community interest, whose conservation requires the designation of a special conservation area under the Habitats Directive Natura 2000. This survey showed that mollusc communities are impacted by several environmental variables that act together at multiple scales. The landscape structure within buffer zones, catchment land use and instream environmental variables were all important and influenced the structure of mollusc communities. Therefore, they should all be taken into consideration in the future restoration of the river, future management projects and programmes for the conservation of biodiversity in running waters. The results of this study may be directly applicable for the rehabilitation of river ecosystems and are recommended to stakeholders in their future decision concerning landscape planning, monitoring species and their habitats, conservation plans and management in accordance with the requirements of sustainable development.     

Motile iridophores of a freshwater goby,Odontobutis obscura

Summary Reflecting chromatophores in the dermis of the skin of a freshwater goby, Odontobutis obscura, are of an iridophore type. These chromatophores contain numerous reflecting platelets, which are similar to those in iridophores of other fish and amphibian species. It was found that these iridophores are motile, i.e., these cells respond to certain stimuli with translocation of the platelets within the cells. K⁺ ions induced dispersion of the platelets in excised scale preparations, but not in excised scales from chemically denervated fish. Norepinephrine and melatonin also induced dispersion of the platelets. Alpha-MSH was effective in aggregating these organelles into the centrospheres of the cells. The conclusions reached are: (1) iridophores of O. obscura are motile; (2) the movement of the iridophores is under nervous and hormonal control.     

Determination of erythromycin, clarithromycin, roxithromycin, and azithromycin in plasma by high-performance liquid chromatography with amperometric detection

In this study, a high-performance liquid chromatographic method was developed for the quantitative determination of erythromycin (EM), roxithromycin (RXM), and azithromycin (AZM) in rat plasma with amperometric detection under a standardized common condition using clarithromycin (CAM) as an internal standard. This method was also proved to be applicable for the determination of CAM by employing RXM as an internal standard. Each drug was extracted from 150 μl of plasma sample spiked with internal standard under an alkaline condition with tert.-butyl methyl ether. The detector cell potential for the oxidation of the drugs was set at +950 mV. The linearity of the calibration curves were preserved over the concentration ranges of 0.1–10 μg/ml for EM and RXM, and 0.03–3.0 μg/ml for CAM and AZM. Coefficients of variation and relative error were less than 9% and ±7%, respectively. The analytical method presented here was proved to be useful for the investigation of the pharmacokinetic characteristics of EM, CAM, RXM, and AZM in rats.     

A locus conferring effective late blight resistance in potato cultivar Sárpo Mira maps to chromosome XI

Late blight of potato, caused by Phytophthora infestans, is one of the most economically important diseases worldwide, resulting in substantial yield losses when not adequately controlled by fungicides. Late blight was a contributory factor in The Great Irish Famine, and breeding for resistance to the disease began soon after. Several disease-resistant cultivars have subsequently been obtained, and amongst them Sárpo Mira is currently one of the most effective. The aim of this work was to extend the knowledge about the genetic basis of the late blight resistance in Sárpo Mira and to identify molecular markers linked to the resistance locus which would be useful for marker-assisted selection. A tetraploid mapping population from a Sárpo Mira × Maris Piper cross was phenotyped for foliar late blight resistance using detached leaflet tests. A locus with strong effect on late blight resistance was mapped at the end of chromosome XI in the vicinity of the R3 locus. Sárpo Mira’s genetic map of chromosome XI contained 11 markers. Marker 45/XI exhibited the strongest linkage to the resistance locus and accounted for between 55.8 and 67.9 % of variance in the mean resistance scores noted in the detached leaflet assays. This marker was used in molecular marker-facilitated gene pyramiding. Ten breeding lines containing a late blight resistance locus from cultivar Sárpo Mira and the Rpi-phu1 gene originating from the late blight resistant accession of Solanum phureja were obtained. These lines have extended the spectrum of late blight resistance compared with Sárpo Mira and it is expected that resistance in plants containing this gene pyramid will have enhanced durability.     

A conserved RxLR effector interacts with host RABA‐type GTPases to inhibit vesicle‐mediated secretion of antimicrobial proteins

Plant pathogens of the oomycete genus Phytophthora produce virulence factors, known as RxLR effector proteins that are transferred into host cells to suppress disease resistance. Here, we analyse the function of the highly conserved RxLR24 effector of Phytophthora brassicae. RxLR24 was expressed early in the interaction with Arabidopsis plants and ectopic expression in the host enhanced leaf colonization and zoosporangia formation. Co‐immunoprecipitation (Co‐IP) experiments followed by mass spectrometry identified different members of the RABA GTPase family as putative RxLR24 targets. Physical interaction of RxLR24 or its homologue from the potato pathogen Phytophthora infestans with different RABA GTPases of Arabidopsis or potato, respectively, was confirmed by reciprocal Co‐IP. In line with the function of RABA GTPases in vesicular secretion, RxLR24 co‐localized with RABA1a to vesicles and the plasma membrane. The effect of RxLR24 on the secretory process was analysed with fusion constructs of secreted antimicrobial proteins with a pH‐sensitive GFP tag. PATHOGENESIS RELATED PROTEIN 1 (PR‐1) and DEFENSIN (PDF1.2) were efficiently exported in control tissue, whereas in the presence of RxLR24 they both accumulated in the endoplasmic reticulum. Together our results imply a virulence function of RxLR24 effectors as inhibitors of RABA GTPase‐mediated vesicular secretion of antimicrobial PR‐1, PDF1.2 and possibly other defence‐related compounds.     

A rapid increase in cAMP in response to 20-hydroxyecdysone in the anterior silk glands of the silkworm, Bombyx mori

In the anterior silk glands (ASGs) of the silkworm, Bombyx mori, intracellular cAMP increases transiently to a very high level shortly after the hemolymph ecdysteroid peak in the prepupal period. In cultured ASGs obtained on the day of gut-purge, cAMP levels were increased by 20-hydroxyecdysone (20E), and this increase was enhanced by an inhibitor of phosphodiesterase, but was not affected by alpha-amanitin, indicating the 20E action may not be mediated via gene expression. The increase in cAMP occurred within 30 seconds of exposure to a physiological concentration of 20E (1 microM), and also by ponasterone A. Our findings indicate a nongenomic action of ecdysteroids in insects, which may be an additional mechanism by which this steroid hormone induces acute responses in tissues and cells.     

Na+-dependent p-aminohippurate transport at the basolateral side of the isolated perfused rat kidney

The uptake of p-amino[3H]hippurate by isolated perfused rat kidney was studied to characterize the mechanism which was responsible for organic anion transport process. A rapid injection multiple indicator dilution technique and the distributed two-compartment model of Sawada et al. (Computer Methods Programs Biomed., 20 (1985) 51) were employed. Some characteristics of a carrier-mediated transport from the antiluminal space to the intracellular space for p-aminohippurate at the basolateral side were demonstrated: the uptake was stimulated by the countertransport effect and showed Na+ dependency. These findings are consistent with p-amino[3H]hippurate's being taken up into the isolated rat basolateral membrane vesicle by Na+-dependent carrier-mediated transport (J. Pharmacol. Exp. Ther. 227 (1983) 122). It is suggested that the multiple indicator dilution technique is a sensitive new method to study the mechanisms of renal tubular transport in the living kidney as an organ.