Kinetic disposition of an aqueous formulation of alphacypermethrin applied to the dorsal mid-line of sheep with long wool and its effect on lice

A group of 5 adult Merino sheep with fleeces about 70 mm long (7-months growth of wool) was treated with a topical formulation of the synthetic pyrethroid insecticide, alphacypermethrin, applied to the dorsal mid-line. Insecticide concentrations at the tip, middle and base of wool staples collected from meridians along the back, upper and lower flanks were measured at intervals from 1 to 98 days after treatment. Some movement of the alphacypermethrin from the back to the lower body occurred within 24h after treatment, but despite careful application of the insecticide there was wide variation in the concentration between and within meridians. The majority of the alphacypermethrin remained close to the dorsal mid-line and near the tip of the staple. There were significant differences in the concentration between the tip, middle and base segments of the staples in the back and lower flank meridians (P < 0.05). Despite exposure of the sheep to normal weathering, there was no significant difference in the concentration of alphacypermethrin between samples collected at day 1 or day 98 after treatment (P > 0.05). Numbers of pyrethroid-susceptible lice surviving exposure in vitro for 20 h differed significantly between samples collected at different times after treatment (P < 0.05). The numbers of lice surviving in samples collected within 28 days after treatment tended to be lower than in those collected from 28 to 98 days but, in some samples, regardless of time after treatment, lice survived for 20 h in wool taken from parts of the fleece that contained high concentrations of alphacypermethrin.     

Characterization of dental pulp stem/stromal cells of Huntington monkey tooth germs

Background

Activation by extracellular ligands of G protein-coupled (GPCRs) and tyrosine kinase receptors (RTKs), results in the generation of second messengers that in turn control specific cell functions. Further, modulation/amplification or inhibition of the initial signalling events, depend on the recruitment onto the plasma membrane of soluble protein effectors. High throughput methodologies to monitor quantitatively second messenger production, have been developed over the last years and are largely used to screen chemical libraries for drug development. On the contrary, no such high throughput methods are yet available for the other aspect of GPCRs regulation, i.e. protein translocation to the plasma membrane, despite the enormous interest of this phenomenon for the modulation of receptor downstream functions. Indeed, to date, the experimental procedures available are either inadequate or complex and expensive.

Results

Here we describe the development of a novel conceptual approach to the study of cytosolic proteins translocation to the inner surface of the plasma membrane. The basis of the technique consists in: i) generating chimeras between the protein of interests and the calcium (Ca²⁺)-sensitive, luminescent photo-protein, aequorin and ii) taking advantage of the large Ca²⁺ concentration [Ca²⁺] difference between bulk cytosolic and the sub-plasma membrane rim.

Conclusion

This approach, that keeps unaffected the translocation properties of the signalling protein, can in principle be applied to any protein that, upon activation, moves from the cytosol to the plasma membrane. Thus, not only the modulation of GPCRs and RTKs can be investigated in this way, but that of all other proteins that can be recruited to the plasma membrane also independently of receptor activation. Moreover, its automated version, which can provide information about the kinetics and concentration-dependence of the process, is also applicable to high throughput screening of drugs affecting the translocation process.     

Methylation of the vitamin D-dependent CaBP gene (calbindin 9 Kd) does not mediate tissue or vitamin D regulation

The vitamin D-dependent intestinal calcium binding protein gene is predominantly expressed in the intestine. In this report we have examined the possibility that methylation of the gene might play a role in its tissue-specific expression employing genomic Southern analysis. None of the Hpa II and Hha I sites examined by the indicated probes in and around the gene were found to be methylated in the intestine, kidney and liver. No change in the methylation of these sites was detected in response to 1,25-dihydroxy-vitamin D3 administration to vitamin D-deficient rats under conditions which stimulate the expression of the gene. These results indicate that the rat intestinal calcium binding protein gene is not methylated in these tissues, at the indicated sites and, therefore, methylation seems not to be involved in the regulation of this gene's expression.     

Private specificities can dominate the humoral response to self-antigens in patients with cryptogenic fibrosing alveolitis

Background  

The pathogenetic mechanisms that underlie the interstitial lung disease cryptogenic fibrosing alveolitis (CFA) may involve an immunological reaction to unidentified antigens in the lung, resulting in tissue damage.     

Sawgrass (Cladium jamaicense) responses as early indicators of low-level phosphorus enrichment in the Florida Everglades

Anthropogenic phosphorus (P) inputs to the Florida Everglades have produced dramatic changes in the wetland vegetation of this otherwise oligotrophic system. While the proliferation of undesirable plant species in response to enrichment has been well documented, nutrient-related changes in the physiological and morphological attributes of existing vegetation, prior to any shifts in species composition or changes in the spatial extent of certain taxa, have yet to be adequately characterized. In this experiment, three sawgrass-dominated areas were enriched with P for 3 years at rates of 0.4 g P/m²/year (HP), 0.1 g P/m²/year (LP), or 0 g P/m²/year (controls) to assess potential impacts of P-enriched discharges from stormwater treatment areas into the Everglades. Elevated concentrations of TP in rhizomes and leaves and reduced ratios of leaf N:P were detected in HP plants within ~1 year at most sites. Live leaf densities, plant heights, and plant densities of the HP groups were generally higher than LP and control groups after 2 years, a pattern that was evident even after major fire events. Total aboveground biomass was significantly elevated in both HP and LP treatments at two of the three sites after 3 years. No change in species composition was detected during the study. Planned hydrologic restoration measures will increase P loads into parts of the Everglades that have not previously experienced anthropogenic P enrichment. Monitoring native vegetation such as sawgrass can be a sensitive and relatively robust means of detecting unintended P enrichment in these areas prior to shifts in vegetation community composition or changes in area cover of key species.