Chlordiazepoxide inhibition of reflex vagal bradycardia in the cat

The effect of chlordiazepoxide (CDZ) on phenylephrine-induced reflex vagal bradycardia was studied in cats anesthetized with chloralose. The sympathetic component of the reflex was eliminated by either pretreating the animals with propranolol (1 mg/kg, i.v.) or sectioning the spinal cord. In animals pretreated with propranolol, CDZ (3, 10 and 20 mg/kg, i.v.) produced a dose-related inhibition of phenylephrine-induced bradycardia. These doses of CDZ had no significant effect on phenylephrine-induced pressor responses. Similar results were obtained with CDZ in animals with spinal cords transected. Chlordiazepoxide did not prevent bradycardia evoked by electrical stimulation of the peripheral cut-end of the right vagus nerve. These results indicate that CDZ can inhibit reflex vagal bradycardia and that the inhibition involves a central action of the drug.     

An overlapping bacterial artificial chromosome system that generates vectorless progeny for channel catfish herpesvirus

Herpesviruses are important pathogens of humans and other animals. Herpesvirus infectious clones that can reconstitute phenotypically wild-type (wt) virus are extremely valuable tools for elucidating the roles of specific genes in virus pathophysiology as well as for making vaccines. Ictalurid herpesvirus 1 (channel catfish herpesvirus [CCV]) is economically very important and is the best characterized of the herpesviruses that occur primarily in bony fish and amphibians. Here, we describe the cloning of the hitherto recalcitrant CCV genome as three overlapping subgenomic bacterial artificial chromosomes (BACs). These clones allowed us to regenerate vectorless wt CCVs with a phenotype that is indistinguishable from that of the wt CCV from which the BACs were derived. To test the recombinogenic systems, we next used the overlapping BACs to construct a full-length CCV BAC by replacing the CCV ORF5 with the BAC cassette and cotransfecting CCO cells. The viral progeny that we used to transform Escherichia coli and the resulting BAC had only one of the 18-kb terminal repeated regions. Both systems suggest that one of the terminal repeat regions is lost during the replicative stage of the CCV life cycle. We also demonstrated the feasibility of introducing a targeted mutation into the CCV BAC infectious clone by constructing a CCV ORF12 deletion mutant and showed that ORF12 encodes a nonessential protein for virus replication. This is the first report of the generation of an infectious BAC clone of a member of the fish and amphibian herpesviruses and its use to generate recombinants.     

Short-term effects of fire frequency on vegetation composition and biomass in mixed prairie in south-western Manitoba

The net effects of one, two, and three spring burns in consecutive yearson the aboveground biomass, species composition, and soil variables wereassessed in two different mixed-grass prairie sites in south-western Manitoba.Precipitation in the first year was greater than the 30-year average but lowerduring the next three years. The first site (Area 6) was characterized byBouteloua gracilis, Stipa spartea,Selaginella densa and lichens, while the second, somewhatdrier site (Area 10) was dominated by B. gracilis andCarex spp. Each burn treatment was applied to 15 plots inarandom block design. Vegetation and soil data collected following the thirdburnare presented. In general, the effects of repeated burning were more pronouncedin the drier Area 10, where litter was significantly reduced with eachadditional burn. In Area 6, litter was significantly reduced only after threeyears of burning. In both sites, the cumulative effect of fire had nosignificant effect on total standing crop, even after three consecutive yearsofburning. In the drier Area 10, however, the biomass of several componentsshifted such that B. gracilis biomass increasedsignificantly and forb biomass decreased significantly after three burns. Plantcover was also affected more in the drier Area 10. The cover of B.gracilis, the dominant C₄ grass, significantly increasedand Carex spp. cover decreased after two burns in thedriersite. In Area 6, the most significant effect of fire was a reduction inSelaginella and lichen cover. Bare ground increased inbothsites as the number of burns increased. There was little change in soilnutrients with burning, but soil moisture somewhat decreased and surfacetemperature significantly increased after three burns.     

Co-translational Processing of Glycoprotein 3 from Equine Arteritis Virus: N-GLYCOSYLATION ADJACENT TO THE SIGNAL PEPTIDE PREVENTS CLEAVAGE*

Signal peptide cleavage and N-glycosylation of proteins are co-translational processes, but little is known about their interplay if they compete for adjacent sites. Here we report two unique findings for processing of glycoprotein 3 of equine arteritis virus. Glycoprotein 3 (Gp3) contains an N-terminal signal peptide, which is not removed, although bioinformatics predicts cleavage with high probability. There is an overlapping sequon, NNTT, adjacent to the signal peptide that we show to be glycosylated at both asparagines. Exchanging the overlapping sequon and blocking glycosylation allows signal peptide cleavage, indicating that carbohydrate attachment inhibits processing of a potentially cleavable signal peptide. Bioinformatics analyses suggest that a similar processing scheme may exist for some cellular proteins. Membrane fractionation and secretion experiments revealed that the signal peptide of Gp3 does not act as a membrane anchor, indicating that it is completely translocated into the lumen of the endoplasmic reticulum. Membrane attachment is caused by the hydrophobic C terminus of Gp3, which, however, does not span the membrane but rather attaches the protein peripherally to endoplasmic reticulum membranes.