The genomes of the animal papillomaviruses European elk papillomavirus, deer papillomavirus, and reindeer papillomavirus contain a novel transforming gene (E9) near the early polyadenylation site.

We report that the genomes of reindeer papillomavirus (RPV), European elk papillomavirus (EEPV), and deer papillomavirus (DPV) contain a short conserved translational open reading frame (ORF), E9, which is located between the E5 ORF and the early polyadenylation site. In RPV, DPV, and EEPV, E9 ORFs have the potential to encode extremely hydrophobic polypeptides of approximately 40 amino acids. In mouse C127 cells transformed by EEPV and RPV, there exists a unique, abundant mRNA species of approximately 700 nucleotides which has the capacity to encode an E9 polypeptide. This mRNA is transcribed from a previously unrecognized promoter at position 4030 in the EEPV genome. The EEPV E9 ORF exhibits weak transforming activity in C127 cells and primary rat embryo fibroblasts. We also show that EEPV E5 is the major oncogene in the EEPV genome when assayed in C127 cells, although it is less efficient in transformation than the E5 genes of bovine papillomavirus type 1, DPV, and RPV.     

Nutrient distribution in a Swedish tree species experiment

The influence of four tree species on the distribution of nutrients between different compartments of the ecosystem was examined. In a randomized block (n=3) experiment in south-western Sweden, Ca, Mg and K were determined as exchangeable amounts in the mineral soil and as total amounts in the O+A₁ horizons (topsoil) and in the aboveground tree biomass. N contents were determined in all compartments as well as P contents of the aboveground tree biomass and the topsoil. The four tree species planted were: silver fir [Abies alba Mill.] (AA), grand fir [Abies grandis Lindl.] (AG), Norway spruce [Picea abies L. Karst.] (PA) and Japanese larch [Larix leptolepis (Sieb. och Zucc.) Endl.] (LL). At the age of 35–36 years, the total stemwood production of the most productive species, AG, was estimated at 471 m³ ha⁻¹. In relation to AG, LL had produced 80%, PA 73% and AA 37%. The system totals [aboveground tree biomass total + topsoil total + exchangeable (Ca, Mg, K) or total (N) in the mineral soil] of Ca, K and N did not differ significantly at the 5% level between the investigated species. For Mg, the system total in LL was significantly higher than for the other species. There was an indication that LL and AA contained higher amounts of Ca, Mg, K and N in the topsoil but less in the biomass than did AG and PA (partly significant). In the mineral soil, there were no significant differences in the exchangeable pools of Ca and K, nor in the total amounts of N. The biomass nutrient concentrations generally decreased in the order: AA > PA > AG > LL. At stem or whole-tree harvest, the Ca export per biomass unit would more than double in the case of PA compared to LL. LL also contained less N in the biomass than the other species. However, the N content in the biomass did not differ between the most (AG) and the least (AA) productive species, although the production of dry weight biomass (standing + harvested) of AG had been twice that of AA. It is concluded that the nutrient budget of a managed forest may vary considerably depending on the choice of tree species.     

Non-motile mutants of Helicobacter pylori and Helicobacter mustelae defective in flagellar hook production

Flagellar hooks were purified from Helicobacter pylori and Helicobacter mustelae. The 70 × 16nm H. pylori hook was composed of FIgE subunits of 78kDa, while the 72 × 16nm H. mustelae hook was composed of 87kDa subunits. N-terminal sequence was obtained for the FIgH proteins of both species, and for an internal H. mustelae FlgE peptide. Degenerate oligonucleotide primers allowed amplification of a 1.2 kb fragment from the H. mustelae chromosome, which carried part of the flgE gene. The corresponding H. pylori gene was cloned by immunoscreening of a genomic library constructed in λZAP Express, The translated H. pylori flgE sequence indicated a protein with limited homology with the hook proteins from Salmonella typhimurium and Treponema phagedenis. Mutants of H. pylori and H. mustelae defective in hook production generated by allele replacement were non-motile and devoid of flagellar filaments but produced both flagellin subunits, which were localized in the soluble fraction of the cell. The level of flagellin production was unchanged in the mutants, indicating that the regulation of flagellin expression in Helicobacter differs from that in the Enterobacteriaceae.     

Effect of lignin-related phenols and their methylated derivatives on the growth of eight white-rot fungi

When various lignin-related para-phenolic benzoic acids, para-phenolic cinnamic acids, para-phenolic phenylpropionic acids, the corresponding unsubstituted and 4-O-methylated derivatives, and 4-hydroxyl substituted benzaldehydes were tested on the growth of eight white-rot fungi, methylation of the 4-hydroxy substituent resulted, in most cases, in increased inhibition of fungal growth. This effect was most notable with monomethoxylated compounds. When the aromatic ring contained additional methoxyl substituents, the toxicity of the 4-O-methylated derivative was less pronounced. Marked inhibition of fungal growth was also observed with aromatic compounds lacking a para-substituent. Higher concentrations of aromatic aldehydes were manifestly more toxic than the corresponding carboxylic acid.J.A. Buswell is with the Department of Biology, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong. K.-E.L. Eriksson is with the Department of Biochemistry, The University of Georgia, Athens, Georgia, 30602, USA.     

Phylogeny of theCyclanthaceae

The affinities ofCyclanthaceae are discussed, and it is concluded that the sister-group to this family is most probablyPandanaceae. A hypothesis of generic relationships inCyclanthaceae, based on cladistic methods, is presented. Bootstrap analysis and Bremer support (decay index) have been used to test the strength of individual clades, and the result is compared with previously made phylogenetic analyses. TheSphaeradenia group (Chorigyne, Stelestylis, Sphaeradenia, andLudovia) is supported as monophyletic and acceptably resolved, while theAsplundia group (remaining genera inCarludovicoideae) may be paraphyletic, with largely uncertain relationships. A formal recognition of these groups is therefore not justified. The probable character evolution inCarludovicoideae is discussed.     

Gene technology-based antimetabolite design: the use of an in vitro protein expression system to facilitate antimetabolite design for virally-induced human diseases and malignant conditions

A precondition for the chemotherapeutic treatment of a variety of virally-induced human diseases and malignant conditions is a highly selective interaction of the drug molecule to be used with it's biological target. To ensure the development of novel, effective drugs, it is essential that the biological target is well characterised with regard to it's structure and activity. Such characterisation relies upon adequate amounts of pure target being available. One of the most important enzymatic importers for antimetabolites is the enzyme thymidine kinase. In this article an in vitro protein expression system is described which facilitates the production of milligram amounts of pure and biologically active thymidine kinase, from a number of important biological sources. Results have shown that the in vitro produced enzyme has the exact biochemical propeties of the in vivo enzyme. Thus the in vitro protein expression system is an ideal vechicle to facilitate an in depth investigation of the enzyme's biological properties.     

Different UmuC requirements for generation of different kinds of UV-induced mutations in Escherichia coli

An Escherichia coli strain bearing the dnaQ49 mutation, which results in a defective s subunit of DNA polymerase III, and carrying the lexA71 mutation, which causes derepression of the SOS regulon, is totally unable to maintain high-copy-number plasmids containing the umuDC operon. The strain is also unable to maintain the pAN4 plasmid containing a partial deletion of the umuD gene but retaining the wild-type umuC gene. These results suggest that a high cellular level of UmuC is exceptionally harmful to the defective DNA polymerase III of the dnaQ49 mutant. We have used this finding as a basis for selection of new plasmid umuC mutants. The properties of two such mutants, bearing the umuC61 or umuC95 mutation, are described in detail. In the umuC122:: Tn 5 strain harbouring the mutant plasmids, UV-induced mutagenesis is severely decreased compared to that observed with the parental umuDC ⁺ plasmid. Interestingly, while the frequency of UV-induced GC → AT transitions is greatly reduced, the frequency of AT → TA transversions is not affected. Both mutant plasmids bear frameshift mutations within the same run of seven A residues present in umuC ⁺; in umuC61 the run is shortened to six A whereas in umuC95 is lengthened to eight A. We have found in both umuC61 and umuC95 that translation is partially restored to the proper reading frame. We propose that under conditions of limiting amounts of UmuC, the protein preferentially facilitates processing of only some kinds of UV-induced lesions.     

Influence of Medetomidine on Acid-base Balance and Urine Excretion in Goats

Seven goats were given medetomidine 5 μg/kg as an iv bolus injection. Venous blood samples were taken repeatedly and urine was collected continuously via a catheter up to 7h after the injection. Medetomidine caused deep clinical sedation. Base excess, pH and PCO2 in venous blood rose after medetomidine administration. There were no significant changes in plasma concentrations of sodium, calcium, magnesium, creatinine or osmolality, whereas potassium and bicarbonate concentrations increased, and phosphate and chloride decreased. Medetomidine increased plasma glucose concentration, and in 4 of 7 goats glucose could also be detected in urine. Medetomidine did not influence urine flow rate, free water clearance, bicarbonate and phosphate excretion or pH, but renal chloride, sodium, potassium, calcium, magnesium and creatinine excretion were reduced. The results suggest that the metabolic alkalosis recorded after medetomidine administration is not caused by increased renal acid excretion.