The high-latitude terrestrial carbon sink: a model analysis

A dynamic, global vegetation model, hybrid v4.1 ( Friend et al. 1997 ), was driven by transient climate output from the UK Hadley Centre GCM (HadCM2) with the IS92a scenario of increasing atmospheric CO₂ equivalent, sulphate aerosols and predicted patterns of atmospheric N deposition. Changes in areas of vegetation types and carbon storage in biomass and soils were predicted for areas north of 50°N from 1860 to 2100. Hybrid is a combined biogeochemical, biophysical and biogeographical model of natural, potential ecosystems. The effect of periodic boreal forest fires was assessed by adding a simple stochastic fire model. Hybrid represents plant physiological and soil processes regulating the carbon, water and N cycles and competition between individuals of parameterized generalized plant types. The latter were combined to represent tundra, temperate grassland, temperate/mixed forest and coniferous forest. The model simulated the current areas and estimated carbon stocks in the four vegetation types. It was predicted that land areas above 50°N (about 23% of the vegetated global land area) are currently accumulating about 0.4 PgC y^?1 (about 30% of the estimated global terrestrial sink) and that this sink could grow to 0.8–1.0 PgC y^?1 by the second half of the next century and persist undiminished until 2100. This sink was due mainly to an increase in forest productivity and biomass in response to increasing atmospheric CO₂, temperature and N deposition, and includes an estimate of the effect of boreal forest fire, which was estimated to diminish the sink approximately by the amount of carbon emitted to the atmosphere during fires. Averaged over the region, N deposition contributed about 18% to the sink by the 2080 s. As expected, climate change (temperature, precipitation, solar radiation and saturation pressure deficit) and N deposition without increasing atmospheric CO₂ produced a carbon source. Forest areas expanded both south and north, halving the current tundra area by 2100. This expansion contributed about 30% to the sink by the 2090 s. Tundra areas which were not invaded by forest fluctuated from sink to source. It was concluded that a high latitude carbon sink exists at present and, even assuming little effect of N deposition, no forest expansion and continued boreal forest fires, the sink is likely to persist at its current level for a century.     

Cytochrome P450105D1 (CYP105D1) from Streptomyces griseus: heterologous expression, activity, and activation effects of multiple xenobiotics.

The open reading frame of CYP105D1, a soluble cytochrome P450 from Streptomyces griseus, was cloned behind the tac promoter of the bacterial expression vector pSPg1910L and expressed in Escherichia coli. The recombinant protein retained normal spectral characteristics having a Soret peak at 448 nm in the reduced carbon monoxide difference spectrum. CYP105D1 was active, obtaining reducing equivalents from endogenous E. coli ferredoxin and ferredoxin reductase redox partners present in E. coli. In vitro activity studies revealed CYP105D1 to catalyse the NADH- and NADPH-dependent oxidation of the xenobiotic substrates benzo[a]pyrene, erythromycin, warfarin, and testosterone. Furthermore, this activity could be stimulated in the presence of either alpha-benzoflavone or beta-benzoflavone in an analogous manner to that reported for mammalian P450 forms including human liver cytochrome P4503A4 (CYP3A4). The system produces an alternative to whole-cell biotransformation of xenobiotic for the production of drug metabolites and an experimental system for probing the structural features of a cytochrome P450 with a broad substrate range.     

Incorporation of 15N from ammonium into the N-linked oligosaccharides of an immunoadhesin glycoprotein expressed in Chinese hamster ovary cells

Elevated ammonium concentrations in the medium of cultivated cells have been shown to increase the intracellular levels of uridine-5'-diphospho- N-acetylglucosamine (UDP-GlcNAc) and uridine-5'-diphospho-N- acetylgalactosamine (UDP-GalNAc; Ryll et al., 1994). These sugar nucleotides are substrates for glycosyltransferases in the glycosylation pathway. In our experiments, recombinant Chinese hamster ovary cells producing an immunoadhesin glycoprotein (GP1-IgG) have been cultivated under controlled cell culture conditions in the presence of different ammonium concentrations.15N-Labeled ammonium chloride (15NH4Cl) was added exogenously to the cell culture media to determine if ammonium was incorporated into UDP-GlcNAc and cytidine-5'- monophospho-N-acetylneuraminic acid (CMP-NANA) pools, and subsequently incorporated into GP1-IgG as N-linked glycans. The intracellular pools of UDP-activated hexosamines (UDP-GNAc) were followed during the time course of the experiment. To assess the extent of15NH4+incorporation into the glycans of GP1-IgG, the glycoprotein was first purified to homogeneity by protein A chromatography. Enzymatically released N- glycans were then analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. N-Glycans synthesized in the presence of15NH4Cl revealed an N-glycan-dependent increase in mass-to-charge of 2.5-4.8 Da. These results indicate that 60-70% of the total nitrogen containing monosaccharides had incorporated15N. Presumably,15NH4+was incorporated into GlcNAc and N- acetylneuraminic acid as proposed earlier (Ryll et al., 1994). This might be a universal and previously not described reaction in mammalian cells when exposed to nonphysiological but in cell culture commonly found concentrations of ammonium. The data presented here are of significance for glycoprotein production in mammalian cell culture, since it has been shown previously that elevated levels of UDP- activated hexosamines affect N-glycan characteristics such as branching and degree of amino sugar incorporation. In addition, our results demonstrate that isotope labeling in combination with MALDI-TOF-MS can be used as an alternate tool to radioactive labeling of sugar substrates in metabolic studies.      

In vivo and in vitro characterization of novel microparticulates based on hyaluronan and chitosan hydroglutamate

This study examined the application of previously characterized microparticles composed of hyaluronan (HA) and chitosan hydroglutamate (CH) as well as novel microparticles consisting of both polymers (HA/CH) to improve the nasal delivery of a model drug. The rabbit bioavailabilities of gentamicin incorporated in HA, CH, and HA/CH microparticles were increased 23-, 31-, and 42-fold, respectively, compared with the control intranasal solution of gentamicin, indicating that all test microparticles were retained for longer periods on the nasal mucosa of the rabbits as supported by previous in vitro dissolution as well as frog palate mucoadhesion studies, thereby improving drug absorption. The higher bioavailabilities of CH-based formulations (CH and HA/CH) suggest the penetration-enhancing effects of CH may also be partially responsible for the improvement. A model was developed, based on a glass impinger device, to deliver dry powder formulations reproducibly onto the surface of cultured cell monolayers. In vitro permeability and fluorescence microscopy studies on the tight junctions of the 16HBE14o- cell lines further confirmed the ability of CH-based formulations to enhance penetration. Furthermore, the in vitro absorption profile from cell culture studies was consistent with those determined from in vivo studies. The complementary effect from the mucoadhesive nature of HA coupled with the penetration-enhancing effects of CH makes the novel HA/CH formulation a promising nasal delivery system.     

Geographic distribution of methyltransferases of Helicobacter pylori: evidence of human host population isolation and migration

Background  

Helicobacter pylori colonizes the human stomach and is associated with gastritis, peptic ulcer, and gastric cancer. This ubiquitous association between H. pylori and humans is thought to be present since the origin of modern humans. The H. pylori genome encodes for an exceptional number of restriction and modifications (R-M) systems. To evaluate if R-M systems are an adequate tool to determine the geographic distribution of H. pylori strains, we typed 221 strains from Africa, America, Asia, and Europe, and evaluated the expression of different 29 methyltransferases.     

Climate change and pollutant impacts on Scottish vegetation

Summary

By the 2050s the UK is projected to be about 1.6°C warmer, when the atmospheric CO₂ concentration will be 525 ppmv. These changes will have profound effects on the Scottish flora and fauna. Vegetation primary productivity will increase, except in dry regions, and the productivity of upland forest plantations may increase by several Yield Classes. The spread of plant species may be less than expected, but a number of slow-growing ‘stress-tolerant’ species, including montane/alpine species, are likely to be lost. Nitrogen deposited as a result of emission of NO_X from vehicles and NH₃ from agriculture is now a major source of acidity, and problems of acidification and eutrophication are linked. Despite reductions in sulphur emissions, critical loads of acid deposition are likely to be exceeded for soils in most of the Scottish uplands until at least 2005. Critical levels affecting tree growth may be exceeded where forests are in cloud for 10% of the time in areas of the Great Glen. Much of the Scottish uplands receives 25–30 kg N ha^-1 yr^-1, which may be causing change in species composition. Background tropospheric ozone concentrations are increasing. Much of the Scottish uplands experiences mean summer ozone concentrations exceeding those in southern England, but with fewer exceedances of critical levels. However, many crops and some sensitive native species are probably being adversely affected.     

Studies on pre-incubation conditions in the seeded agar plate well inhibition assay

Pre-incubation at 4°C increases the zone of bacterial growth inhibition produced by methanolic extracts of freshwater algae in the agar diffusion assay. Preloading of wells with methanol increases the inhibition zone produced by water-insoluble antibiotics.     

A computer program for processing data from amino acid analysis and for the calculation of molecular weights from those data.

A computer program is described for the calculation of the complete amino acid composition of a protein from the analytical data. The program also derives a molecular weight on the basis of the amino acid composition. The use of the program for the determination of the molecular weights of the liver carboxylesterases of chicken, horse, ox, and sheep is described.