Genetics of the peroxidase isoenzymes in Petunia

Summary By starch gel electrophoresis three mobility variants of a cathodic moving doublet of bands, encoded by the structural gene prxC, were detected in all organs of flowering petunias. In root tissue two of the variants showed a lower electrophoretic mobility than in other organs. During development of flower buds the PRXc enzymes showed an increase in mobility. The gene prxC was located on chromosome IV by showing linkage to the genes An3 and Dw1, by trisomic segregation, and by the construction of triply heterozygous trisomics IV. The gene order on chromosome IV is B1-An3/Dw1-prxC. It was concluded that the temporal programming difference in the expression of the alleles prxC2 and prxC3 is caused by internal site mutation. Analysis of progeny obtained by crossing of lines to the trisomic IV with genotype prxC1/C1/C2 showed differential expression of the two prxC1 alleles of the trisomic IV.     

Chromium oxalate: a new spin label broadening agent for use with thylakoids.

Potassium tris(oxalato)chromate(III) trihydrate (chromium ixalate) has been shown to be a more useful broadening agent than potassium ferricyanide for the spin label 2,2,6,6-tetramethylpiperidine-N-oxyl-4-amine (Tempamine) in thylakoid suspensions. Our data show that chromium oxalate is less permeable than ferricyanide, does not inhibit thylakoid electron transport or photophosphorylation, and is not photoreduced by thylakoids.     

A new species of Polystoma (Monogenea: Polystomatidae) parasitic in Hyperolius marmoratus (Anura: Hyperoliidae) in South Africa

Polystoma marmorati n. sp. is described as a new polystomatid species parasitic in the urinary bladder of the painted reed frog Hyperolius marmoratus marmoratus collected in southern Natal, South Africa. Parasite prevalence varied from 14.5 to 47.4% and the mean intensity from 1.0 to 1.1. The hamuli of the new species have a characteristic, partly fragmented appearance, a feature also seen in Polystoma batchvarivi, the only other polystome described from a species of the genus Hyperolius. The similarity between the two species, in spite of their geographical separation, is commented upon.     

Present and Future Developments in Hepatic Tissue Engineering for Liver Support Systems

The liver is the most important organ for the biotransformation of xenobiotics, and the failure to treat acute or acute-on-chronic liver failure causes high mortality rates in affected patients. Due to the lack of donor livers and the limited possibility of the clinical management there has been growing interest in the development of extracorporeal liver support systems as a bridge to liver transplantation or to support recovery during hepatic failure. Earlier attempts to provide liver support comprised non-biological therapies based on the use of conventional detoxification procedures, such as filtration and dialysis. These techniques, however, failed to meet the expected efficacy in terms of the overall survival rate due to the inadequate support of several essential liver-specific functions. For this reason, several bioartificial liver support systems using isolated viable hepatocytes have been constructed to improve the outcome of treatment for patients with fulminant liver failure by delivering essential hepatic functions. However, controlled trials (phase I/II) with these systems have shown no significant survival benefits despite the systems’ contribution to improvements in clinical and biochemical parameters. For the development of improved liver support systems, critical issues, such as the cell source and culture conditions for the long-term maintenance of liver-specific functions in vitro, are reviewed in this article. We also discuss aspects concerning the performance, biotolerance and logistics of the selected bioartificial liver support systems that have been or are currently being preclinically and clinically evaluated.     

Litter quality in a north European transect versus carbon storage potential

Newly shed foliar plant litter often has a decomposition rate of ca 0.1–0.2% day^–1, which decreases greatly with time and may reach 0.0001 to 0.00001% day^–1 or lower in litter material in the last stages of decay. The decrease in decomposability (substrate quality) varies among species and is complex, involving both direct chemical changes in the substrate itself and the succession in microorganisms able to compete for substrate with a given chemical composition. In late stages, the decomposition appears very little affected by climate, suggesting that climate change will have little effect on late-stages decomposition rates. Here, we apply a model for the late stages of litter decomposition to address the question of climate-change effects on soil-C storage. Decomposition of litter turning into soil organic matter (SOM) is determined by the degradation rate of lignin. In the last phases of decay, raised N concentrations have a rate-retarding effect on lignin degradation and thus on the decomposition of far-decomposed litter and litter in near-humus stages. The retardation of the decomposition rate in late stages may be so strong that decomposition reaches a limit value at which total mass losses virtually stop. At such a stage the remaining litter would be close to that of stabilized SOM. The estimated limit values for different species range from about 45 to 100% decomposition indicating that between 0 and 55% should either be stabilized or decompose extremely slowly. For no less than 106 long-term studies on litter decomposition, encompassing 21 litter types, limit values were significantly and negatively related to N concentration, meaning that the higher the N concentration in the newly shed litter (the lower the C/N ratio) the more litter was left when it reached its limit value. Trees growing under warmer and wetter climates (higher actual evapotranspiration, AET) tend to shed foliar litter more rich in N than those growing under colder and drier climates. A change in climate resulting in higher AET would thus mean that within species, e.g., Scots pine, a higher N level in the foliar litter may result. Further, within the boreal system deciduous species appear to have foliar litter richer in N than have conifers and within the conifers group, Norway spruce has needle litter more rich in N than, e.g., Scots pine. Thus, a change of species (e.g., by planting) from pine to spruce or from spruce to a deciduous species such as birch may result in a higher N level in the litter fall at a given site. In both cases the result would be a lower limit value for decomposition. The paper presents an hypothesis, largely based on available data that a change in climate of 4° higher annual average temperature and 40% higher precipitation in the Baltic basin would result in higher N levels in litter, lower decomposition and thus a considerable increase in humus accumulation.     

Amplification of multicistronic plasmids in the human 293 cell line and secretion of correctly processed recombinant human protein C

We have constructed multicistronic vectors containing the cDNAs for murine dihydrofolate reductase (DHFR), hygromycin phosphotransferase (HyPR), and human protein C (HPC), an antithrombotic factor. Using a sequential selection protocol with hygromycin (Hy) and methotrexate (MTX), we demonstrate the selective amplification of the murine dhfr cDNA in the adenovirus-transformed human kidney cell line 293, and the coamplification of the cDNA for HPC. Such recombinant 293 cell lines secreted HPC at levels as high as 25 micrograms/10(6) cells/day. In addition, we found that the complex vitamin K-dependent posttranslational modification of gamma-carboxylation of glutamate was not limiting at these high secretion levels, although the proteolytic processing of the protein was slightly reduced. Further, the HPC secreted from the gene-amplified cell lines had full anticoagulant activity when compared to plasma-derived HPC.     

Differentially oriented populations of actin filaments generated in lamellipodia collaborate in pushing and pausing at the cell front

Eukaryotic cells advance in phases of protrusion, pause and withdrawal. Protrusion occurs in lamellipodia, which are composed of diagonal networks of actin filaments, and withdrawal terminates with the formation of actin bundles parallel to the cell edge. Using correlated live-cell imaging and electron microscopy, we have shown that actin filaments in protruding lamellipodia subtend angles from 15-90 degrees to the front, and that transitions from protrusion to pause are associated with a proportional increase in filaments oriented more parallel to the cell edge. Microspike bundles of actin filaments also showed a wide angular distribution and correspondingly variable bilateral polymerization rates along the cell front. We propose that the angular shift of filaments in lamellipodia serves in adapting to slower protrusion rates while maintaining the filament densities required for structural support; further, we suggest that single filaments and microspike bundles contribute to the construction of the lamella behind and to the formation of the cell edge when protrusion ceases. Our findings provide an explanation for the variable turnover dynamics of actin filaments in lamellipodia observed by fluorescence speckle microscopy and are inconsistent with a current model of lamellipodia structure that features actin filaments branching at 70 degrees in a dendritic array.