Response of fine roots to an experimental gap in a boreal Picea abies forest

We examined the initial response of the quantity and distribution of fine roots to the creation of an experimental canopy gap with a diameter of 50 m in a mature managed Norway spruce forest. Under the canopy, the fine root length densities of trees, shrubs, and grasses and herbs were 3207, 707 and 2738 m m^–2, respectively. The fine root biomass of trees, shrubs, and grasses and herbs were 182, 47 and 52 g m^–2, respectively. Two growing seasons after gap creation hardly any fine tree roots were found in the middle part of the gap. The living tree roots in the gap edge zone were mainly located within a 5-m distance from the standing edge trees. The indices developed here to show the influence of trees on fine root lenght density clearly revealed the effect of the vicinity of living trees on fine root lenght density. The root densities of grasses, herbs and dwarf shrubs did not show a clear response to gap creation despite the increase of their foliage. Our results suggest that in boreal spruce forests a gap disturbance creates a distinct tree root gap and that the gap edge trees do not extend their root systems rapidly into the formed root gap.     

Aspects of the ecology and natural history of Paraergasilus rylovi (Copepoda, Ergasilidae) parasitic in unionids of Finland

The distribution of Paraergasilus rylovi in 17 populations of unionids was investigated. In 1 unionid population, the parasite was studied regarding host age, size, sex, and the reproductive period (occurrence of egg sacs). Results from pooled material from the years 1987--1989 and 1996 (southern Finland, 11 populations) indicated that Anodonta piscinalis (n = 1,359) is the main host (total mean prevalence 71% and intensity +/-SE of infection 16.4+/-0.6). Pseudanodonta complanata (n = 106) was infected occasionally (3% and 1.3+/-0.3), whereas Unio pictorum (n = 108) and U. tumidus (n = 17) were not infected. Results from 17 A. piscinalis populations showed that P. rylovi occurs in southern Finland but not in northern Finland. In A. piscinalis, the mean intensity of infection was higher in lake populations than in river populations. Both host age and length had a negative relationship with the intensity of P. rylovi infection. Host sex did not affect the intensity of infection. Egg sacs of P. rylovi were found from June to August. There was a tendency for higher intensities of infection in autumn. Infection by the digenean Rhipidocotyle fennica had no effect on the intensity of P. rylovi infection.     

Differential localization of two acid proteinases in germinating barley (Hordeum vulgare) seed

A cathepsin D-like aspartic proteinase (EC 3.4.23) is abundant in ungerminated barley ( Hordeum vulgare ) seed while a 30 kDa cysteine endoproteinase (EC 3.4.22) is one of the proteinases synthesized de novo in the germinating seed. In this work, the localization of these two acid proteinases was studied at both the tissue and subcellular levels by immunomicroscopy. The results confirm that they have completely different functions. The aspartic proteinase was present in the ungerminated seed and, during germination, it appeared in all the living tissues of the grain, including the shoot and root. Contrary to previous suggestions, it was not observed in the starchy endosperm. By immunoblotting, the high molecular mass form of the enzyme (32 + 16 kDa) was found in all the living tissues, whereas the low molecular mass form (29 + 11 kDa) was not present in the shoot or root, indicating that the two enzyme forms have different physiological roles. The aspartic proteinase was localized first in the scutellar protein bodies of germinating seed, and later in the vacuoles which are formed by fusion of the protein bodies. In contrast to the aspartic proteinase, the expression of the 30 kDa cysteine proteinase began during the first germination day, and it was secreted into the starchy endosperm; first from the scutellum and later from the aleurone layer. It was not found in either shoots or roots. The 30 kDa cysteine proteinase was detected in the Golgi apparatus and in the putative secretory vesicles of the scutellar epithelium. These results suggest that the aspartic proteinase functions only in the living tissues of the grain, as opposed to the 30 kDa cysteine proteinase which is apparently one of the proteases initiating the hydrolysis of storage proteins in the starchy endosperm.     

Determination of nitecapone and (13C6)nitecapone in human plasma by gas chromatography/mass spectrometry.

A gas chromatographic/mass spectrometric method is developed and validated for simultaneous determination of nitecapone and its 13C6-labelled analogue in human plasma using (2H6,13C6)nitecapone as internal standard. The method involves extraction of the analytes from plasma to ethyl acetate-hexane mixture (20:80) and conversion to bis(trimethylsilyl) ethers prior to determination by gas chromatography/mass spectrometry using selected ion monitoring. The quantification range is 0.5-2000 ng ml-1. Precision ranges from 11.3% (coefficient of variation) at low levels to 2.4% at high levels. Recovery is about 50% in the whole range. The method is applied to a pharmacokinetic study where nitecapone diluted with 14C-labelled nitecapone is given intravenously concomitantly with an oral dose of (13C6)nitecapone.     

Immunolocalization of the saposin-like insert of plant aspartic proteinases exhibiting saposin C activity. Expression in young flower tissues and in barley seeds

The plant-specific insert (PSI) of cypro11 gene-encoding cyprosin, an aspartic proteinase from Cynara cardunculus , has been cloned by polymerase chain reaction (PCR) into a bacterial expression vector. A rearranged form of this PSI in which the N- and C-terminal sequences were permutated to make it more similar to the structural arrangement observed in saposins was also cloned and expressed in the same system. The biological activities of the two purified recombinant proteins were compared to those of human saposins B and C. The proteins showed similar activity to saposin C, i.e. capacity to activate human glucosylceramidase. At a concentration of 5 µ M , wild-type PSI, saposin C, and rearranged PSI activated human glucosylceramidase two-, three-, and five-fold, respectively. The K_m for 4-methylumbelliferyl β-glucopyranoside was around 7 µ M in the presence of any of the three activators (5 µ M ). The neurotropic activity using NS20Y cells and lipid-binding properties of the plant recombinant proteins were tested. The two plant proteins showed lipid-binding properties similar to those of saposins but did not have any effect on neurite outgrowth. Immunolocalization of PSI showed its expression in protective tissues in flower meristem – protodermis, in C. cardunculus and embryonic root cap and coleorhiza in mature barley grains – as well as husk, pericarp, and the aleurone layer. Possible biological functions suggested for the plant homologue to saposins besides the general activation of enzymes involved in lipid metabolism would be involvement in plant defence.     

Modelling the interplay between pest movement and the physical design of trap crop systems

Abstract 1 The interplay between pest movement and trap crop physical design is modelled in a situation where the pest moves by a random walk with spatially variable mobility. Questions addressed are: (i) how does the proportion of trap crop area of the total field area influence the equilibrium distribution of pests among the crop and the trap crop and (ii) how do crop patch size and shape influence the speed of pest redistribution from the crop to the trap crop. 2 When pest mobility in the trap crop is clearly lower than that in the crop, the pest population in the crop decreases very sharply for small trap crop proportions. When mobility in the trap crop is slightly closer to that in the crop, the pest population in the crop decreases much more gradually with increasing trap crop proportion. Thus finding a trap crop that the pest distinctly prefers over the crop appears to be crucial for developing efficient trap crop systems. 3 The rate of decay in the pest population in the crop increases with increasing perimeter to area ratio of the crop patch. Hence, designing field layouts to increase the perimeter to area ratio of crop patches may be beneficial.     

Role of apolipoproteins E and C in type V hyperlipoproteinemia

Type V hyperlipoproteinemia is characterized by elevations of chylomicron (CM) and very low density lipoprotein (VLDL) triglycerides. The development of this lipid disorder involves a multitude of metabolic derangements including deficient clearance of triglycerides and/or their increased output aggravated by obesity, diabetes, alcohol intake, or use of some hormones. Some studies have suggested that the apolipoprotein E4 phenotype is involved in this dyslipoproteinemia but this concept is still a matter of controversy. Therefore, we determined the apoE phenotype in 21 patients with severe hypertriglyceridemia classified as type V. Their apoE4 gene frequency was 0.595 which is 2.6-fold higher (P less than 0.001) than that in the Finnish population. Correspondingly, their apoE3 gene frequency was lower than that in the normal population. No differences were noted in plasma lipoproteins of the apoE4 phenotypes and the other type V subjects. The apolipoprotein C-II and C-III distribution was similar to that in normolipidemic subjects. The results suggest that apoE4 may be involved in the development of type V hyperlipoproteinemia.     

Polymorphisms of the gene encoding cholesterol ester transfer protein and serum lipoprotein levels in subjects with and without coronary heart disease

Summary We determined TaqI-A, TaqI-B and EcoNI genotypes at the cholesteryl ester transfer protein (CETP) locus in 111 healthy volunteers and in 187 hyperlipidemic men of whom 72 had suffered a myocardial infarction. There were no significant differences in the allele distributions at these polymorphic loci either between the population sample and the hyperlipidemic subjects, or between patients with and without previous myocardial infarction. To detect the associations between the CETP polymorphisms and serum lipid and apoprotein levels, we determined the serum concentrations of total cholesterol, triglycerides, high density lipoprotein (HDL)-cholesterol, apoA-I, apoA-II and apoB in the subjects studied and correlated them to the 3 RFLPs. No significant differences were observed in the serum levels of apoproteins and lipid parameters between subjects with different genotypes in any of these polymorphic CETP loci, either in the population sample or in hyperlipidemic men. Multivariate analyses did not reveal a significant independent role for any of the 3 polymorphisms in determining serum HDL-cholesterol or apoA-I levels after adjusting for triglyceride and low density lipoprotein cholesterol concentrations. This was evident for the group of healthy volunteers and for hyperlipidemic subjects, including those who had survived a myocardial infarction. We conclude that, in Finns, the CETP RFLPs are not useful markers for the risk of coronary heart disease.     

Radiochemical high-performance liquid chromatographic assay for the determination of catechol O-methyltransferase activity towards various substrates

A new chromatographic catechol O-methyltransferase (COMT) assay based on S-adenosyl- -[methyl-¹⁴C]methionine and on-line radioactivity detection was developed. With minor modifications in the mobile phase composition the methylation velocities for 30 structurally diverse compounds including simple catechols, neurotransmitters, catecholestrogens and catecholic drugs could be measured using human and rat recombinant soluble COMT. The enzymes showed very similar substrate selectivities. The radiochemical method was validated using 3,4-dihydroxybenzoic acid as a model substrate and it was shown that accurate and reproducible methylation velocity values could be achieved for both of the catecholic hydroxyls. The method proved to be suited for determining the enzyme kinetic parameters and can probably be further used for gathering enzyme kinetic data on differentially substituted catechols in order to construct proper structure-activity relationships for COMT.