Atomic resolution structure of the HFBII hydrophobin, a self-assembling amphiphile

Hydrophobins are proteins specific to filamentous fungi. Hydrophobins have several important roles in fungal physiology, for example, adhesion, formation of protective surface coatings, and the reduction of the surface tension of water, which allows growth of aerial structures. Hydrophobins show remarkable biophysical properties, for example, they are the most powerful surface-active proteins known. To this point the molecular basis of the function of this group of proteins has been largely unknown. We have now determined the crystal structure of the hydrophobin HFBII from Trichoderma reesei at 1.0 A resolution. HFBII has a novel, compact single domain structure containing one alpha-helix and four antiparallel beta-strands that completely envelop two disulfide bridges. The protein surface is mainly hydrophilic, but two beta-hairpin loops contain several conserved aliphatic side chains that form a flat hydrophobic patch that makes the molecule amphiphilic. The amphiphilicity of the HFBII molecule is expected to be a source for surface activity, and we suggest that the behavior of this surfactant is greatly enhanced by the self-assembly that is favored by the combination of size and rigidity. This mechanism of function is supported by atomic force micrographs that show highly ordered arrays of HFBII at the air water interface. The data presented show that much of the current views on structure function relations in hydrophobins must be re-evaluated.     

Determination of lignans in human plasma by liquid chromatography with coulometric electrode array detection

The presence of mammalian lignans, mainly enterolactone, in human plasma has been related to lower incidence of certain cancers and cardiovascular disease. The plant lignans secoisolariciresinol and matairesinol have been reported to be precursors of mammalian lignans, but recently other plant lignans relatively abundant in the diet have also been identified as precursors. To evaluate the importance and contribution of these new dietary precursors to the mammalian lignan formation in vivo, metabolic studies in human subjects must be carried out. For this purpose a method based on high-performance liquid chromatography using coulometric electrode array detection for the simultaneous determination of nine plant lignans and two mammalian lignans in human plasma was developed, validated, and shown to fulfill the reliability criteria.     

Oestradiol potentiates the effects of certain pyrethroid compounds in the MCF7 human breast carcinoma cell line

Pyrethroids are the most widely used insecticides for indoor pest control, so human exposure to them is common. The main target of pyrethroids is the nervous system, but their endocrine disrupting capabilities may also be of toxicological concern. In the present study, the proliferation of the breast cancer cell line, MCF7, was studied after a 7-day exposure to various concentrations of pyrethrin, permethrin and cypermethrin. The effects of oestradiol and the combined effects of oestradiol (0.10 nM) and pyrethroids (0.1-100 microM) on MCF7 cell proliferation were also evaluated. Proliferation and cell toxicity were studied by measuring the ATP content with a luminescence method, and mitochondrial metabolic enzyme activity with the WST-1 test. In the ATP test, low concentrations (0.1-1 microM) of pyrethroids in co-exposure with oestradiol caused a clear statistically significant increase in the proliferation of MCF7 cells. This was evident when compared to the proliferative effect caused by 0.1 nM oestradiol alone. High concentrations were cytotoxic, and the greatest cell toxicity was that of cypermethrin, which has a cyano group in its molecular structure.     

Distribution of hyaluronan in articular cartilage as probed by a biotinylated binding region of aggrecan

The proportion of total tissue hyaluronan involved in interactions with aggrecan and link protein was estimated from extracts of canine knee articular cartilages using a biotinylated hyaluronan binding region-link protein complex (bHABC) of proteoglycan aggregate as a probe in an ELISA-like assay. Microscopic sections were stained with bHABC to reveal free hyaluronan in various sites and zones of the cartilages. Articular cartilage, cut into 20 m-thick sections, was extracted with 4 M guanidinium chloride (GuCl). Aliquots of the extract (after removing GuCl) were assayed for hyaluronan, before and after papain digestion. The GuCl extraction residues were analyzed after solubilization by papain. It was found that 47–51% of total hyaluronan remained in the GuCl extraction residue, in contrast to the 8–15% of total proteoglycans. Analysis of the extract revealed that 24–50% of its hyaluronan was directly detecable with the probe, while 50–76% became available only after protease digestion. The extracellular matrix in cartilage sections was stained with the bHABC probe only in the superficial zone and the periphery of the articular surfaces, both sites known to have a relatively low proteoglycan concentration. Trypsin pretreatment of the sections enhanced the staining of the intermediate and deep zones, presumably by removing the steric obstruction caused by the chondroitin sulfate binding region of aggrecans. Enhanced matrix staining in these zones was also obtained by a limited digestion with chondroitinase ABC. The results indicate that a part of cartilage hyaluronan is free from endogenous binding proteins, such as aggrecan and link protein, but that the chondroitin sulfate-rich region of aggrecan inhibits its probing in intact tissue sections. Therefore, hyaluronan staining was more intense in cartilage areas with lower aggrecan content. A large proportion of hyaluronan resists GuCl extraction, even from 20-m-thick tissue sections.     

Production of endo-1,4-β-glucanase and xylanase by Trichoderma reesei immobilized on polyurethane foam

Summary Endo-1,4--glucanase and xylanase were produced by Trichoderma reesei immobilized on polyurethane foam using lactose as the main carbon source. The most porous carrier was found to be the best of those tested. The nitrogen source and KH₂PO₄ concentration of the production medium had a marked effect on culture pH during the course of fermentation and, consequently, on xylanase activity. An increase in lactose concentration from 7 to 27 g/l resulted in an increase in endoglucanase activity (max. 730 U/ml), xylanase activity (max. 3350 U/ml) and filter paper activity (max. 3.0 FPU/ml).     

Production of Hormoconis resinae glucoamylase P by a stable industrial strain of Saccharomyces cerevisiae

A stable strain of Saccharomyces cerevisiae secreting glucoamylase (EC 3.2.1.3) with high debranching activity was constructed using recombinant DNA technology. An expression cassette without bacterial sequences, containing Hormoconis resinae glucoamylase P cDNA and the dominant selection marker MEL1 was integrated into the yeast chromosome using ARS1 homology. The glucoamylase expression level of the integrant yeast strain was increased by chemical mutagenesis. The yeast strains secreting glucoamylase were able to grow on soluble starch (5%, w/v) and ferment it to ethanol.Correspondence to: A. Vainio     

Predator–rodent–plant interactions along a coast–inland gradient in Fennoscandian tundra

Spatial variation in the strength of trophic cascades in arctic tundra has been related to flows of subsidies across ecosystem boundaries. Here, we ask whether the input of marine subsidies in tundra systems would cause spatial variation in the strength of rodent–plant interactions between coastal areas, where predators have access to marine‐derived resources, and non‐subsidized inland areas of northern Fennoscandia. We present a detailed evaluation of predator–rodent–vegetation interactions along a coast‐inland gradient, during the 2011 rodent outbreak and the two following decline years, by using direct assessments of rodent impacts and tracing of marine‐derived nutrients in the food web. Our results revealed that the main rodent predator during summer, the long‐tailed jaeger Stercorarius longicaudus, did not benefit from marine resources while breeding (relative dietary proportion in chicks’ diet = 0–3%). Contrary to this pattern, parasitic jaegers S. parasiticus, bred exclusively near the coast and preyed effectively on both marine resources (41% of chicks’ diet) and rodents (12%). Mammalian predators also showed a higher activity during winter near the coast. Despite overall higher predator numbers, no evidence was found for lower rodent population growth rates during the three monitoring summers and for weaker rodent grazing impacts in the coastal area. Instead, we documented pronounced damages caused by lemmings and voles on bryophytes and vascular plants, especially dwarf shrubs (e.g. Vaccinum myrtillus) all along the coast–inland gradient. Taken together, our results did not support the hypothesis that marine subsidies would trigger a trophic cascade in coastal tundra areas of northern Fennoscandia during a major rodent outbreak, probably due to a relatively low diversity of marine‐subsidized predators in the region. Comparative observational and experimental studies at large spatial scales in various arctic regions are absolutely necessary for a better understanding of factors causing regional variations in the functioning of arctic food webs.     

A N-acetyllactosamine-specific cell-binding activity in a plant pathogen, Erwinia rhapontici

A strain of the phytopathogenic bacterial species, Erwinia rhapontici, was found to cause hemagglutination of human erythrocytes that was specifically inhibited by beta-galactosides. Of the monosaccharides tested, N-acetyl galactosamine and galactose efficiently inhibited the hemagglutination. The most potent inhibitor identified was Ga1 beta 1-4GlcNAc that was 30-100-fold more potent than Ga1 beta 1-3GlcNac or Ga1 beta 1-3GalNAc. Fetuin had no effect on the hemagglutination whereas asialofetuin was inhibitory. No blood group specificity was found for the hemagglutinin. These findings indicate that the E. rhapontici strain possesses a novel bacterial cell-binding activity with specificity for terminal N-acetyllactosamine residues.     

Serum fatty acid and lipoprotein subclass concentrations and their associations in prepubertal healthy Norwegian children

Introduction

The lipid metabolism is one of the most important and complex processes in the body. Serum concentrations of 18 fatty acids (FAs) and 24 lipoprotein features, i.e. concentrations of lipoprotein main and subclasses and average particle size in main classes, in 195 ethnic Norwegian children from the rural Fjord region were quantified by chromatography.

Objectives

To assess gender differences in prepubertal children and reveal predictive FA patterns for lipoprotein features.

Methods

Lipoprotein features were modelled from FA profiles using multivariate regression.

Results

Contrary to observations for adults from the same region, gender differences in prepubertal children were generally small. However, higher concentrations of C16–C18 FAs for girls compared to boys correlated to higher concentrations of triglycerides (TG) and very low density lipoprotein (VLDL) particles and larger average size of VLDL particles. Concentrations of high density lipoprotein (HDL) and its subclass of medium particle size were higher in boys than in girls. These findings are opposite to observations in adults from the same region, but reflect that prepubertal boys are more physically active than girls. Furthermore, children possessed only half the serum levels of eicosapentaenoic acid and docosahexaenoic acid measured in adults. Since sampling was done after 12 h of fasting, these differences may reflect higher rate of utilization of these crucial FAs in children.

Conclusion

Good predictive models were obtained for TGs, VLDL and chylomicrons with C14–C18 FAs as major contributors. Weak predictive associations were observed for HDL and Apolipoprotein A1 (ApoA1) with C20–C24 FAs as contributors.