Domains in the S-layer protein CbsA of Lactobacillus crispatus involved in adherence to collagens,laminin and lipoteichoic acids and in self-assembly

The protein regions in the S-layer protein CbsA of Lactobacillus crispatus JCM 5810, needed for binding to collagens and laminin, anchoring to bacterial cell wall, as well as self-assembly, were mapped by deletion analysis of His-tagged peptides isolated from Escherichia coli and by heterologous expression on Lactobacillus casei. Mature CbsA is 410 amino acids long, and stepwise genetic truncation at both termini revealed that the region 32-271 carries the infor-mation for self-assembly of CbsA into a periodic structure. The lactobacillar S-layer proteins exhibit sequence variation in their assembly domain, but the border regions 30-34 and 269-274 in CbsA are conserved in valine-rich short sequences. Short deletions or substitutions at these regions affected the morphology of His-CbsA polymers, which varied from sheet-like to cylindrical tubular polymers, and further truncation beyond the DNA encoding residues 32 and 271 leads to a non-periodic aggregation. The self-assembly of the truncated peptides, as seen by electron microscopy, was correlated with their behaviour in a cross-linking study. The shorter peptides not forming a regular polymer were observed by the cross-linking study and mass spectrometry to form dimers, trimers and tetramers, whereas the other peptides were cross-linked to large multimers only. Binding of solubilized type I and IV collagens was observed with the His-CbsA peptides 1-274 and 31-287, but not with the smaller peptides regardless of their ability to form regular polymers. Strain JCM 5810 also adheres to immobilized laminin and, in order to analyse the possible laminin binding by CbsA, cbsA and its fragments were expressed on the surface of L. casei. Expression of the CbsA peptides 1-274, 1-287, 28-287 and 31-287 on L. casei conferred adhesiveness to both laminin and collagen immobilized on glass as well as to laminin- and collagen-containing regions in chicken colon and ileum. The C-terminal peptides 251-410 and 288-410 bound to L. crispatus JCM 5810 cells from which the S-layer had been depleted by chemical extraction, whereas no binding was seen with the His-CbsA peptides 1-250 or 1-269 or to cells with an intact S-layer. The His-CbsA peptides 251-410 and 288-410 bound to teichoic acids of several bacterial species. The results show that CbsA is an adhesive complex with an N-terminal assembly domain exhibiting affinity for pericellular tissue components and a cationic C-terminal domain binding to negatively charged cell wall components.     

Expression of progesterone receptor isoforms A and B is differentially regulated by estrogen in different breast cancer cell lines

Progesterone action in target tissues is mediated through two progesterone receptor (PR) isoforms, PR-A and PR-B, which display different regulatory functions in target cells. Relative expression ratio of these isoforms varies depending on cell and tissue types. Here, we studied the regulation of PR isoform expression by estradiol (E(2)), insulin, IGF-1 and cAMP in different breast cancer cell lines. Although, E(2) induced PR expression in all cell lines studied, the expression ratio of PR-A/PR-B induced by E(2) was dependent on the cell line. The differential regulation of the isoforms was also seen at the mRNA level suggesting that the PR-A and PR-B promoters are differentially regulated by E(2) in different breast cancer cells. Insulin, IGF-1 or cAMP previously reported to induce PR expression however failed to alter the PR expression in our study. This is the first report describing that in different breast cancer cell lines the expression of PR-A and PR-B is regulated by E(2) in a distinct way.     

The effects of a two-week reflection-intention training program on lucid dream recall.

Previous research in the field of lucid dreams has produced several techniques which are claimed to be a means of inducing lucid dreams, but there have been only a small number of reported studies which have evaluated their effectiveness. The present study investigated the effects of one of the most promising of these, the reflection-intention technique, which is specifically designed to increase the frequency of dream and lucid dream recall. The sample here consisted of 20 participants whose frequencies of normal dream recall and lucid dream recall were recorded before, during and after a two week training program by using a dream questionnaire and a dream journal: A one-way repeated measures analysis of variance showed a significant increase in both dream recall frequency (F2,38 = 22.09, p2,38 = 5.96, p     

The structure of bovine mitochondrial F1-ATPase: an example of rotary catalysis

There is now compelling evidence in support of a rotary catalytic mechanism in F1-ATPase, and, by extension, in the intact ATP synthase. Although models have been proposed to explain how protein translocation in F0 results in rotation of the gamma-subunit relative to the alpha 3/beta 3 assembly in F1 [22], these are still speculative. It seems likely that a satisfactory explanation of this mechanism will ultimately depend on structural information on the intact ATP synthase.     

Effect of Plasterboard Composition on Stachybotrys chartarum Growth and Biological Activity of Spores

The effects of plasterboard composition on the growth and sporulation of Stachybotrys chartarum as well as on the inflammatory potential of the spores were studied. S. chartarum was grown on 13 modified plasterboards under saturated humidity conditions. The biomass was estimated by measuring the ergosterol content of the S. chartarum culture while the spore-induced cytotoxicity and production of nitric oxide (NO), tumor necrosis factor alpha (TNF-α), and interleukin-6 in mouse macrophages was used to illustrate the bioactivity of spores. The ergosterol content of S. chartarum correlated with the number of spores collected from plasterboards. The growth and sporulation decreased compared to that of the reference board in those cases where (i) the liner was treated with biocide, (ii) starch was removed from the plasterboard, or (iii) desulfurization gypsum was used in the core. Spores collected from all the plasterboards were toxic to the macrophages. The biocide added to the core did not reduce the growth; in fact, the spores collected from that board evoked the highest cytotoxicity. The conventional additives used in the core had inhibitory effects on growth. Recycled plasterboards used in the core and the board lacking the starch triggered spore-induced TNF-α production in macrophages. In summary, this study shows that the growth of a strain of S. chartarum on plasterboard and the subsequent bioactivity of spores were affected by minor changes to the composition of the core or liners, but it could not be totally prevented without resorting to the use of biocides. However, incomplete prevention of microbial growth by biocides even increased the cytotoxic potential of the spores.     

Stemwood volume increment changes in European forests due to climate change—a simulation study with the EFISCEN model

This paper presents the results of a modelling study of future net annual increment changes in stemwood of European forests owing to climate change. Seven process‐based growth models were applied to 14 representative forest sites across Europe under one climate change scenario. The chosen scenario was the HadCM2 run, based on emission scenario IS92a, and resulted in an increase in mean temperature of 2.5 °C between 1990 and 2050, and an increase in annual precipitation of 5–15%. The information from those runs was incorporated in a transient way in a large‐scale forest resource scenario model, EFISCEN (European forest information scenario). European scale forest resource projections were made for 28 countries covering 131.7 million ha of forest under two management scenarios for the period until 2050. The results showed that net annual increments in stemwood of European forests under climate change will further increase with an additional 0.9 m³ ha^?1 y^?1 in 2030 compared to the ongoing increase under a current climate scenario, i.e. an extra 18% increase. After 2030 the extra increment increase is reduced to 0.79 m³ ha^?1 y^?1 in 2050. Under climate change, absolute net annual increments will increase from the present 4.95, on average for Europe, to 5.93 m³ ha^?1 y^?1 in 2025. After 2025, increments in all scenarios start to decline owing to ageing of the forest and the high growing stocks being reached. The results of the present study are surrounded by large uncertainties. These uncertainties are caused by unknown emissions in the future, unknown extent of climate change, uncertainty in process‐based models, uncertainty in inventory data, and uncertainty in inventory projection. Although the results are thus not conclusive, climate change may lead to extra felling opportunities in European forests of 87 million m³y^?1. Because Europe's forests are intensively managed already, management may adapt to climate change relatively easily. However, this study also indicates that climate change may lead to a faster build‐up of growing stocks. That may create a less stable forest resource in terms of risks to storm damage.     

Raised atmospheric CO2 levels and increased N deposition cause shifts in plant species composition and production in Sphagnum bogs

Part of the missing sink in the global CO₂ budget has been attributed to the positive effects of CO₂ fertilization and N deposition on carbon sequestration in Northern Hemisphere terrestrial ecosystems. The genus Sphagnum is one of the most important groups of plant species sequestrating carbon in temperate and northern bog ecosystems, because of the low decomposability of the dead material it produces. The effects of raised CO₂ and increased atmospheric N deposition on growth of Sphagnum and other plants were studied in bogs at four sites across Western Europe. Contrary to expectations, elevated CO₂ did not significantly affect Sphagnum biomass growth. Increased N deposition reduced Sphagnum mass growth, because it increased the cover of vascular plants and the tall moss Polytrichum strictum. Such changes in plant species composition may decrease carbon sequestration in Sphagnum‐dominated bog ecosystems.     

In vitro experimental studies of sialyl Lewis x and sialyl Lewis a on endothelial and carcinoma cells: crucial glycans on selectin ligands

Extravasation from the blood of malignant tumour cells that form metastasis and leukocytes that go into tissues require contact between selectins and their sialyl Lewis x and sialyl Lewis a (sLex and sLea respectively) decorated ligands. Endothelial cells have been shown to express sLex epitopes in lymph nodes and at sites of inflammation, and this is crucial for the selectin-dependent leukocyte traffic. Besides the ability to synthesize sLex on sialylated N-acetyllactosamine via the action of α(1,3)fucosyltransferase(s), endothelial cells can also degrade sLex to Lewis x through the action of α(2,3)sialidase(s). In addition, several epithelial tumors possess the machinery to synthesize sLex, which facilitates their adhesion to endothelial E- and P-selectin. This revised version was published online in November 2006 with corrections to the Cover Date.     

Intracellular resistance correlates with initial stage of frost hardening in willow (Salix viminalis)

Distributed model parameters of shoots of five clones of willow ( Salix viminalis ) were examined with electrical impedance analysis at the end of the growing season and with cold acclimation. The parameters were compared with regard to frost hardiness, linoleic (18:2) and linolenic (18:3) fatty acids, unsaturation/saturation ratio of fatty acids and dry weight content. The intracellular resistance (r_i) correlated best with changes in frost hardiness. The value of r_i rose from 1–2 Ω m in non-hardened to about 12 Ω m in hardened samples. In the initial stages of frost hardening, a linear relationship was found between r_i and frost hardiness and levels of 18:2 fatty acid, and an inverse relationship between r_i and levels of 18:3 fatty acid. The unsaturation/saturation ratio of fatty acids rose fairly rapidly in the initial stage of hardening. The dry weight content increased stepwise during the experimental period, and less steadily than r_i. In addition, equivalent circuit parameters changed in the prehardening phase, and were probably connected with cell differentiation and lignification. Frost hardiness by the visual method and by extracellular resistance, determined after controlled freezing tests, correlated well in the initial stages of hardening until about − 10°C but deviated upon further hardening.     

Adhesion of fimbriated nitrogen-fixing enteric bacteria to roots of grasses and cereals

Summary The role of fimbriae in enterobacterial adhesion to roots of grasses and cereals is discussed. All nitrogen-fixing enteric bacteria isolated in Finland had fimbriae. AllEnterobacter isolates had mannose-binding type-1 fimbriae, whereas most of theKlebsiella isolates had both type-1 and type-3 fimbriae. The strains were isolated from a total of ten different grass species, and no specific association was found between grass species and bacterial fimbriation, biogroup or serogroup. Purified, radiolabeled fimbriae bound to roots ofPoa pratensis in vitro, and bacterial adhesion was inhibited by Fab fragments specific for fimbriae.Klebsiella strains carrying type-3 fimbriae adhered to roots of various grasses and cereals more efficiently than type-1- or nonfimbriated strains, and it was concluded that type-3 fimbriae are the major adhesions ofKlebsiella. Immunofluorescence studies revealed that the bacteria preferentially adhered to root hairs, and to a lesser extent, to the zone of elongation and the root cap mucilage. No strict host specificity in enterobacterial adhesion was observed.