Formation and Characterization of Supported Lipid Bilayers Composed of Hydrogenated and Deuterated Escherichia coli Lipids

Supported lipid bilayers are widely used for sensing and deciphering biomolecular interactions with model cell membranes. In this paper, we present a method to form supported lipid bilayers from total lipid extracts of Escherichia coli by vesicle fusion. We show the validity of this method for different types of extracts including those from deuterated biomass using a combination of complementary surface sensitive techniques; quartz crystal microbalance, neutron reflection and atomic force microscopy. We find that the head group composition of the deuterated and the hydrogenated lipid extracts is similar (approximately 75% phosphatidylethanolamine, 13% phosphatidylglycerol and 12% cardiolipin) and that both samples can be used to reconstitute high-coverage supported lipid bilayers with a total thickness of 41 ± 3 Å, common for fluid membranes. The formation of supported lipid bilayers composed of natural extracts of Escherichia coli allow for following biomolecular interactions, thus advancing the field towards bacterial-specific membrane biomimics.     

On the effect of misclassification on bias of perfectly measured covariates in regression

This note clarifies under what conditions a naive analysis using a misclassified predictor will induce bias for the regression coefficients of other perfectly measured predictors in the model. An apparent discrepancy between some previous results and a result for measurement error of a continuous variable in linear regression is resolved. We show that similar to the linear setting, misclassification (even when not related to the other predictors) induces bias in the coefficients of the perfectly measured predictors, unless the misclassified variable and the perfectly measured predictors are independent. Conditional and asymptotic biases are discussed in the case of linear regression, and explored numerically for an example relating birth weight to the weight and smoking status of the mother.     

Random amplification of polymorphic DNA (RAPD) typing of carnobacteria isolated from hindgut chamber and large intestine of Atlantic cod (Gadus morhua l.)

Autochthonous and allochthonous bacteria were isolated from hindgut chamber and large intestine of fed and starved Atlantic cod (Gadus morhua L.). All bacterial strains isolated from hindgut chamber belong to carnobacteria. However, only 10.2% of the bacteria strains from the large intestine belong to carnobacteria. Random amplification of polymorphic DNA (RAPD) analysis using three selective primers, was performed to further identify the carnobacteria strains. Nine of these were isolated from hindgut chamber contents, ten associated with epithelial cells of the hindgut chamber, and six isolated from the large intestines of fed and starved fish. The 25 isolates segregated into eight clusters. The major cluster comprised nine strains isolated from the hindgut chamber of both fed and starved fish showing low similarity with the reference strains. The other strains isolated from the hindgut were located in clusters showing high similarity with Carnobacterium gallinarum or Carnobacterium piscicola. Strains isolated from large intestine appeared more divergent and were located in five different clusters. Autochthonous (indigenous) bacteria were clearly demonstrated in the hindgut chamber as transmission electron microscopy revealed rod-shaped bacteria between adjacent microvilli. Endocytosis of bacteria by epithelial cells was observed in the hindgut chamber.     

Self-assembly and biphasic iron-binding characteristics of Mms6, a bacterial protein that promotes the formation of superparamagnetic magnetite nanoparticles of uniform size and shape

Highly ordered mineralized structures created by living organisms are often hierarchical in structure with fundamental structural elements at nanometer scales. Proteins have been found responsible for forming many of these structures, but the mechanisms by which these biomineralization proteins function are generally poorly understood. To better understand its role in biomineralization, the magnetotactic bacterial protein, Mms6, which promotes the formation in vitro of superparamagnetic magnetite nanoparticles of uniform size and shape, was studied for its structure and function. Mms6 is shown to have two phases of iron binding: one high affinity and stoichiometric and the other low affinity, high capacity, and cooperative with respect to iron. The protein is amphipathic with a hydrophobic N-terminal domain and hydrophilic C-terminal domain. It self-assembles to form a micelle, with most particles consisting of 20-40 monomers, with the hydrophilic C-termini exposed on the outside. Studies of proteins with mutated C-terminal domains show that the C-terminal domain contributes to the stability of this multisubunit particle and binds iron by a mechanism that is sensitive to the arrangement of carboxyl/hydroxyl groups in this domain.     

Do Personalized Feedback Messages about Activity Patterns Stimulate Patients with Chronic Low Back Pain to Change their Activity Behavior on a Short Term Notice?

The aim of this study was to explore whether patients responded to personalized messages on top of continuous visual feedback by changes in activity patterns and whether this response is related to the stages of change and the pain intensity levels. Patients wore a movement sensor and a PDA for 2 weeks and received continuously feedback and time-related messages to influence activity behavior. The response was calculated by calculating the activity 30 min before and after a message. In addition, the readiness to change was measured with the Stage of Change questionnaire and pain intensity was measured on a visual analogue scale. Sixteen patients participated, receiving a total of 517 messages. Overall, patients responded to personalized messages (p < .049), with a higher response in the morning. Patients in different stages of change responded differently to the messages (p = .009) and the response was significantly related to the pain intensity levels (Pearson correlation −.226) in the second week of feedback. This study suggests that personalized messages have the potential to influence activity behavior. It seems relevant to take time of the day, the stages of change and pain intensity levels of the patient into account to further optimize the feedback strategy used.     

Electrophysiological responses from neurons of antennal taste sensilla in the polyphagous predatory ground beetle Pterostichus oblongopunctatus (Fabricius 1787) to plant sugars and amino acids

The responses of antennal contact chemoreceptors, in the polyphagous predatory ground beetle Pterostichus oblongopunctatus, to twelve 1-1000mmoll(-1) plant sugars and seven 10-100mmoll(-1) amino acids were tested. The disaccharides with an alpha-1.4-glycoside linkage, sucrose and maltose, were the two most stimulatory sugars for the sugar-sensitive neuron innervating these contact chemosensilla. The firing rates they evoked were concentration dependent and reached up to 70impulses/s at 1000mmoll(-1). The stimulatory effect of glucose on this neuron was approximately two times lower. This can be partly explained by the fact that glucose exists in at least two anomeric forms, alpha and beta. These two forms interconvert over a timescale of hours in aqueous solution, to a final stable ratio of alpha:beta 36:64, in a process called mutarotation. So the physiologically active alpha-anomere forms only 36% of the glucose solution which was reflected in its relatively low dose/response curve. Due to the partial herbivory of P. oblongopunctatus these plant sugars are probably involved in its search for food, for example, for conifer seeds. Several carbohydrates, in addition to glucose, such as cellobiose, arabinose, xylose, mannose, rhamnose and galactose are known as components of cellulose and hemicelluloses. They are released by brown-rot fungi during enzymatic wood decay. None of them stimulated the antennal sugar-sensitive neuron. They are therefore not implicated in the search for hibernation sites, which include rotting wood, by this beetle. The weak stimulating effect (below 3impulses/s) of some 100mmoll(-1) amino acids (methionine, serine, alanine, glutamine) to the 4th chemosensory neuron of these sensilla was characterized as non-specific, or modulating the responses of non-target chemosensory neurons.     

Phylogenetic and functional conservation of the NKR-P1F and NKR-P1G receptors in rat and mouse

Two clusters of rat Nkrp1 genes can be distinguished based on phylogenetic relationships and functional characteristics. The proximal (centromeric) cluster encodes the well-studied NKR-P1A and NKR-P1B receptors and the distal cluster, the largely uncharacterized, NKR-P1F and NKR-P1G receptors. The inhibitory NKR-P1G receptor is expressed only by the Ly49s3⁺ NK cell subset as detected by RT-PCR, while the activating NKR-P1F receptor is detected in both Ly49s3⁺ and NKR-P1B⁺ NK cells. The mouse NKR-P1G ortholog is expressed by both NKR-P1D⁻ and NKR-P1D⁺ NK cells in C57BL/6 mice. The rat and mouse NKR-P1F and NKR-P1G receptors demonstrate a striking, cross-species conservation of specificity for Clr ligands. NKR-P1F and NKR-P1G reporter cells reacted with overlapping panels of tumour cell lines and with cells transiently transfected with rat Clr2, Clr3, Clr4, Clr6 and Clr7 and mouse Clrc, Clrf, Clrg and Clrd/x, but not with Clr11 or Clrb, which serve as ligands for NKR-P1 from the proximal cluster. These data suggest that the conserved NKR-P1F and NKR-P1G receptors function as promiscuous receptors for a rapidly evolving family of Clr ligands in rodent NK cells.     

Mycorrhizal plants of traditionally managed boreal grasslands in Norway

This paper reports on the mycorrhizal status of 82 plant species growing in traditionally managed grasslands in three different locations in the boreal and boreo-nemoral vegetation zone in the eastern part of Norway. Seventy-four species were found to have arbuscular mycorrhiza (AM). To our knowledge, we report AM for the first time in Achillea ptarmica, Ajuga pyramidalis, Alchemilla glaucescens, Carex brunnescens, Carex pallescens, Crepis praemorsa, Hieracium lactucella, Rumex longifolius, Scorzonera humilis, Trifolium aureum and Trifolium spadiceum. The rare and threatened species Arnica montana, S. humilis, C. praemorsa, Gentianella campestris, Parnassia palustris, T. aureum and T. spadiceum, all confined to grasslands, were found to possess AM fungi.