Monitoring cell‐specific neutral lipid accumulation in Phaeodactylum tricornutum (Bacillariophyceae) with Nile Red staining – a new method for FlowCAM

With the fluorescent stain Nile Red (NR), phytoplankton lipid accumulation can be monitored quickly and in situ. In the light of recent results in phytoplankton diversity research, there is also a need for cell‐ and species‐specific lipid measurement techniques. The objective of this work was to investigate whether cell‐specific phytoplankton lipid accumulation could be monitored with the image‐based particle analyzer FlowCAM? and NR staining. Applying Phaeodactylum tricornutum as a model species, we compared the FlowCAM method to two established lipid quantification methods: spectrofluorometric NR fluorescence measurement and total lipid analysis by gas chromatography. The experiment was carried out in batch cultures under nitrogen limitation to induce lipid accumulation. We showed significant correlation between the three different lipid quantification methods confirming the applicability of the novel FlowCAM method in cell‐specific and near real‐time lipid quantification. Furthermore, with the method described here, the lipid content of taxonomically distinguished cells can eventually be measured from multispecies cultures, opening several new possibilities to study species‐specific responses to stress conditions and the complementarity effect.     

Open tundra persist,but arctic features decline—Vegetation changes in the warming Fennoscandian tundra

In the forest‐tundra ecotone of the North Fennoscandian inland, summer and winter temperatures have increased by two to three centigrades since 1965, which is expected to result in major vegetation changes. To document the expected expansion of woodlands and scrublands and its impact on the arctic vegetation, we repeated a vegetation transect study conducted in 1976 in the Darju, spanning from woodland to a summit, 200 m above the tree line. Contrary to our expectations, tree line movement was not detected, and there was no increase in willows or shrubby mountain birches, either. Nevertheless, the stability of tundra was apparent. Small‐sized, poorly competing arctic species had declined, lichen cover had decreased, and vascular plants, especially evergreen ericoid dwarf shrubs, had gained ground. The novel climate seems to favour competitive clonal species and species thriving in closed vegetation, creating a community hostile for seedling establishment, but equally hostile for many arctic species, too. Preventing trees and shrubs from invading the tundra is thus not sufficient for conserving arctic biota in the changing climate. The only dependable cure is to stop the global warming.     

1H NMR at 800 MHz facilitates detailed phospholipid follow-up during atherogenic modifications in low density lipoproteins

The structure of low density lipoprotein (LDL) particles and, particularly, the enzymatic and oxidative modifications of their surface is crucial in the initiation of atherosclerosis. Due to the structural complexity of LDL, there is a lack of suitable methods for dynamic follow-up studies of the molecular mechanisms in native and modified particles in physiological conditions. Here, we report that phosphatidylcholine (PC), lysophosphatidylcholine (lyso-PC), and sphingomyelin (SM) can all be identified and quantified in LDL particles by (1)H NMR spectroscopy at 800 MHz. The signal assignment for the lyso-PC is novel and we illustrate the applicability of the methodology in the case of lipid peroxidation that is generally considered as one of the key proatherogenic modifications of LDL. It was found, somewhat surprisingly, that the LDL-associated phospholipase A(2) is activated in the very beginning of the formation of PC-hydroperoxides. The (patho)physiological rationale of the resulting lyso-PC generation is also briefly discussed.     

Root morphology,mycorrhizal roots and extramatrical mycelium growth in silver birch (Betula pendula Roth) genotypes exposed to experimental warming and soil moisture manipulations

Plant and Soil - In the future, boreal forests will be growing in a warmer climate with more fluctuating soil moisture conditions. However, the knowledge about the effects of simultaneous warming...     

Differentially methylated regions in maternal and paternal uniparental disomy for chromosome 7

DNA methylation is a hallmark of genomic imprinting and differentially methylated regions (DMRs) are found near and in imprinted genes. Imprinted genes are expressed only from the maternal or paternal allele and their normal balance can be disrupted by uniparental disomy (UPD), the inheritance of both chromosomes of a chromosome pair exclusively from only either the mother or the father. Maternal UPD for chromosome 7 (matUPD7) results in Silver-Russell syndrome (SRS) with typical features and growth retardation, but no gene has been conclusively implicated in SRS. In order to identify novel DMRs and putative imprinted genes on chromosome 7, we analyzed eight matUPD7 patients, a segmental matUPD7q31-qter, a rare patUPD7 case and ten controls on the Infinium HumanMethylation450K BeadChip with 30 017 CpG methylation probes for chromosome 7. Genome-scale analysis showed highly significant clustering of DMRs only on chromosome 7, including the known imprinted loci GRB10, SGCE/PEG10, and PEG/MEST. We found ten novel DMRs on chromosome 7, two DMRs for the predicted imprinted genes HOXA4 and GLI3 and one for the disputed imprinted gene PON1. Quantitative RT-PCR on blood RNA samples comparing matUPD7, patUPD7, and controls showed differential expression for three genes with novel DMRs, HOXA4, GLI3, and SVOPL. Allele specific expression analysis confirmed maternal only expression of SVOPL and imprinting of HOXA4 was supported by monoallelic expression. These results present the first comprehensive map of parent-of-origin specific DMRs on human chromosome 7, suggesting many new imprinted sites.     

Two distinct regions in the model protein Peb1 are critical for its heterologous transport out of <Emphasis Type="Italic">Escherichia coli</Emphasis>

Background  

Escherichia coli is frequently the first-choice host organism in expression of heterologous recombinant proteins in basic research as well as in production of commercial, therapeutic polypeptides. Especially the secretion of proteins into the culture medium of E. coli is advantageous compared to intracellular production due to the ease in recovery of the recombinant protein. Since E. coli naturally is a poor secretor of proteins, a few strategies for optimization of extracellular secretion have been described. We have previously reported efficient secretion of the diagnostically interesting model protein Peb1 of Campylobacter jejuni into the growth medium of Escherichia coli strain MKS12 (ΔfliCfliD). To generate a more detailed understanding of the molecular mechanisms behind this interesting heterologous secretion system with biotechnological implications, we here analyzed further the transport of Peb1 in the E. coli host.     

Remnant population dynamics in the facultative biennial Carum carvi in fragmented semi-natural grasslands

Transition matrix models were used to examine the population dynamics in the facultative biennial Carum carvi L. in semi-natural grasslands, specifically to assess what life cycle stages are important for population development and to evaluate the effects of environmental stochasticity on population persistence and, hence, the ability to develop remnant populations. The demographic studies were conducted over a 4-year period in three moderately grazed grasslands that differed in onset and duration of grazing. Experimental seed-sowing was also conducted in disturbed and undisturbed plots in the populations. Deterministic and stochastic models yielded overall negative population growth (λ < 1) for the populations. λ was sensitive to transitions in the most frequent vegetative stage classes. Elasticity analysis indicated that a large proportion of population growth could be ascribed to the stasis of individuals in the largest vegetative stage class. Life-table response experiment (LTRE) analyses showed also that progression to larger stage classes was important in explaining the between-population variation in λ. The expected time to extinction was on the order of several decades for the study populations. Seed-sowing indicated that seedling establishment was limited by both seed and micro-site availability. The populations of C. carvi seem to be able to persist for a rather long time in moderately grazed semi-natural grasslands, even in cases where populations are destined to become extinct. The results, thus, indicate that “biennials” are able to maintain remnant populations in managed semi-natural grasslands.     

Experiments on adhesive dispersal by wood mouse: seed shadows and dispersal distances of 13 plant species from cultivated areas in southern Sweden

This study on epizoochory offers experimental data on retention times and potential dispersal distances of propagules of 13 plant species that commonly inhabit cultivated areas in the south of Sweden. Wood mouse Apodemus flavicollis was used as dispersal vector. Seven of the investigated species produce fruits that carry obvious hooks, barbs or bristles and for comparison the remaining six species, lacking such features, were also included. Excised fruits that initially carried appendages were used in a complementary comparison. Propagules were applied by hand to the back of the animals, which were which released in enclosures to move about freely. Observations were made continually until the seeds were dropped. The distance covered by the animals while moving was measured and the potential dispersal distances calculated. Small fruits were in general found to be more efficiently transported than large ones, but large propagules carrying appendages sometimes remained in place for considerable periods. The potential dispersal distances ranged up to nearly 30 m. The retention time was significantly enhanced by the presence of dispersal attributes. The behavioural pattern of the animals (grooming, movements) plays a significant part in the successful transportation of propagules. By comparing the result with similar experiments using fallow deer Dama dama and domestic cattle Bos taurus as dispersal vectors it can be stated that mice are surprisingly effective as seed dispersers. Large mammals disperse propagules on a larger scale (<1 km) compared with small mammals, that affect plant distribution more locally (<100 m). We consider adhesive dispersal a most significant component in plant metapopulation dynamics, considerably enhancing the probability of occasional propagules reaching suitable sites in a fragmented landscape.